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Vitellogenin (VTG), an egg yolk precursor, is abnormally produced by male and juvenile oviparous species after exposure to estrogens. Plasma VTG in loggerhead sea turtles (Caretta caretta) helped us understand their reproductive maturation and investigate it as a biomarker of contaminant exposure. The presence of VTG was screened in plasma from 404 loggerheads from the northwestern Atlantic Ocean using a freshwater turtle antibody in western blots. The concentrations of VTG were semiquantified using band intensities calibrated to results from a loggerhead antibody enzyme-linked immunoassay. The detection and concentrations of VTG were in (from highest to lowest): nesting females, in-water adult females, subadult females, smaller females, unknown sex, and males. Loggerheads from this region begin vitellogenesis at ≅77 cm straight carapace length. We classified VTG expression as abnormal in nine male or juvenile turtles. Organochlorine contaminant (OC) concentrations were measured in blood and/or fat biopsies of some turtles. One abnormal VTG female had the second highest fat polychlorinated biphenyl (PCB) and 4,4'-dichlorodiphenyldichloroethylene concentrations compared among 43 VTG-negative juveniles. The nine VTG-abnormal turtles had average blood PCB concentrations 8.5% higher, but not significantly different, than 46 VTG-negative juveniles (p = 0.453). In turtles less than 77 cm, blood PCB concentrations were significantly, but weakly, correlated with semiquantified VTG concentrations (tau = 0.1, p = 0.004). Greater blood OC concentrations were found in adult females than in males, which motivated the creation of a conceptual model of OC, VTG, and hormone concentrations across a reproductive cycle. A decision tree is also provided incorporating VTG as a sexing tool. Abnormal VTG expression cannot conclusively be linked to endocrine disruption caused by these OC concentrations. Studies should further investigate causes of abnormal VTG expression in wild sea turtles. Environ Toxicol Chem 2023;42:1309-1325. © 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA., (© 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Objective: Medical schools are charged with assisting medical students to acquire the confidence, knowledge and skills for behavior change conversations in primary healthcare. The present study evaluated teaching brief motivational interviewing (MI) to pre-clinical medical students., Methods: Forty-six students participated in an educational intervention premised on the Learn, See, Practice, Prove, Do, Maintain pedagogical framework, comprising 2 × 2-h lectures, a 2-h role-play triad session, and 3 × 2-h small group simulated patient encounters supported by scaffolding strategies. Measures of brief MI knowledge (MI Knowledge and Attitudes Test & Multiple-Choice Knowledge Test) and confidence (MI Confidence Scale) were taken at baseline, post-training, and 3-month follow-up, and skills (Behavior Change Counseling Index) were assessed at three intervals during simulated patient encounters., Results: Students who received brief MI training improved in knowledge and confidence from baseline to post-training and gains remained at 3-months. Brief MI skills improved across the simulation sessions., Conclusion: Pre-clinical medical students can attain knowledge, confidence and skills in brief MI after participation in a short intervention and improvements are sustainable., Practice Implications: Our results support the use of an evidence-based pedagogical framework for teaching brief MI in pre-clinical years of medical curricula and our scaffolding strategy affords promise., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: It is well recognised that medical students need to acquire certain procedural skills during their medical training, however, agreement on the level and acquisition of competency to be achieved in these skills is under debate. Further, the maintenance of competency of procedural skills across medical curricula is often not considered. The purpose of this study was to identify core procedural skills competencies for Australian medical students and to establish the importance of the maintenance of such skills., Methods: A three-round, online Delphi method was used to identify consensus on competencies of procedural skills for graduating medical students in Australia. In Round 1, an initial structured questionnaire was developed using content identified from the literature. Respondents were thirty-six experts representing medical education and multidisciplinary clinicians involved with medical students undertaking procedural skills, invited to rate their agreement on the inclusion of teaching 74 procedural skills and 11 suggested additional procedures. In Round 2, experts re-appraised the importance of 85 skills and rated the importance of maintenance of competency (i.e., Not at all important to Extremely important). In Round 3, experts rated the level of maintenance of competence (i.e., Observer, Novice, Competent, Proficient) in 46 procedures achieving consensus., Results: Consensus, defined as > 80% agreement, was established with 46 procedural skills across ten categories: cardiovascular, diagnostic/measurement, gastrointestinal, injections/intravenous, ophthalmic/ENT, respiratory, surgical, trauma, women's health and urogenital procedures. The procedural skills that established consensus with the highest level of agreement included cardiopulmonary resuscitation, airway management, asepsis and surgical scrub, gown and gloving. The importance for medical students to demonstrate maintenance of competency in all procedural skills was assessed on the 6-point Likert scale with a mean of 5.03., Conclusions: The findings from the Delphi study provide critical information about procedural skills for the Clinical Practice domain of Australian medical curricula. The inclusion of experts from medical faculty and clinicians enabled opportunities to capture a range of experience independent of medical speciality. These findings demonstrate the importance of maintenance of competency of procedural skills and provides the groundwork for further investigations into monitoring medical students' skills prior to graduation., (© 2022. The Author(s).)

Introduction: In 2013, the U.S. Preventive Services Task Force again recommended alcohol misuse screening and provision of brief behavioral counseling interventions to those engaged in risky drinking for all adults aged ≥18 years in primary care. This report presents national estimates of the provision of alcohol screening and brief intervention by U.S. primary care physicians, the screening methods, and the resources they identified as helpful in implementing alcohol/substance screening and intervention in primary care settings., Methods: Data included 876 self-identified primary care physicians from the Physician Induction Interview portion of the 2015-2016 National Ambulatory Medical Care Survey, an annual nationally representative sample survey of nonfederal, office-based physicians in the U.S., encompassing all the 50 states and the District of Columbia. Descriptive estimates (annualized percentages) of alcohol misuse screening were generated for selected primary care physician characteristics. Estimates of how primary care physicians reported screening, the frequency of brief intervention, and resources identified as helpful in the implementation of screening/intervention procedures were also generated. Two-tailed significance tests were used to determine the differences between the compared groups. Data analyses were conducted in 2019-2021., Results: In total, 71.7% of office-based primary care physicians reported screening patients for alcohol misuse. Statistically significant differences in screening were observed geographically and by provider specialty., Conclusions: Less than 40% of primary care physicians who screened patients for alcohol misuse reported always intervening with patients who screened positive for risky alcohol use. Collection of data on resources that primary care physicians report as being helpful for alcohol/substance screening and intervention implementation may be useful in continuous improvement efforts., (Published by Elsevier Inc.)

Background: In 2014, the Centers for Disease Control and Prevention funded a four-year partnership effort between university and health care professional associations (HCPAs) to reach health care providers (HCPs) nationally in six health disciplines and engage them to adopt evidence-based practices for the prevention, identification, and treatment of fetal alcohol spectrum disorders (FASDs). The aim of this project was to evaluate partnerships for their (1) structure and formation, (2) collaboration process, and (3) outcomes with regard to resources and strategies developed for FASD prevention and management., Methods: We used quantitative and qualitative data from quarterly progress reports, a semi-annual collaboration survey, and annual interviews with each discipline's members., Results: Partnerships in each discipline varied in the number of members and organizations, expertise in the discipline, and access to HCPs. Assigned partnerships with limited researchers' expertise in the discipline or the inability of HCPAs to reach priority audiences created challenges in the development and dissemination of resources. Two partnerships showed challenges in the collaboration process regarding understanding respective responsibilities, sharing similar ideas, and resolving disagreements despite efforts at facilitated discussion. Messaging and resource dissemination by HCPAs and the use of provider champions developed through HCPAs' national network emerged as promising approaches to engage HCPs., Conclusion: Circumstances under which partnerships are formed can facilitate or challenge collaboration and outcome efforts. Discipline-specific partnerships between researchers and HCPAs provide a model for evidence-based resources to be developed and disseminated widely for adoption by HCPs in their practice., Competing Interests: Conflicts of Interest None.

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Following the explosion of the Deepwater Horizon MC252 oil rig in 2010, 319 live sea turtles exposed to crude oil and oil-dispersant (Corexit) combinations were admitted to rehabilitation centers for decontamination and treatment. Treatment of oiled sea turtles was guided by expected physiological and pathological effects of crude oil exposure extrapolated from studies in other species and from a single loggerhead sea turtle ( Caretta caretta ) study. While invaluable starting points, inherent limitations to extrapolation, and small sample size of the experimental exposure study, reduce their utility for clinical guidance and for assessing oil spill impacts. Effects of dispersants were not included in the previous experimental exposure study, and cannot be effectively isolated in the analysis of field data from actual spills. A terminal study of pivotal temperature of sex determination using eggs salvaged from doomed loggerhead nests provided an opportunity for an ancillary exposure study to investigate the acute effects of crude oil, dispersant, and a crude oil/dispersant combination in sea turtle hatchlings. Eggs were incubated at 27.2-30.8°C, and hatchlings were randomly assigned to control, oil, dispersant, and combined oil/dispersant exposures for 1 or 4 days. Contaminant exposures were started after a 3 day post-hatching period simulating nest emergence. Turtles were placed in individual glass bowls containing aged seawater and exposed to oil (Gulf Coast-Mixed Crude Oil Sweet, CAS #8002-05-9, 0.833 mL/L) and/or dispersant (Corexit 9500A, 0.083 mL/L), replicating concentrations encountered during oil spills and subsequent response. Statistically significant differences between treatments and non-exposed controls were detected for PCV, AST, uric acid, glucose, calcium, phosphorus, total protein, albumin, globulin, potassium, and sodium. The principal dyscrasias reflected acute osmolar, electrolyte and hydration challenges that were more numerous and greater in combined oil/dispersant exposures at 4 days. Clinicopathological findings were supported by a failure to gain weight (associated with normal hatchling hydration in seawater) in dispersant and combination exposed hatchlings. These findings can help guide clinical response for sea turtles exposed to crude oil and crude oil/dispersant combinations, and indicate potential impacts on wildlife to consider when deploying dispersants in an oil spill response., (Copyright © 2019 Harms, McClellan-Green, Godfrey, Christiansen, Broadhurst and Godard-Codding.)

The metalloproteinase ADAM17 plays a pivotal role in initiating inflammation by releasing TNF from its precursor. Prolonged TNF release causes many chronic inflammatory diseases, indicating that tight regulation of ADAM17 activity is essential for resolution of inflammation. In this study, we report that the endogenous ADAM17 inhibitor TIMP-3 inhibits ADAM17 activity only when it is bound to the cell surface and that cell surface levels of TIMP-3 in endotoxin-activated human macrophages are dynamically controlled by the endocytic receptor LRP1. Pharmacological blockade of LRP1 inhibited endocytic clearance of TIMP-3, leading to an increase in cell surface levels of the inhibitor that blocked TNF release. Following LPS stimulation, TIMP-3 levels on the surface of macrophages increased 4-fold within 4 h and continued to accumulate at 6 h, before a return to baseline levels at 8 h. This dynamic regulation of cell surface TIMP-3 levels was independent of changes in TIMP-3 mRNA levels, but correlated with shedding of LRP1. These results shed light on the basic mechanisms that maintain a regulated inflammatory response and ensure its timely resolution., (Copyright © 2019 by The American Association of Immunologists, Inc.)

Pathologists are faced with a variety of problems when considering placental tissue in cases of stillbirth. It is recognized that there are changes which occur following fetal demise and which can complicate the assessment and may coexist with other morphological changes. It is recognized that up to 25% of stillbirths may have a recognizable abnormality causing fetal demise. A systematic review of placental tissue allows many of these disorders to be identified. This review considers macroscopic and microscopic features of placental pathology in stillbirth together with clinicopathological correlation. Stillbirth definitions, general aspects of macroscopic assessment of placentas, placental changes after fetal demise, and some recognizable causes of fetal demise are considered., Competing Interests: Disclosures & Declaration of Conflicts of Interest: The authors, reviewers, editors, and publication staff do not report any relevant conflicts of interest, (© 2019 The Authors.)

We used nuclear magnetic spectroscopy (NMR) to evaluate the metabolic impacts of crude oil, Corexit 5900A, a dispersant, and a crude oil Corexit 5900A mixture exposure on skeletal muscle, heart, and liver physiology of hatchling loggerhead sea turtles ( Caretta caretta ). Tissue samples were obtained from 22 seven-day-old hatchlings after a four day cutaneous exposure to environmentally relevant concentrations of crude oil, Corexit 5900A, a combination of crude oil and Corexit 9500A, or a seawater control. We identified 38 metabolites in the aqueous extracts of the liver, and 30 metabolites in both the skeletal and heart muscle aqueous extracts, including organic acids/osmolytes, energy compounds, amino acids, ketone bodies, nucleosides, and nucleotides. Skeletal muscle lactate, creatines, and taurine concentrations were significantly lower in hatchlings exposed to crude oil than in control hatchlings. Lactate, taurine, and cholines appeared to be the basis of some variation in hatchling heart samples, and liver inosine, uracil, and uridine appeared to be influenced by Corexit and crude oil exposure. Observed decreases in concentrations of lactate and creatines may reflect energy depletion in skeletal muscle of oil-exposed animals, while decreased taurine concentrations in these animals may reflect higher oxidative stress.

Aims: Atherosclerosis is characterized by the abundant infiltration of myeloid cells starting at early stages of disease. Myeloid cells are key players in vascular immunity during atherogenesis. However, the subsets of vascular myeloid cells have eluded resolution due to shared marker expression and atypical heterogeneity in vascular tissues. We applied the high-dimensionality of mass cytometry to the study of myeloid cell subsets in atherosclerosis., Methods and Results: Apolipoprotein E-deficient (ApoE-/-) mice were fed a chow or a high fat (western) diet for 12 weeks. Single-cell aortic preparations were probed with a panel of 35 metal-conjugated antibodies using cytometry by time of flight (CyTOF). Clustering of marker expression on live CD45+ cells from the aortas of ApoE-/- mice identified 13 broad populations of leucocytes. Monocyte, macrophage, type 1 and type 2 conventional dendritic cell (cDC1 and cDC2), plasmacytoid dendritic cell (pDC), neutrophil, eosinophil, B cell, CD4+ and CD8+ T cell, γδ T cell, natural killer (NK) cell, and innate lymphoid cell (ILC) populations accounted for approximately 95% of the live CD45+ aortic cells. Automated clustering algorithms applied to the Lin-CD11blo-hi cells revealed 20 clusters of myeloid cells. Comparison between chow and high fat fed animals revealed increases in monocytes (both Ly6C+ and Ly6C-), pDC, and a CD11c+ macrophage subset with high fat feeding. Concomitantly, the proportions of CD206+ CD169+ subsets of macrophages were significantly reduced as were cDC2., Conclusions: A CyTOF-based comprehensive mapping of the immune cell subsets within atherosclerotic aortas from ApoE-/- mice offers tools for myeloid cell discrimination within the vascular compartment and it reveals that high fat feeding skews the myeloid cell repertoire toward inflammatory monocyte-macrophage populations rather than resident macrophage phenotypes and cDC2 during atherogenesis.

We used proton nuclear magnetic resonance spectroscopy ( 1 H-NMR) to evaluate metabolic impacts of environmentally relevant crude oil and Corexit exposures on the physiology of hatchling loggerhead sea turtles ( Caretta caretta ). Sample extraction and data acquisition methods for very small volume whole blood samples and sources of variation between individual hatchlings were assessed. Sixteen unclotted, whole blood samples were obtained from 7-day-old hatchlings after a 4-day cutaneous exposure to either control seawater, crude oil, Corexit 9500A or a combination of crude oil and Corexit 9500A. After extraction, one- and two-dimensional 1 H-NMR spectra of the samples were obtained, and 17 metabolites were identified and confirmed in the whole blood spectra. Variation among samples due to the concentrations of metabolites 3-hydroxybutyrate, lactate, trimethylamine oxide and propylene glycol did not statistically correlate with treatment group. However, the characterization of the hatchling loggerhead whole blood metabolome provides a foundation for future metabolomic research with sea turtles and a basis for the study of tissues from exposed hatchling sea turtles., Competing Interests: We declare we have no competing interests.

Background: Myeloid cells are central to atherosclerotic lesion development and vulnerable plaque formation. Impaired ability of arterial phagocytes to uptake apoptotic cells (efferocytosis) promotes lesion growth and establishment of a necrotic core. The transcription factor interferon regulatory factor (IRF)-5 is an important modulator of myeloid function and programming. We sought to investigate whether IRF5 affects the formation and phenotype of atherosclerotic lesions., Methods: We investigated the role of IRF5 in atherosclerosis in 2 complementary models. First, atherosclerotic lesion development in hyperlipidemic apolipoprotein E-deficient (ApoE -/- ) mice and ApoE -/- mice with a genetic deletion of IRF5 (ApoE -/- Irf5 -/- ) was compared and then lesion development was assessed in a model of shear stress-modulated vulnerable plaque formation., Results: Both lesion and necrotic core size were significantly reduced in ApoE -/- Irf5 -/- mice compared with IRF5-competent ApoE -/- mice. Necrotic core size was also reduced in the model of shear stress-modulated vulnerable plaque formation. A significant loss of CD11c + macrophages was evident in ApoE -/- Irf5 -/- mice in the aorta, draining lymph nodes, and bone marrow cell cultures, indicating that IRF5 maintains CD11c + macrophages in atherosclerosis. Moreover, we revealed that the CD11c gene is a direct target of IRF5 in macrophages. In the absence of IRF5, CD11c - macrophages displayed a significant increase in expression of the efferocytosis-regulating integrin-β3 and its ligand milk fat globule-epidermal growth factor 8 protein and enhanced efferocytosis in vitro and in situ., Conclusions: IRF5 is detrimental in atherosclerosis by promoting the maintenance of proinflammatory CD11c + macrophages within lesions and controlling the expansion of the necrotic core by impairing efferocytosis., (© 2017 The Authors.)

Excessive or risky alcohol use is a preventable cause of significant morbidity and mortality in the U.S. and worldwide. Alcohol use is a common preventable cancer risk factor among young adults; it is associated with increased risk of developing at least six types of cancer. Alcohol consumed during early adulthood may pose a higher risk of female breast cancer than alcohol consumed later in life. Reducing alcohol use may help prevent cancer. Alcohol misuse screening and brief counseling or intervention (also called alcohol screening and brief intervention among other designations) is known to reduce excessive alcohol use, and the U.S. Preventive Services Task Force recommends that it be implemented for all adults aged ≥18 years in primary healthcare settings. Because the prevalence of excessive alcohol use, particularly binge drinking, peaks among young adults, this time of life may present a unique window of opportunity to talk about the cancer risk associated with alcohol use and how to reduce that risk by reducing excessive drinking or misuse. This article briefly describes alcohol screening and brief intervention, including the Centers for Disease Control and Prevention's recommended approach, and suggests a role for it in the context of cancer prevention. The article also briefly discusses how the Centers for Disease Control and Prevention is working to make alcohol screening and brief intervention a routine element of health care in all primary care settings to identify and help young adults who drink too much., (Published by Elsevier Inc.)

In their journey to becoming doctors, students engage with a range of teachers and trainers. Among these are simulated patients (SPs), who, through role-playing, assist students to develop their communication and physical examination skills, in contexts of formative and summative assessments. This paper explores the teaching and learning relationship between medical students and SPs, and considers how this might affect feedback and assessment. 14 SPs were interviewed on the subject of medical students' professional identity development in 2014. Data were examined using narrative analysis in conjunction with positioning theory to identify the positions that SPs assigned to themselves and to students. Narrative analysis yielded three interpretative positioning themes: Occupational, familial and cultural and discursive and embodied positioning. The interview process revealed that SPs adopt different positions intra-and interpersonally. SPs appear to hold dissonant perceptions of students in terms relating to their emerging professional identities, which may confound assessment and feedback. Training should include reflections on the SP/student relationship to uncover potential biases and positions, giving SPs the opportunity to reflect on and manage their individual and occupational selves.

Introduction: The U.S. Preventive Services Task Force recommends for adults alcohol screening and brief behavioral counseling interventions in primary care settings. However, there is a paucity of population-based data on the prevalence of alcohol screening. This study examines adherence to this U.S. Preventive Services Task Force recommendation by estimating the prevalence of alcohol screening by demographic characteristics and binge drinking., Methods: A cross-sectional analysis was conducted in 2013 and 2014 on data from the 2013 fall wave of the ConsumerStyles survey. ConsumerStyles is drawn from an Internet panel randomly recruited by probability-based sampling to be representative of the U.S., Population: Data from 2,592 adult respondents who visited primary care physicians in the last year were analyzed to determine the prevalence of alcohol screening., Results: Only 24.7% of respondents reported receiving alcohol screening. The prevalence of screening was similar among women (24.9%) and men (24.5%). Black non-Hispanics reported a significantly lower prevalence of screening than white non-Hispanics (16.2% vs 26.9%, prevalence ratio=0.60, 95% CI=0.40, 0.90). College graduates reported a significantly higher prevalence of screening than respondents with a high school degree or less (28.1% vs 20.8%, prevalence ratio=1.35, 95% CI=1.08, 1.69)., Conclusions: Only about one in four respondents who visited a primary care physician in the last year reported being screened for alcohol misuse. Therefore, many men and women who misuse alcohol are unlikely to be identified. Increased screening may help reduce alcohol misuse and related negative health outcomes., (Published by Elsevier Inc.)

Background: Alcohol is a teratogen.* Prenatal alcohol exposure is associated with a range of adverse reproductive outcomes and can cause fetal alcohol spectrum disorders (FASDs) characterized by lifelong physical, behavioral, and intellectual disabilities. FASDs are completely preventable if a woman does not drink alcohol while pregnant., Methods: CDC analyzed data from the 2011-2013 National Survey of Family Growth to generate U.S. prevalence estimates of risk for an alcohol-exposed pregnancy for 4,303 nonpregnant, nonsterile women aged 15-44 years, by selected demographic and behavioral factors. A woman was considered at risk for an alcohol-exposed pregnancy during the past month if she had sex with a male, drank any alcohol, and did not (and her partner did not with her) use contraception in the past month; was not sterile; and had a partner (or partners) not known to be sterile., Results: The weighted prevalence of alcohol-exposed pregnancy risk among U.S. women aged 15-44 years was 7.3%. During a 1-month period, approximately 3.3 million women in the United States were at risk for an alcohol-exposed pregnancy., Conclusions and Implications for Public Health Practice: Alcohol use in pregnancy is associated with low birthweight, preterm birth, birth defects, and developmental disabilities. Women of reproductive age should be informed of the risks of alcohol use during pregnancy, and contraception should be recommended, as appropriate, for women who do not want to become pregnant. Women wanting a pregnancy should be advised to stop drinking at the same time contraception is discontinued. Health care providers should advise women not to drink at all if they are pregnant or there is any chance they might be pregnant. Alcohol misuse screening and behavioral counseling (also known as alcohol screening and brief intervention) is recommended for all adults in primary care, including reproductive-aged and pregnant women, as an evidenced-based approach to reducing alcohol consumption among persons who consume alcohol in excess of the recommended guidelines.

Atherosclerosis is the major cause of cardiovascular disease (CVD), the leading cause of death worldwide. Despite much focus on lipid abnormalities in atherosclerosis, it is clear that the immune system also has important pro- and antiatherogenic functions. The enzyme indoleamine-2,3-dioxygenase (IDO) catalyses degradation of the essential amino acid tryptophan into immunomodulatory metabolites. How IDO deficiency affects immune responses during atherogenesis is unknown and we explored potential mechanisms in models of murine and human atherosclerosis. IDO deficiency in hypercholesterolemic ApoE(-/-) mice caused a significant increase in lesion size and surrogate markers of plaque vulnerability. No significant changes in cholesterol levels were observed but decreases in IL-10 production were found in the peripheral blood, spleen and lymph node B cells of IDO-deficient compared with IDO-competent ApoE(-/-) mice. 3,4,-Dimethoxycinnamoyl anthranilic acid (3,4-DAA), an orally active synthetic derivative of the tryptophan metabolite anthranilic acid, but not l-kynurenine, enhanced production of IL-10 in cultured splenic B cells. Finally, 3,4-DAA treatment reduced lesion formation and inflammation after collar-induced arterial injury in ApoE(-/-) mice, and reduced cytokine and chemokine production in ex vivo human atheroma cell cultures. Our data demonstrate that endogenous production of tryptophan metabolites via IDO is an essential feedback loop that controls atherogenesis and athero-inflammation. We show that the IDO pathway induces production of IL-10 in B cells in vivo and in vitro, suggesting that IDO may induce immunoregulatory functions of B cells in atherosclerosis. The favorable effects of anthranilic acid derivatives in atherosclerosis indicate a novel approach toward therapy of CVD.

Background. Acute bronchiolitis infection during infancy is associated with an increased risk of asthma later in life. The objective of this study was to determine if inhaled steroids are effective in preventing the development of recurrent wheeze or asthma following acute bronchiolitis. Methods. Multiple databases and bibliographies of selected references were searched. Inclusion required (a) a randomized controlled trial of inhaled steroids and control group, (b) at least 2 weeks duration of therapy started during the acute phase of disease, and (c) identification of the rate of recurrent wheeze or asthma at least 6 months after therapy. Results. Of 1410 studies reviewed, 8 reports were included in this meta-analysis (748 patients). The overall odds ratio for developing recurrent wheeze or asthma with treatment versus without treatment was 1.02 (95% confidence interval = 0.58-1.81). Conclusions. A course of inhaled steroids after acute bronchiolitis is not effective in preventing recurrent wheeze or asthma.

Scombrotoxin fish poisoning remains the primary cause of seafood poisoning outbreaks despite preventive guidelines. The purpose of this study was to investigate the use of pH for the control of growth and histamine formation by histamine-producing bacteria in fish muscle. We examined pH effects on growth and histamine formation in tuna fish infusion broth and in inoculated tuna and mahi-mahi fish muscle. Histamine production was significantly less for all bacterial strains at pH 8.5 compared to pH 5.5 in tuna fish infusion broth with no significant difference in growth. Elevated pH due to phosphate treatment of fish muscle tissues significantly reduced histamine formation with no effect on the growth of histamine-producing bacteria. This study revealed that phosphate treatment of mahi-mahi and tuna fish muscle resulted in significantly lower histamine production over 4 d of storage at 10 °C. Phosphate treatment of fish muscle may serve as a secondary barrier in addition to FDA recommended time and temperature controls for reducing public health concerns of scombrotoxin fish poisoning., (© 2015 Institute of Food Technologists®)

Objective: We have previously shown, in a cohort of untreated rheumatoid arthritis (RA) patients, that the suppressive function of Treg cells is defective. However, other studies in cohorts of patients with established RA have shown that Treg cell function is normal. We hypothesized that treatment may restore Treg cell function and lead to reduced disease activity. The aim of this study was to investigate whether treatment with methotrexate (MTX) can result in epigenetic changes that lead to restoration of the Treg cell suppressive function in RA., Methods: Peripheral blood samples from RA patients were assessed using (3) H-thymidine incorporation to measure Treg cell suppression of T cell proliferation, and by enzyme-linked immunosorbent assay to determine Treg cell suppression of interferon-γ production. CTLA-4 and FoxP3 expression was measured by flow cytometry and quantitative polymerase chain reaction (qPCR) in Treg cells from healthy individuals and RA patients. CD4+ T cells isolated from healthy individuals were cultured with interleukin-2 (IL-2), IL-6, and tumor necrosis factor α in the presence or absence of MTX, and FoxP3 expression was determined using qPCR and flow cytometry. Methylation of the FOXP3 upstream enhancer was analyzed by bisulfite sequencing PCR., Results: Defective Treg cell function was observed only in RA patients who had not been treated with MTX, whereas Treg cells from MTX-exposed RA patients had restored suppressive function. This restored suppression was associated with increased expression of FoxP3 and CTLA-4 in Treg cells. Bisulfite sequencing PCR of Treg cells cultured in MTX revealed a significant reduction in methylation of the FOXP3 upstream enhancer., Conclusion: This study identifies a novel mechanism of action of MTX, in which treatment of RA patients with MTX restores defective Treg cell function through demethylation of the FOXP3 locus, leading to a subsequent increase in FoxP3 and CTLA-4 expression., (© 2015, American College of Rheumatology.)

Background: On their journey to "becoming" doctors, medical students encounter a range of health professionals who contribute to their socialisation into clinical practice. Amongst these individuals are registered nurses (RNs) in clinical practice who are often employed by medical schools as clinical tutors. These RNs will encounter medical students on campus and later in the clinical setting., Aims: This qualitative study explored RNs' perceptions of their contribution to medical students' developing professional identities in order to provide a greater understanding of this process and ultimately inform future curriculum., Methods: This qualitative study took place in 2012 at one Australian medical school as part of a broader study exploring medical students' professional identity development from the perspectives of their teachers and trainers. Eight of the nine RNs involved in teaching procedural skills were interviewed. Recorded interviews were transcribed verbatim. Data were analysed inductively by the research team., Results: Two major themes emerged: RNs as change agents and RNs as facilitators of medical students' transition to the clinical environment. RNs as change agents related to their role modelling good practice, being patient-centred, and by emphasising factors contributing to good teamwork such as recognising and respecting individual professional roles. They facilitated students' transition to the clinical environment often through personal narratives, by offering advice on how to behave and work with members of the healthcare team, and by being a point of contact in the hospital., Conclusion: Based on their descriptions of how they role modelled good practice and how they facilitated students' transition to clinical practice, we believe that RN clinical tutors do have the experience and expertise in clinical practice and a professional approach to patients to contribute to medical students' developing professional identities as future doctors.

Non-pregnant women can avoid alcohol-exposed pregnancies (AEPs) by modifying drinking and/or contraceptive practices. The purpose of this study was to estimate the number and characteristics of women in the United States who are at risk of AEPs. We analyzed data from in-person interviews obtained from a national probability sample (i.e., the National Survey of Family Growth) of reproductive-aged women conducted from January 2002 to March 2003. To be at risk of AEP, a woman had to have met the following criteria in the last month: (1) was drinking; (2) had vaginal intercourse with a man; and (3) did not use contraception. During a 1-month period, nearly 2 million U.S. women were at risk of an AEP (95 % confidence interval 1,760,079-2,288,104), including more than 600,000 who were binge drinking. Thus, 3.4 %, or 1 in 30, of all non-pregnant women were at risk of an AEP. Most demographic and behavioral characteristics were not clearly associated with AEP risk. However, pregnancy intention was strongly associated with AEP risk (prevalence ratio = 12.0, P < 0.001) because women often continued to drink even after they stopped using contraception. Nearly 2 million U.S. women are at AEP risk and therefore at risk of having children born with fetal alcohol spectrum disorders. For pregnant women and women intending a pregnancy, there is an urgent need for wider implementation of prevention programs and policy approaches that can reduce the risk for this serious public health problem.

Introduction: Much has been written about medical students' professional identity formation, the process of "becoming" a doctor. During their training, medical students interact with a range of teachers and trainers. Among these are simulated patients (SPs) who role-play patients, assisting students with their communication, procedural, and physical examination skills. With SPs regularly interacting with students, this qualitative study explored their views of students' emerging professional identities at one Australian medical school. SPs' contributions to developing professional identities were also explored., Methods: Fourteen SPs were interviewed individually or in pairs. After template analysis of the transcripts using a priori themes, a follow-up focus group (n = 7) was arranged., Findings: Although being older (implying maturity and more life experience) and exposure to real patients and previous health care experience were identified as contributing to developing an identity as a doctor, SPs recognized that for some, an existing professional identity might impede the development of a new identity. Simulated patients were of the opinion that they contributed to students' professional identities by creating a supportive environment for honing skills, which they did by realistically role-playing patient scripts, by making their bodies available, and by providing feedback as "patients.", Conclusions: Through their authentic portrayal of patients and through their feedback, we are of the opinion that our SPs can contribute to students' developing identities as doctors. As lay individuals who often encounter students longitudinally, we believe that SPs offer a particular lens through which to view students' emerging identities as future doctors.

Although strong binding interactions between protein receptor and ligand do not require the participation of a large number of amino acids in either site, short peptide chains are generally poor at recreating the types of protein-protein interactions which take place during cell recognition and signalling process, probably because their flexible backbones prevent the side chains from forming sufficiently rigid and stable epitopes, which can take part in binding with the desired strength and specificity. In a recently-reported study, it was shown that a proto-epitope containing F, R and S amino acids has the ability to down-regulate TNF secretion by macrophages. This paper extends these findings, putting those amino acids into a short cyclic peptide scaffold, and determining the optimal configuration required to overcome the problems of conformational instability, and give rise to molecules which have potential as therapeutic agents in human disease, such as rheumatoid arthritis.

Treg-cell function is compromised in rheumatoid arthritis (RA). As the master regulator of Treg cells, FOXP3 controls development and suppressive function. Stable Treg-cell FOXP3 expression is epigenetically regulated; constitutive expression requires a demethylated Treg-specific demethylated region. Here, we hypothesised that methylation of the FOXP3 locus is altered in Treg cells of established RA patients. Methylation analysis of key regulatory regions in the FOXP3 locus was performed on Treg cells from RA patients and healthy controls. The FOXP3 Treg-specific demethylated region and proximal promoter displayed comparable methylation profiles in RA and healthy-donor Treg cells. We identified a novel differentially methylated region (DMR) upstream of the FOXP3 promoter, with enhancer activity sensitive to methylation-induced silencing. In RA Treg cells we observed significantly reduced DMR methylation and lower DNA methyltransferase (DNMT1/3A) expression compared with healthy Treg cells. Furthermore, DMR methylation negatively correlated with FOXP3 mRNA expression, and Treg cells isolated from rheumatoid factor negative RA patients were found to express significantly higher levels of FOXP3 than Treg cells from RhF-positive patients, with an associated decrease in DMR methylation. In conclusion, the novel DMR is involved in the regulation of Treg-cell FOXP3 expression, but this regulation is lost post-transcriptionally in RA Treg cells., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: Functionally impaired Treg cells expressing abnormally low levels of CTLA-4 have been well documented in rheumatoid arthritis (RA). However, the molecular defect underlying this reduced expression is unknown. The aims of this study were to assess the role of DNA methylation in regulating CTLA-4 expression in Treg cells isolated from RA patients and to elucidate the mechanism of their reduced suppressor function., Methods: CTLA-4 expression in Treg cells from RA patients and healthy controls was measured by quantitative polymerase chain reaction (PCR) and flow cytometry. Methylation of the CTLA-4 gene promoter was analyzed by bisulfite-specific PCR, followed by sequencing. Methylation-dependent transcriptional activity of the CTLA-4 gene promoter was measured by luciferase assay, and NF-AT binding to the CTLA-4 gene promoter was determined by chromatin immunoprecipitation. The role of CTLA-4 expression in controlling Teff cells was analyzed using an autologous mixed lymphocyte reaction., Results: Down-regulation of CTLA-4 expression in Treg cells from RA patients was caused by methylation of a previously unidentified NF-AT binding site within the CTLA-4 gene promoter. As a consequence, Treg cells were unable to induce expression and activation of the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO), which in turn resulted in a failure to activate the immunomodulatory kynurenine pathway., Conclusion: We show for the first time that epigenetic modifications contribute to defective Treg cell function in RA through an inability to activate the IDO pathway. Therefore, this study sets a precedent for investigating potential therapeutic strategies aimed at reinforcing the IDO pathway in RA patients., (Copyright © 2014 by the American College of Rheumatology.)

Continued development, use, and disposal of quantum dots (QDs) ensure their entrance into aquatic environments where they could pose a risk to biological organisms as whole nanoparticles or as degraded metal constituents. Reproductive Fundulus heteroclitus were fed a control diet with lecithin, diets containing 1 or 10 μg of lecithin-encapsulated CdSe/ZnS QD/day, or a diet containing 5.9 μg CdCl2/day for 85 days. Cadmium concentrations in liver, intestine, and eggs were quantified with inductively coupled plasma mass spectrometry. In fish fed 10 μg QD/day, QDs or their degradation products traversed the intestinal epithelia and accumulated in the liver. Less than 0.01% of the QD's cadmium was retained in the liver or intestinal tissues. This compares to 0.9% and 0.5% of the cadmium in the intestine and liver, respectively of fish fed a CdCl2 diet. Cadmium was also detected in the eggs from parents fed 10 μg QD/day. No significant changes in hepatic total glutathione, lipid peroxidation, or expression of genes involved in metal metabolism or oxidative stress were observed. While QDs in the diet are minimally bioavailable, unusual levels of vitellogenin transcription in male fish as well as declining fecundity require further investigation to determine if endocrine disruption is of environmental concern., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Alcohol-exposed pregnancy (AEP) is a significant public health problem in the United States. Sexually transmitted disease (STD) clinics serve female clients with a high prevalence of heavy alcohol consumption coupled with ineffective contraceptive use. Project CHOICES (Changing High-Risk AlcOhol Use and Increasing Contraception Effectiveness) is an evidence-based, brief intervention to lower risk of AEP by targeting alcohol and contraceptive behaviors through motivational interviewing and individualized feedback. We describe our experience integrating and implementing CHOICES in STD clinics. This endeavor aligns with CDC's National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention's program collaboration and service integration strategic priority to strengthen collaborative work across disease areas and integrate services provided by related programs at the client level.

Objective: To determine whether selective blockade of tumor necrosis factor receptor I (TNFRI) affects spontaneous proinflammatory cytokine and chemokine production in ex vivo-cultured human rheumatoid arthritis synovial membrane mononuclear cells (MNCs) and to compare this response to that of TNF ligand blockade using etanercept., Methods: A bispecific, single variable-domain antibody (anti-TNFRI moiety plus an albumin binding moiety [TNFRI-AlbudAb]) was used to selectively block TNFRI. Inhibition of TNFα-mediated responses in cell lines expressing TNFRI/II confirmed TNFRI-AlbudAb potency, human rhabdomyosarcoma cell line KYM-1D4 cytotoxicity, and human umbilical vein endothelial cell (HUVEC) vascular cell adhesion molecule 1 (VCAM-1) upregulation. Eighteen RA synovial membrane MNC suspensions were cultured for 2 days or 5 days, either alone or in the presence of TNFRI-AlbudAb, control-AlbudAb, or etanercept. Proinflammatory cytokines and chemokines in culture supernatants were measured by enzyme-linked immunosorbent assays. A mixed-effects statistical analysis model was used to assess the extent of TNFRI selective blockade, where the results were expressed as the percentage change with 95% confidence intervals (95% CIs)., Results: TNFRI-AlbudAb inhibited TNFα-induced KYM-1D4 cell cytotoxicity (50% inhibition concentration [IC50 ] 4 nM) and HUVEC VCAM-1 up-regulation (IC50 12 nM) in a dose-dependent manner. In ex vivo-cultured RA synovial membrane MNCs, selective blockade of TNFRI inhibited the production of proinflammatory cytokines and chemokines to levels similar to those obtained with TNF ligand blockade, without inducing cellular toxicity. Changes in cytokine levels were as follows: -23.5% (95% CI -12.4, -33.2 [P = 0.004]) for granulocyte-macrophage colony-stimulating factor, -33.4% (95% CI -20.6, -44.2 [P ≤ 0.0001]) for interleukin-10 (IL-10), -17.6% (95% CI 3.2, -34.2 [P = 0.0880]) for IL-1β, and -19.0% (95% CI -3.4, -32.1 [P = 0.0207]) for IL-6. Changes in chemokine levels were as follows: -34.2% (-14.4, -49.4 [P = 0.0030]) for IL-8, -56.6% (-30.7, -72.9 [P = 0.0011]) for RANTES, and -24.9% (2, -44.8 [P = 0.0656]) for monocyte chemotactic protein 1., Conclusion: In ex vivo-cultured RA synovial membrane MNCs, although a limited role of TNFRII cannot be ruled out, TNFRI signaling was found to be the dominant pathway leading to proinflammatory cytokine and chemokine production. Thus, selective blockade of TNFRI could potentially be therapeutically beneficial over TNF ligand blockade by retaining the beneficial TNFRII signaling., (Copyright © 2013 by the American College of Rheumatology.)

Background: Research is limited regarding the adequacy of preparation of medical students for their placement in obstetrics and gynecology. The aim of this study was to determine the perceptions of a cohort of undergraduate medical students from an Australian university and their clinical supervisors of the on-campus preparation of students for their clinical rotation in obstetrics and gynecology., Methods: We used a descriptive exploratory qualitative research approach and purposive sampling to address the aim of the study. Ten undergraduate medical students and 4 of their supervisors participated in the study. Data were collected from focus group discussions, follow-up interviews, and individual semistructured interviews. Interview transcripts were analyzed using an inductive coding approach., Results: Students and their clinical supervisors who participated in the study agreed that students should be as well prepared as possible by the university prior to their placement in obstetrics and gynecology because adequate preparation would provide a solid clinical framework upon which the discipline's knowledge and skills could be built. Overall, participants considered that the on-campus preparation was adequate in many aspects; however, they identified some specific areas in which preparation could be enhanced. These preparation enhancements included specific skills related to examining pregnant women, interpreting cardiotocography, conversing with patients and their families, and improving students' understanding of the hospital culture., Conclusion: These findings provide an increased understanding of the factors a cohort of medical students and their clinical supervisors consider essential for student preparation for the clinical rotation in obstetrics and gynecology.

The goal of the current article is to present the results of a randomized pilot investigation of a brief dynamic psychotherapy compared with treatment-as-usual (TAU) in the treatment of moderate-to-severe depression in the community mental health system. Forty patients seeking services for moderate-to-severe depression in the community mental health system were randomized to 12 weeks of psychotherapy, with either a community therapist trained in brief dynamic psychotherapy or a TAU therapist. Results indicated that blind judges could discriminate the dynamic sessions from the TAU sessions on adherence to dynamic interventions. The results indicate moderate-to-large effect sizes in favor of the dynamic psychotherapy over the TAU therapy in the treatment of depression. The Behavior and Symptom Identification Scale-24 showed that 50% of patients treated with dynamic therapy moved into a normative range compared with only 29% of patients treated with TAU., (PsycINFO Database Record (c) 2012 APA, all rights reserved.)

Objective: At-risk drinking, cigarette smoking, obesity, diabetes, and frequent mental distress, as well as their co-occurrence in childbearing aged women, are risk factors for adverse pregnancy outcomes. This study estimated the prevalence of these five risk factors individually and in combination among nonpregnant women aged 18-44 years by demographic and psychosocial characteristics, with a focus on racial and ethnic disparities., Methods: Data from the 2008 Behavioral Risk Factor Surveillance System (BRFSS) on nonpregnant women aged 18-44 years (n=54,612) were used to estimate the prevalences of five risk factors, pairs of co-occurring risk factors, and multiple risk factors for poor pregnancy outcomes., Results: The majority of women had at least one risk factor, and 18.7% had two or more risk factors. Having two or more risk factors was highest among women who were American Indian and Alaska Native (34.4%), had less than a high school education (28.7%), were unable to work (50.1%), were unmarried (23.3%), and reported sometimes, rarely, or never receiving sufficient social and emotional support (32.8%). The most prevalent pair of co-occurring risk factors was at-risk drinking and smoking (5.7%)., Conclusions: The high proportion of women of childbearing age with preconception risk factors highlights the need for preconception care. The common occurrence of multiple risk factors suggests the importance of developing screening tools and interventions that address risk factors that can lead to poor pregnancy outcomes. Increased attention should be given to high-risk subgroups.

Increasing evidence suggests that regulatory T cell (Treg) function is impaired in chronic inflammatory diseases such as rheumatoid arthritis (RA). Here we demonstrate that Tregs are unable to modulate the spontaneous production of TNF-α from RA synovial cells cultured from the diseased synovium site. Cytokine (IL-2, IL-6, TNF-α) activated T cells (Tck), cells we previously demonstrated to mimic the effector function of pathogenic RA synovial T cells, contained Tregs that survived and divided in this cytokine environment; however, the up-regulation of key molecules associated with Treg function (CTLA-4 and LFA-1) was impaired. Furthermore, Tregs were unable to suppress the function of Tcks, including contact-dependent induction of TNF-α from macrophages, supporting the concept that impaired Treg function/responsiveness contributes to chronicity of RA. However, ectopic foxp3 expression in both Tcks and pathogenic RA synovial T cells attenuated their cytokine production and function, including contact-dependent activation of macrophages. This diminished response to cytokine activation after ectopic foxp3 expression involved inhibited NF-κB activity and differed mechanistically from that displayed endogenously in conventional Tregs. These results suggest that diseases such as RA may perpetuate owing to the inability of Tregs to control cytokine-activated T-cell function. Understanding the mechanism whereby foxp3 attenuates the pathogenic function of synovial T cells may provide insight into the mechanisms of chronicity in inflammatory disease and potentially reveal new therapeutic candidates.

Computerized administration of mental health-related questionnaires has become relatively common, but little research has explored this mode of assessment in "real-world" settings. In the current study, 200 consumers at a community mental health center completed the BASIS-24 via handheld computer as well as paper and pen. Scores on the computerized BASIS-24 were compared with scores on the paper BASIS-24. Consumers also completed a questionnaire which assessed their level of satisfaction with the computerized BASIS-24. Results indicated that the BASIS-24 administered via handheld computer was highly correlated with pen and paper administration of the measure and was generally acceptable to consumers. Administration of the BASIS-24 via handheld computer may allow for efficient and sustainable outcomes assessment, adaptable research infrastructure, and maximization of clinical impact in community mental health agencies.

Background: In the context of a National Institutes of Mental Health-funded Interventions and Practice Research Infrastructure Programs (IP-RISP) grant for the treatment of depression, a partnership was developed between a community mental health organization and a team of researchers., Objectives: This paper describes the collaborative process, key challenges, and strategies employed to meet the goals of the first phase of the grant, which included development of a working and sustainable partnership and building capacity for recruitment and research., Methods: This paper was developed through the use of qualitative interviews and discussion with a variety of IP-RISP partners., Lessons Learned: Communication with multiple stakeholders through varied channels, feedback from stakeholders on research procedures, and employing a research liaison at the clinic have been key strategies in the first phase of the grant., Conclusion: The strategies we employed allowed multiple stakeholders to contribute to the larger mission of the IP-RISP and helped to establish an ongoing research program within the mental health organization.

This study was conducted to provide nationally representative findings on the prevalence and distribution of concurrent alcohol use or heavier use of alcohol and cigarette smoking among women of childbearing age with accessible health care. For the years 2003-2005, a total of 20,912 women 18-44 years of age who participated in the National Health Interview Survey (NHIS) reported that during the study period, there was a place where they would usually go for health care when sick or in need of advice about their health. The prevalence and distribution of concurrent alcohol use or heavier use of alcohol and cigarette smoking reported by such women was calculated. Logistic regression analysis was used to evaluate the "most often visited health care place" among concurrent users who reported having seen or talked to a health care provider during the previous 12 months. Among surveyed women with accessible health care, 12.3% reported concurrent alcohol use and cigarette smoking, and 1.9% reported concurrent heavier use of alcohol and cigarette smoking during the study period. Of women who reported either type of concurrent use, at least 84.4% also indicated having seen or talked to one or more health care providers during the previous 12 months. Such women were more likely than non-concurrent users to indicate that the "most often visited health care place" was a "hospital emergency room or outpatient department or some other place" or a "clinic or health center," as opposed to an "HMO or doctor's office." Concurrent alcohol use or heavier use of alcohol and cigarette smoking among women of childbearing age is an important public health concern in the United States. The findings of this study highlight the importance of screening and behavioral counseling interventions for excessive drinking and cigarette smoking by health care providers in both primary care and emergency department settings.

Histamine (or scombroid) fish poisoning is a significant cause of food borne disease in the United States. In this study, we describe the development of a molecular-based technique which uses digoxigenin (DIG) labeled DNA probes for the detection of gram negative bacteria producing high amounts of histamine (>1000 ppm). A cocktail of PCR amplification fragments corresponding to a 709 bp fragment of the histidine decarboxylase (hdc) gene of four high producing bacteria (Morganella morganii, Enterobacter aerogenes, Raoultella planticola and Photobacterium damselae) was DIG-labeled and screened against a strain bank of 152 gram negative bacteria isolated from scrombroid fish and their harvest environment. The probe cocktail reacted specifically (100%) with the high histamine producing strains but failed to react with low histamine producers and non-producers. To further evaluate the feasibility of the approach, fish homogenate inoculated with known concentrations of four high histamine producing bacterial strains was plated on modified Niven's medium (culture method) and trypticase soy agar supplemented with 2% NaCl (for colony lift hybridization). The colony lift hybridization counts did not differ significantly from the level of the initial inoculum (p>0.05), while the modified Niven's counts were significantly lower (p<0.05) than either inoculum or colony lift counts. The use of digoxigenin (DIG) labeled DNA probes with colony lift hybridization shows promise for accurate and specific enumeration of histamine producing bacteria in scombroid fish., (2010 Elsevier B.V. All rights reserved.)

Inter- and intra-population variation in the toxicity of the antifouling biocide copper pyrithione (CuPT) was examined for nauplius larvae of the barnacle Balanus amphitrite. Nauplii were collected from brooding adults from four sites within the Newport River estuary (NC), chosen based on an initial estimation of recent and historical human activities that affect local contamination levels. Each site was characterized for the presence of polycyclic aromatic hydrocarbons and for the frequency of gastropod imposex, an indicator of contamination by organotins. Sensitivity of nauplii to CuPT varied significantly across the sites/populations, with LC(50) values ranging from 4.0 microg l(-1) to 6.1 microg l(-1). Larvae from the most contaminated site were the most sensitive to CuPT. Intrapopulation variation in toxicity was investigated by exposing nauplius larvae from 15 maternal families to a fixed concentration of CuPT (6.1 microg l(-1)). Variation in larval mortality among the families was significant, ranging from 15.1% to 98.9%.

Antileishmanial therapy is suboptimal due to toxicity, high cost, and development of resistance to available drugs. Pyrazinamide (PZA) is a constituent of short-course tuberculosis chemotherapy. We investigated the effect of PZA on Leishmania major promastigote and amastigote survival. Promastigotes were more sensitive to the drug than amastigotes, with concentrations at which 50% of parasites were inhibited (MIC(50)) of 16.1 and 8.2 microM, respectively (48 h posttreatment). Moreover, 90% of amastigotes were eliminated at 120 h posttreatment, indicating that longer treatments will result in parasite elimination. Most strikingly, PZA treatment of infected C57BL/6 mice resulted in protection against disease and in a 100-fold reduction in the parasite burden. PZA treatment of J774 cells and bone marrow-derived dendritic cells and macrophages increased interleukin 12, tumor necrosis factor alpha, and activation marker expression, as well as nitric oxide production, suggesting that PZA enhances effective immune responses against the parasite. PZA treatment also activates dendritic cells deficient in Toll-like receptor 2 and 4 expression to initiate a proinflammatory response, confirming that the immunostimulatory effect of PZA is directly caused by the drug and is independent of Toll-like receptor stimulation. These results not only are strongly indicative of the promise of PZA as an alternative antileishmanial chemotherapy but also suggest that PZA causes collateral immunostimulation, a phenomenon that has never been reported for this drug.

Poisoning due to ingestion of foods with elevated levels of biogenic amines (histamine, putrescine, cadaverine, and tyramine) is well documented. Histamine fish poisoning largely is due to growth of naturally occurring bacteria associated with scombroid fish species. A rapid and reliable method is needed to screen for the presence of histamine-forming bacteria in fish. This study included a comparison of three methods for the detection of histamine-producing bacteria. A total of 152 histamine-producing and non-histamine-producing bacteria from multiple sources were screened using a modified Niven's agar method, a potentiometric method, and a PCR-based assay targeting a 709-bp fragment of the histidine decarboxylase gene. Histamine production by bacterial isolates was confirmed by high-performance liquid chromatography (HPLC). Bacterial strains were categorized as producing high amounts of histamine, low amounts of histamine, or no histamine. Of the 152 strains tested, 128 (84%) were positive with the Niven's agar method, 73 (48%) were positive with the potentiometric technique, and 74 (49%) were positive with the PCR assay. Overall, a 38% false-positive rate was observed with the modified Niven's agar method, although this method detected both low-histamine and high-histamine strains. There was a high degree of concordance (> 99%) between results of the potentiometric and PCR methods, but neither of these methods detected low-histamine bacteria. These observations support the need for a simple and straightforward yet sensitive method for detecting histamine-producing bacteria in seafood and environmental samples.

Aqueous suspensions of fullerene aggregates (aqua-nC60) were used to investigate the movement of carbon-based nanomaterials in a marine water column and to determine their effects on different life stages of a marine teleost. Fullerene aggregates formed precipitates as a result of mixing in natural seawater, and levels of aqua-nC60 were significantly increased in bottom waters after 24 h. Exposure of Fundulus heteroclitus embryos, larvae, and adults to increasing concentrations of aqua-nC60 resulted in very little mortality, and no median lethal concentrations could be calculated at < or = 10 mg/L. Aggregates of aqua-nC60 did adhere to the chorion but did not affect development of the embryos or their hatching success. Movements of aqua-nC60 through the chorion and into the embryo tended to increase with higher exposure levels; however, the concentrations were extremely low and did not differ significantly. Larvae exposed to increasing concentrations of aqua-nC60 exhibited a significant dose-dependent increase in total glutathione (GSH). This was accompanied by a decreasing trend in lipid peroxidation (LPO), but LPO was not statistically different between treatments. Adult F. heteroclitus exposed to increasing concentrations of aqua-nC60 exhibited an increase in total GSH in liver tissue but not in the gill. No significant effects on LPO were observed in either tissue. Thus, we conclude that aqua-nC60 affects the oxidative stress response of F. heteroclitus and that increased antioxidant defenses provide some physiological tolerance for these materials. Environmental factors influencing uptake, metabolism, and physiological response following exposure, however, need further investigation.

IL-10 is a potent anti-inflammatory molecule, which regulates TNF-alpha at multiple levels. We investigated whether IL-10 also modulated the activity of the TNF-alpha-converting enzyme (TACE). Using an ex vivo fluorogenic assay we observed that LPS rapidly induced TACE activity in monocytes coinciding with release of soluble TNF-alpha. In the presence of IL-10, TNF-alpha production and activation of surface TACE was significantly inhibited. Paradoxically, both LPS with or without IL-10 led to accumulation of surface TACE (albeit catalytically inactive) over a 24 h period. We investigated whether this was mediated through induction of endogenous tissue inhibitor metalloproteinase-3 (TIMP-3). We found that the inhibition of TACE activity at 2 h by IL-10 was not TIMP-3 dependent but that the late accumulation of surface TACE was prevented with TIMP-3 antibodies. Furthermore, induction of endogenous TIMP-3 was observed by western blotting in both LPS- and in LPS with IL-10-treated monocytes from 6 to 8 h of culture. These results indicate that IL-10 further regulates TNF-alpha by modulating TACE activation at early time points and by contributing to the induction of TIMP-3, the natural inhibitor of active TACE, at later time points. These observations add to our understanding of inflammation and the importance of homeostatic regulators of these events.

Background: Previously we described a system whereby human peripheral blood T cells stimulated for 8 days in a cytokine cocktail acquired effector function for contact-dependent induction of proinflammatory cytokines from monocytes. We termed these cells cytokine-activated (Tck) cells and found that the signalling pathways elicited in the responding monocytes were identical whether they were placed in contact with Tck cells or with T cells isolated from rheumatoid arthritis (RA) synovial tissue., Methods: Here, using magnetic beads and fluorescence-activated cell sorting, we extensively phenotype the Tck effector cells and conclude that effector function resides within the CD4+CD45RO+, CCR7-, CD49dhigh population, and that these cells are derived from the effector memory CD4+ T cells in resting blood., Results: After stimulation in culture, these cells produce a wide range of T-cell cytokines, undergo proliferation and differentiate to acquire an extensively activated phenotype resembling RA synovial T cells. Blocking antibodies against CD69, CD18, or CD49d resulted in a reduction of tumour necrosis factor-alpha production from monocytes stimulated with CD4+CD45RO+ Tck cells in the co-culture assay. Moreover, blockade of these ligands also resulted in inhibition of spontaneous tumour necrosis factor-alpha production in RA synovial mononuclear cell cultures., Conclusion: Taken together, these data strengthen our understanding of T-cell effector function, highlight the multiple involvement of different cell surface ligands in cell-cell contact and, provide novel insights into the pathogenesis of inflammatory RA disease.

Objectives: Maternal alcohol use is a leading preventable cause of neurobehavioral and developmental abnormalities in children. This study examines the patterns and average volume of alcohol use among U.S. women of childbearing age in order to identify subgroups of high-risk women for selective intervention., Methods: A sample of 188,290 women aged 18-44 years participated in the Centers for Disease Controls and Prevention (CDC)'s Behavioral Risk Factor Surveillance System (BRFSS) survey during the period of 2001-2003. Reported alcohol use patterns and average volume were examined for pregnant and nonpregnant women. Efforts were made to evaluate and characterize women who practiced various levels of binge drinking., Results: The results showed that approximately 2% of pregnant women and 13% of nonpregnant women in the United States engaged in binge drinking during the period of 2001-2003. Among the estimated average of 6.7 million women of childbearing age overall who engaged in binge drinking during the period, approximately 28.5% women also reported consuming an average of 5 drinks or more on typical drinking days, or about 21.4% women consumed at least 45 drinks on average in a month. Larger proportions of binge drinkers with high usual quantity of consumption were found among women of younger ages (18-24 years) or current smokers., Conclusions: Future prevention efforts should include strategies that combine health messages and encourage women of childbearing age, with particular emphasis on women 18-24 years, to avoid alcohol and tobacco use, and take multivitamins and folic acid daily for better pregnancy outcomes. Other efforts must also include broad-based implementation of screening and brief intervention for alcohol misuse in primary and women's health care settings.

Exposure to contaminants in the environment is indiscriminate and multiple species/populations of all sexes are potentially at risk. In this paper we examine the current information available on gender specific differences in invertebrates following exposure to environmental contaminants. Because of their close association with the environment and diversity of habitats, invertebrates are uniquely at risk for adverse responses to pollutants. Since 97% of all animal species are invertebrates, it would be impossible to cover each of the phyla in this review. Instead, this paper discusses major invertebrate species including insects (Periplaneta americana, Panorpa vulgaris, Lycosa hilaris, Haematobia irritans irritans (L.), and Drosophilia melanogaster), nematodes (Caenorhabditis elegans), crustaceans (Streptocephalus dichotomus, Amphiascus tenuiremis, Microarthridion littorale, Tisbe bulbisetosa, Acartia tonsa, and Palaemonetes pugio), mollusks (Pinctada fucata martensii, Ilyanassa obsoleta, Nucella lapillus, Hinia reticulata, Thais clavigera, and Mercenaria mercenaria), corals (Euphyllia ancora and Montipara capitata), and echinoderms (Asterias rubens) that have been used in studies examining the differences between males and females. Our discussion focuses on gender differences that occur in both toxicokinetic mechanisms (uptake and elimination, metabolism and physiology) and other toxicological endpoints (survival and behavior as well as morphology and development). It will become evident that the endocrine systems of invertebrates have many traits and/or pathways that are comparable to those observed in higher organisms. Yet the sensitivity of some elements of the invertebrate endocrine system, e.g., disruption of neuropeptide hormone signaling following TBT exposure, highlights the uniqueness of their systems and their potential for disruption.

Male and female organisms may have significant differences in their exposure, toxicokinetics, and response to chemicals, but gender effects have received relatively little attention, often viewed as a confounder rather than of primary importance. In this paper, we examine some of the key issues and methodologies for incorporating gender in studies of the effects of chemicals on wildlife, and explore bioindicators and biomarkers of gender effects. Examining gender-related differences in response to chemicals is complicated in wildlife because of the vast array of species, and differences in niches, lifespans, reproductive cycles and modes, and population dynamics. Further, organisms are more at risk in some ecosystems than others, which may increase the magnitude of effects. Only by studying wild animals, especially native species, can we truly understand the potential impact of gender-specific effects of chemical exposure on populations. Several factors affect gender-related differences in responses to chemicals, including exposure, age, size, seasonality, and genetic and phenotypic variation. There are clear examples where gender-related differences have had significant effects on reproductive success and population stability, including destabilization of gamete release in invertebrates, and alterations of endocrine and neuroendocrine system functioning in vertebrates. A wide range of new technologies and methods are available for examining gender-related differences in responses to chemicals. We provide examples that show that there are gender-related differences in responses to chemicals that have significant biological effects, and these gender-related differences should be taken into account by scientists, regulators, and policy makers, as well as the public.

Introduction: This study examined birth outcomes in five towns in Shoshone County, Idaho, where residents were exposed to high levels of lead in air emissions during a 6-month period after a fire had damaged the main baghouse (pollution-control device) of a local lead smelter plant in September 1973., Methods: We studied birth certificate data of 169,878 live singleton infants born to mothers who resided in Idaho at the time of delivery. The outcomes evaluated were preterm infants, small-for-gestational-age (SGA) infants, low birthweight among term infants (TLBW), and mean birthweight among term infants (TMBW). The study compared births in the five towns in Shoshone County (exposed group) to births in the rest of Idaho during three exposure periods: "pre-fire," January 1, 1970-August 31, 1973; "high exposure," September 1, 1973-December 31, 1974; and "post-fire," January 1, 1975-December 31, 1981., Results: During the high-exposure period, the exposed group had an increased prevalence of TLBW (OR=2.4; 90% CI: 1.6-3.6) and SGA (OR=1.9; 90% CI: 1.3-2.8) compared with the rest of Idaho. During the pre- and post-fire periods, the ORs for TLBW were 0.8 and 1.3, respectively, and for SGA, 1.0, and 1.3, respectively. During the high-exposure period, TMBW for the exposed group was 71 g lower than in the comparison group. The TMBW in the exposed group was 8 g lower in the pre-fire period and 26 g lower in the post-fire period than in the comparison group. The study found no increased risk for preterm birth in the exposed group., Conclusions: Maternal exposures to airborne lead emissions appeared to be associated with increased risks for SGA, TLBW, and reduced TMBW.