Degrowth - Taking Stock and Reviewing an Emerging Academic Paradigm.
Degrowth has evolved within a decade from an activist movement into a multi-disciplinary academic paradigm. However, an overview taking stock of the peer-refereed degrowth literature is yet missing. Here, we review 91 articles that were published between 2006 and 2015. We find that the academic degrowth discourse occupies a small but expanding niche at the intersection of social and applied environmental sciences. The discourse is shaped by authors from high-income, mainly Mediterranean, countries. Until 2012, articles largely constitute conceptual essays endorsed by normative claims. More recently, degrowth has branched out into modelling, empirical assessments, and the study of concrete implementations. Authors tend to agree in that economic growth cannot be sustained ad infinitum on a resource constraint planet and that degrowth requires far reaching societal change. Whether degrowth should be considered as a collectively consented choice or an environmentally-imposed inevitability constitutes a major debate among degrowth thinkers. We argue that the academic discourse could benefit from rigid hypotheses testing through input-output modelling, material flow analysis, life-cycle assessments, or social surveys. By analyzing the potentials for non-market value creation and identifying concrete well-being benefits, the degrowth discourse could receive wider public support and contribute to a paradigmatic change in the social sciences.


Cannabidiol as a Novel Candidate Alcohol Use Disorder Pharmacotherapy: A Systematic Review.
There is substantial interest in the therapeutic potential of cannabidiol (CBD), a nonpsychoactive cannabinoid found in plants of the genus Cannabis. The goal of the current systematic review was to characterize the existing literature on this topic and to evaluate the credibility of CBD as a candidate pharmacotherapy for alcohol use disorder (AUD). Using a comprehensive search strategy, 303 unique potential articles were identified and 12 ultimately met criteria for inclusion (8 using rodent models, 3 using healthy adult volunteers, and 1 using cell culture). In both rodent and cell culture models, CBD was found to exert a neuroprotective effect against adverse alcohol consequences on the hippocampus. In rodent models, CBD was found to attenuate alcohol-induced hepatotoxicity, specifically, alcohol-induced steatosis. Finally, findings from preclinical rodent models also indicate that CBD attenuates cue-elicited and stress-elicited alcohol seeking, alcohol self-administration, withdrawal-induced convulsions, and impulsive discounting of delayed rewards. In human studies, CBD was well tolerated and did not interact with the subjective effects of alcohol. Collectively, given its favorable effects on alcohol-related harms and addiction phenotypes in preclinical models, CBD appears to have promise as a candidate AUD pharmacotherapy. This is further bolstered by the absence of abuse liability and its general tolerability. A clear limitation to the literature is the paucity of human investigations. Human preclinical and clinical studies are needed to determine whether these positive effects in model systems substantively translate into clinically relevant outcomes.


Zerumbone-Induced Analgesia Modulated via Potassium Channels and Opioid Receptors in Chronic Constriction Injury-Induced Neuropathic Pain.
Zerumbone, a monocyclic sesquiterpene from the wild ginger plant Zingiber zerumbet (L.) Smith, attenuates allodynia and hyperalgesia. Currently, its mechanisms of action in neuropathic pain conditions remain unclear. This study examines the involvement of potassium channels and opioid receptors in zerumbone-induced analgesia in a chronic constriction injury (CCI) neuropathic pain mice model. Male Institute of Cancer Research (ICR) mice were subjected to CCI and behavioral responses were tested on day 14. Responses toward mechanical allodynia and thermal hyperalgesia were tested with von Frey's filament and Hargreaves' tests, respectively. Symptoms of neuropathic pain were significantly alleviated following treatment with zerumbone (10 mg/kg; intraperitoneal, i.p.). However, when the voltage-dependent K+ channel blocker tetraethylammonium (TEA, 4 mg/kg; i.p.), ATP-sensitive K+ channel blocker, glibenclamide (GLIB, 10 mg/kg; i.p.); small-conductance Ca2+-activated K+ channel inhibitor apamin (APA, 0.04 mg/kg; i.p.), or large-conductance Ca2+-activated K+ channel inhibitor charybdotoxin (CHAR, 0.02 mg/kg; i.p.) was administered prior to zerumbone (10 mg/kg; i.p.), the antiallodynic and antihyperalgesic effects of zerumbone were significantly reversed. Additionally, non-specific opioid receptors antagonist, naloxone (NAL, 10 mg/kg; i.p.), selective µ-, δ- and κ-opioid receptor antagonists; β-funaltrexamine (β-FN, 40 mg/kg; i.p.), naltrindole (20 mg/kg; s.c.), nor-binaltorphamine (10 mg/kg; s.c.) respectively attenuated the antiallodynic and antihyperalgesic effects of zerumbone. This outcome clearly demonstrates the participation of potassium channels and opioid receptors in the antineuropathic properties of zerumbone. As various clinically used neuropathic pain drugs also share this similar mechanism, this compound is, therefore, a highly potential substitute to these therapeutic options.


Thymic dendritic cells traffic to thymi of allogeneic recipients and prolong graft survival.
We have demonstrated that murine thymic dendritic cells (DCs) isolated from donor mice have the capability to home to thymi of fully allogeneic recipients after intravenous injections, where they induce T cell deletions and prolong donor-strain airway and skin graft survival. In contrast, infused splenic DCs immigrated poorly to thymi, and did not affect graft survival. These findings suggest that preferential homing may be an important mechanistic difference among subpopulations of DCs that mediate immune functions and illustrate a novel methodology that could have utility for induction of specific immunologic nonreactivity to allografts, or other disease-associated antigens.


Dogs attending primary-care practice in England with clinical signs suggestive of Chiari-like malformation/syringomyelia.
Chiari-like malformation/syringomyelia (CM/SM) in dogs describes a developmental disorder that can cause pain and reduced quality of life. This retrospective study aimed to report the period prevalence, clinical signs and risk factors for diagnosis of symptomatic CM/SM in the veterinary primary-care setting using a cross-sectional design. The study population included all dogs within the VetCompass Programme (September 1, 2009-June 13, 2014). Overall, the period prevalence of symptomatic CM/SM was 0.05 per cent (95 per cent CI 0.04 per cent to 0.06 per cent) for all breeds. The period prevalence in the Cavalier King Charles Spaniel (CKCS) was 1.6 per cent (95 per cent CI 1.2 to 2.06). Other breeds at increased odds included the King Charles Spaniel (KCS), Affenpinscher, Chihuahua and Pomeranian. Insured dogs had 4.6 times the odds (95 per cent CI 2.95 to 7.17) of having a diagnosis of CM/SM compared with uninsured dogs. Pain was the most commonly associated clinical sign (67 dogs, 72 per cent). Analgesics were prescribed to 72 (77.4 per cent) of the symptomatic dogs. Despite its low overall period prevalence, the high proportion of affected dogs identified with chronic pain suggests a significant welfare issue. Financial implications could impede the diagnostic process and lead to underestimation of the true prevalence. This study may help to inform clinicians about the clinical relevance and the need for improved awareness of clinical signs, particularly in high-risk breeds, to optimise the management of CM/SM in primary-care practice.


Evening and morning plasma levels of protein S100B in patients with obstructive sleep apnea.
Responsible for sleep brain perfusion changes, obstructive sleep apnea (OSA) constitutes a cardiovascular risk. To find out about any diffuse damage to the brain tissue, we studied the S100B protein, whose serum level is known to rise in stroke and craniocerebral trauma. 60 men (mean age 51.7+/-11.8 years) referred to us for OSA without any major comorbidity were examined polygraphically. S100B was determined with electrochemiluminiscence immunoassay (ECLIA) from evening and morning blood samples. All sixty men were diagnosed with OSA. The difference between the evening level of S100B 0.068+/-0.030 microg/l and the morning level 0.059+/-0.029 microg/l was significant (p=0.0004). Patients with mild OSA were found to have the evening S100B 0.063+/-0.023 microg/l, the morning level 0.042+/-0.012 microg/l, the difference being significant (p=0.00051). In moderate OSA the difference between the evening -0.070+/-0.017 microg/l and morning levels -0.055+/-0.025 microg/l was less significant (p=0.043). In severe OSA no difference was found between the evening and morning concentrations of S100B (0.070+/-0.036 microg/l and 0.070+/-0.031 microg/l respectively). The difference between the evening and morning S100B levels correlated negatively with AHI and ODI and positively with basal saturation and average minimal oxygen saturation. Sleep with signs of severe OSA influences S100B protein release.


Unrecognized left main coronary artery disease in patients undergoing interventional procedures.
Selective, coronary arteriographic, catheter-based, intravascular ultrasound images were obtained to determine the presence and extent of angiographically undetected or underestimated left main (LM) coronary arterial narrowing in patients receiving coronary interventional therapy. Coronary arteriograms were determined to be either normal or abnormal by visual inspection. Abnormal arteriograms were digitized and quantitated using a semiautomated edge-detection algorithm. Thirty-eight patients receiving percutaneous treatment of stenoses in the left coronary artery system were studied. Optimal LM coronary angiograms were obtained in 2 views, and intravascular ultrasound images were obtained after the coronary interventional procedure. Intravascular ultrasound detected plaque in 24 of 27 angiographically normal LM arteries (89%), whereas narrowing was observed in 11 of 11 angiographically abnormal LM arteries (100%). Eight of 38 patients (21%) had > 40% area stenosis by intravascular ultrasound. In patients with angiographic disease, there was no correlation between quantitative angiographic and ultrasound percent area stenosis (r = 0.12; p = 0.72; SEE 19%). The median plaque area was not different between angiographically normal (0.05 cm2; 0.03, 0.08 [25th, 75th percentile]) and abnormal (0.06 cm2; 0.03, 0.1) patients. The median percent area stenosis in arteriographically normal subjects (26%; 14, 32%) was less than that in abnormal ones (37%; 20, 46%) (p = 0.03). Unrecognized LM disease is widespread and often underestimated in patients with normal LM angiograms undergoing interventional procedures. Plaque area is similar for angiographically normal and insignificantly abnormal vessels. This study suggests that intravascular ultrasound overcomes the limitations of silhouette imaging and can be a clinically useful, adjunctive method to evaluate LM coronary artery disease.


The twin-arginine translocation (Tat) protein export pathway.
The twin-arginine translocation (Tat) protein export system is present in the cytoplasmic membranes of most bacteria and archaea and has the highly unusual property of transporting fully folded proteins. The system must therefore provide a transmembrane pathway that is large enough to allow the passage of structured macromolecular substrates of different sizes but that maintains the impermeability of the membrane to ions. In the Gram-negative bacterium Escherichia coli, this complex task can be achieved by using only three small membrane proteins: TatA, TatB and TatC. In this Review, we summarize recent advances in our understanding of how this remarkable machine operates.


Antibacterial activity of cefprozil in vitro.
Cefprozil is a new orally active cephalosporin which is undergoing in vitro and in vivo evaluations. Using the standard agar dilution method we compared the in vitro activity of this drug with other oral cephalosporins and ciprofloxacin against 637 recent clinical isolates from the Kaohsiung Veterans General Hospital, Taiwan. Against Escherichia coli and Klebsiella pneumoniae, cefprozil showed good activity, inhibiting over 80% of these isolates at 8 mg/l. Like other oral drugs of its class, it had little activity against Pseudomonas aeruginosa and Acinetobacter spp. Against Haemophilus influenzae, irrespective of beta-lactamase production, its activity was similar to comparable drugs. Against methicillin-susceptible Staphylococcus aureus, cefprozil showed high activity, inhibiting 90% of these isolates at 4 mg/l, whereas against methicillin-resistant S. aureus, its activity was higher than that of other oral cephalosporins tested and was similar to that of ciprofloxacin. Of enterococci tested, 57.7% were inhibited by 8 mg/l of cefprozil. Against beta-hemolytic streptococci, its activity was superior to all other drugs tested. The results of this in vitro study indicate that oral administration of cefprozil might be efficacious in the treatment of community-acquired cutaneous, respiratory and urinary tract infections.


Participatory modelling to support decision making in water management under uncertainty: two comparative case studies in the Guadiana river basin, Spain.
A participatory modelling process has been conducted in two areas of the Guadiana river (the upper and the middle sub-basins), in Spain, with the aim of providing support for decision making in the water management field. The area has a semi-arid climate where irrigated agriculture plays a key role in the economic development of the region and accounts for around 90% of water use. Following the guidelines of the European Water Framework Directive, we promote stakeholder involvement in water management with the aim to achieve an improved understanding of the water system and to encourage the exchange of knowledge and views between stakeholders in order to help building a shared vision of the system. At the same time, the resulting models, which integrate the different sectors and views, provide some insight of the impacts that different management options and possible future scenarios could have. The methodology is based on a Bayesian network combined with an economic model and, in the middle Guadiana sub-basin, with a crop model. The resulting integrated modelling framework is used to simulate possible water policy, market and climate scenarios to find out the impacts of those scenarios on farm income and on the environment. At the end of the modelling process, an evaluation questionnaire was filled by participants in both sub-basins. Results show that this type of processes are found very helpful by stakeholders to improve the system understanding, to understand each other's views and to reduce conflict when it exists. In addition, they found the model an extremely useful tool to support management. The graphical interface, the quantitative output and the explicit representation of uncertainty helped stakeholders to better understand the implications of the scenario tested. Finally, the combination of different types of models was also found very useful, as it allowed exploring in detail specific aspects of the water management problems.


Cannabidiol: Influence on B Cells, Peripheral Blood Mononuclear Cells, and Peripheral Blood Mononuclear Cell/Rheumatoid Arthritis Synovial Fibroblast Cocultures.
Background: Cannabidiol (CBD), one major nonintoxicating phytocannabinoid from Cannabis sativa demonstrated anti-inflammatory effects in animal models of several inflammatory conditions, including arthritis. However, it is still unknown which cell types mediate these anti-inflammatory effects of CBD, and, since CBD binds to a plethora of receptors and enzymes, it is complicated to pinpoint its mechanism of action. In this study, we elucidate the effects of CBD on B cells and peripheral blood mononuclear cells (PBMCs) in respect to survival, calcium mobilization, drug uptake, and cytokine (IL-6, IL-10, and TNF) and immunoglobulin production. Methods: Modulation of intracellular calcium and drug uptake in B cells was determined by using the fluorescent dyes Cal-520 and PoPo3, respectively. Cytokine and immunoglobulin production was evaluated by enzyme-linked immunosorbent assay. PBMC composition and B cell survival after CBD treatment was assessed by flow cytometry. Results: B cells express two major target receptors for CBD, TRPV2 (transient receptor potential vanilloid 2) and TRPA1 (transient receptor potential ankyrin 1), which are not regulated by B cell activation. CBD increased intracellular calcium levels in mouse and human B cells, which was accompanied by enhanced uptake of PoPo3. These effects were not dependent on transient receptor potential channel activation. CBD increased the number of early apoptotic B cells at the expense of viable cells and diminished interleukin (IL)-10 and tumor necrosis factor (TNF) production when activated T cell independently. In PBMCs, CBD increased IL-10 production when B cells were activated T cell dependent, while decreasing TNF levels when activated T cell independently. In PBMC/rheumatoid synovial fibroblast cocultures, CBD reduced IL-10 production when B cells were activated T cell independently. Immunoglobulin M production was augmented by CBD when B cells were activated with CpG. Conclusion: CBD is able to provide pro- and anti-inflammatory effects in isolated B cells and PBMCs. This is dependent on the activating stimulus (T cell dependent or independent) and concentration of CBD. Therefore, CBD might be used to dampen B cell activity in autoimmune conditions such as rheumatoid arthritis, in which B cells are activated by specific autoantigens.


Molecular basis of sidekick-mediated cell-cell adhesion and specificity.
Sidekick (Sdk) 1 and 2 are related immunoglobulin superfamily cell adhesion proteins required for appropriate synaptic connections between specific subtypes of retinal neurons. Sdks mediate cell-cell adhesion with homophilic specificity that underlies their neuronal targeting function. Here we report crystal structures of Sdk1 and Sdk2 ectodomain regions, revealing similar homodimers mediated by the four N-terminal immunoglobulin domains (Ig1-4), arranged in a horseshoe conformation. These Ig1-4 horseshoes interact in a novel back-to-back orientation in both homodimers through Ig1:Ig2, Ig1:Ig1 and Ig3:Ig4 interactions. Structure-guided mutagenesis results show that this canonical dimer is required for both Sdk-mediated cell aggregation (via trans interactions) and Sdk clustering in isolated cells (via cis interactions). Sdk1/Sdk2 recognition specificity is encoded across Ig1-4, with Ig1-2 conferring the majority of binding affinity and differential specificity. We suggest that competition between cis and trans interactions provides a novel mechanism to sharpen the specificity of cell-cell interactions.


Eosinophil infiltration and degranulation in normal human tissue.
Localization of eosinophil granule major basic protein by immunofluorescence permits recognition of both eosinophil infiltration and degranulation. Over the past decade and a half, our laboratory has shown that eosinophil infiltration and degranulation occur in many diseased tissues in humans; among normal tissues studied as controls, only the gut showed striking eosinophil infiltration and degranulation. Using an indirect immunofluorescence procedure for the detection of major basic protein, we extended our analyses of normal human tissues to include tissues from essentially all body organs; a total of 117 biopsy/autopsy specimens were analyzed. To determine whether the method of tissue procurement affected the level of eosinophil degranulation in the normal gastrointestinal tract, normal proximal jejunum from six patients was biopsied using either an endoscopic forceps or a scalpel at the time of elective surgery and examined by immunofluorescence. Spleen, lymph node, and thymus tissues showed eosinophil infiltration with scant evidence of degranulation, but the only organ showing both eosinophil infiltration and remarkable degranulation was the gastrointestinal tract. Eosinophil degranulation was significantly increased in specimens obtained by endoscopic forceps compared to those obtained by scalpel (P = 0.021). These results indicate that tissue procurement methods affect the degree of eosinophil degranulation in the gut. Thus, among normal human body organs, both eosinophil infiltration and appreciable degranulation consistently occur only in the gut.


Carbon assimilation profiles as a tool for identification of zygomycetes.
Identification of Zygomycetes is difficult and time-consuming by standard microbiological procedures. Carbon assimilation profiles are commonly used for yeast-and bacterial-species identification but rarely for filamentous-fungus identification. Carbon assimilation profiles were evaluated using the commercialized kits ID32C and API 50 CH, which contain 31 and 49 tests, respectively, to serve as simple tools for species identification of Zygomycetes in clinical microbiology laboratories. Fifty-seven strains belonging to 15 species and varieties of Zygomycetes, including Rhizopus, Absidia, Mucor, and Rhizomucor species, were tested for intra- and interspecies variability based on their carbon assimilation profiles. Using ID32C strips, 6 tests were always positive, 7 were never positive, and 18 showed consistently different results between species. With API 50 CH strips, 15 tests were positive for all species, 13 were never positive, and 21 showed different results between species. Nevertheless, assimilation patterns were highly variable among Rhizopus oryzae isolates, and it was not possible to define a specific carbon assimilation profile. With both ID32C and API CH 50 strips, intraspecies variation was found to be low, while large differences were found between genera and species. The clustering of isolates based on their carbon assimilation profiles was in accordance with DNA-based phylogeny of Zygomycetes. In conclusion, carbon assimilation profiles allowed precise and accurate identification of most Zygomycetes to the species level.


Implementing a standardized community-based cardiovascular risk assessment program in 20 Ontario communities.
The aim of the study is to describe the implementation of the Cardiovascular Health Awareness Program (CHAP) in 20 mid-sized communities across Ontario, Canada, and identify key factors in the successful multi-site delivery of a collaborative cardiovascular risk assessment and management program. Lead organizations were identified and contracted following a request for proposals. An Implementation Guide detailed steps in community mobilization and delivery of volunteer-led pharmacy-based cardiovascular risk assessment sessions. Process data were collected through final reports; a debriefing meeting; and interviews with program staff. All 20 communities successfully implemented CHAP. Overall, 99% (338/341) of family physicians agreed to receive assessment results and 89% (129/145) of pharmacies held sessions. Five hundred and seventy-seven volunteers conducted 27,358 risk assessments for 15,889 unique participants. Essential program components were consistently included, however, variations in materials, processes and support occurred. Factors in program success included: local expertise, centralized support, identification and engagement of local physician and pharmacist opinion leaders and a balance of standardization and flexibility. Monitoring delivery of a multi-community cardiovascular risk assessment program yielded key factors in program success to inform development of a sustainable and transferable model.


Access to knowledge and the Global Abortion Policies Database.
Research shows that women, healthcare providers, and even policy makers worldwide have limited or inaccurate knowledge of the abortion law and policies in their country. These knowledge gaps sometimes stem from the vague and broad terms of the law, which breed uncertainty and even conflict when unaccompanied by accessible regulation or guidelines. Inconsistency across national law and policy further impedes safe and evidence-based practice. This lack of transparency creates a crisis of accountability. Those seeking care cannot know their legal entitlements, service providers cannot practice with legal protection, and governments can escape legal responsibility for the adverse effects of their laws. This is the context for the newly launched Global Abortion Policies Database-an open-access repository that seeks to promote transparency and state accountability by providing clear and comprehensive information about national laws, policies, health standards, and guidelines, and by creating the capacity for comparative analysis and cross-referencing to health indicators, WHO recommendations, and human rights standards.


S-Metric calculation by considering dominated hypervolume as Klee's measure problem.
The dominated hypervolume (or S-metric) is a commonly accepted quality measure for comparing approximations of Pareto fronts generated by multi-objective optimizers. Since optimizers exist, namely evolutionary algorithms, that use the S-metric internally several times per iteration, a fast determination of the S-metric value is of essential importance. This work describes how to consider the S-metric as a special case of a more general geometric problem called Klee's measure problem (KMP). For KMP, an algorithm exists with runtime O(n log n + n(d/2) log n), for n points of d >or= 3 dimensions. This complex algorithm is adapted to the special case of calculating the S-metric. Conceptual simplifications realize the algorithm without complex data structures and establish an upper bound of O(n(d/2) log n) for the S-metric calculation for d >or= 3. The performance of the new algorithm is studied in comparison to another state of the art algorithm on a set of academic test functions.


Biophysical analyses of synthetic amyloid-beta(1-42) aggregates before and after covalent cross-linking. Implications for deducing the structure of endogenous amyloid-beta oligomers.
A neuropathological hallmark of Alzheimer's disease (AD) is the presence of large numbers of senile plaques in the brain. These deposits are rich in fibrils that are composed of 40- and 42-residue amyloid-beta (Abeta) peptides. Several lines of evidence indicate that soluble Abeta aggregates as well as fibrils are important in the etiology of AD. Low levels of endogenous soluble Abeta aggregates make them difficult to characterize, but several species in extracts of AD brains have been detected by gel electrophoresis in sodium dodecyl sulfate (SDS) and immunoblotting. Individual Abeta oligomers ranging in size from dimers through dodecamers of 4 kDa monomeric Abeta have been resolved in other laboratories as discrete species by size exclusion chromatography (SEC). In an effort to reconstitute soluble Abeta aggregates in vitro that resemble the endogenous soluble Abeta aggregates, we previously found that monomeric Abeta(1-42) rapidly forms soluble oligomers in the presence of dilute SDS micelles. Here we extend this work in two directions. First, we contrast the size and secondary structure of these oligomers with those of synthetic Abeta(1-42) fibrils. SEC and multiangle light scattering were used to obtain a molecular mass of 150 kDa for the isolated oligomers. The oligomers partially dissociated to monomers through nonamers when incubated with SDS, but in contrast to endogenous oligomers, we saw no evidence of these discrete species prior to SDS treatment. One hypothesis to explain this difference is that endogenous oligomers are stabilized by covalent cross-linking induced by unknown cellular agents. To explore this hypothesis, optimal mass spectrometry (MS) analysis procedures need to be developed for Abeta cross-linked in vitro. In our second series of studies, we began this process by treating monomeric and aggregated Abeta(1-42) with three cross-linking agents: transglutaminase, glutaraldehyde, and Cu(II) with peroxide. We compared the efficiency of covalent cross-linking with these agents, the effect of cross-linking on peptide secondary structure, the stability of the cross-linked structures to thermal unfolding, and the sites of peptide cross-linking obtained from proteolysis and MS.


Factors associated with number of duodenal samples obtained in suspected celiac disease.
Many people with celiac disease are undiagnosed and there is evidence that insufficient duodenal samples may contribute to underdiagnosis. The aims of this study were to investigate whether more samples leads to a greater likelihood of a diagnosis of celiac disease and to elucidate factors that influence the number of samples collected. We identified patients from two community hospitals who were undergoing duodenal biopsy for indications (as identified by International Classification of Diseases code) compatible with possible celiac disease. Three cohorts were evaluated: no celiac disease (NCD, normal villi), celiac disease (villous atrophy, Marsh score 3), and possible celiac disease (PCD, Marsh score < 3). Endoscopic features, indication, setting, trainee presence, and patient demographic details were evaluated for their role in sample collection. 5997 patients met the inclusion criteria. Patients with a final diagnosis of celiac disease had a median of 4 specimens collected. The percentage of patients diagnosed with celiac disease with one sample was 0.3 % compared with 12.8 % of those with six samples ( P = 0.001). Patient factors that positively correlated with the number of samples collected were endoscopic features, demographic details, and indication ( P = 0.001). Endoscopist factors that positively correlated with the number of samples collected were absence of a trainee, pediatric gastroenterologist, and outpatient setting ( P < 0.001). Histological diagnosis of celiac disease significantly increased with six samples. Multiple factors influenced whether adequate biopsies were taken. Adherence to guidelines may increase the diagnosis rate of celiac disease.


Effect of UK policy on medical migration: a time series analysis of physician registration data.
Economically developed countries have recruited large numbers of overseas health workers to fill domestic shortages. Recognition of the negative impact this can have on health care in developing countries led the United Kingdom Department of Health to issue a Code of Practice for National Health Service (NHS) employers in 1999 providing ethical guidance on international recruitment. Case reports suggest this guidance had limited influence in the context of other NHS policy priorities. The temporal association between trends in new professional registrations from doctors qualifying overseas and relevant United Kingdom government policy is reported. Government policy documents were identified by a literature review; further information was obtained, when appropriate, through requests made under the Freedom of Information Act. Data on new professional registration of doctors were obtained from the General Medical Council (GMC). New United Kingdom professional registrations by doctors trained in Africa and south Asia more than doubled from 3105 in 2001 to 7343 in 2003, as NHS Trusts sought to achieve recruitment targets specified in the 2000 NHS Plan; this occurred despite ethical guidance to avoid active recruitment of doctors from resource-poor countries. Registration of such doctors declined subsequently, but in response to other government policy initiatives. A fall in registration of South African-trained doctors from 3206 in 2003 to 4 in 2004 followed a Memorandum of Understanding with South Africa signed in 2003. Registrations from India and Pakistan fell from a peak of 4626 in 2004 to 1169 in 2007 following changes in United Kingdom immigration law in 2005 and 2006. Since 2007, registration of new doctors trained outside the European Economic Area has remained relatively stable, but in 2010 the United Kingdom still registered 722 new doctors trained in Africa and 1207 trained in India and Pakistan. Ethical guidance was ineffective in preventing mass registration by doctors trained in resource-poor countries between 2001 and 2004 because of competing NHS policy priorities. Changes in United Kingdom immigration laws and bilateral agreements have subsequently reduced new registrations, but about 4000 new doctors a year continue to register who trained in Africa, Asia and less economically developed European countries.


Student placement and skill ranking predictors for programming classes using class attitude, psychological scales, and code metrics.
In some situations, it is necessary to measure personal programming skills. For example, often students must be divided according to skill level and motivation to learn or companies recruiting employees must rank candidates by evaluating programming skills through programming tests, programming contests, etc. This process is burdensome because teachers and recruiters must prepare, implement, and evaluate a placement examination. This paper tries to predict the placement and ranking results of programming contests via machine learning without such an examination. Explanatory variables used for machine learning are classified into three categories: Psychological Scales, Programming Tasks, and Student-answered Questionnaires. The participants are university students enrolled in a Java programming class. One target variable is the placement result based on an examination by a teacher of a class and the ranking results of the programming contest. Our best classification model with a decision tree has an F-measure of 0.912, while our best ranking model with an SVM-rank has an nDCG of 0.962. In both prediction models, the best explanatory variable is from the Programming Task followed in order by Psychological Sale and Student-answered Questionnaire. Our classification model uses 9 explanatory variables, while our ranking model uses 20 explanatory variables. These include all three types of explanatory variables. The source code complexity, which is a source code metrics from Programming Task, shows best performance when the prediction uses only one explanatory variable. Contribution (1), this method can automate some of the teacher's workload, which may improve educational quality and increase the number of acceptable students in the course. Contribution (2), this paper shows the potential of using difficult-to-formulate information for an evaluation such as a Psychological Scale is demonstrated. These are the contributions and implications of this paper.


Genomic structure and transcript variants of the bovine DAZL gene.
The Deleted in AZoospermia Like (DAZL) gene is a member of the DAZ family and encodes an RNA-binding protein that is expressed in prenatal and postnatal germ cells of males and females. In the human, there are five highly-related members in the DAZ family, four (DAZ1-4) on the Y chromosome and one (DAZL) on an autosome (HSA3). Mutations in these genes have been linked to severe spermatogenic failure and infertility in men. In the present study, we have cloned and characterized the bovine DAZL (bDAZL) gene. The full-length bDAZL cDNA is predicted to encode a protein of 295 amino acids with an RNA recognition motif. The deduced protein sequence of bDAZL is 96 and 97% similar to human and mouse DAZL, respectively. Fluorescence in situ hybridization (FISH) maps bDAZL to the distal region on BTA1q. The bDAZL gene consists of 11 exons and 10 introns. A bDAZL pseudogene was identified on BTA16. Expression analysis of bDAZL in 13 different tissues by RT-PCR shows that two transcripts, variant 1 (2,996 bp) and variant 2 (1,373 bp), of the bDAZL gene are detected only in testis mRNA. The variants probably result from alternative RNA splicing as variant 1 contains an additional 1,623-bp insertion in the 3' UTR. Our results lay the groundwork for possible single nucleotide polymorphism (SNP) and functional studies of the DAZL gene in cattle.


Curcumin reverses T cell-mediated adaptive immune dysfunctions in tumor-bearing hosts.
Immune dysfunction is well documented during tumor progression and likely contributes to tumor immune evasion. CD8(+) cytotoxic T lymphocytes (CTLs) are involved in antigen-specific tumor destruction and CD4(+) T cells are essential for helping this CD8(+) T cell-dependent tumor eradication. Tumors often target and inhibit T-cell function to escape from immune surveillance. This dysfunction includes loss of effector and memory T cells, bias towards type 2 cytokines and expansion of T regulatory (Treg) cells. Curcumin has previously been shown to have antitumor activity and some research has addressed the immunoprotective potential of this plant-derived polyphenol in tumor-bearing hosts. Here we examined the role of curcumin in the prevention of tumor-induced dysfunction of T cell-based immune responses. We observed severe loss of both effector and memory T-cell populations, downregulation of type 1 and upregulation of type 2 immune responses and decreased proliferation of effector T cells in the presence of tumors. Curcumin, in turn, prevented this loss of T cells, expanded central memory T cell (T(CM))/effector memory T cell (T(EM)) populations, reversed the type 2 immune bias and attenuated the tumor-induced inhibition of T-cell proliferation in tumor-bearing hosts. Further investigation revealed that tumor burden upregulated Treg cell populations and stimulated the production of the immunosuppressive cytokines transforming growth factor (TGF)-beta and IL-10 in these cells. Curcumin, however, inhibited the suppressive activity of Treg cells by downregulating the production of TGF-beta and IL-10 in these cells. More importantly, curcumin treatment enhanced the ability of effector T cells to kill cancer cells. Overall, our observations suggest that the unique properties of curcumin may be exploited for successful attenuation of tumor-induced suppression of cell-mediated immune responses.


Conformational biosensors reveal GPCR signalling from endosomes.
A long-held tenet of molecular pharmacology is that canonical signal transduction mediated by G-protein-coupled receptor (GPCR) coupling to heterotrimeric G proteins is confined to the plasma membrane. Evidence supporting this traditional view is based on analytical methods that provide limited or no subcellular resolution. It has been subsequently proposed that signalling by internalized GPCRs is restricted to G-protein-independent mechanisms such as scaffolding by arrestins, or GPCR activation elicits a discrete form of persistent G protein signalling, or that internalized GPCRs can indeed contribute to the acute G-protein-mediated response. Evidence supporting these various latter hypotheses is indirect or subject to alternative interpretation, and it remains unknown if endosome-localized GPCRs are even present in an active form. Here we describe the application of conformation-specific single-domain antibodies (nanobodies) to directly probe activation of the β2-adrenoceptor, a prototypical GPCR, and its cognate G protein, Gs (ref. 12), in living mammalian cells. We show that the adrenergic agonist isoprenaline promotes receptor and G protein activation in the plasma membrane as expected, but also in the early endosome membrane, and that internalized receptors contribute to the overall cellular cyclic AMP response within several minutes after agonist application. These findings provide direct support for the hypothesis that canonical GPCR signalling occurs from endosomes as well as the plasma membrane, and suggest a versatile strategy for probing dynamic conformational change in vivo.


Simvastatin impairs exercise training adaptations.
This study sought to determine if simvastatin impairs exercise training adaptations. Statins are commonly prescribed in combination with therapeutic lifestyle changes, including exercise, to reduce cardiovascular disease risk in patients with metabolic syndrome. Statin use has been linked to skeletal muscle myopathy and impaired mitochondrial function, but it is unclear whether statin use alters adaptations to exercise training. This study examined the effects of simvastatin on changes in cardiorespiratory fitness and skeletal muscle mitochondrial content in response to aerobic exercise training. Sedentary overweight or obese adults with at least 2 metabolic syndrome risk factors (defined according to National Cholesterol Education Panel Adult Treatment Panel III criteria) were randomized to 12 weeks of aerobic exercise training or to exercise in combination with simvastatin (40 mg/day). The primary outcomes were cardiorespiratory fitness and skeletal muscle (vastus lateralis) mitochondrial content (citrate synthase enzyme activity). Thirty-seven participants (exercise plus statins: n = 18; exercise only: n = 19) completed the study. Cardiorespiratory fitness increased by 10% (p < 0.05) in response to exercise training alone, but was blunted by the addition of simvastatin resulting in only a 1.5% increase (p < 0.005 for group by time interaction). Similarly, skeletal muscle citrate synthase activity increased by 13% in the exercise-only group (p < 0.05), but decreased by 4.5% in the simvastatin-plus-exercise group (p < 0.05 for group-by-time interaction). Simvastatin attenuates increases in cardiorespiratory fitness and skeletal muscle mitochondrial content when combined with exercise training in overweight or obese patients at risk of the metabolic syndrome. (Exercise, Statins, and the Metabolic Syndrome; NCT01700530).


A Nationwide Survey of Hepatitis E Virus Infection and Chronic Hepatitis in Heart and Kidney Transplant Recipients in Japan.
Recently, chronic hepatitis E has been reported in solid organ transplant (SOT) recipients in European countries. Previously, we clarified the prevalence of hepatitis E virus (HEV) infection in Japanese liver transplant recipients and identified 2 chronic hepatitis E patients infected by blood transfusion. However, the rate of HEV infection in recipients of SOTs other than liver in Japan remains unclear, so we conducted a nationwide survey to clarify the prevalence of chronic HEV infection in Japanese heart and kidney transplant recipients. A total of 99 heart and 2526 kidney transplant recipients in 17 hospitals in Japan were examined for the presence of the IgG class of anti-HEV antibodies as well as for serum HEV RNA. The prevalence of anti-HEV IgG among heart and kidney transplant recipients was 7.07% (7/99) and 4.08% (103/2526), respectively. One heart transplant patient (1.01%) and 11 kidney transplant patients (0.44%) were found to be positive for HEV RNA. The HEV isolates from all viremic patients were typed as genotype 3. Four patients developed chronic hepatitis E after transplantation. Three patients were treated with ribavirin; their liver enzymes normalized, and HEV RNA became negative immediately. Sustained virologic response was achieved in all cases. This is the first nationwide survey of HEV infection in Japanese heart and kidney transplant recipients. The prevalence of anti-HEV IgG and HEV RNA in heart and kidney transplant recipients in Japan was lower than that in European countries. Of note, 42% of viremic transplant patients developed chronic hepatitis.


HER2-associated radioresistance of breast cancer stem cells isolated from HER2-negative breast cancer cells.
To understand the role of HER2-associated signaling network in breast cancer stem cells (BCSC) using radioresistant breast cancer cells and clinical recurrent breast cancers to evaluate HER2-targeted therapy as a tumor eliminating strategy for recurrent HER2(-/low) breast cancers. HER2-expressing BCSCs (HER2(+)/CD44(+)/CD24(-/low)) were isolated from radiation-treated breast cancer MCF7 cells and in vivo irradiated MCF7 xenograft tumors. Tumor aggressiveness and radioresistance were analyzed by gap filling, Matrigel invasion, tumor-sphere formation, and clonogenic survival assays. The HER2/CD44 feature was analyzed in 40 primary and recurrent breast cancer specimens. Protein expression profiling in HER2(+)/CD44(+)/CD24(-/low) versus HER2(-)/CD44(+)/CD24(-/low) BCSCs was conducted with two-dimensional difference gel electrophoresis (2-D DIGE) and high-performance liquid chromatography tandem mass spectrometry (HPLC/MS-MS) analysis and HER2-mediated signaling network was generated by MetaCore program. Compared with HER2-negative BCSCs, HER2(+)/CD44(+)/CD24(-/low) cells showed elevated aldehyde dehydrogenase (ALDH) activity and aggressiveness tested by Matrigel invasion, tumor sphere formation, and in vivo tumorigenesis. The enhanced aggressive phenotype and radioresistance of the HER2(+)/CD44(+)/CD24(-/low) cells were markedly reduced by inhibition of HER2 via siRNA or Herceptin treatments. Clinical breast cancer specimens revealed that cells coexpressing HER2 and CD44 were more frequently detected in recurrent (84.6%) than primary tumors (57.1%). In addition, 2-D DIGE and HPLC/MS-MS of HER2(+)/CD44(+)/CD24(-/low) versus HER2(-)/CD44(+)/CD24(-/low) BCSCs reported a unique HER2-associated protein profile including effectors involved in tumor metastasis, apoptosis, mitochondrial function, and DNA repair. A specific feature of HER2-STAT3 network was identified. This study provides the evidence that HER2-mediated prosurvival signaling network is responsible for the aggressive phenotype of BCSCs that could be targeted to control the therapy-resistant HER2(-/low) breast cancer.


Short-term effects of school-based weight gain prevention among adolescents.
To determine whether a multicomponent health promotion intervention for Dutch adolescents would be successful in influencing body composition and aerobic fitness. Randomized controlled trial. Ten intervention and 8 control prevocational secondary schools. A total of 978 adolescents (mean age, 12.7 years). An interdisciplinary multicomponent intervention program with an adapted curriculum for 11 biology and physical education lessons and environmental change options, including additional lessons on physical education and advice on the school canteen selection. Body height and weight, hip and waist circumference, 4 skinfold thickness measurements, and aerobic fitness. Multilevel analyses showed significant differences in changes after the 8-month intervention period in favor of the intervention group with regard to hip circumference (mean difference, 0.53 cm; 95% confidence interval, 0.07 to 0.98) and sum of skinfolds among girls (mean difference, -2.31 mm; 95% confidence interval, -4.34 to -0.28). In boys, the intervention resulted in a significant difference in waist circumference (mean difference, -0.57 cm; 95% confidence interval, -1.10 to -0.05). No significant intervention effects were found related to aerobic fitness. The multicomponent Dutch Obesity Intervention in Teenagers program positively influenced several measures of body composition among both girls and boys. Our results indicate that secondary prevocational school curriculum changes may contribute to excessive weight gain prevention among adolescents.


D1 and D2 dopamine receptor distribution in the neuroleptic nonresponsive and neuroleptic responsive lines of mice, a quantitative receptor autoradiographic study.
The present study uses quantitative receptor autoradiography to examine D1 and D2 receptor binding in the neuroleptic responsive and neuroleptic nonresponsive (NNR) lines of mice. The neuroleptic responsive and NNR mice have differed for at least eight generations by an order of magnitude in their sensitivity (ED50s) to catalepsy induced by neuroleptics with a high D2/D1 receptor activity profile. Across the entire rostral-caudal dimensions of the lateral caudate-putamen (CPu), of the dorsomedial CPu, the nucleus accumbens and substantia nigra zona reticulata, we found no difference in the density of [3H]SCH 23390 binding sites. [3H]Spiroperidol binding sites were not different in the dorsomedial CPu or nucleus accumbens but were significantly decreased (20-30%) in the NNR line in the caudal aspect of the lateral CPu. The NNR line also showed a significant elevation of D2 autoreceptors across all the midbrain dopamine cell groups (A8, A9 and A10); the increases ranged from 20 to 50%. Overall, the data show that selection of mice for response and nonresponse to neuroleptic-induced catalepsy is associated with significant changes in D2 but not D1 receptor density.


An examination of PTSD symptoms and relationship functioning in U.S. soldiers of the Iraq war over time.
We examined associations between overall posttraumatic stress disorder (PTSD) symptoms, symptom clusters of PTSD (reexperiencing, avoidance, dysphoria, and arousal), and relationship adjustment cross sectionally and longitudinally using self-report measures from a dyadic sample of U.S. National Guard soldiers from the Iraq war and their intimate partners (N = 49 couples). Results of multilevel modeling revealed that Time 1 PTSD symptom severity significantly predicted lower relationship adjustment as rated by partners at Time 2 after controlling for baseline relationship adjustment (β = -.20, p = .025). Total PTSD symptoms did not significantly predict soldiers' ratings of relationship adjustment at Time 2. For soldiers, the PTSD symptom cluster of dysphoria was uniquely and significantly related to relationship adjustment ratings both at Time 1 and at Time 2, controlling for Time 1 adjustment. For partners, none of the soldiers' PTSD symptoms clusters was uniquely associated with Time 1 relationship adjustment or with change in adjustment over time. In contrast, findings regarding the effect of relationship adjustment on changes in PTSD over time found that Time 1 relationship adjustment was not associated with changes in PTSD symptoms at Time 2.


Neural tube, skeletal and body wall defects in mice lacking transcription factor AP-2.
The retinoic acid-inducible transcription factor AP-2 is expressed in epithelial and neural crest cell lineages during murine development. AP-2 can regulate neural and epithelial gene transcription, and is associated with overexpression of c-erbB-2 in human breast-cancer cell lines. To ascertain the importance of AP-2 for normal development, we have derived mice containing a homozygous disruption of the AP-2 gene. These AP-2-null mice have multiple congenital defects and die at birth. In particular, the AP-2 knockout mice exhibit anencephaly, craniofacial defects and thoraco-abdominoschisis. Skeletal defects occur in the head and trunk region, where many bones are deformed or absent. Analysis of these mice earlier in embryogenesis indicates a failure of cranial neural-tube closure and defects in cranial ganglia development. We have shown that AP-2 is a fundamental regulator of mammalian craniofacial development.


Anatomical variation of celiac axis, superior mesenteric artery, and hepatic artery: Evaluation with multidetector computed tomography angiography.
The celiac axis, superior mesenteric artery (SMA), and hepatic artery are the most important branches of abdominal aorta due to their vascularization field. The aim of our study was to evaluate the prevalence of different anatomical variation of celiac axis, SMA, hepatic artery, and its branches with multidetector computed tomography (MDCT) angiography of upper abdomen arteries. MDCT of 607 kidney donor and traumatic patients that referred to MDCT unit at Al Zahra Hospital in Isfahan from 2012 to 2015 were retrospectively evaluated. We excluded patients with history of abdominal vascular surgery and hepatic or pancreatic surgery. Computed tomography images of the patient were obtained with 64-row MDCT scanner and anatomical variations were analyzed. Three hundred and eighty-eight (63.9%) of the 607 patients had classic arterial anatomy and 219 (36.1%) patients had variant types. The most common type of variation was the origin of the right hepatic artery (RHA) from SMA (9.6%), and the next common variation was the origin of the left hepatic artery (LHA) from the left gastric artery (6.9%). Variations in the origin of the common hepatic artery (CHA) were seen in 16 (2.6%) patients. Buhler arc was identified in two patients. The RHA originated from the celiac axis in 11 (1.8%) patients and from the aorta in 8 (1.3%) patients. Trifurcation of CHA into gastroduodenal artery, RHA, and LHA was detected in 11 (1.8%) patients. The results of the present study showed that anatomical variation occurs in a high percentage of patients. Detection of these variations can guide surgical and radiological interventional planning.


Kidney growth and hypertrophy: the role of mTOR and vesicle trafficking.
The kidney, like other organs, grows in constant proportion to the rest of the body. When one kidney is removed, the remaining one hypertrophies. In a comprehensive series of studies, Chen et al. show that growth during maturation is mediated by the mTORC1 signaling pathway, which is induced by EGF-like peptides, and requires PI3K, PDK, AKT, mTORC2, and activation of mTORC1 through the combined effects of TSC and RHEB as part of a multiprotein complex localized on lysosomes. However, compensatory growth is mediated by amino acids, which act on mTORC1 independently of the previous pathway, and requires a class III PI3K (VPS34) that is known to be involved in vesicle trafficking to the lysosomes.


Tunable synthetic phenotypic diversification on Waddington's landscape through autonomous signaling.
Phenotypic diversification of cells is crucial for developmental and regenerative processes in multicellular organisms. The diversification concept is described as the motion of marbles rolling down Waddington's landscape, in which the number of stable states changes as development proceeds. In contrast to this simple concept, the complexity of natural biomolecular processes prevents comprehension of their design principles. We have constructed, in Escherichia coli, a synthetic circuit with just four genes, which programs cells to autonomously diversify as the motion on the landscape through cell-cell communication. The circuit design was based on the combination of a bistable toggle switch with an intercellular signaling system. The cells with the circuit diversified into two distinct cell states, "high" and "low," in vivo and in silico, when all of the cells started from the low state. The synthetic diversification was affected by not only the shape of the landscape determined by the circuit design, which includes the synthesis rate of the signaling molecule, but also the number of cells in the experiments. This cell-number dependency is reminiscent of the "community effect": The fates of developing cells are determined by their number. Our synthetic circuit could be a model system for studying diversification and differentiation in higher organisms. Prospectively, further integrations of our circuit with different cellular functions will provide unique tools for directing cell fates on the population level in tissue engineering.


Prospective study of POLG mutations presenting in children with intractable epilepsy: prevalence and clinical features.
To assess the frequency and clinical features of childhood-onset intractable epilepsy caused by the most common mutations in the POLG gene, which encodes the catalytic subunit of mitochondrial DNA polymerase gamma. Children presenting with nonsyndromic intractable epilepsy of unknown etiology but without documented liver dysfunction at presentation were eligible for this prospective, population-based study. Blood samples were analyzed for the three most common POLG mutations. If any of the three tested mutations were found, all the exons and the exon-intron boundaries of the POLG gene were sequenced. In addition, we retrospectively reviewed the notes of patients presenting with intractable epilepsy in which we had found POLG mutations. All available clinical data were collected by questionnaire and by reviewing the medical records. We analyzed 213 blood DNA samples from patients fulfilling the inclusion criteria of the prospective study. Among these, five patients (2.3%) were found with one of the three common POLG mutations as homozygous or compound heterozygous states. In addition, three patients were retrospectively identified. Seven of the eight patients had either raised cerebrospinal fluid (CSF) lactate (n = 3) or brain magnetic resonance imaging (MRI) changes (n = 4) at presentation with intractable epilepsy. Three patients later developed liver dysfunction, progressing to fatal liver failure in two without previous treatment with sodium valproate (VPA). Furthermore, it is worth mentioning that one patient presented first with an autism spectrum disorder before seizures emerged. Mutations in POLG are an important cause of early and juvenile onset nonsyndromic intractable epilepsy with highly variable associated manifestations including autistic features. This study emphasizes that genetic testing for POLG mutations in patients with nonsyndromic intractable epilepsies is very important for clinical diagnostics, genetic counseling, and treatment decisions because of the increased risk for VPA-induced liver failure in patients with POLG mutations. We recommend POLG gene testing for patients with intractable seizures and at least one elevated CSF lactate or suggestive brain MRI changes (predominantly abnormal T2 -weighted thalamic signal) with or without status epilepticus, epilepsia partialis continua, or liver manifestations typical for Alpers disease, especially when the disease course is progressive.


Chronic calf pain in athletes due to sural nerve entrapment. A report of 18 cases.
We retrospectively analyzed the charts of 13 athletes (18 limbs) who had sural nerve entrapment localized in the passage of the nerve through the superficial sural aponeurosis. There were 11 men and 2 women (average age, 43 years; range, 31 to 59). All patients reported chronic calf pain that was exacerbated during physical exertion. Delay to diagnosis averaged 9 months (range, 5 to 24). Tenderness in the calf was identified along the course of the sural nerve in all cases. In 10 patients (15 limbs) electrodiagnostic testing before surgery was positive. After failure of nonoperative treatment, surgery was conducted under local anesthesia. Neurolysis was performed by incising the superficial sural aponeurosis and the fibrous band in it through which the nerve passes. The results of the operation were evaluated in terms of residual symptoms, ability to return to the former sport, and degree of patient satisfaction. A final follow-up examination was performed an average of 14 months (range, 6 to 30) after the operation. The final result was excellent in 9 limbs (2 bilateral), good in 8 limbs (2 bilateral), and fair in 1 case. The differential diagnosis of sural nerve entrapment in athletes is discussed. Increase in sural muscle mass or development of local fibrous scar tissue compromised the sural nerve in its course through the unyielding and inextensible superficial sural aponeurosis.


Antenatal corticosteroids: the good, the bad and the unknown.
The purpose of this review was to summarize recently published clinical and animal-based studies of antenatal corticosteroids, describe current recommendations for practice, and suggest priority areas for future research. A single course of antenatal corticosteroids is known to improve newborn lung function after preterm birth and to reduce the risk of newborn death. Current evidence from animal and clinical studies suggests that additional numbers of courses do not further improve lung function and are associated with risks of adverse consequences. These risks principally include restricted intrauterine growth, altered brain development, postnatal insulin resistance and changes to the hypothalamic-pituitary-adrenal axis. The research challenges in this field are now centred on uncovering the mechanisms by which cortisol is involved in programming the fetus for its future metabolic life, and discovering ways in which the effectiveness and safety of antenatal glucocorticoids can be enhanced.


Immunoglobulin recognition of fecal bacteria in stunted and non-stunted children: findings from the Afribiota study.
Child undernutrition is a global health issue that is associated with poor sanitation and an altered intestinal microbiota. Immunoglobulin (Ig) A mediates host-microbial homeostasis in the intestine, and acutely undernourished children have been shown to have altered IgA recognition of the fecal microbiota. We sought to determine whether chronic undernutrition (stunting) or intestinal inflammation were associated with antibody recognition of the microbiota using two geographically distinct populations from the Afribiota project. Fecal bacteria from 200 children between 2 and 5 years old in Antananarivo, Madagascar, and Bangui, Central African Republic (CAR), were sorted into IgA-positive (IgA+) and IgA-negative (IgA-) populations by flow cytometry and subsequently characterized by 16S rRNA gene sequencing to determine IgA-bacterial targeting. We additionally measured IgG+ fecal bacteria by flow cytometry in a subset of 75 children. Stunted children (height-for-age z-score ≤ -2) had a greater proportion of IgA+ bacteria in the fecal microbiota compared to non-stunted controls. This trend was consistent in both countries, despite the higher overall IgA-targeting of the microbiota in Madagascar, but lost significance in each country individually. Two of the most highly IgA-recognized bacteria regardless of nutritional status were Campylobacter (in CAR) and Haemophilus (in both countries), both of which were previously shown to be more abundant in stunted children; however, there was no association between IgA-targeting of these bacteria and either stunting or inflammatory markers. IgG-bound intestinal bacteria were rare in both stunted and non-stunted children, similar to levels observed in healthy populations. Undernourished children carry a high load of intestinal pathogens and pathobionts. Our data suggest that stunted children have a greater proportion of IgA-recognized fecal bacteria. We moreover identify two putative pathobionts, Haemophilus and Campylobacter, that are broadly targeted by intestinal IgA. This study furthers our understanding of host-microbiota interactions in undernutrition and identifies immune-recognized microbes for future study.


Effect of copper-doped silicate 13-93 bioactive glass scaffolds on the response of MC3T3-E1 cells in vitro and on bone regeneration and angiogenesis in rat calvarial defects in vivo.
The release of inorganic ions from biomaterials could provide an alternative approach to the use of growth factors for improving tissue healing. In the present study, the release of copper (Cu) ions from bioactive silicate (13-93) glass scaffolds on the response of cells in vitro and on bone regeneration and angiogenesis in vivo was studied. Scaffolds doped with varying concentrations of Cu (0-2.0wt.% CuO) were created with a grid-like microstructure by robotic deposition. When immersed in simulated body fluid in vitro, the Cu-doped scaffolds released Cu ions into the medium in a dose-dependent manner and converted partially to hydroxyapatite. The proliferation and alkaline phosphatase activity of pre-osteoblastic MC3T3-E1 cells cultured on the scaffolds were not affected by 0.4 and 0.8wt.% CuO in the glass but they were significantly reduced by 2.0wt.% CuO. The percent new bone that infiltrated the scaffolds implanted for 6weeks in rat calvarial defects (46±8%) was not significantly affected by 0.4 or 0.8wt.% CuO in the glass whereas it was significantly inhibited (0.8±0.7%) in the scaffolds doped with 2.0wt.% CuO. The area of new blood vessels in the fibrous tissue that infiltrated the scaffolds increased with CuO content of the glass and was significantly higher for the scaffolds doped with 2.0wt.% CuO. Loading the scaffolds with bone morphogenetic protein-2 (1μg/defect) significantly enhanced bone infiltration and reduced fibrous tissue in the scaffolds. These results showed that doping the 13-93 glass scaffolds with up to 0.8wt.% CuO did not affect their biocompatibility whereas 2.0wt.% CuO was toxic to cells and detrimental to bone regeneration.


A new aporocotylid (Digenea) species from blood vascular system of gag grouper, Mycteroperca microlepis (Perciformes: Serranidae), off Alabama, with an emendation of Pearsonellum Overstreet & Køie, 1989.
Pearsonellum lemusi n. sp. (Digenea: Aporocotylidae) infects the blood vascular system of the gag grouper, Mycteroperca microlepis (Perciformes: Serranidae), in the north central Gulf of Mexico, approximately 80 km south of Dauphin Island, Alabama (29°34'09″N, 88°22'16″W). The new species can be most easily differentiated from its only congeners Pearsonellum corventum Overstreet and Køie, 1989 (type species) and Pearsonellum pygmaeus Nolan and Cribb, 2004 , both of which infect Australian serranids, by the combination of having a large adult body (3,237 × 570 µm), a cecal intersection comprising an elongated medial channel, anterior ceca >10% of total body length, ovary narrower than testis, and pre-ovarian uterus not looping between testis and ovary. The embryonated eggs of the new species infect gill epithelium, are spheroid, and measure 25-30 µm in diameter. Sympatric Gulf of Mexico serranids were negative for aporocotylid infections: coney, Cephalopholis fulva (n = 1); Nassau grouper, Epinephelus striatus (3); red grouper, Epinephelus morio (32); yellowedge grouper, Epinephelus flavolimbatus (1); rock hind, Epinephelus adscensionis (1); red hind, Epinephelus guttatus (2); Warsaw grouper, Epinephelus nigritus (3); graysby, Cephalopholis cruentata (1); black grouper, Mycteroperca bonaci (1), and tattler, Serranus phoebe (2). The new species is the first aporocotylid described from a serranid outside of the southwestern Pacific Ocean. The diagnosis of Pearsonellum Overstreet and Køie, 1989 is herein emended to include anterior sucker having concentric rows of spines anterior to mouth, pharynx absent, esophagus length <1/2 total body length, vas deferens connecting with cirrus sac anteromedially, ovary occupying posterior 1/4-1/3 of body, primary vitelline duct dextral, and oviducal seminal receptacle extending posteriad in parallel with lateral body margin, not transverse nor constricted anteriorly or posteriorly by sharp bends or kinks.


Mitochondrial fission, fusion, and stress.
Mitochondrial fission and fusion play critical roles in maintaining functional mitochondria when cells experience metabolic or environmental stresses. Fusion helps mitigate stress by mixing the contents of partially damaged mitochondria as a form of complementation. Fission is needed to create new mitochondria, but it also contributes to quality control by enabling the removal of damaged mitochondria and can facilitate apoptosis during high levels of cellular stress. Disruptions in these processes affect normal development, and they have been implicated in neurodegenerative diseases, such as Parkinson's.


Cancer risk assessment using genetic panel testing: considerations for clinical application.
With the completion of the Human Genome Project and the development of high throughput technologies, such as next-generation sequencing, the use of multiplex genetic testing, in which multiple genes are sequenced simultaneously to test for one or more conditions, is growing rapidly. Reflecting underlying heterogeneity where a broad range of genes confer risks for one or more cancers, the development of genetic cancer panels to assess these risks represents just one example of how multiplex testing is being applied clinically. There are a number of issues and challenges to consider when conducting genetic testing for cancer risk assessment, and these issues become exceedingly more complex when moving from the traditional single-gene approach to panel testing. Here, we address the practical considerations for clinical use of panel testing for breast, ovarian, and colon cancers, including the benefits, limitations and challenges, genetic counseling issues, and management guidelines.


Familial incidence of insomnia.
This study evaluated the familial incidence of sleep disturbances among individuals with insomnia complaints. The sample consisted of 285 patients evaluated for insomnia at a sleep disorders clinic. All patients completed a sleep survey and underwent a semistructured clinical interview as part of their initial evaluation of insomnia. Information on the presence and nature of sleep disturbances among their family members (first- and second-degree relatives) was obtained from a sleep survey. The findings indicate that 35% of patients consulting for insomnia had a positive family history of sleep disturbances. Insomnia was the most common type of sleep disturbance identified (76%) and the mother was the most frequently afflicted family member. Reports of sleep disturbances among a family member were more prevalent when the onset of insomnia was before 40-years-old than when it was later in life. A positive family history was slightly higher when the insomnia complaint involved sleep-onset difficulties relative to sleep-maintenance or mixed insomnias. Although the present findings suggest that a positive family history of insomnia may be a potential risk factor for insomnia, it is unclear whether this reflects a genetic predisposition or a social learning phenomenon.


The reliability of the cervical relocation test on people with and without a history of neck pain.
Physical therapy intervention is often sought to treat cervical spine conditions and a comprehensive physical therapy examination has been associated with more favourable outcomes. The cervical relocation test (CRT) is one method used to assess joint position sense (PS) integrity of the cervical spine. Previous research has found significant differences in the CRT between symptomatic and asymptomatic subjects. Impaired kinaesthetic awareness in the cervical spine may be associated with degenerative joint disease, chronicity of the complaint and increased susceptibility to re-injury. The purpose of this study was to determine the intertester and intratester reliability of cervical relocation using the cervical range of motion instrument (CROM) and an affixed laser (AL) device among subjects with and without a history of neck pain. In addition, it was hypothesised that those individuals with a history of neck pain would have greater difficulty on the CRT. A total of 50 asymptomatic subjects (n = 50) were assigned to two researchers. The CRT was performed for each tester by the subject rotating the cervical spine for three trials to the right and left for the CROM and AL. The results indicate a significant intertester reliability of the CROM (interclass correlation coefficient (ICC) = 0.717[0.502-0.839]; 0.773[0.595-0.873]) for the subjects in this sample. This study demonstrated that the CROM is a reliable device for measuring cervical relocation between different testers. Future research should investigate if the CRT is predictive of prognosis in patients with cervical pathology.


Dissociation of bovine cytochrome c1 subcomplex and the status of cysteine residues in the subunits.
Purified bovine heart two-band cytochrome c1 subcomplex was dissociated by treatment with p-chloromercuribenzoic acid (pCMB) into its heme subunit and a colorless subunit called hinge protein, which is essential for the formation of cytochrome c1-c complex. The subcomplex was found by titration to react with 4 mol of pCMB per mol of cytochrome c1. The contents of mercury of the dissociated heme subunit and the hinge protein were 3 and 1 mol per mol of polypeptide, respectively. These results, together with the sequence analysis, indicated that the three cysteine residues in cytochrome c1 heme subunit not involved in heme-binding existed in free thiol form. One of the five cysteine residues in the hinge protein was in free form and four in two disulfide bonds. The dissociated hinge protein was digested with staphylococcal protease and the cysteine-containing peptides were separated by reversed-phase high-performance liquid chromatography (HPLC). The content of mercury and the result of performic acid oxidation of cystine peptides revealed that Cys-30 existed in free thiol form and two disulfide bridges were formed between Cys-24 and Cys-68 and between Cys-40 and Cys-54. The conformation of the hinge protein was predicted to be composed largely of either two-alpha-helical or four-alpha-helical conformation with the amino (N)-terminal 20 residues being in a random structure.


Treatment of Periradicular Bone Defect by Periosteal Pedicle Graft as a Barrier Membrane and Demineralized Freeze-Dried Bone Allograft.
The purpose of this case report is to describe the usefulness of Periosteal Pedicle Graft (PPG) as a barrier membrane and Demineralized Freeze-Dried Bone Allograft (DFDBA) for bone regeneration in periradicular bone defect. A patient with intraoral discharging sinus due to carious exposed pulp involvement was treated by PPG and DFDBA. Clinical and radiological evaluations were done immediately prior to surgery, three months, six months and one year after surgery. Patient was treated using split-thickness flap, PPG, apicoectomy, defect fill with DFDBA and lateral displacement along with suturing of the PPG prior to suturing the flap, in order to close the communication between the oral and the periapical surroundings through sinus tract opening. After one year, successful healing of periradicular bone defect was achieved. Thus, PPG as a barrier membrane and DFDBA have been shown to have the potential to stimulate bone formation when used in periradicular bone defect.


Global migration and the changing distribution of sickle haemoglobin: a quantitative study of temporal trends between 1960 and 2000.
Changes in the geographical distribution of genetic disorders are often thought to happen slowly, especially when compared with infectious diseases. Whereas mutations, genetic drift, and natural selection take place over many generations, epidemics can spread through large populations within a few days or weeks. Nevertheless, population movements can interfere with these processes, and few studies have been done of their eff ect on genetic disorders. We aimed to investigate the eff ect of global migration on the distribution of the sickle-cell gene-the most common and clinically significant haemoglobin structural variant. For each country, we extracted data from the World Bank's Global Bilateral Migration Database about international human migrations between 1960 and 2000. We combined this information with evidence-based estimates of national HbS allele frequencies, generated within a Bayesian geostatistical framework, to analyse temporal changes in the net numbers of migrants, and classified countries with an index summarising these temporal trends. The number of international migrants increased from 92.6 million in 1960, to 165.2 million in 2000. The estimated global number of migrants with HbS increased from about 1.6 million in 1960, to 3.6 million in 2000. This increase was largely due to an increase in the number of migrants from countries with HbS allele frequencies higher than 10%, from 3.1 million in 1960, to 14.2 million in 2000. Additionally, the mean number of countries of origin for each destination country increased from 70 (SE 46) in 1960, to 98 (48) in 2000, showing an increasing diversity in the network of international migrations between countries. Our index of change map shows a patchy distribution of the magnitude of temporal changes, with the highest positive and negative values scattered across all continents. Global human population movements have had a substantial eff ect on the distribution of the HbS gene. Population movements can create a long-term burden on health-care systems. Our findings, which emphasise countries in which migration fluxes are changing the most, should increase awareness about the global burden of haemoglobinopathies and encourage policy makers to implement specific public health interventions, such as screening programmes and genetic counselling. Wellcome Trust, European Research Council, Bill & Melinda Gates Foundation, National Institute of Allergy and Infectious Diseases-National Institutes of Health, the Research and Policy for Infectious Disease Dynamics program, Fogarty International Center.


Melanosome maturation defect in Rab38-deficient retinal pigment epithelium results in instability of immature melanosomes during transient melanogenesis.
Pathways of melanosome biogenesis in retinal pigment epithelial (RPE) cells have received less attention than those of skin melanocytes. Although the bulk of melanin synthesis in RPE cells occurs embryonically, it is not clear whether adult RPE cells continue to produce melanosomes. Here, we show that progression from pmel17-positive premelanosomes to tyrosinase-positive mature melanosomes in the RPE is largely complete before birth. Loss of functional Rab38 in the "chocolate" (cht) mouse causes dramatically reduced numbers of melanosomes in adult RPE, in contrast to the mild phenotype previously shown in skin melanocytes. Choroidal melanocytes in cht mice also have reduced melanosome numbers, but a continuing low level of melanosome biogenesis gradually overcomes the defect, unlike in the RPE. Partial compensation by Rab32 that occurs in skin melanocytes is less effective in the RPE, presumably because of the short time window for melanosome biogenesis. In cht RPE, premelanosomes form but delivery of tyrosinase is impaired. Premelanosomes that fail to deposit melanin are unstable in both cht and tyrosinase-deficient RPE. Together with the high levels of cathepsin D in immature melanosomes of the RPE, our results suggest that melanin deposition may protect the maturing melanosome from the activity of lumenal acid hydrolases.


Intraperitoneal immunotherapy of epithelial ovarian carcinoma with Corynebacterium parvum.
Corynebacterium parvum was administered intraperitoneally to 21 patients with epithelial ovarian cancer. Nineteen patients had surgically measurable disease and two received adjuvant therapy. Surgically confirmed responses were documented in six of 19 patients (31.6%), with two complete responses (10.5%) and four partial responses (21.1%). Three patients (15.8%) had stable disease, and 10 patients (52.6%) had disease progression. The mean survival of the patients who had a complete response was 35.5 months; the four patients who had a partial response the mean survival was 26.6 months, and of the nonresponders the mean survival was 12.6 months (p less than 0.02). The mean survival of the entire group was 18.2 months. Initial response and patient survival correlated with the amount of disease pretreatment. Thus six responding patients had less than or equal to 5 mm maximum diameter tumors, that is, minimal residual disease. Toxicity in the 86 courses of therapy included abdominal pain in 78% of cases, fever in 56%, nausea in 40%, and vomiting in 22%. Stimulation of cytotoxic lymphocytes resulted from the administration of C. parvum, which induced a significant increase of both intraperitoneal natural killer lymphocyte cytotoxicity and antibody-dependent cell-mediated cytotoxicity in six of nine patients tested; these two types of cytotoxicity correlated with response to therapy and may be partially responsible for the surgically documented tumor regression. While the clinical usefulness of intraperitoneal C. parvum is limited because of its toxicity, intraperitoneal immunotherapy may prove useful in patients with minimal residual ovarian cancer when more refined agents become available.


Perspective. How many women have osteoporosis?
Osteoporosis is widely viewed as a major public health concern, but the exact magnitude of the problem is uncertain and likely to depend on how the condition is defined. Noninvasive bone mineral measurements can be used to define a state of heightened fracture risk (osteopenia), or the ultimate clinical manifestation of fracture can be assessed (established osteoporosis). If bone mineral measurements more than 2 standard deviations below the mean of young normal women represent osteopenia, then 45% of white women aged 50 years and over have the condition at one or more sites in the hip, spine, or forearm on the basis of population-based data from Rochester, Minnesota. A smaller proportion is affected at each specific skeletal site: 32% have bone mineral values this low in the lumbar spine, 29% in either of two regions in the proximal femur, and 26% in the midradius. Although this overall estimate is substantial, some other serious chronic diseases are almost as common. More importantly, low bone mass is associated with adverse health outcomes, especially fractures. The lifetime risk of any fracture of the hip, spine, or distal forearm is almost 40% in white women and 13% in white men from age 50 years onward. If the enormous costs associated with these fractures are to be reduced, increased attention must be given to the design and implementation of control programs directed at this major health problem.


First Report of Bursaphelenchus yongensis on Pinus rigida in Korea.
During a pinewood nematode survey in 2011, a dead pitch pine (Pinus rigida) was found in Daejeon city, Korea. Samples were taken from several regions of the trunk and nematodes were extracted from the wood samples by a modified Baermann funnel method (1). Recovered nematodes were then cultured on Botrytis cinerea grown in potato dextrose agar (PDA) medium. Males and females (n = 10) were examined under an inverted light microscope. Morphologically, these nematodes were identified as Bursaphelenchus yongensis sharing the characters of the genus Bursaphelenchus including a large median bulb, offset cephalic region, a well-developed stylet with small basal knobs, female with vulva at 70% of body length, and males with terminal bursa. Females of B. yongensis were similar to B. muconatus and B. fraudulentus by presence of tail mucron, but they were differentiated by a smaller vulval flap. Males had spicules with small cucullus, pointed rostrum, and with strongly dorsally bent condylus. The identity of the nematode was confirmed through molecular diagnosis. DNA was extracted from each of 10 nematodes using commercial DNeasy kits (Qiagen, Valencia, CA) and the ITS regions were amplified with a primer set by PCR (Bio Rad, US/PTC-0220) (3,5). Amplified ITS DNA was cloned and sequenced, and the sequence data was deposited to GenBank (Accession No. KJ857070). The ITS sequence of Korean isolate (KJ857070) showed 99% similarity to that from Chinese isolate in GenBank (AM180513). ITS-RFLP patterns using five different restriction enzymes, RsaI, HaeIII, MspI, HinfI, and AluI, matched 100% with those previously reported for B. yongensis (2,4). Host pathogenicity tests were conducted from June to August on saplings of five native and introduced conifer species found in Korea (Abies sp., Larix leptolepsis, P. koraiensis, P. strobus, and P. thunbergii). All saplings were 5 to 6 years old and 15 saplings were inoculated for each species. B. yongensis was cultures on B. cinerea for 10 days at 25°C and the inoculum concentration was calibrated at 5,000 nematodes/50 μl sterile water. Nematodes were artificially inoculated by making a 1-cm-long incision into the stem using a sterile razor blade. B. yongensis was pathogenic on L. leptolepsis, causing wilt and death in all inoculated saplings; however, the nematode had no effect on P. thunbergii, P. koraiensis, Abies sp., or P. strobus. To our knowledge, this is the first report of B. yongensis in Korea. This nematode is a potential threat to L. leptolepsis; however, the role of B. yongensis in the mortality of P. rigida is uncertain at this time. References: (1) S. M. Ayoub. Plant Nematology in Agricultural Training Aid. Department of Food and Agriculture, Division of Plant Industry, Sacramento, CA, 1977. (2) W. Burgermeister et al. Nematology 11:649, 2009. (3) V. R. Ferris et al. Fund. Appl. Nematol. 16:177, 1993. (4) J. Gu et al. Russian J. Nematol. 14:91, 2006. (5) T. C. Vrain. J. Nematol. 25:361, 1993.


Neuroprotective effects of triterpene glycosides from glycine max against glutamate induced toxicity in primary cultured rat cortical cells.
To examine the neuroprotective effects of Glycine max, we tested its protection against the glutamate-induced toxicity in primary cortical cultured neurons. In order to clarify the neuroprotective mechanism(s) of this observed effect, isolation was performed to seek and identify active fractions and components. From such fractionation, two triterpene glycosides, 3-O-[α-l-rhamnopyranosyl(1-2)-β-d-glucopyranosyl(1-2)-β-d-glucuronopyranosyl]olean-12-en-3β,22β,24-triol (1) and 3-O-[β-d-glucopyranosyl(1-2)-β-d-galactopyranosyl(1-2)-β-d-glucuronopyranosyl]olean-12-en-3β,22β,24-triol (2) were isolated with the methanol extracts with of air-dried Glycine max. Among these compounds, compound 2 exhibited significant neuroprotective activities against glutamate-induced toxicity, exhibiting cell viability of about 50% at concentrations ranging from 0.1 μM to 10 μM. Therefore, the neuroprotective effect of Glycine max might be due to the inhibition of glutamate-induced toxicity by triterpene glycosides.


Formation of an interphase layer during deposition of cobalt onto tetraphenylporphyrin: a hard X-ray photoelectron spectroscopy (HAXPES) study.
The interface formation upon vapor deposition of a metal onto a molecular organic semiconductor was studied using a well-defined complexation reaction between a metal and a porphyrin. Specifically, metallic cobalt (Co) was vapor deposited onto a thin film of 2H-tetraphenylporphyrin (2HTPP) at room temperature. The resulting interface was probed with Hard X-ray Photoelectron Spectroscopy (HAXPES) using photon energies between 2 and 6 keV to obtain a detailed depth profile of the chemical composition. Characteristic changes in the N 1s core level signals reveal the formation of a cobalt tetraphenylporphyrin (CoTPP) layer between the Co and 2HTPP layers. Assuming an abrupt interface between CoTPP and 2HTPP (layer-by-layer model), analysis of the XPS data results in a thickness of the CoTPP reaction layer of 1.6 nm. However, a more advanced numerical analysis allowed us to reconstruct details of the actual depth distribution of the CoTPP interphase layer: up to a depth of 1.5 nm, all 2HTPP molecules were converted into CoTPP. Beyond this depth, the CoTPP concentration decreases sharply within 0.15 nm to zero.


Association of retirement age with mortality: a population-based longitudinal study among older adults in the USA.
Retirement is an important transitional process in later life. Despite a large body of research examining the impacts of health on retirement, questions still remain regarding the association of retirement age with survival. We aimed to examine the association between retirement age and mortality among healthy and unhealthy retirees and to investigate whether sociodemographic factors modified this association. On the basis of the Health and Retirement Study, 2956 participants who were working at baseline (1992) and completely retired during the follow-up period from 1992 to 2010 were included. Healthy retirees (n=1934) were defined as individuals who self-reported health was not an important reason to retire. The association of retirement age with all-cause mortality was analysed using the Cox model. Sociodemographic effect modifiers of the relation were examined. Over the study period, 234 healthy and 262 unhealthy retirees died. Among healthy retirees, a 1-year older age at retirement was associated with an 11% lower risk of all-cause mortality (95% CI 8% to 15%), independent of a wide range of sociodemographic, lifestyle and health confounders. Similarly, unhealthy retirees (n=1022) had a lower all-cause mortality risk when retiring later (HR 0.91, 95% CI 0.88 to 0.94). None of the sociodemographic factors were found to modify the association of retirement age with all-cause mortality. Early retirement may be a risk factor for mortality and prolonged working life may provide survival benefits among US adults.


Characterization of bovine and human neuronal nicotinic acetylcholine receptors using monoclonal antibodies.
Neuronal acetylcholine receptors (AChRs), which bind nicotine with high affinity but do not bind alpha-bungarotoxin (alpha Bgt), have recently been immunoaffinity-purified from chicken (Whiting and Lindstrom, 1986a) and rat (Whiting and Lindstrom, 1987a) brain using monoclonal antibodies (mAbs). Here we report the characterization of nicotinic AChRs of bovine and human brain using as probes mAbs prepared to AChRs from rat brain. Both the human and bovine brain AChRs exhibit high affinity for L-nicotine (Ki = 16 nM for bovine AChR and Ki = 6.5 nM for human AChR) and other cholinergic agonists, relatively lower affinity for cholinergic antagonists, and do not bind alpha Bgt. These AChRs are affinity-labeled with bromoacetylcholine and 4-(N-maleimido)benzyltrimethylammonium iodide (MBTA) after reduction with dithiothreitol, indicating that amino acid residues homologous to cysteines 192 and 193 of alpha subunits of Torpedo electric organ AChRs are conserved. Immunoaffinity-purified bovine brain AChR consists of 2 types of subunit, Mr 50,600 and Mr 74,400. The Mr 74,400 subunit was affinity-labeled with 3H-MBTA, indicating that it contains the ACh binding site. Thus, mAbs have proven to be excellent probes for these proteins, and have been used to show that neuronal nicotinic AChRs of chickens, rats, and cattle are macromolecules approximately 10 S in size and composed of only 2 kinds of subunits: an ACh-binding subunit and a structural subunit.


Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle.
The COP9-Signalosome (CSN) regulates cullin-RING ubiquitin ligase (CRL) activity and assembly by cleaving Nedd8 from cullins. Free CSN is autoinhibited, and it remains unclear how it becomes activated. We combine structural and kinetic analyses to identify mechanisms that contribute to CSN activation and Nedd8 deconjugation. Both CSN and neddylated substrate undergo large conformational changes upon binding, with important roles played by the N-terminal domains of Csn2 and Csn4 and the RING domain of Rbx1 in enabling formation of a high affinity, fully active complex. The RING domain is crucial for deneddylation, and works in part through conformational changes involving insert-2 of Csn6. Nedd8 deconjugation and re-engagement of the active site zinc by the autoinhibitory Csn5 glutamate-104 diminish affinity for Cul1/Rbx1 by ~100-fold, resulting in its rapid ejection from the active site. Together, these mechanisms enable a dynamic deneddylation-disassembly cycle that promotes rapid remodeling of the cellular CRL network.


The measurement and prevalence of depression in patients with chronic heart failure.
Chronic heart failure is a common disease accompanied by poor mental health and depression. The diagnosis of depression is based on the presence of affective, cognitive, and somatic symptoms assessed by categoric or dimensional instruments. Depression is prominent and high rates are found with dimensional instruments in hospitalized heart failure patients. Categoric instruments seem to be more reliable but are more complex to use. Because of poor outcomes, attention should also be paid to subthreshold depression. In screening for such cases, dimensional instruments might be preferable because of the risk for underestimation by categoric instruments. Dimensional instruments might also be easier to implement in daily practice but, to reduce bias, cutoff scores might need refinement according to the clinical setting. Therefore, studies that evaluate different cutoff values are needed to find a critical level of burden from a depressive symptomatology on outcomes such as mortality, hospitalizations, and quality of life.


First pass cable artefact correction for cardiac C-arm CT imaging.
Cardiac C-arm CT imaging delivers a tomographic region-of-interest reconstruction of the patient's heart during image guided catheter interventions. Due to the limited size of the flat detector a volume image is reconstructed, which is truncated in the cone-beam (along the patient axis) and the fan-beam (in the transaxial plane) direction. To practically address this local tomography problem correction methods, like projection extension, are available for first pass image reconstruction. For second pass correction methods, like metal artefact reduction, alternative correction schemes are required when the field of view is limited to a region-of-interest of the patient. In classical CT imaging metal artefacts are corrected by metal identification in a first volume reconstruction and generation of a corrected projection data set followed by a second reconstruction. This approach fails when the metal structures are located outside the reconstruction field of view. When a C-arm CT is performed during a cardiac intervention pacing leads and other cables are frequently positioned on the patients skin, which results in propagating streak artefacts in the reconstruction volume. A first pass approach to reduce this type of artefact is introduced and evaluated here. It makes use of the fact that the projected position of objects outside the reconstruction volume changes with the projection perspective. It is shown that projection based identification, tracking and removal of high contrast structures like cables, only detected in a subset of the projections, delivers a more consistent reconstruction volume with reduced artefact level. The method is quantitatively evaluated based on 50 simulations using cardiac CT data sets with variable cable positioning. These data sets are forward projected using a C-arm CT system geometry and generate artefacts comparable to those observed in clinical cardiac C-arm CT acquisitions. A C-arm CT simulation of every cardiac CT data set without cables served as a ground truth. The 3D root mean square deviation between the simulated data set with and without cables could be reduced for 96% of the simulated cases by an average of 37% (min -9%, max 73%) when using the first pass correction method. In addition, image quality improvement is demonstrated for clinical whole heart C-arm CT data sets when the cable removal algorithm was applied.


Inotropic selectivity of dobutamine enantiomers in the pithed rat.
The inotropic selectivities of the (-)- and (+)-enantiomers of dobutamine were assessed in pithed rat by comparing the relative ability of each enantiomer to increase left ventricular contractility (left ventricular dp/dt) and heart rate. The (-)-enantiomer of dobutamine, which is predominantly an alpha-1 adrenoceptor agonist, displayed greater inotropic selectivity than the (+)-enantiomer, which is predominantly a beta-1 and beta-2 adrenoceptor agonist. Pretreatment with the alpha-1 adrenoceptor antagonist prazosin significantly inhibited the effect of (-)-dobutamine on left ventricular dp/dt, but did not affect the chronotropic activity of this enantiomer. As such, pretreatment with prazosin decreased the inotropic selectivity of (-)-dobutamine. In contrast, the inotropic activity and selectivity of (+)-dobutamine were not affected by prazosin pretreatment. These results indicate that the inotropic effects of (-)-dobutamine are mediated, at least in part, by alpha-1 adrenoceptors. We conclude, based on the marked inotropic activity of (-)-dobutamine and the greater inotropic selectivity of (-)-dobutamine over (+)-dobutamine, that alpha-1 adrenoceptors may play a role in the inotropic activity and selectivity of racemic dobutamine used clinically. The possible involvement of both myocardial alpha-1 and beta-1 adrenoceptors in the inotropic activity of dobutamine must be considered.


The contribution of benzene to smoking-induced leukemia.
Cigarette smoking is associated with an increased risk of leukemia; benzene, an established leukemogen, is present in cigarette smoke. By combining epidemiologic data on the health effects of smoking with risk assessment techniques for low-dose extrapolation, we assessed the proportion of smoking-induced total leukemia and acute myeloid leukemia (AML) attributable to the benzene in cigarette smoke. We fit both linear and quadratic models to data from two benzene-exposed occupational cohorts to estimate the leukemogenic potency of benzene. Using multiple-decrement life tables, we calculated lifetime risks of total leukemia and AML deaths for never, light, and heavy smokers. We repeated these calculations, removing the effect of benzene in cigarettes based on the estimated potencies. From these life tables we determined smoking-attributable risks and benzene-attributable risks. The ratio of the latter to the former constitutes the proportion of smoking-induced cases attributable to benzene. Based on linear potency models, the benzene in cigarette smoke contributed from 8 to 48% of smoking-induced total leukemia deaths [95% upper confidence limit (UCL), 20-66%], and from 12 to 58% of smoking-induced AML deaths (95% UCL, 19-121%). The inclusion of a quadratic term yielded results that were comparable; however, potency models with only quadratic terms resulted in much lower attributable fractions--all < 1%. Thus, benzene is estimated to be responsible for approximately one-tenth to one-half of smoking-induced total leukemia mortality and up to three-fifths of smoking-related AML mortality. In contrast to theoretical arguments that linear models substantially overestimate low-dose risk, linear extrapolations from empirical data over a dose range of 10- to 100-fold resulted in plausible predictions.