Connective tissue disorders in domestic animals.
Though soft tissue disorders have been recognized and described to some detail in several types of domestic animals and small mammals for some years, not much progress has been made in our understanding of the biochemical basis and pathogenesis of these diseases in animals. Ehlers-Danlos syndrome described in dogs already in 1943 and later in cats affects mainly skin in these animals. The involved skin is thin and hyperextensible with easily inflicted injuries resulting in hemorrhagic wounds and atrophic scars. Joint laxity and dislocation common in people are less frequently found in dogs. No systemic complications, such as organ rupture or cardiovascular problems which have devastating consequences in people have been described in cats and dogs. The diagnosis is based on clinical presentation and on light or electron microscopic features of disorganized and fragmented collagen fibrils. Several cases of bovine and ovine dermatosparaxis analogous to human Ehlers-Danlos syndrome type VIIC were found to be caused by mutations in the procollagen I N-proteinase (pnPI) or ADAMTS2 gene, though mutations in other sites are likely responsible for other types of dermatosparaxis. Cattle suffering from a form of Marfan syndrome were described to have aortic dilatation and aneurysm together with ocular abnormalities and skeletal involvement. As in people mutations at different sites of bovine FBN1 may be responsible for Marfan phenotype. Hereditary equine regional dermal asthenia (HERDA), or hyperelastosis cutis, has been recognized in several horse breeds as affecting primarily skin, and, occasionally, tendons. A mutation in cyclophilin B, a chaperon involved in proper folding of collagens, has been identified in some cases. Degenerative suspensory ligament desmitis (DSLD) affects primarily tendons and ligaments of certain horse breeds. New data from our laboratory showed excessive accumulation of proteoglycans in organs with high content of connective tissues. We have identified an abnormal form of decorin with altered biological activity in these proteoglycan deposits, and more recently changes in processing of aggrecan were found by us and other investigators.The naturally occurring diseases of soft tissues in domestic animals described here have a potential to serve as good models for analogous human diseases. This is the case particularly relevant to dogs as a half out of the more than 400 naturally occurring hereditary canine diseases has the potential to serve as a model for human disease.


Docetaxel plus oxaliplatin with or without fluorouracil or capecitabine in metastatic or locally recurrent gastric cancer: a randomized phase II study.
Docetaxel/cisplatin/infusional 5-fluorouracil (5-FU; DCF) is a standard chemotherapy regimen for patients with advanced gastric cancer (GC). This phase II study evaluated docetaxel/oxaliplatin (TE), docetaxel/oxaliplatin/5-FU (TEF), and docetaxel/oxaliplatin/capecitabine (TEX) in patients with advanced GC. Patients with metastatic or locally recurrent gastric adenocarcinoma (including carcinoma of the gastro-oesophageal junction) were randomly assigned (1 : 1 : 1) to TE, TEF, or TEX. Each regimen was tested at two doses before full evaluation at optimized dose levels. The primary end point was progression-free survival (PFS). Overall survival (OS), tumour response, and safety were also assessed. A therapeutic index (median PFS relative to the incidence of febrile neutropenia) was calculated for each regimen and compared with DCF (historical data). Overall, 248 patients were randomly assigned to receive optimized dose treatment. Median PFS was longer with TEF (7.66 [95% confidence interval (CI): 6.97-9.40] months) versus TE (4.50 [3.68-5.32] months) and TEX (5.55 [4.30-6.37] months). Median OS was 14.59 (95% CI: 11.70-21.78) months for TEF versus 8.97 (7.79-10.87) months for TE and 11.30 (8.08-14.03) months for TEX. The rate of tumour response (complete or partial) was 46.6% (95% CI 35.9-57.5) for TEF versus 23.1% (14.3-34.0) for TE and 25.6% (16.6-36.4) for TEX. The frequency and type of adverse events (AEs) were similar across the three arms. Common grade 3/4 AEs were fatigue (21%), sensory neuropathy (14%), and diarrhoea (13%). Febrile neutropenia was reported in 2% (TEF), 14% (TE), and 9% (TEX) of patients. The therapeutic index was improved with TEF versus TEX, TE, or DCF. These results suggest that TEF is worthy of evaluation as an arm in a phase III trial or as a backbone regimen for new targeted agents in advanced GC. CLINICALTRIALS.GOV: Identifier Trial registration number: NCT00382720.


High blood glutamate oxaloacetate transaminase levels are associated with good functional outcome in acute ischemic stroke.
The capacity of the blood enzyme glutamate oxaloacetate transaminase (GOT) to remove glutamate from the brain by means of blood glutamate degradation has been shown in experimental models to be an efficient and novel neuroprotective tool against ischemic stroke; however, the beneficial effects of this enzyme should be tested in patients with stroke to validate these results. This study aims to investigate the association of GOT levels in blood with clinical outcome in patients with acute ischemic stroke. In two clinical independent studies, we found that patients with poor outcome show higher glutamate and lower GOT levels in blood at the time of admission. Lower GOT levels and higher glutamate levels were independently associated with poorer functional outcome at 3 months and higher infarct volume. These findings show a clear association between high blood glutamate levels and worse outcome and vice versa for GOT, presumably explained by the capacity of this enzyme to metabolize blood glutamate.


Parallel reaction monitoring using quadrupole-Orbitrap mass spectrometer: Principle and applications.
Targeted mass spectrometry-based approaches are nowadays widely used for quantitative proteomics studies and more recently have been implemented on high resolution/accurate mass (HRAM) instruments resulting in a considerable performance improvement. More specifically, the parallel reaction monitoring technique (PRM) performed on quadrupole-Orbitrap mass spectrometers, leveraging the high resolution and trapping capabilities of the instrument, offers a clear advantage over the conventional selected reaction monitoring (SRM) measurements executed on triple quadrupole instruments. Analyses performed in HRAM mode allow for an improved discrimination between signals derived from analytes and those resulting from matrix interferences translating in the reliable quantification of low abundance components. The purpose of the study defines various implementation schemes of PRM, namely: (i) exploratory experiments assessing the detectability of very large sets of peptides (100-1000), (ii) wide-screen analyses using (crude) internal standards to obtain statistically meaningful (relative) quantitative analyses, and (iii) precise/accurate quantification of a limited number of analytes using calibrated internal standards. Each of the three implementation schemes requires specific acquisition methods with defined parameters to appropriately control the acquisition during the actual peptide elution. This tutorial describes the different PRM approaches and discusses their benefits and limitations in terms of quantification performance and confidence in analyte identification.


Comparison of CPP uptake methods.
In the last 15 years, an ever expanding pool of cell-penetrating peptides (CPPs) has been discovered and recently focus has shifted towards improving already existing CPPs by different modifications. Since the number of published peptide sequences with cell-penetrating ability is now reaching several hundreds, the consensus methods to compare the efficacy of these is clearly needed. Many research groups are evaluating the applicability of CPPs as drug delivery vectors, all having their preferred methods of assessing uptake and intracellular distribution. Even when applying the same method, the use of different cell lines, peptide concentrations, exposure conditions, etc. are complicating comparison of data between different groups. This book is a welcome contribution to the CPP research field, hopefully paving the way for standardized protocols to be used in the future. Some of the most common methods used to this date are presented and compared in this chapter.


Does body satisfaction matter? Five-year longitudinal associations between body satisfaction and health behaviors in adolescent females and males.
This study addresses the question, "Does body satisfaction matter?" by examining longitudinal associations between body satisfaction and weight-related health-promoting and health-compromising behaviors five years later among adolescents. Project EAT-II followed an ethnically and socioeconomically diverse sample of 2516 adolescents from 1999 (Time 1) to 2004 (Time 2). Associations between body satisfaction at Time 1 and health behaviors at Time 2 were examined, adjusting for sociodemographic characteristics and Time 1 health behaviors, with and without adjustment for body mass index (BMI). In females, lower body satisfaction predicted higher levels of dieting, unhealthy and very unhealthy weight control behaviors and binge eating, and lower levels of physical activity and fruit and vegetable intake. After adjusting for BMI, associations between body satisfaction and dieting, very unhealthy weight control behaviors, and physical activity remained statistically significant. In males, lower body satisfaction predicted higher levels of dieting, healthy, unhealthy, and very unhealthy weight control behaviors, binge eating, and smoking, and lower levels of physical activity. After adjusting for BMI, associations between body satisfaction and dieting, unhealthy weight control behavior, and binge eating remained statistically significant. The study findings indicate that, in general, lower body satisfaction does not serve as a motivator for engaging in healthy weight management behaviors, but rather predicts the use of behaviors that may place adolescents at risk for weight gain and poorer overall health. Interventions with adolescents should strive to enhance body satisfaction and avoid messages likely to lead to decreases in body satisfaction.


Improving Research Practice in Rat Orthotopic and Partial Orthotopic Liver Transplantation: A Review, Recommendation, and Publication Guide.
Due to a worldwide shortage of donor organs for liver transplantation, alternative approaches, such as split and living donor liver transplantations, were introduced to increase the donor pool and reduce mortality on liver transplant waiting lists. Numerous details concerning the mechanisms and pathophysiology of liver regeneration, small-for-size syndrome, rejection, and tolerance in partial liver transplantation facilitated the development of various animal models. The high number of preclinical animal studies contributed enormously to our understanding of many clinical aspects of living donor and partial liver transplantations. Microsurgical rat models of partial orthotopic liver transplantation are well established and widely used. Nevertheless, several issues regarding this procedure are controversial, not clarified, or not yet properly standardized (graft rearterialization, size reduction techniques, etc.). The major aim of this literature review is to give the reader a current overview of rat orthotopic liver transplantation models with a special focus on partial liver transplantation. The aspects of model evolution, microsurgical training, and different technical problems are analyzed and discussed in detail. Our further aim in this paper is to elaborate a detailed publication guide in order to improve the quality of reporting in the field of rat liver transplantation according to the ARRIVE guidelines and the 3R principle. Key Messages: Partial orthotopic liver transplantation in rats is an indispensable, reliable, and cost-efficient model for transplantation research. A certain consensus on different technical issues and a significant improvement in scientific reporting are essential to improve transparency and comparability in this field as well as to foster refinement.


Electromyogenic Artifacts and Electroencephalographic Inferences Revisited.
Recent years have witnessed a renewed interest in using oscillatory brain electrical activity to understand the neural bases of cognition and emotion. Electrical signals originating from pericranial muscles represent a profound threat to the validity of such research. Recently, McMenamin et al (2010) examined whether independent component analysis (ICA) provides a sensitive and specific means of correcting electromyogenic (EMG) artifacts. This report sparked the accompanying commentary (Olbrich, Jödicke, Sander, Himmerich & Hegerl, in press), and here we revisit the question of how EMG can alter inferences drawn from the EEG and what can be done to minimize its pernicious effects. Accordingly, we briefly summarize salient features of the EMG problem and review recent research investigating the utility of ICA for correcting EMG and other artifacts. We then directly address the key concerns articulated by Olbrich and provide a critique of their efforts at validating ICA. We conclude by identifying key areas for future methodological work and offer some practical recommendations for intelligently addressing EMG artifact.


Ready for Change? The Role of Physician and Staff Engagement, Burnout, and Workplace Attributes.
We examined factors associated with change readiness among 343 primary care physicians and 590 nonphysician staff undergoing "Lean"-based process improvements. Baseline levels of engagement were associated with greater readiness for change across all measured domains. Job-related burnout correlated with greater need for change, but lower self-efficacy and perceived support, whereas a personal sense of accomplishment was associated with higher efficacy to implement changes. At a department level, teamwork, participation in decision making, and change history were associated with higher engagement and lower burnout among physicians and staff; conversely, a busy or stressful department correlated with lower engagement and higher burnout.


Antagonistic co-contraction can minimize muscular effort in systems with uncertainty.
Muscular co-contraction of antagonistic muscle pairs is often observed in human movement, but it is considered inefficient and it can currently not be predicted in simulations where muscular effort or metabolic energy are minimized. Here, we investigated the relationship between minimizing effort and muscular co-contraction in systems with random uncertainty to see if muscular co-contraction can minimize effort in such system. We also investigated the effect of time delay in the muscle, by varying the time delay in the neural control as well as the activation time constant. We solved optimal control problems for a one-degree-of-freedom pendulum actuated by two identical antagonistic muscles, using forward shooting, to find controller parameters that minimized muscular effort while the pendulum remained upright in the presence of noise added to the moment at the base of the pendulum. We compared a controller with and without feedforward control. Task precision was defined by bounding the root mean square deviation from the upright position, while different perturbation levels defined task difficulty. We found that effort was minimized when the feedforward control was nonzero, even when feedforward control was not necessary to perform the task, which indicates that co-contraction can minimize effort in systems with uncertainty. We also found that the optimal level of co-contraction increased with time delay, both when the activation time constant was increased and when neural time delay was added. Furthermore, we found that for controllers with a neural time delay, a different trajectory was optimal for a controller with feedforward control than for one without, which indicates that simulation trajectories are dependent on the controller architecture. Future movement predictions should therefore account for uncertainty in dynamics and control, and carefully choose the controller architecture. The ability of models to predict co-contraction from effort or energy minimization has important clinical and sports applications. If co-contraction is undesirable, one should aim to remove the cause of co-contraction rather than the co-contraction itself.


Photostable polymorphic organic cages for targeted live cell imaging.
Fluorescent microscopy is a powerful tool for studying the cellular dynamics of biological systems. Small-molecule organic fluorophores are the most commonly used for live cell imaging; however, they often suffer from low solubility, limited photostability and variable targetability. Herein, we demonstrate that a tautomeric organic cage, OC1, has high cell permeability, photostability and selectivity towards the mitochondria. We further performed a structure-activity study to investigate the role of the keto-enol tautomerization, which affords strong and consistent fluorescence in dilute solutions through supramolecular self-assembly. Significantly, OC1 can passively diffuse through the cell membrane directly targeting the mitochondria without going through the endosomes or the lysosomes. We envisage that designing highly stable and biocompatible self-assembled fluorophores that can passively diffuse through the cell membrane while selectively targeting specific organelles will push the boundaries of fluorescent microscopy to visualize intricate cellular processes at the single molecule level in live samples.


Early Venous Stent Failure Predicted by Platelet Count and Neutrophil/Lymphocyte Ratio.
Inflammation and platelet activation have been shown to be involved in acute thromobogenicity following venous occlusive conditions. The aim of this study was to identify the association of baseline platelet count and neutrophil/lymphocyte ratio (NLR) with venous stent failure. Methods and Results: Patients who underwent technically successful iliocaval venous stent placement with available baseline complete blood count and follow-up stent patency data were selected (n=50). Stent failure was defined as >50% stenosis or occlusion at follow-up angiography, contrast-enhanced CT, MRI or duplex US. Median patient age was 49.5 years (range, 13-76 years), and 62% were female. Median follow-up time was 10.2 months (range, 0.1-76.4 months). Stent failure occurred in 13 patients (26%) after a median of 1.2 months (range, 1 day-76.4 months). On multivariable-adjusted Cox modeling, baseline platelets (HR, 2.28; P=0.004) and WBC count (HR, 2.03; P=0.013) were significantly associated with stent failure on follow-up; neutrophils (HR, 16.10; P=0.050); and NLR (HR, 12.19; P=0.050) had borderline significance. Compared with patients without stent failure, those with early, but not late, stent failure had higher baseline platelets (P=0.031) and neutrophils (P=0.025), and NLR (P=0.026). Baseline platelet count and NLR are associated with early but not late failure of iliocaval venous stents. This suggests different pathophysiologic mechanisms and a role for both platelet activation and inflammatory mechanisms in early rather than late stent thrombosis. Future research is needed to better explain this novel finding.


Thymidine kinase from Tetrahymena thermophila. Purification and immunological analysis.
Thymidine kinase is an enzyme involved in DNA precursor metabolism and DNA replication. The synthesis of this enzyme is highly regulated during the cell cycle and the activity of the enzyme is also regulated by feedback inhibition. Genes encoding thymidine kinase have been extremely useful as selectable markers for introducing DNA into a number of cells. In order to study cell cycle regulation of thymidine kinase, the gene which encodes this enzyme, as well as aspects of DNA replication in the ciliated protozoan Tetrahymena thermophila, we have purified thymidine kinase from Tetrahymena. Two forms of thymidine kinase with native molecular masses of 59 kDa and 80 kDa have been identified and purified 6800- and 4600-fold, respectively. The 59-kDa enzyme, a homodimer of 30-kDa subunits, has been purified to near homogeneity and polyclonal antibodies have been raised against the 30-kDa subunit. Serological studies indicate that the two enzymes are antigenically distinct. The antibody against the Tetrahymena protein cross-reacts with a polypeptide in Chinese hamster ovary (CHO) cell extracts of 26 kDa which corresponds to the reported size of Chinese hamster thymidine kinase protein.


Knowledge, attitudes, and normative beliefs as predictors of hookah smoking initiation: a longitudinal study of university students.
While cross-sectional studies have shown that hookah tobacco smoking (HTS) is an increasingly popular behavior among university students, little is known about factors associated with initiation. This study sought to determine associations between knowledge, attitudes, and normative beliefs and initiation of HTS among university students. Data were from a prospective longitudinal cohort study of 569 randomly selected first- and second-year university students. Online questionnaires that were developed in accordance with our composite theoretical model were completed in September 2010 and April 2011. About one-seventh (13%) of participants initiated HTS by follow-up. Positive attitudes and favorable normative beliefs were associated with increased adjusted odds of initiation (AOR = 4.12, 95% CI = 2.56, 6.59; and AOR = 2.01, 95% CI = 1.35, 2.99, respectively), while negative attitudes were associated with decreased adjusted odds (AOR = 0.62, 95% CI = 0.48, 0.80). Correct knowledge regarding toxicants associated with HTS was not significantly associated with initiation. While positive attitudes and favorable normative beliefs are associated with initiation of HTS in a cohort of never-users, increased knowledge about toxins is not associated with lower initiation. It may be particularly valuable for educational interventions to attempt to alter positive attitudes and normative beliefs related to HTS.


Differing mechanisms underlie sexual size-dimorphism in two populations of a sex-changing fish.
Variability in the density of groups within a patchy environment lead to differences in interaction rates, growth dynamics and social organization. In protogynous hermaphrodites there are hypothesised trade-offs among sex-specific growth, reproductive output and mortality. When differences in density lead to changes to social organization the link between growth and the timing of sex-change is predicted to change. The present study explores this prediction by comparing the social organisation and sex-specific growth of two populations of a protogynous tropical wrasse, Halichoeres miniatus, which differ in density. At a low density population a strict harem structure was found, where males maintained a tight monopoly of access and spawning rights to females. In contrast, at a high density population a loosely organised system prevailed, where females could move throughout multiple male territories. Otolith microstructure revealed the species to be annual and deposit an otolith check associated with sex-change. Growth trajectories suggested that individuals that later became males in both populations underwent a growth acceleration at sex-change. Moreover, in the high density population, individuals that later became males were those individuals that had the largest otolith size at hatching and consistently deposited larger increments throughout early larval, juvenile and female life. This study demonstrates that previous growth history and growth rate changes associated with sex change can be responsible for the sexual dimorphism typically found in sex-changing species, and that the relative importance of these may be socially constrained.


The origin and role of the renal stroma.
The postnatal kidney is predominantly composed of nephron epithelia with the interstitial components representing a small proportion of the final organ, except in the diseased state. This is in stark contrast to the developing organ, which arises from the mesoderm and comprises an expansive stromal population with distinct regional gene expression. In many organs, the identity and ultimate function of an epithelium is tightly regulated by the surrounding stroma during development. However, although the presence of a renal stromal stem cell population has been demonstrated, the focus has been on understanding the process of nephrogenesis whereas the role of distinct stromal components during kidney morphogenesis is less clear. In this Review, we consider what is known about the role of the stroma of the developing kidney in nephrogenesis, where these cells come from as well as their heterogeneity, and reflect on how this information may improve human kidney organoid models.


Identification of bacteria infecting Ixodes ricinus ticks by 16S rDNA amplification and denaturing gradient gel electrophoresis.
Ticks harbor a complex microbial population, which they acquire while feeding on a variety of mammalians and birds. Zoonotic diseases transferred by ticks are an increasing problem and have become a burden to the community. 16S rDNA amplification and denaturing gradient gel electrophoresis (DGGE) enables detection of the broad spectrum of bacteria that settles in the ticks. Profiling the complete microbial population in ticks may provide a better understanding of the ticks' potential to harbor and disperse pathogens. Separation of pathogenic species by DGGE is based on variation in %GC content within the 16S rDNA genetic region. Sequencing of these fragments allows identification of bacterial species. Present study identified some well-known tick-infecting bacteria, such as members of genus Borrelia, Rickettsiales, and Pseudomonas, but also less described tick-infecting bacteria such as Rhodococcus erythropolis, Spiroplasma spp., and an endosymbiont of the microarthropod Folsomia candida. This is the first report of Segniliparus rugosus-infected Ixodes ricinus ticks. Also, it is the first report of several of these pathogens in the Norwegian tick population.


Genome-scale dynamic modeling of the competition between Rhodoferax and Geobacter in anoxic subsurface environments.
The advent of rapid complete genome sequencing, and the potential to capture this information in genome-scale metabolic models, provide the possibility of comprehensively modeling microbial community interactions. For example, Rhodoferax and Geobacter species are acetate-oxidizing Fe(III)-reducers that compete in anoxic subsurface environments and this competition may have an influence on the in situ bioremediation of uranium-contaminated groundwater. Therefore, genome-scale models of Geobacter sulfurreducens and Rhodoferax ferrireducens were used to evaluate how Geobacter and Rhodoferax species might compete under diverse conditions found in a uranium-contaminated aquifer in Rifle, CO. The model predicted that at the low rates of acetate flux expected under natural conditions at the site, Rhodoferax will outcompete Geobacter as long as sufficient ammonium is available. The model also predicted that when high concentrations of acetate are added during in situ bioremediation, Geobacter species would predominate, consistent with field-scale observations. This can be attributed to the higher expected growth yields of Rhodoferax and the ability of Geobacter to fix nitrogen. The modeling predicted relative proportions of Geobacter and Rhodoferax in geochemically distinct zones of the Rifle site that were comparable to those that were previously documented with molecular techniques. The model also predicted that under nitrogen fixation, higher carbon and electron fluxes would be diverted toward respiration rather than biomass formation in Geobacter, providing a potential explanation for enhanced in situ U(VI) reduction in low-ammonium zones. These results show that genome-scale modeling can be a useful tool for predicting microbial interactions in subsurface environments and shows promise for designing bioremediation strategies.


Overexposure to Bisphenol A and Its Chlorinated Derivatives of Patients with End-Stage Renal Disease during Online Hemodiafiltration.
The health safety conditions governing the practice of online hemodiafiltration (OL-HDF) do not yet incorporate the risks related to the presence of endocrine disruptors such as bisphenol A (BPA). The aim of this study was to assess, for the first time, the exposure to BPA but also to its chlorinated derivatives (ClxBPA) (100 times more estrogenic than BPA) during OL-HDF. We demonstrated that BPA is transmitted by the different medical devices used in OL-HDF: ultrafilters, dialysis concentrate cartridges (and not only dialyzers, as previously described). Moreover, BPA has been found in dialysis water as well as in ultrapure dialysate and replacement fluid due to contamination of water coming from municipal network. Indeed, due to contaminations provided by both ultrafilters and water, high levels of BPA were determined in the infused replacement fluid (1033 ng.L-1) from the beginning of the session. Thus, our results demonstrate that dialysis water must be considered as an important exposure source to endocrine disruptors, especially since other micropollutants such as ClxBPA have also been detected in dialysis fluids. While assessment of the impact of this exposure remains to be done, these new findings should be taken into account to assess exposure risks in end-stage renal disease patients.


Synchrotron microradiography study on acute lung injury of mouse caused by PM(2.5) aerosols.
In order to investigate FeSO(4), ZnSO(4) (the two of main metal compositions of Shanghai PM(2.5) (particle matter with those aerodynamical diameter <2.5 microm)) effects on acute lung injury, six solutions contained PM(2.5) aerosol particles, FeSO(4), ZnSO(4) and their mixtures were instilled intratracheally into mouse lungs for experiment. By 2 days after instillation, the live mice were checked in vivo by synchrotron refractive index microradiography. In addition after extracted and examined by dissection, the right lobes of lung were fixed by formalin, then imaged by synchrotron microradiography again. Corresponding parts of those lung tissues were embedded in paraffin for histopathologic study. The synchrotron X-ray microradiographs of live mouse lung showed different lung texture changes after instilled with different toxic solutions. Hemorrhage points in lung were observed more from those mice instilled by FeSO(4) contained toxin solutions groups. Bronchial epithelial hyperplasia can be observed in ZnSO(4) contained solution-instilled groups from histopathologic analysis. It was found that the acute lung injury of mice caused by solution of PM(2.5)+FeSO(4)+ZnSO(4) was more serious than other toxin solutions. Results suggested that FeSO(4) mainly induced hemorrhage and ZnSO(4) mainly induced inflammation and bronchiolar epithelial hyperplasia in the early toxicological effects of PM(2.5).


Trade liberalisation and the nutrition transition: mapping the pathways for public health nutritionists.
To describe pathways through which trade liberalisation affects the food environment, relevant to the nutrition transition, in order to enable public health nutritionists to understand trade policy as a macro-level influence on food consumption. The pathways mapped in the present paper are based on the agreements of the World Trade Organization, which shape national policy. Implications for nutrition are presented based on a comprehensive literature review, and case studies are used to illustrate the various pathways. Developing countries are currently experiencing a nutrition transition, resulting in dietary patterns associated with chronic disease. Chronic diseases are amenable to prevention, and trade policy has been highlighted as a potential avenue for nutrition-related prevention. Trade liberalisation influences the food environment through facilitating trade in goods and services, enabling investment and decreasing support/protection for domestic industry. These policy outcomes facilitate the nutrition transition particularly through increasing the availability and affordability of processed foods and animal products. The framework highlights the complex relationship between trade policy and the nutrition transition, with both negative and positive outcomes arising from different aspects of trade liberalisation. Policy change associated with trade liberalisation has created incentives for consumption patterns associated with the nutrition transition, but has also had some positive nutritional outcomes. As a result, it is important for public health nutritionists to consider the implications of trade policy decisions in their efforts to prevent and control diet-related chronic diseases.


Evaluation of a novel haematology analyser for use with feline blood.
A novel haematology analyser was evaluated for its use with feline samples. Complete blood cell counts, a five-part differential count, and reticulocyte numbers were determined, and the results compared with reference data. Coefficients of correlation, Passing-Bablok regression analysis and Bland-Altmann difference plots with biases and 95% limits of agreement are reported. Precision and linearity were also studied. The instrument demonstrated very low imprecision, and the tested range of linearity exceeded the reference ranges provided by the manufacturer. For all parameters except monocytes (r = 0.65), the analyser results correlated well with reference methods. Compared with the manual count of aggregated reticulocytes, the instrument showed good agreement with a positive bias. The optical platelet count correlated well with the manual chamber count. In conclusion the analyser was found to be highly reliable for the analysis of feline blood samples in a large veterinary laboratory.


Hormone replacement therapy in a risk-benefit perspective.
The relative cost-effectiveness of different treatment strategies for hormone replacement therapy (HRT) was assessed within the framework of a computer model. Where data were lacking, it was necessary to make assumptions about the effects of HRT, particularly in relation to combined oestrogen-progestogen therapy and cardiovascular disease; however, sensitivity analyses were performed to assess the impact of changing these assumptions on the cost-effectiveness equation. It appears that net expenditure by the NHS will depend critically on the direct costs of treatment, rather than on any indirect costs incurred or saved as a result of side-effects. In terms of mortality, a reduction in cardiovascular disease risk would have greatest impact and would overshadow any small increase in breast cancer risk which may be associated with long-term use. If the cardioprotective effect of oestrogen is real, our results suggest that long-term prophylactic treatment of hysterectomised women would be relatively cost-effective. Treatment of symptomatic menopausal women for any period of time appears to offer very good value for money. The lack of data relating to combined oestrogen-progestogen therapy and cardioprotection, and the major importance of the latter in the equation of benefits and risks, make it more difficult to draw conclusions about the cost-effectiveness of treating non-hysterectomised asymptomatic women for prophylactic reasons.


A protocol for a randomised double-blind placebo-controlled feasibility study to determine whether the daily consumption of flavonoid-rich pure cocoa has the potential to reduce fatigue in people with relapsing and remitting multiple sclerosis (RRMS).
Dietary interventions including consumption of flavonoids, plant compounds found in certain foods, may have the ability to improve fatigue. However, to date, no well-designed intervention studies assessing the role of flavonoid consumption for fatigue management in people with MS (pwMS) have been performed. The hypothesis is that the consumption of a flavonoid-rich pure cocoa beverage will reduce fatigue in pwMS. The aim of this study is to determine the feasibility and potential outcome of running a trial to evaluate this hypothesis. Using a randomised (1:1) double-blind placebo-controlled feasibility study, 40 men and women (20 in each trial arm) with a recent diagnosis (< 10 years) of relapsing and remitting MS (RRMS) and who are over 18 years of age will be recruited from neurology clinics and throughout the Thames Valley community. During a 6-week nutrition intervention period, participants will consume the cocoa beverage, high flavonoid or low flavonoid content, at breakfast daily. At baseline, demographic factors and disease-related factors will be assessed. Fatigue, activity and quality of life, in addition to other measures, will be taken at three visits (baseline, week 3 and week 6) in a university setting by a researcher blinded to group membership. Feasibility and fidelity will be assessed through recruitment and retention, adherence and a quantitative process evaluation at the end of the trial.We will describe demographic factors (age, gender, level of education) as well as disease-related factors (disease burden scores, length of time diagnosed with MS) and cognitive assessment, depression and quality of life and general physical activity in order to characterise participants and determine possible mediators to identify the processes by which the intervention may bring about change. Feasibility (recruitment, safety, feasibility of implementation of the intervention and evaluation, protocol adherence and data completion) and potential for benefit (estimates of effect size and variability) will be determined to inform future planned studies. Results will be presented using point estimates, 95% confidence intervals and p values. Primary statistical analysis will be on an intention-to-treat basis and will use the complete case data set. We propose that a flavonoid-enriched cocoa beverage for the management of fatigue will be well received by participants. Further, if it is implemented early in the disease course of people diagnosed with RRMS, it will improve mobility and functioning by modifying fatigue. Registered with ISRCTN Registry. Trial registration No: ISRCTN69897291; Date April 2016.


A study of the mechanisms involved in the neurotoxic action of 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy') on dopamine neurones in mouse brain.
1. Administration of 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy') to mice produces acute hyperthermia and long-term degeneration of striatal dopamine nerve terminals. Attenuation of the hyperthermia decreases the neurodegeneration. We have investigated the mechanisms involved in producing the neurotoxic loss of striatal dopamine. 2. MDMA produced a dose-dependent loss in striatal dopamine concentration 7 days later with 3 doses of 25 mg kg(-1) (3 h apart) producing a 70% loss. 3. Pretreatment 30 min before each MDMA dose with either of the N-methyl-D-aspartate antagonists AR-R15896AR (20, 5, 5 mg kg(-1)) or MK-801 (0.5 mg kg(-1)x3) failed to provide neuroprotection. 4. Pretreatment with clomethiazole (50 mg kg(-1)x3) was similarly ineffective in protecting against MDMA-induced dopamine loss. 5. The free radical trapping compound PBN (150 mg kg(-1)x3) was neuroprotective, but it proved impossible to separate neuroprotection from a hypothermic effect on body temperature. 6. Pretreatment with the nitric oxide synthase (NOS) inhibitor 7-NI (50 mg kg(-1)x3) produced neuroprotection, but also significant hypothermia. Two other NOS inhibitors, S-methyl-L-thiocitrulline (10 mg kg(-1)x3) and AR-R17477AR (5 mg kg(-1)x3), provided significant neuroprotection and had little effect on MDMA-induced hyperthermia. 7. MDMA (20 mg kg(-1)) increased 2,3-dihydroxybenzoic acid formation from salicylic acid perfused through a microdialysis tube implanted in the striatum, indicating increased free radical formation. This increase was prevented by AR-R17477AR administration. Since AR-R17477AR was also found to have no radical trapping activity this result suggests that MDMA-induced neurotoxicity results from MDMA or dopamine metabolites producing radicals that combine with NO to form tissue-damaging peroxynitrites.


Biomarkers of exposure in environment-wide association studies - Opportunities to decode the exposome using human biomonitoring data.
The European Union's 7th Framework Programme (EU's FP7) project HEALS - Health and Environment-wide Associations based on Large Population Surveys - aims a refinement of the methodology to elucidate the human exposome. Human biomonitoring (HBM) provides a valuable tool for understanding the magnitude of human exposure from all pathways and sources. However, availability of specific biomarkers of exposure (BoE) is limited. The objective was to summarize the availability of BoEs for a broad range of environmental stressors and exposure determinants and corresponding reference and exposure limit values and biomonitoring equivalents useful for unraveling the exposome using the framework of environment-wide association studies (EWAS). In a face-to-face group discussion, scope, content, and structure of the HEALS deliverable "Guidelines for appropriate BoE selection for EWAS studies" were determined. An expert-driven, distributed, narrative review process involving around 30 individuals of the HEALS consortium made it possible to include extensive information targeted towards the specific characteristics of various environmental stressors and exposure determinants. From the resulting 265 page report, targeted information about BoE, corresponding reference values (e.g., 95th percentile or measures of central tendency), exposure limit values (e.g., the German HBM I and II values) and biomonitoring equivalents (BEs) were summarized and updated. 64 individual biological, chemical, physical, psychological and social environmental stressors or exposure determinants were included to fulfil the requirements of EWAS. The list of available BoEs is extensive with a number of 135; however, 12 of the stressors and exposure determinants considered do not leave any measurable specific substance in accessible body specimens. Opportunities to estimate the internal exposure stressors not (yet) detectable in human specimens were discussed. Data about internal exposures are useful to decode the exposome. The paper provides extensive information for EWAS. Information included serves as a guideline - snapshot in time without any claim to comprehensiveness - to interpret HBM data and offers opportunities to collect information about the internal exposure of stressors if no specific BoE is available.


Grading the level of evidence of neonatal pharmacotherapy: midazolam and phenobarbital as examples.
Many drugs are used off-label or unlicensed in neonates. This does not mean they are used without evidence or knowledge. We aimed to apply and evaluate the Grading and Assessment of Pharmacokinetic-Pharmacodynamic Studies (GAPPS) scoring system for the level of evidence of two commonly used anti-epileptic drugs. Midazolam and phenobarbital as anti-epileptics were evaluated with a systematic literature search on neonatal pharmacokinetic (PK) and/or pharmacodynamic [PD, (amplitude-integrated) electroencephalography effect] studies. With the GAPPS system, two evaluators graded the current level of evidence. Inter-rater agreement was assessed for dosing evidence score (DES), quality of evidence (QoE), and strength of recommendation (REC). Seventy-two studies were included. DES scores 4 and 9 were most frequently used for PK, and scores 0 and 1 for PD. Inter-rater agreements on DES, QoE, and REC ranged from moderate to very good. A final REC was provided for all PK studies, but only for 25% (midazolam) and 33% (phenobarbital) of PD studies. There is a reasonable level of evidence concerning midazolam and phenobarbital PK in neonates, although using a predefined target without integrated PK/PD evaluation. Further research is needed on midazolam use in term neonates with therapeutic hypothermia, and phenobarbital treatment in preterms. There is a reasonable level of evidence concerning pharmacotherapy of midazolam and phenobarbital in neonates. Most evidence is however based on PK studies, using a predefined target level or concentration range without integrated, combined PK/PD evaluation. Using the GAPPS system, final strength of recommendation could be provided for all PK studies, but only for 25% (midazolam) to 33% (phenobarbital) of PD studies. Due to the limited PK observations of midazolam in term neonates with therapeutic hypothermia, and of phenobarbital in preterm neonates these subgroups can be identified for further research.


Are clinically insignificant prostate cancers really insignificant among Korean men?
We aimed to determine whether 12 core-extended biopsies of the prostate could predict insignificant prostate cancer (IPCa) in Koreans reliably enough to recommend active surveillance. Two hundred and ninety-seven patients who underwent radical prostatectomy after 12 core-extended prostate biopsies were retrospectively reviewed. 38 cases (12.8%) were shown to be IPCa. The average age was 65.2 years, serum PSA was 5.49 ng/dL, and the PSA density was 0.11. The Gleason scores (GS) were 6 (3+3) in 31, 5 (3+2) in 4, and 4 (2+2) in 3. After radical prostatectomy, higher GS was given in 16 (42.1%), whereas lower GS was given in 1 case (2.6%), as compared with the GS obtained from biopsy. 11 (28.9%) had GS of 7 (3+4) and 5 (13.2%) had GS of 7 (4+3). 6 in GS 7 (4+3) and 1 in GS 7 (3+4) showed prostate capsule invasion and 1 in GS 7 (4+3) had seminal vesicle invasion. Prostate capsule invasion was observed in 1 with GS 6 (3+3). The rate of inaccuracy of the contemporary Epstein criteria was 42.1%. Only PSA density was a reliable indicator of clinically IPCa (odds ratio=1.384, 95% CI, 1.103 to 2.091). Diagnosis of IPCa from a prostate biopsy underestimated the true nature of prostate cancer in as many as 42.1% of Koreans.


Keeping those telomeres short! an innovative intratumoral long-term drug delivery system.
Telomerase activation and an alternative lengthening of telomeres (ALT) mechanism are two telomere-lengthening cancer cell survival mechanisms elicited by both chemo- and/or radiotherapy. Telomere lengthening interferes with cell lethality and results in the immortalization of cancer cells. To counteract these mechanisms, we developed a drug delivery system (DDS) consisting of a polymeric implant that is inserted directly into tumors. The DDS releases, continuously and gradually, a cationic porphyrin (PdTMPyP4) for >30 days after a single application, and inhibits telomerase activation. The PdTMPyP4 porphyrin is incorporated into a poly(co-glycolic lactic)acid (PLGA) polymer, solidified and cut into small rods. PdTMPyP4 release from the rods was measured spectrophotometrically over time. Uptake of Pd in the DNA of in L428 Hodgkins lymphoma cells was measured by ICP-MS, and telomerase activation by the TRAP assay. The rods were placed into the growth medium of cells whose growth rate was measured for 11 and 19 days. The cylinders were also inserted directly into KHJJ murine mammary tumors borne on the thighs of BALB/c mice and the tumor growth rate measured. In vitro, >10(9)Pd atoms were measured in the DNA of each L428 cell and telomerase activity was reduced by ~15% within 24 h. A one-time application of the rod in the cell medium induced a factor of >5 greater lethality compared to a blank rod or untreated controls. In vivo, a one-time insertion of the rod into tumors resulted in the retardation of the growth rate by factors of 3-5 compared to untreated controls. Systemic uptake after intratumoral insertion of the rod was negligible. The results suggest that the direct intratumoral insertion of a PdTMPyP4-containing polymeric rod would be of benefit as an adjuvant treatment for patients undergoing chemo- or radiotherapy. By preventing the lengthening of telomeres and therefore the unrestricted growth of cancer cells, our DDS will provide a significant therapeutic advantage to these treatments without affecting normal tissues.


Angiotensin II-dependent persistent podocyte loss from destabilized glomeruli causes progression of end stage kidney disease.
Podocyte depletion is a major mechanism driving glomerulosclerosis. Progression is the process by which progressive glomerulosclerosis leads to end stage kidney disease (ESKD). In order to determine mechanisms contributing to persistent podocyte loss, we used a human diphtheria toxin transgenic rat model. After initial diphtheria toxin-induced podocyte injury (over 30% loss in 4 weeks), glomeruli became destabilized, resulting in continued autonomous podocyte loss causing global podocyte depletion (ESKD) by 13 weeks. This was monitored by urine mRNA analysis and by quantitating podocytes in glomeruli. Similar patterns of podocyte depletion were found in the puromycin aminonucleoside and 5/6 nephrectomy rat models of progressive end-stage disease. Angiotensin II blockade (combined enalapril and losartan) restabilized the glomeruli, and prevented continuous podocyte loss and progression to ESKD. Discontinuing angiotensin II blockade resulted in recurrent glomerular destabilization, podocyte loss, and progression to ESKD. Reduction in blood pressure alone did not reduce proteinuria or prevent podocyte loss from destabilized glomeruli. The protective effect of angiotensin II blockade was entirely accounted for by reduced podocyte loss. Thus, an initiating event resulting in a critical degree of podocyte depletion can destabilize glomeruli and initiate a superimposed angiotensin II-dependent podocyte loss process that accelerates progression resulting in eventual global podocyte depletion and ESKD. These events can be monitored noninvasively in real-time through urine mRNA assays.


Microbial sensing by the intestinal epithelium in the pathogenesis of inflammatory bowel disease.
Recent years have raised evidence that the intestinal microbiota plays a crucial role in the pathogenesis of chronic inflammatory bowels diseases. This evidence comes from several observations. First, animals raised under germ-free conditions do not develop intestinal inflammation in several different model systems. Second, antibiotics are able to modulate the course of experimental colitis. Third, genetic polymorphisms in a variety of genes of the innate immune system have been associated with chronic intestinal inflammatory diseases. Dysfunction of these molecules results in an inappropriate response to bacterial and antigenic stimulation of the innate immune system in the gastrointestinal tract. Variants of pattern recognition receptors such as NOD2 or TLRs by which commensal and pathogenic bacteria can be detected have been shown to be involved in the pathogenesis of IBD. But not only pathways of microbial detection but also intracellular ways of bacterial processing such as autophagosome function are associated with the risk to develop Crohn's disease. Thus, the "environment concept" and the "genetic concept" of inflammatory bowel disease pathophysiology are converging via the intestinal microbiota and the recognition mechanisms for an invasion of members of the microbiota into the mucosa.


Effect of early life exposure to air pollution on development of childhood asthma.
There is increasing recognition of the importance of early environmental exposures in the development of childhood asthma. Outdoor air pollution is a recognized asthma trigger, but it is unclear whether exposure influences incident disease. We investigated the effect of exposure to ambient air pollution in utero and during the first year of life on risk of subsequent asthma diagnosis in a population-based nested case-control study. We assessed all children born in southwestern British Columbia in 1999 and 2000 (n = 37,401) for incidence of asthma diagnosis up to 34 years of age using outpatient and hospitalization records. Asthma cases were age- and sex-matched to five randomly chosen controls from the eligible cohort. We estimated each individual's exposure to ambient air pollution for the gestational period and first year of life using high-resolution pollution surfaces derived from regulatory monitoring data as well as land use regression models adjusted for temporal variation. We used logistic regression analyses to estimate effects of carbon monoxide, nitric oxide, nitrogen dioxide, particulate matter <or= 10 microm and <or= 2.5 microm in aerodynamic diameter (PM10 and PM2.5), ozone, sulfur dioxide, black carbon, woodsmoke, and proximity to roads and point sources on asthma diagnosis. A total of 3,482 children (9%) were classified as asthma cases. We observed a statistically significantly increased risk of asthma diagnosis with increased early life exposure to CO, NO, NO2, PM10, SO2, and black carbon and proximity to point sources. Traffic-related pollutants were associated with the highest risks: adjusted odds ratio = 1.08 (95% confidence interval, 1.041.12) for a 10-microg/m3 increase of NO, 1.12 (1.071.17) for a 10-microg/m3 increase in NO2, and 1.10 (1.061.13) for a 100-microg/m3 increase in CO. These data support the hypothesis that early childhood exposure to air pollutants plays a role in development of asthma.


Constructing monsters: correctional discourse and nursing practice.
This article presents the results of a nursing research that aimed at describing the practice of nursing in an extreme environment where social control and psychiatric nursing care are inextricably enmeshed with one another. The study results indicate that the Correctional Psychiatric Centre (CPC) is a site where two antagonistic discourses (that of the hospital and that of the prison) are contending for the human resources in place. Given that the asylum and the prison are as such two distinct institutions, the correctional psychiatric centre constitutes an ideological space that results from the fusion of both the psychiatric and penal apparatuses. However, characteristics commonly attributed to prisoners, such as 'lying', 'dangerous', 'monstrous' and 'manipulative' are superimposed on the nurses' common theoretical representation that a patient is a person to whom care is provided. Monstrosity was a term regularly employed to describe particular types of inmates. The literature on monsters in quite informative in order to understand the impact of such a representation of forensic psychiatric nursing practice.


Isolation and preliminary enzymatic characterization of a novel PLA2 from Crotalus durissus collilineatus venom.
A crotoxin homolog was purified from the Crotalus durissus collilineatus venom using molecular exclusion and reverse-phase HPLC. This crotoxin contained one PLA2 (Cdcolli III F6) and four crotapotin isoforms, whereas crotoxin from Crotalus durissus terrificus venom had three PLA2 iso forms and two crotapotin isoforms. SDS-PAGE showed that the C. d. collilineatus PLA2 and crotapotin had relative molecular mass of 15 and 9 kDa, respectively. Neither the PLA2 (Cdcolli III F6) nor the crotapotins (Cdcolli III F3 and F4) had any neurotoxicity in mouse phrenic nerve-diaphragm preparations when tested alone. However, when PLA2 and crotapotin were coincubated before testing, the neurotoxicity was restored to a level similar to test in the venom in native crotoxin. The two crotapotins (Cdcolli III F3 and F4) differed in their ability to inhibit PLA2 activity, perhaps because of variations in their affinities for this enzyme. Cdcolli III F6 showed allosteric enzymatic behavior, with maximal activity at pH 8.3 and 36 degrees C. Full PLA2 activity required the presence of a low Ca2+ concentration and was inhibited by Cu2+ and Zn2+ and by Cu2+ and Mg2+ in the presence and absence of Ca2+, respectively. These results indicate that crotoxin from C. d. collineatus venom is very similar enzymatically to crotoxin from C. d. terrificus.


Intracellular Target-Specific Accretion of Cell Penetrating Peptides and Bioportides: Ultrastructural and Biological Correlates.
Cell penetrating peptide (CPP) technologies provide a viable strategy to regulate the activities of intracellular proteins that may be intractable to other biological agents. In particular, the cationic helical domains of proteins have proven to be a reliable source of proteomimetic bioportides, CPPs that modulate the activities of intracellular proteins. In this study we have employed live cell imaging confocal microscopy to determine the precise intracellular distribution of a chemically diverse set of CPPs and bioportides. Our findings indicate that, following efficient cellular entry, peptides are usually accreted at intracellular sites rather than being freely maintained in an aqueous cytosolic environment. The binding of CPPs to proteins in a relatively stable manner provides a molecular explanation for our findings. By extension, it is probable that many bioportides influence biological processes through a dominant-negative influence upon discrete protein-protein interactions. As an example, we report that bioportides derived from the leucine-rich repeat kinase 2 discretely influence the biology and stability of this key therapeutic target in Parkinson's disease. The intracellular site-specific accretion of CPPs and bioportides can also be readily modulated by the attachment of larger cargoes or, more conveniently, short homing motifs. We conclude that site-specific intracellular targeting could be further exploited to expand the scope of CPP technologies.


Strategic implications of changing rule of halves in hypertension: A cross-sectional observational study.
The burden of non-communicable diseases will sooner overwhelm the health system of the country and could curtail future economic development. Hypertension causes highest cardiovascular morbidities. To attain target Blood Pressure (BP), different strategies are required, which are area specific. The validity of rule of halves of hypertension in various settings needs to be studied for developing strategies for that particular geographical area. To utilize the resources in more efficient way, strategies need to differ in rural-urban and underserved- well-served areas. The present study aims at exploring whether the rule of halves holds good in the city of Mumbai as the findings of such research may have implications on strategies to control hypertension in communities. Hence, the objectives of this study are to ascertain the proportion of persons with hypertension with respect to the status of diagnosis, treatment, adequate treatment, and validate it with existing rule of halves. Study was conducted in the urban field practice area of a teaching hospital and medical college of Mumbai that comprises 42 chawl (housing structures); study design- cross-sectional observational study. This observational cross-sectional study was conducted over a period of 18 months in the field practice area of a teaching hospital and medical college of Mumbai with a total population of 43,069. Sample size was 667. After ethical clearance, participants were interviewed using pretested semi-structured interview schedule that included socio-demographic factors, risk factor evaluation, clinical examination, and anthropometry. Subjects with diagnosed hypertension were questioned thoroughly about hypertension adherence. Descriptive statistics was applied on quantitative data. A total of 667 people from five chawls of age more than 35 years were included into the study. The mean age of study subjects was 50.84 ± 11.47 years. Out of total 667 respondents, 20.3% respondents were normotensive, 50.3% respondents were pre-hypertensive, 25.3% respondents were in stage 1 hypertension, and 4% respondents were in stage 2 hypertension. In contrary to the classical rule of halves, the present study shows only 31% of the diagnosed hypertensives are adequately treated. The strategies in the national programs are from the traditional understanding of rule of halves and emphasize increasing awareness and treatment. Family physician should actively involve in regular screening, treatment, and adherence of hypertension treatment. As a part of primary care, community participation is required for effectively achieving the target BP goal of the population.


Nationwide prospective study of outcomes after elective incisional hernia repair.
Incisional hernia repair is a frequent surgical procedure, but perioperative risk factors and outcomes have not been prospectively assessed in large-scale studies. The aim of this nationwide study was to analyze surgical risk factors for early and late outcomes after incisional hernia repair. We conducted a prospective nationwide study on all elective incisional hernia repairs registered in the Danish Ventral Hernia Database between January 1, 2007 and December 31, 2010. Main outcomes measures were surgical risk factors for 30-day readmission, reoperation (excluding recurrence), and mortality after incisional hernia repair. Late outcomes included reoperation for recurrence during the follow-up period. Follow-up was obtained by merging the Danish Ventral Hernia Database with the Danish National Patient Register. Results were evaluated by multivariate analyses. The study included 3,258 incisional hernia repairs. Median follow-up was 21 months (interquartile range 10 to 35 months). The 30-day readmission, reoperation, and mortality rates were 13.3%, 2.2%, and 0.5%, respectively. Advanced age, open repair, large hernia defect, and vertical incision at the primary laparotomy were significant independent risk factors for poor early outcomes (p < 0.05). The cumulated risk of recurrence repair after open and laparoscopic repair was 21.1% and 15.5%, respectively (p = 0.03). Younger age, open repair, hernia defects >7 cm, and onlay or intraperitoneal mesh positioning in open repair were significant risk factors for poor late outcomes (p < 0.05). Elective incisional hernia repair were beset with high rates of readmission and reoperation for recurrence. Readmission and reoperation for recurrence were most pronounced after open repair and repair for hernia defects up to 20 cm. Additionally, sublay mesh position reduced the risk of reoperation for recurrence after open repairs.


'Risking enchantment': how are we to view the smoking person?
The idea of the smoking person portrayed in public health has been criticised as being based on too narrow a view of human nature. This article discusses that view: that of a person with a stable core and epiphenomenal 'behaviours' that can be removed by rational persuasion or Pavlovian manipulation, and examines social scientific critiques of it. The social sciences explore the meanings smoking has for individuals and portray human identity as malleable, the result of ongoing interactions with human and non-human others. Aspects of smokers' experience revealed in qualitative research - descriptions of cigarettes as 'companions' or 'friends', deep reliance, sensual pleasure - are sometimes difficult to articulate but can be given full voice in the context of the literary arts. We explore some examples of these sources and argue that a complete picture of smoking meanings is impossible without reference to them. We take a pragmatic approach, following the philosopher William James, who argued that emotional and spiritual experiences contribute to the truth of human existence as much as material explanations, to suggest that this understanding should be part of a critical but supportive engagement with public health research in order to develop more nuanced and humane approaches to smoking cessation.


Admixture into and within sub-Saharan Africa.
Similarity between two individuals in the combination of genetic markers along their chromosomes indicates shared ancestry and can be used to identify historical connections between different population groups due to admixture. We use a genome-wide, haplotype-based, analysis to characterise the structure of genetic diversity and gene-flow in a collection of 48 sub-Saharan African groups. We show that coastal populations experienced an influx of Eurasian haplotypes over the last 7000 years, and that Eastern and Southern Niger-Congo speaking groups share ancestry with Central West Africans as a result of recent population expansions. In fact, most sub-Saharan populations share ancestry with groups from outside of their current geographic region as a result of gene-flow within the last 4000 years. Our in-depth analysis provides insight into haplotype sharing across different ethno-linguistic groups and the recent movement of alleles into new environments, both of which are relevant to studies of genetic epidemiology.


Study toward resolving the controversy over the definition of allergic fungal rhinosinusitis.
Dysbiosis of the microbiome on the airway mucosa leads to the development of chronic inflammatory and allergic disorders. The aim of this study was to consider the potential diagnostic criteria for allergic fungal rhinosinusitis (AFRS) and nonallergic fungal rhinosinusitis (FRS), and the role of fungal presence in an environment for the development of AFRS. In this study, 136 patients were divided into two groups: patients with positive specific immunoglobulin E (sIgE) and fungal finding (AFRS group), and patients with negative sIgE and positive fungal finding (FRS group). The study design included: anamnesis data, sIgE, eosinophil count and skin-prick test, rhinology and computerized tomography (CT) observation and mycological finding. Our results showed: (i) the prevalence in Serbia is: AFRS 1.3%, FRS 2.8%; (ii) 30.4% patients with sIgE+ had more often severe and recurrent chronic rhinosinusitis (CRS) (P = .005) and the presence of polyps (P = .025); (iii) 46.4% patients with sIgE+ had positive fungi on the sinonasal mucosa and were considered as AFRS; (iv) patients with AFRS had more frequent asthma (P = .024) and chronicity of CRS >10 years (P = .000). The persistent fungal presence and prolonged duration of CRS could be a silent threat for the progression of inflammation and development of FRS. Lavage with hypertonic-NaCl should be included in the everyday hygiene routine in an effort to decrease fungal load and antigenic exposure. The presence of allergological parameters and better response to corticosteroid therapy in AFRS patients should be considered as crucial diagnostic criteria for AFRS.


Comparative glycan profiling of Ceratopteris richardii 'C-Fern' gametophytes and sporophytes links cell-wall composition to functional specialization.
Innovations in vegetative and reproductive characters were key factors in the evolutionary history of land plants and most of these transformations, including dramatic changes in life cycle structure and strategy, necessarily involved cell-wall modifications. To provide more insight into the role of cell walls in effecting changes in plant structure and function, and in particular their role in the generation of vascularization, an antibody-based approach was implemented to compare the presence and distribution of cell-wall glycan epitopes between (free-living) gametophytes and sporophytes of Ceratopteris richardii 'C-Fern', a widely used model system for ferns. Microarrays of sequential diamino-cyclohexane-tetraacetic acid (CDTA) and NaOH extractions of gametophytes, spores and different organs of 'C-Fern' sporophytes were probed with glycan-directed monoclonal antibodies. The same probes were employed to investigate the tissue- and cell-specific distribution of glycan epitopes. While monoclonal antibodies against pectic homogalacturonan, mannan and xyloglucan widely labelled gametophytic and sporophytic tissues, xylans were only detected in secondary cell walls of the sporophyte. The LM5 pectic galactan epitope was restricted to sporophytic phloem tissue. Rhizoids and root hairs showed similarities in arabinogalactan protein (AGP) and xyloglucan epitope distribution patterns. The differences and similarities in glycan cell-wall composition between 'C-Fern' gametophytes and sporophytes indicate that the molecular design of cell walls reflects functional specialization rather than genetic origin. Glycan epitopes that were not detected in gametophytes were associated with cell walls of specialized tissues in the sporophyte.


Epidermal-growth-factor receptor status as predictor of early recurrence of and death from breast cancer.
In 135 primary breast cancers, there was a significant inverse relation between epidermal-growth-factor receptor (EGFR) and oestrogen receptor (ER) status, and a significant association with tumour size and poor differentiation. The relapse-free survival and overall survival were significantly worse for patients with EGFR+ tumours compared with EGFR- tumours. Relapse-free survival and overall survival were also worse for patients with ER- tumours compared with ER+ tumours. Of the 71 ER- patients 28 were EGFR+ and 43 were EGFR-. The relapse-free and overall survival for ER- but EGFR+ patients were significantly worse than for "double-negative" patients. Moreover, relapse-free survival and overall survival for "double-negative" patients were similar to those for ER+ patients. Thus EGFR status divides the ER- population into good and poor prognosis subgroups. The presence of EGFR was the most important variable in the primary tumours for predicting relapse-free and overall survival. Multivariate analysis showed that EGFR status was the most important variable in predicting relapse-free and overall survival in lymph-node-negative patients, and the second most important variable in lymph-node positive patients.


Intervention reporting of clinical trials published in high-impact cardiology journals: effect of the TIDieR checklist and guide.
Randomised controlled trials (RCTs) provide the highest-level of evidence among primary research in cardiovascular medicine. Yet, even the best trial may be less useful if it fails to provide an accurate means of reproducibility. Unfortunately, discrepancies in the standards of trial reporting have been persistent in previous trials. The Template for Intervention Description and Replication (TIDieR) checklist aims to improve research efficacy by setting standards for quality intervention reporting and reproducibility. The goal of this study was to assess adherence to the TIDieR checklist among RCTs published in cardiovascular health journals. We also compared the quality of intervention reporting before and after the publication of TIDieR. This cross-sectional, methodological study analysed 101 trials published within high-impact cardiology journals. Our primary objective was to assess overall adherence to the TIDieR checklist. Our secondary objective was to use an interrupted time-series analysis to determine if intervention reporting increased following the publication of TIDieR in March 2014. Additionally, we used generalised estimating equations to identify trial characteristics associated with intervention reporting. Trials in our sample reported 8.6/12 TIDieR checklist items, on average. The most under-reported items were those for describing the expertise of the interventionists and for describing the location of the intervention. Improved outcome reporting and intervention reproducibility among RCTs are greatly needed in cardiovascular medicine. Clinicians and researchers should advocate for the ethical publication of complete, translatable and replicable clinical research results.


Periodic segmental anomalies induced by heat shock in the chick embryo are associated with the cell cycle.
This study provides evidence that cells destined to segment together into somites have a degree of cell division synchrony. We have measured the duration of the cell division cycle in somite and segmental plate cells of the chick embryo as 9.5 h using [3H]thymidine pulse- and-chase. Treatment of embryos with any of a variety of inhibitors known to affect the cell division cycle causes discrete periodic segmental anomalies: these anomalies appear about 6-7 somites after treatment and, in some cases, a second anomaly is observed 6 to 7 somites after the first. Since somites take 1.5 h to form, the 6- to 7- somite interval corresponds to about 9-10 h, which is the duration of the cell cycle as determined in these experiments. The anomalies are similar to those seen after heat shock of 2-day chick embryos. Heat shock and some of the other treatments induce the expression of heat-shock proteins (hsp); however, since neither the expression nor the distribution of these proteins relate to the presence or distribution of anomalies seen, we conclude that hsps are not responsible for the pattern of segmental anomalies observed. The production of periodic segmental anomalies appears to be linked to the cell cycle. A simple model is proposed, in which we suggest that the cell division cycle is involved directly in gating cells that will segment together.


Emergency care practitioners' barriers to mental health assessment, treatment, and referral of post-injury patients.
Nearly half of all severely injured patients suffer some form of post-trauma mental stress, but little is known about factors that influence emergency care practitioners' decisions to refer injured patients to mental health care services. This study aimed to: (1) advance our understanding of the practice barriers that hinder mental health assessment, treatment, and referral of injured patients in emergency care settings, and (2) determine the preferred learning format of emergency care practitioners who desire to gain knowledge about mental health problems after injury. Using a mail survey research design, data were collected from a random sample of emergency medicine physicians and nurses in Wisconsin and New York. Data was provided by 108 respondents with an average of 16 years of emergency care experience. More than half indicated they never refer trauma patients for mental health follow-up. Primary reasons for not dealing with trauma-related mental health issues were insufficient time and lack of symptoms. Providers who were most satisfied with their hospital's capacity to support mental health care were significantly more likely to refer patients. The top preference for receiving additional training related to mental health needs of trauma patients was on-site lectures. Injured patients rely on emergency care practitioners to provide multidimensional care. However, few practitioners facilitate mental health referrals for post-trauma victims, despite their known value.


An international effort towards developing standards for best practices in analysis, interpretation and reporting of clinical genome sequencing results in the CLARITY Challenge.
There is tremendous potential for genome sequencing to improve clinical diagnosis and care once it becomes routinely accessible, but this will require formalizing research methods into clinical best practices in the areas of sequence data generation, analysis, interpretation and reporting. The CLARITY Challenge was designed to spur convergence in methods for diagnosing genetic disease starting from clinical case history and genome sequencing data. DNA samples were obtained from three families with heritable genetic disorders and genomic sequence data were donated by sequencing platform vendors. The challenge was to analyze and interpret these data with the goals of identifying disease-causing variants and reporting the findings in a clinically useful format. Participating contestant groups were solicited broadly, and an independent panel of judges evaluated their performance. A total of 30 international groups were engaged. The entries reveal a general convergence of practices on most elements of the analysis and interpretation process. However, even given this commonality of approach, only two groups identified the consensus candidate variants in all disease cases, demonstrating a need for consistent fine-tuning of the generally accepted methods. There was greater diversity of the final clinical report content and in the patient consenting process, demonstrating that these areas require additional exploration and standardization. The CLARITY Challenge provides a comprehensive assessment of current practices for using genome sequencing to diagnose and report genetic diseases. There is remarkable convergence in bioinformatic techniques, but medical interpretation and reporting are areas that require further development by many groups.


The aquaglyceroporin AQP9 contributes to the sex-specific effects of in utero arsenic exposure on placental gene expression.
Sex-specific factors play a major role in human health and disease, including responses to environmental stresses such as toxicant exposure. Increasing evidence suggests that such sex differences also exist during fetal development. In a previous report using the resources of the New Hampshire Birth Cohort Study (NHBCS), we found that low-to-moderate in utero exposure to arsenic, a highly toxic and widespread pollutant, was associated with altered expression of several key developmental genes in the fetal portion of the placenta. These associations were sex-dependent, suggesting that in utero arsenic exposure differentially impacts male and female fetuses. In the present study, we investigated the molecular basis for these sex-specific responses to arsenic. Using NanoString technology, we further analyzed the fetal placenta samples from the NHBCS for the expression of genes encoding arsenic transporters and metabolic enzymes. Multivariable linear regression analysis was used to examine their relationship with arsenic exposure and with key developmental genes, after stratification by fetal sex. We found that maternal arsenic exposure was strongly associated with expression of the AQP9 gene, encoding an aquaglyceroporin transporter, in female but not male fetal placenta. Moreover, AQP9 expression associated with that of a subset of female-specific arsenic-responsive genes. Our results suggest that AQP9 is upregulated in response to arsenic exposure in female, but not male, fetal placenta. Based on these results and prior studies, increased AQP9 expression may lead to increased arsenic transport in the female fetal placenta, which in turn may alter the expression patterns of key developmental genes that we have previously shown to be associated with arsenic exposure. Thus, this study suggests that AQP9 may play a role in the sex-specific effects of in utero arsenic exposure.


Deletion of genes encoding cytochrome oxidases and quinol monooxygenase blocks the aerobic-anaerobic shift in Escherichia coli K-12 MG1655.
The constitutive activation of the anoxic redox control transcriptional regulator (ArcA) in Escherichia coli during aerobic growth, with the consequent production of a strain that exhibits anaerobic physiology even in the presence of air, is reported in this work. Removal of three terminal cytochrome oxidase genes (cydAB, cyoABCD, and cbdAB) and a quinol monooxygenase gene (ygiN) from the E. coli K-12 MG1655 genome resulted in the activation of ArcA aerobically. These mutations resulted in reduction of the oxygen uptake rate by nearly 98% and production of d-lactate as a sole by-product under oxic and anoxic conditions. The knockout strain exhibited nearly identical physiological behaviors under both conditions, suggesting that the mutations resulted in significant metabolic and regulatory perturbations. In order to fully understand the physiology of this mutant and to identify underlying metabolic and regulatory reasons that prevent the transition from an aerobic to an anaerobic phenotype, we utilized whole-genome transcriptome analysis, (13)C tracing experiments, and physiological characterization. Our analysis showed that the deletions resulted in the activation of anaerobic respiration under oxic conditions and a consequential shift in the content of the quinone pool from ubiquinones to menaquinones. An increase in menaquinone concentration resulted in the activation of ArcA. The activation of the ArcB/ArcA regulatory system led to a major shift in the metabolic flux distribution through the central metabolism of the mutant strain. Flux analysis indicated that the mutant strain had undetectable fluxes around the tricarboxylic acid (TCA) cycle and elevated flux through glycolysis and anaplerotic input to oxaloacetate. Flux and transcriptomics data were highly correlated and showed similar patterns.


Decline in male circumcision in South Korea.
To investigate the changing circumcision rate in South Korea in the last decade and to propose underlying causes for this change, in the context of the present fluctuating world-wide trends in circumcision. From 2009 to 2011, 3,296 South Korean males (or their parents) aged 0-64 years were asked about their circumcision status, their age at circumcision, and their information level regarding circumcision. We employed non-probability sampling considering the sensitive questions on the study theme. Currently the age-standardized circumcision rate for South Korean males aged 14-29 is found to be 75.8%. In an earlier study performed in 2002, the rate for the same age group was 86.3%. Of particular interest, males aged 14-16 show a circumcision rate of 56.4%, while the same age group 10 years ago displayed a much higher percentage, at 88.4%. In addition, the extraordinarily high circumcision rate of 95.2% found 10 years ago for the 17-19 age group is now reduced to 74.4%. Interestingly, of the circumcised males, the percentage circumcised in the last decade was only 25.2%; i.e., the majority of the currently circumcised males had undergone the operation prior to 2002, indicating that the actual change in the last decade is far greater. Consistent with this conjecture, the 2002 survey showed that the majority of circumcised males (75.7%) had undergone the operation in the decade prior to that point. Focusing on the flagship age group of 14-16, this drop suggests that, considering the population structure of Korean males, approximately one million fewer circumcision operations have been performed in the last decade relative to the case of non-decline. This decline is strongly correlated with the information available through internet, newspapers, lectures, books, and television: within the circumcised population, both the patients and their parents had less prior knowledge regarding circumcision, other than information obtained from person to person by oral communication. Within the uncircumcised population, the prior knowledge was far greater, suggesting that information discouraging circumcision played an important role. South Korean male circumcision is likely to be undergoing a steep decline. The cause for this decline seems to be the increase in information available on the pros and cons of circumcision.


Physical activity parenting: a systematic review of questionnaires and their associations with child activity levels.
Insufficient physical activity (PA) is considered a critical contributor to childhood overweight. Parents are a key in influencing their child's PA through various mechanisms of PA parenting, including support, restriction of PA and facilitation of enrolment in PA classes or activities. However, study findings are difficult to compare because instruments vary in terms of constructs, psychometric assessment and type of PA assessed. The goal of the current review was to identify existing PA parenting questionnaires and report on the validation of these measures through findings of their psychometric performance and correlation to youth's PA. The search of eligible studies was restricted to instruments with multiple items. Eleven unique PA parenting questionnaires were identified, and 46 studies that used these instruments were included. Extracted data include sample characteristics, as well as type and assessment methods of parental influence and PA. Findings highlight the tremendous variation in the conceptualization and measurement of PA parenting, common use of non-validated instruments and lack of comprehensive measures. The development of theory-based PA parenting measures (preferably multidimensional) should be prioritized to guide the study of the parental role in promoting child's PA as well as the design of family-based PA interventions.


Intravoxel Incoherent Motion Metrics as Potential Biomarkers for Survival in Glioblastoma.
Intravoxel incoherent motion (IVIM) is an MRI technique with potential applications in measuring brain tumor perfusion, but its clinical impact remains to be determined. We assessed the usefulness of IVIM-metrics in predicting survival in newly diagnosed glioblastoma. Fifteen patients with glioblastoma underwent MRI including spin-echo echo-planar DWI using 13 b-values ranging from 0 to 1000 s/mm2. Parametric maps for diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) were generated for contrast-enhancing regions (CER) and non-enhancing regions (NCER). Regions of interest were manually drawn in regions of maximum f and on the corresponding dynamic susceptibility contrast images. Prognostic factors were evaluated by Kaplan-Meier survival and Cox proportional hazards analyses. We found that fCER and D*CER correlated with rCBFCER. The best cutoffs for 6-month survival were fCER>9.86% and D*CER>21.712 x10-3mm2/s (100% sensitivity, 71.4% specificity, 100% and 80% positive predictive values, and 80% and 100% negative predictive values; AUC:0.893 and 0.857, respectively). Treatment yielded the highest hazard ratio (5.484; 95% CI: 1.162-25.88; AUC: 0.723; P = 0.031); fCER combined with treatment predicted survival with 100% accuracy. The IVIM-metrics fCER and D*CER are promising biomarkers of 6-month survival in newly diagnosed glioblastoma.


Pathological processing techniques and final diagnosis of breast cancer sentinel lymph nodes.
Recommendations for intraoperative and postoperative breast sentinel lymph node (SLN) processing differ widely. Micrometastases and isolated tumor cells (ITC) have recently been proposed as prognostically and therapeutically relevant. We compared 3 SLN protocols with regard to intraoperative and postoperative diagnosis. SLN in cohort I (270 patients) were intraoperatively assessed by stereomicroscopy. Intraoperative frozen section (IFS) was used only in stereomicroscopically suspicious SLN. In cohort II (197 patients), all SLN were examined with only 1 IFS. Final SLN workup in cohorts I and II consisted of complete step sectioning with immunohistochemistry. In cohort III (268 patients) 2 or more IFS were performed followed by 3 step sections and immunohistochemistry. pN1 stages were significantly higher in cohorts I and II (33.3% and 34.0% respectively) than in cohort III (24.6%). Intraoperative false negativity for the detection of metastases (pN1) ranged from 54.4% (cohort I) and 35.8% (cohort II) to 21.2% (cohort III). In contrast, ITC were detected significantly more frequently in cohort I (9.3%) and cohort II (14.7%) than in cohort III (1.9%). Higher rates of SLN metastases and ITC in cohort I/II compared to cohort III suggest that IFS may result in tissue loss thus increasing the risk of missing metastases. Sparse IFS but complete postoperative SLN workup with step sectioning and immunohistochemistry provides more accurate information regarding minimal disease in SLN, but often results in delayed axillary lymph node dissection. This is important for preoperative patient information and recommendations in SLN processing protocols.


Phosphotyrosine signaling analysis in human tumors is confounded by systemic ischemia-driven artifacts and intra-specimen heterogeneity.
Tumor protein phosphorylation analysis may provide insight into intracellular signaling networks underlying tumor behavior, revealing diagnostic, prognostic or therapeutic information. Human tumors collected by The Cancer Genome Atlas program potentially offer the opportunity to characterize activated networks driving tumor progression, in parallel with the genetic and transcriptional landscape already documented for these tumors. However, a critical question is whether cellular signaling networks can be reliably analyzed in surgical specimens, where freezing delays and spatial sampling disparities may potentially obscure physiologic signaling. To quantify the extent of these effects, we analyzed the stability of phosphotyrosine (pTyr) sites in ovarian and colon tumors collected under conditions of controlled ischemia and in the context of defined intratumoral sampling. Cold-ischemia produced a rapid, unpredictable, and widespread impact on tumor pTyr networks within 5 minutes of resection, altering up to 50% of pTyr sites by more than 2-fold. Effects on adhesion and migration, inflammatory response, proliferation, and stress response pathways were recapitulated in both ovarian and colon tumors. In addition, sampling of spatially distinct colon tumor biopsies revealed pTyr differences as dramatic as those associated with ischemic times, despite uniform protein expression profiles. Moreover, intratumoral spatial heterogeneity and pTyr dynamic response to ischemia varied dramatically between tumors collected from different patients. Overall, these findings reveal unforeseen phosphorylation complexity, thereby increasing the difficulty of extracting physiologically relevant pTyr signaling networks from archived tissue specimens. In light of this data, prospective tumor pTyr analysis will require appropriate sampling and collection protocols to preserve in vivo signaling features.


Genetic diversity and population structure of wheat in India and Turkey.
Genetic diversity among plant species offers prospects for improving the plant characteristics. Its assessment is necessary to help tackle the threats of environmental fluctuations and for the effective exploitation of genetic resources in breeding programmes. Although wheat is one of the most thoroughly studied crops in terms of genetic polymorphism studies, phylogenetic affinities of Indian and Turkish Triticum species have not been assessed to date. In this study, genetic association of 95 tetraploid and hexaploid wheat genotypes originating from India and Turkey was determined for the first time. Combined analysis of random amplified polymorphic DNA and inter-simple sequence repeat markers disclosed 177 polymorphic bands, and both the dendrogram and two-dimensional scatterplot showed similar groupings of the wheat genotypes. Turkish hexaploid varieties were basically divided into two clusters, one group showed its close association with Indian hexaploid varieties and the other with Indian tetraploid varieties. Analysis of molecular variance revealed high (77 %) genetic variation within Indian and Turkish populations. Population structure analysis elucidated distinct clustering of wheat genotypes on the basis of both geographical origin and ploidy. The results revealed in this study will support worldwide wheat breeding programmes and assist in achieving the target of sustainable wheat production.


Merchandising of cigarettes in San Francisco pharmacies: 27 years later.
To estimate changes since 1976 in the proportion of San Francisco pharmacies that sell cigarettes and to characterise the advertising of cigarettes and the merchandising of non-prescription nicotine replacement therapy (NRT) products in these retail establishments. 100 randomly selected San Francisco pharmacies were visited in 2003. Pharmacies were characterised based on the sale of cigarettes, advertising for cigarettes, and the merchandising of non-prescription NRT products. In 2003, 61% of pharmacies sold cigarettes, a significant decrease compared to 89% of pharmacies selling cigarettes in 1976 (p < 0.001); 84% of pharmacies selling cigarettes also displayed cigarette advertising. Non-prescription NRT products were stocked by 78% of pharmacies, and in 55% of pharmacies selling cigarettes, the NRT products were stocked immediately adjacent to the cigarettes. Since 1976, there has been a decline in the overall proportion of pharmacies in San Francisco that sell cigarettes yet most pharmacies, particularly traditional chain pharmacies, continue to merchandise the primary known risk factor for death in the USA.


Detection of stratified microbial populations related to Chlorobium and Fibrobacter species in the Atlantic and Pacific oceans.
A gene lineage (SAR406) related to Chlorobium and Fibrobacter species was found in 16S rRNA gene clone libraries prepared from samples from two oceans. The clone libraries were constructed from total picoplankton genomic DNA to assess bacterial diversity in the lower surface layer. The samples were collected by filtration from a depth of 80 m at a site in the western Sargasso Sea and from a depth of 120 m at a site in the Pacific Ocean, approximately 70 km from the Oregon coast. The PCR and primers which amplified nearly full-length 16S rRNA genes were used to prepare the clone libraries. Among the diverse gene clones in these libraries were two related clones (SAR406 and OCS307) which could not be assigned to any of the major bacterial phyla. Phylogenetic analyses demonstrated that these genes were distant relatives of the genus Fibrobacter and the green sulfur bacterial phylum, which includes the genus Chlorobium. The inclusion of SAR406 in phylogenetic trees inferred by several methods resulted in support from bootstrap replicates for the conclusion that Fibrobacter and Chlorobium species and SAR406 are a monophyletic group. An oligonucleotide probe that selectively hybridized to clone SAR406 was used to examine the distribution of this gene lineage in vertical profiles from the Atlantic and Pacific Oceans and in monthly time series at 0 and 200 m in the Atlantic Ocean. During stratified periods, the genes were most abundant slightly below the deep chlorophyll layer. Seasonal changes in the surface abundance of SAR406 rDNA were highly correlated with chlorophyll a levels (r = 0.75).


Enantioselective CuH-catalyzed anti-Markovnikov hydroamination of 1,1-disubstituted alkenes.
Enantioselective synthesis of β-chiral amines has been achieved via copper-catalyzed hydroamination of 1,1-disubstituted alkenes with hydroxylamine esters in the presence of a hydrosilane. This mild process affords a range of structurally diverse β-chiral amines, including β-deuterated amines, in excellent yields with high enantioselectivities. Furthermore, catalyst loading as low as 0.4 mol% could be employed to deliver product in undiminished yield and selectivity, demonstrating the practicality of this method for large-scale synthesis.


pOp6/LhGR: a stringently regulated and highly responsive dexamethasone-inducible gene expression system for tobacco.
We describe pOp/LhGR, a dexamethasone-inducible derivative of the pOp/LhG4 transcription activation system, and its use in tobacco to regulate expression of uidA (encoding beta-glucuronidase; GUS) and the cytokinin-biosnythetic gene ipt. The pOp/LhGR system exhibited stringent regulation and strong induced phenotypes in soil and tissue culture. In conjunction with an improved target promoter, pOp6, that carries six copies of an optimized lac operator sequence the pOp6/LhGR system directed induced GUS activities that exceeded those obtained with pOp/LhG4 or the CaMV 35S promoter but without increased uninduced activity. A single dose of dexamethasone was sufficient to direct cytotoxic levels of ipt expression in soil-grown plants although uninduced plants grew normally throughout a complete life cycle. In vitro, induced transcripts were detectable within an hour of dexamethasone application and 1 nM dexamethasone was sufficient for half maximal induction of GUS activity. Various methods of dexamethasone application were successfully applied under tissue culture and greenhouse conditions. We observed no inhibitory effects of dexamethasone or LhGR on plant development even with the highest concentrations of inducer, although tobacco seedlings were adversely affected by ethanol used as a solvent for dexamethasone stock solutions. The pOp/LhGR system provides a highly sensitive, efficient, and tightly regulated chemically inducible transgene expression system for tobacco plants.


Smoking cessation and desire to stop smoking in nine countries of the former Soviet Union.
Smoking rates and corresponding levels of premature mortality from smoking-related diseases in the former Soviet Union (fSU) are among the highest in the world. To reduce this health burden, greater focus on smoking cessation is needed, but little is currently known about rates and characteristics of cessation in the fSU. Nationally representative household survey data from a cross-sectional study of 18,000 respondents in Armenia, Azerbaijan, Belarus, Georgia, Kazakhstan, Kyrgyzstan, Moldova, Russia, and Ukraine were analyzed to describe patterns of desire and action taken to stop smoking, quit ratios (former ever-smokers as a percent of ever-smokers, without a specified recall period), and help used to stop smoking. Multivariate logistic regression was used to analyze characteristics associated with smoking cessation and desire to stop smoking. Quit ratios varied from 10.5% in Azerbaijan to 37.6% in Belarus. About 67.2% of respondents expressed a desire to quit, and 64.9% had taken action and tried to stop. The use of help to quit was extremely low (12.6%). Characteristics associated with cessation included being female, over 60, with higher education, poorer health, lower alcohol dependency, higher knowledge of tobacco's health effects, and support for tobacco control. Characteristics associated with desire to stop smoking among current smokers included younger age, poorer health, greater knowledge of tobacco's health effects, and support for tobacco control. Quit ratios are low in the fSU but there is widespread desire to stop smoking. Stronger tobacco control and cessation support are urgently required to reduce smoking prevalence and associated premature mortality.