Bradycardia during therapeutic hypothermia is associated with good neurologic outcome in comatose survivors of out-of-hospital cardiac arrest.
Comatose patients resuscitated after out-of-hospital cardiac arrest receive therapeutic hypothermia. Bradycardia is frequent during therapeutic hypothermia, but its impact on outcome remains unclear. We explore a possible association between bradycardia during therapeutic hypothermia and neurologic outcome in comatose survivors of out-of-hospital cardiac arrest. Retrospective cohort study, from January 2009 to January 2011. University hospital medical and cardiac ICUs. One hundred eleven consecutive comatose out-of-hospital cardiac arrest patients treated with therapeutic hypothermia. Patients treated with standardized treatment protocol after cardiac arrest. All out-of-hospital cardiac arrest patients' records were reviewed. Hemodynamic data were obtained every fourth hour during the first days. The patients were in temperature target range (32-34°C) 8 hours after out-of-hospital cardiac arrest and dichotomized into bradycardia and nonbradycardia groups depending on their actual heart rate less than or equal to 60 beats/min or more than 60 beats/min at that time. Primary endpoint was Cerebral Performance Category score at hospital discharge. More nonbradycardia group patients received epinephrine during resuscitation and epinephrine and norepinephrine in the early in-hospital period. They also had lower base excess at admission. Survival rate with favorable outcome was significantly higher in the bradycardia than the nonbradycardia group (60% vs 37%, respectively, p = 0.03). For further heart rate quantification, patients were divided into quartiles: less than or equal to 49 beats/min, 50-63 beats/min, 64-77 beats/min, and more than or equal to 78 beats/min, with respective proportions of patients with good outcome at discharge of 18 of 27 (67%), 14 of 25 (56%), 12 of 28 (43%), and 7 of 27 (26%) (p = 0.002). Patients in the lowest quartile had significantly better outcome than the higher groups (p = 0.027), whereas patients in the highest quartile had significantly worse outcome than the lower three groups (p = 0.013). Bradycardia during therapeutic hypothermia was associated with good neurologic outcome at hospital discharge. Our data indicate that bradycardia should not be aggressively treated in this period if mean arterial pressure, lactate clearance, and diuresis are maintained at acceptable levels. Studies, both experimental and clinical, are warranted.


An experimental medicine study of the effects of simvastatin on emotional processing, reward learning, verbal memory, and inflammation in healthy volunteers.
Clinical studies suggest that the highly lipophilic, anti-inflammatory molecule, simvastatin, might be an ideal candidate for drug repurposing in the treatment of depression. The neuropsychological effects of simvastatin are not known, but their ascertainment would have significant translational value about simvastatin's influence on mood and cognition. We aimed to investigate the effects of simvastatin on a battery of psychological tests and inflammatory markers in healthy volunteers. Fifty-three healthy subjects were randomly assigned to 7 days of either simvastatin (N = 27) or sucrose-based placebo (N = 26) given in a double-blind fashion. Then, participants were administered questionnaires measuring subjective rates of mood and anxiety, and a battery of tasks assessing emotional processing, reward learning, and verbal memory. Blood samples for C-reactive protein were also collected. Compared to placebo, participants on simvastatin showed a higher number of positively valenced intrusions in the emotional recall task (F1,51 = 4.99, p = 0.03), but also an increase in anxiety scores (F1,51 = 5.37, p = 0.02). An exploratory analysis of the females' subgroup (N = 27) showed lower number of misclassifications as sad facial expression in the simvastatin arm (F1,25 = 6.60, p = 0.02). No further statistically significant changes could be observed on any of the other outcomes measured. We found limited evidence that 7-day simvastatin use in healthy volunteer induces a positive emotional bias while also being associated with an increase in anxiety, potentially reflecting the early effects of antidepressants in clinical practice. Such effect might be more evident in female subjects. Different drug dosages, treatment lengths, and sample selection need consideration in further experimental medicine and clinical studies. Clinicaltrials.gov: NCT04652089.


Envisioning Future Directions: Conversations With Leaders in Domestic and Sexual Assault Advocacy, Policy, Service, and Research.
This article delves into the views of 72 leaders in domestic violence and sexual assault advocacy, policy, service, and research to determine their vision for the future direction of the field. Through discussions with experts, we identified numerous strategies necessary to best meet the needs of domestic violence and sexual assault victims. Common themes focused on the need to (a) examine the context of victims' and offenders' experiences; (b) increase cultural competence to adequately provide appropriate victim services and criminal justice responses for underserved, marginalized, and culturally specific populations; (c) increase reliance on victims' voices; (d) continue to develop partnerships at both the community and the state levels and ensure the role of local communities; (e) expand the concept of successful outcomes that can be reliably and validly assessed; (f) emphasize mixed-methods approaches to address these questions, in recognition that various methods complement each other; and (g) be open to novel or emerging approaches to intervention.


Understanding and improving aggregated gold nanoparticle/dsDNA interactions by molecular spectroscopy and deconvolution methods.
It is well known that single-stranded DNA (ssDNA) is easily able to adsorb on citrate-capped, non-functionalized gold nanoparticles (AuNPs). However, the affinity of double-stranded DNA (dsDNA) for them is much more limited. The present work demonstrates that long dsDNA suffers from a bending conformational change when anionic nanoparticles are present in solution. A striking decrease in the persistence length of the double helix in the absence of salt is observed through dynamic light scattering (DLS), viscometric, and atomic force microscopy (AFM) methods. Long dsDNA is therefore shown to be able to interact with anionic gold nanoparticles. To date, only ssDNA detection has been described by making use of interparticle cross-linking aggregation mechanisms; however, the data shown in this work allow for the development of new methods for detecting dsDNA in solution by using aggregated AuNPs as a starting point. The aggregation state is induced by the controlled addition of an inert electrolyte. A deconvolution procedure of the experimental plasmon shows how individual bands corresponding to aggregated nanoclusters diminish as the DNA concentration increases in the presence of 0.075 M NaCl.


Changing reimbursement models and private equity ownership in spine surgery.
Rising costs of healthcare have been a problem in the United States in the past decade, with unnecessary strain on both providers and patients. The fee-for-service reimbursement model has been proposed as one of the biggest contributors to this phenomenon; as such, newer reimbursement model-including the bundled payments model-have been proposed and are becoming more widely adopted. This review will first discuss the current payment models in practice and then overview private equity ownership as a new avenue in this particular domain.


Integrated glycoprotein immobilization method for glycopeptide and glycan analysis of cardiac hypertrophy.
Post-translational modifications of proteins can have a major role in disease initiation and progression. Incredible efforts have recently been made to study the regulation of glycoproteins for disease prognosis and diagnosis. It is essential to elucidate glycans and intact glycoproteins to understand the role of glycosylation in diseases. Sialylated N-glycans play crucial roles in physiological and pathological processes; however, it is laborious to study sialylated glycoproteins due to the labile nature of sialic acid residues. In this study, an integrated platform is developed for the analysis of intact glycoproteins and glycans using a chemoenzymatic approach for immobilization and derivatization of sialic acids. N-Glycans, deglycosylated proteins, and intact glycoproteins from heart tissues of wild type (WT) and transverse aortic constriction (TAC) mouse models were analyzed. We identified 291 unique glycopeptides from 195 glycoproteins; the comparative studies between WT and TAC mice indicate the overexpression of extracellular proteins for heart matrix remodeling and the down-regulation of proteins associated with energy metabolism in cardiac hypertrophy. The integrated platform is a powerful tool for the analysis of glycans and glycoproteins in the discovery of potential cardiac hypertrophy biomarkers.


A gene complex controlling segmentation in Drosophila.
The bithorax gene complex in Drosophila contains a minimum of eight genes that seem to code for substances controlling levels of thoracic and abdominal development. The state of repression of at least four of these genes is controlled by cis-regulatory elements and a separate locus (Polycomb) seems to code for a repressor of the complex. The wild-type and mutant segmentation patterns are consistent with an antero-posterior gradient in repressor concentration along the embryo and a proximo-distal gradient along the chromosome in the affinities for repressor of each gene's cis-regulatory element.


Supervision to prevent childhood unintentional injury: developmental knowledge and self-efficacy count.
Caregiver developmental knowledge was tested as a moderator of the association between caregiver-perceived self-efficacy and perceived supervision to prevent childhood unintentional injury. Caregivers (N = 123; 95 mothers and 28 fathers) of children (M = 3.5 ± 1.2 years, 49.6% female, 80.8% Caucasian) were recruited from pediatric primary care offices and local message boards. All caregivers completed self-report questionnaires on perceived self-efficacy, developmental knowledge, and perceived supervision to prevent injury. Separate hierarchical linear regression models were conducted to test moderation for mothers and fathers. The interaction effect of perceived self-efficacy and developmental knowledge significantly predicted maternal-perceived supervision, R (2) change = .06, F(1, 86) = 6.76, p < .01. No significant findings were detected for fathers. Models of studying injury prevention that consider complex cognitive-behavioral interactions and their potential modifiable role in the development of injury-prevention practices may elucidate upon the attitude-practice gap currently identified in the literature.


Endpoints in stem cell trials in ischemic heart failure.
Despite multimodal regimens and diverse treatment options alleviating disease symptoms, morbidity and mortality associated with advanced ischemic heart failure remain high. Recently, technological innovation has led to the development of regenerative therapeutic interventions aimed at halting or reversing the vicious cycle of heart failure progression. Driven by the unmet patient need and fueled by encouraging experimental studies, stem cell-based clinical trials have been launched over the past decade. Collectively, these trials have enrolled several thousand patients and demonstrated the clinical feasibility and safety of cell-based interventions. However, the totality of evidence supporting their efficacy in ischemic heart failure remains limited. Experience from the early randomized stem cell clinical trials underscores the key points in trial design ranging from adequate hypothesis formulation to selection of the optimal patient population, cell type and delivery route. Importantly, to translate the unprecedented promise of regenerative biotherapies into clinical benefit, it is crucial to ensure the appropriate choice of endpoints along the regulatory path. Accordingly, we here provide considerations relevant to the choice of endpoints for regenerative clinical trials in the ischemic heart failure setting.


From computers to cultivation: reconceptualizing evolutionary psychology.
Does evolutionary theorizing have a role in psychology? This is a more contentious issue than one might imagine, given that, as evolved creatures, the answer must surely be yes. The contested nature of evolutionary psychology lies not in our status as evolved beings, but in the extent to which evolutionary ideas add value to studies of human behavior, and the rigor with which these ideas are tested. This, in turn, is linked to the framework in which particular evolutionary ideas are situated. While the framing of the current research topic places the brain-as-computer metaphor in opposition to evolutionary psychology, the most prominent school of thought in this field (born out of cognitive psychology, and often known as the Santa Barbara school) is entirely wedded to the computational theory of mind as an explanatory framework. Its unique aspect is to argue that the mind consists of a large number of functionally specialized (i.e., domain-specific) computational mechanisms, or modules (the massive modularity hypothesis). Far from offering an alternative to, or an improvement on, the current perspective, we argue that evolutionary psychology is a mainstream computational theory, and that its arguments for domain-specificity often rest on shaky premises. We then go on to suggest that the various forms of e-cognition (i.e., embodied, embedded, enactive) represent a true alternative to standard computational approaches, with an emphasis on "cognitive integration" or the "extended mind hypothesis" in particular. We feel this offers the most promise for human psychology because it incorporates the social and historical processes that are crucial to human "mind-making" within an evolutionarily informed framework. In addition to linking to other research areas in psychology, this approach is more likely to form productive links to other disciplines within the social sciences, not least by encouraging a healthy pluralism in approach.


Allele surfing promotes microbial adaptation from standing variation.
The coupling of ecology and evolution during range expansions enables mutations to establish at expanding range margins and reach high frequencies. This phenomenon, called allele surfing, is thought to have caused revolutions in the gene pool of many species, most evidently in microbial communities. It has remained unclear, however, under which conditions allele surfing promotes or hinders adaptation. Here, using microbial experiments and simulations, we show that, starting with standing adaptive variation, range expansions generate a larger increase in mean fitness than spatially uniform population expansions. The adaptation gain results from 'soft' selective sweeps emerging from surfing beneficial mutations. The rate of these surfing events is shown to sensitively depend on the strength of genetic drift, which varies among strains and environmental conditions. More generally, allele surfing promotes the rate of adaptation per biomass produced, which could help developing biofilms and other resource-limited populations to cope with environmental challenges.


Setting mastery learning standards.
Mastery learning is an instructional approach in which educational progress is based on demonstrated performance, not curricular time. Learners practice and retest repeatedly until they reach a designated mastery level; the final level of achievement is the same for all, although time to mastery may vary. Given the unique properties of mastery learning assessments, a thoughtful approach to establishing the performance levels and metrics that determine when a learner has demonstrated mastery is essential.Standard-setting procedures require modification when used for mastery learning settings in health care, particularly regarding the use of evidence-based performance data, the determination of appropriate benchmark or comparison groups, and consideration of patient safety consequences. Information about learner outcomes and past performance data of learners successful at the subsequent level of training can be more helpful than traditional information about test performance of past examinees. The marginally competent "borderline student" or "borderline group" referenced in traditional item-based and examinee-based procedures will generally need to be redefined in mastery settings. Patient safety considerations support conjunctive standards for key knowledge and skill subdomains and for items that have an impact on clinical outcomes. Finally, traditional psychometric indices used to evaluate the quality of standards do not necessarily reflect critical measurement properties of mastery assessments. Mastery learning and testing are essential to the achievement and assessment of entrustable professional activities and residency milestones. With careful attention, sound mastery standard-setting procedures can provide an essential step toward improving the effectiveness of health professions education, patient safety, and patient care.


Cysteine supplementation reverses methionine restriction effects on rat adiposity: significance of stearoyl-coenzyme A desaturase.
Stearoyl-CoA desaturase-1 (SCD1) is a key enzyme in fatty acid and energy metabolism, but little is known about its nutritional regulation. Dietary methionine restriction in rats decreases hepatic Scd1 mRNA and protein, increases energy expenditure, and decreases fat-pad mass/body-weight% (FM/BW%). In humans, plasma concentrations of the methionine product, cysteine, are associated with obesity. To determine which consequences of methionine-restriction are mediated by decreased cysteine availability, we monitored obesity-related variables in 4 dietary groups for 12 weeks: control-fed (CF), methionine-restricted (MR), MR supplemented with 0.5% l-cysteine (MR+Cys) and CF+Cys rats. MR lowered weight gain and FM/BW% despite higher food intake/weight than CF, and lowered serum cysteine. Hepatic Scd1 expression was decreased, with decreased serum SCD1 activity indices (calculated from serum fatty acid profile), decreased serum insulin, leptin and triglycerides, and higher adiponectin. Cysteine supplementation (MR+Cys) essentially reversed all these phenotypes and raised serum cysteine but not methionine to CF levels. Adding extra cysteine to control diet (CF+Cys) increased serum taurine but did not affect serum cysteine, lipids, proteins, or total weight gain. FM/BW% and serum leptin were modestly decreased. Our results indicate that anti-obesity effects of MR are caused by low cysteine and that dietary sulfur amino acid composition contributes to SCD1 regulation.


Mineralocorticoid receptor in the NTS stimulates saline intake during fourth ventricular infusions of aldosterone.
The purpose of this study was to determine whether neurons within the nucleus tractus solitarius (NTS) that express the mineralocorticoid receptor (MR) play a role in aldosterone stimulation of salt intake. Adult Wistar-Kyoto (WKY) rats received microinjections into the NTS of a short-hairpin RNA (shRNA) for the MR, to site specifically reduce levels of the MR by RNA interference (shRNA; n = 9) or scrambled RNA as a control (scRNA; n = 8). After injection of the viral construct, aldosterone-filled osmotic minipumps were implanted subcutaneously and connected to a cannula extending into the fourth ventricle to infuse aldosterone at a rate of 25 ng/h. Before and after surgeries, rats had ad libitum access to normal sodium (0.26%) rat chow and two graduated drinking bottles filled with either distilled water or 0.3 M NaCl. Before the surgeries, basal saline intake was 1.6 ± 0.6 ml in the scRNA group and 1.56 ± 0.6 ml in the shRNA group. Twenty-four days postsurgery, saline intake was elevated to a greater extent in the scRNA group (5.9 ± 1.07 ml) than in the shRNA group (2.41 ± 0.6 ml). Post mortem immunohistochemistry revealed a significant reduction in the number of NTS neurons exhibiting immunoreactivity for MR in shRNA-injected rats (23 ± 1 cells/section) versus scRNA-injected rats (33 ± 2 cells/section; P = 0.008). shRNA did not alter the level of 11-β-hydroxysteroid dehydrogenase type II (HSD2) protein in the NTS as judged by the number of HSD2 immunoreactive neurons. These results suggest that fourth ventricular infusions of aldosterone stimulate saline intake, and that this stimulation is at least in part mediated by hindbrain NTS neurons that express MR.


SU-E-I-114: Clinical Ultrasound Transducer Degradation Effects on the Accuracy of Spectral Doppler Velocity Measurements.
Ultrasound Doppler velocity measurements are routinely used to determine the severity of a stenosis in the carotid, renal or peripheral arteries. The objective of this study is to investigate and demonstrate the relationship between Doppler velocity measurements and transducer degradation conditions encountered in a clinical environment. Assessing transducer performance was accomplished using the First Call aPerio Test System transducer analyzer (Sonora Medical Systems, Longmont, CO). This system was used to conduct 1,145 semi-annual transducer assessments at 7 clinical sites from September 2007 to February 2012 as part of a comprehensive quality control program. The results were evaluated to determine the degree of transducer degradation encountered in the clinical environment. A Siemens Acuson S2000 (Siemens AG, Erlangen, Germany) ultrasound system was employed with the 1425A LE Doppler Flow System (Gammex, Middleton, WI) to determine Doppler velocity measurements. A transducer fixation device was fabricated to provide accurate, repeatable velocity measurements. Transducers with the most severe degradation were evaluated by comparing velocity measurements to those without defects. Using several matched transducer pairs, simulated failure modes were tested. The Doppler measured time-average mean (TAMn) and time-average maximum (TAMx) velocities were obtained and reported. This investigation found that 4.5% of the transducers surveyed during this 5-year period of time failed at least one acceptance criteria. Many (346 or 30%) had at least one defect. Typical findings include dead elements, lens delamination, wire cuts, and capacitance shorts. Only the most severe transducer defects and degradation conditions resulted in a noticeable deviation in the velocity measurements. Simulated transducer degradation testing confirmed this finding. The relationship between Doppler velocity measurements and transducer degradation conditions is limited, becoming significant for only the most severe degradation conditions. Based on our experience, this level of transducer degradation is rarely encountered in the clinical environment.


Indeterminate pulmonary nodules: risk for having or for developing lung cancer?
This perspective discusses the report by Pinsky and colleagues, which addresses whether noncalcified pulmonary nodules identified on CT screening carry short- and long-term risk for lung cancer. We are facing challenges related to distinguishing a large majority of benign nodules from malignant ones and among those a majority of aggressive from indolent cancers. Key questions in determining individual probabilities of disease, given their history, findings on CT, and upcoming biomarkers of risk, remain most challenging. Reducing the false positives associated with current low-dose computed tomography practices and identification of individuals who need therapy and at what time during tumor surveillance could reduce costs and morbidities associated with unnecessary interventions.


Self-Optimization in Continuous-Time Recurrent Neural Networks.
A recent advance in complex adaptive systems has revealed a new unsupervised learning technique called self-modeling or self-optimization. Basically, a complex network that can form an associative memory of the state configurations of the attractors on which it converges will optimize its structure: it will spontaneously generalize over these typically suboptimal attractors and thereby also reinforce more optimal attractors-even if these better solutions are normally so hard to find that they have never been previously visited. Ideally, after sufficient self-optimization the most optimal attractor dominates the state space, and the network will converge on it from any initial condition. This technique has been applied to social networks, gene regulatory networks, and neural networks, but its application to less restricted neural controllers, as typically used in evolutionary robotics, has not yet been attempted. Here we show for the first time that the self-optimization process can be implemented in a continuous-time recurrent neural network with asymmetrical connections. We discuss several open challenges that must still be addressed before this technique could be applied in actual robotic scenarios.


Effectiveness of dietary intervention in general practice.
The scope for dietary intervention in general practice is substantial. The three most prevalent conditions for which general practitioners are likely to give dietary advice are hypertension, functional digestive disorders, and ischemic heart disease. As well as clinical diseases, risk factors such as raised plasma cholesterol concentrations also provide opportunities for dietary intervention. But resources are limited. If a general practitioner or nurse spends 5 min of a 10-min consultation on dietary advice, there is 5 min less to spend on the rest of the consultation. Research studies in general practice show that small changes in plasma cholesterol concentrations can be achieved by dietary interventions. Intensive intervention can also influence salt intake to a small extent. However, the most important, potentially cost-effective roles for the general practitioner in health promotion are the legitimization and reinforcement of public health information by brief advice and the distribution of written material. Secondary and tertiary prevention is a priority in general practice and may entail use of drugs, but drugs are not a desirable solution for the unhealthy diets of healthy people.


Lessons from families and communities about interpersonal violence, victimization, and seeking help.
Despite significant incidence and physical and mental health consequences, most college-age women do not tell anyone about experiences of interpersonal violence. Limited research explores the sociocultural context of seeking help related to violence in young women. The overall purpose of this research was to understand socially and culturally relevant factors associated with violence help seeking in college women. Eight focus groups were held with 64 participants. Narrative analysis was the primary method of analysis. Four qualitative categories emerged from the data: "Learning from one's mother"; "We're strong women; we fight"; "We didn't talk about it"; and "Where I'm from." Findings suggest that help seeking is influenced by the messages from and experiences of mothers and extended family members. An understanding of familial and cultural determinants of help seeking is essential for relevant and effective prevention efforts.


In situ solution-phase Raman spectroscopy under forced convection.
In situ Raman spectra of solution-phase electrogenerated species have been recorded in a channel-type electrochemical cell incorporating a flat optically transparent window placed parallel to the channel plane that contains the embedded working electrode. A microscope objective with its main axis (Z) aligned normal to the direction of flow was used to focus the excitation laser beam (lambda exc = 532 nm) in the solution and also to collect the Raman scattered light from species present therein. Judicious adjustment of the cell position along Z allowed the depth of focus to overlap the diffusion boundary layer to achieve maximum detection sensitivity. Measurements were performed using a Au working electrode in iron hexacyanoferrate(II), [Fe(CN)6]4-, and nitrite, NO2-, containing aqueous solutions as a function of the applied potential, E. Linear correlations were found between both the gain and the loss of the integrated Raman intensity, IR, of bands, attributed to [Fe(CN)6]3- and [Fe(CN)6]4-, respectively, recorded downstream from the edge of the working electrode, and the current measured at the Au electrode as a function of E. The same overall trend was found for the gain in the IR of the NO3- band in the nitrite solution. Also included in this work is a ray trace analysis of the optical system.


Physical activities during pregnancy and type of delivery in nulliparae.
The aim of this study was to investigate changes in physical activity across pregnancy and the relationship between trimester-specific physical activity and unplanned caesarean sections (CSs). A cohort study design was carried out. A cohort of 150 pregnant women was established when they received prenatal care at 29-40 weeks of gestation at a medical centre in northern Taiwan. Participants were asked to recall the amounts of physical activity in which they had engaged in the three trimesters as assessed by the Pregnancy Physical Activity Questionnaire (PPAQ). Overall self-reported physical activity for the cohort decreased by 31% in the first trimester compared to the pre-gravid period, then increased in the second trimester and remained stable until delivery. A repeated measures analysis of variance was used to evaluate the data and revealed significantly more physical activity during the second trimester than in the first and third trimesters (F = 36.471, P = 0.000). In addition, there was a significant difference between normal spontaneous delivery and unplanned CS groups (F = 4.770, P = 0.031). Logistic regression determined that the odds ratio of undergoing a CS increased by 0.644 (95% confidence interval: 0.429-0.968) for women in the third trimester who performed low levels of physical activity. Results support the benefits of physical activity, and professionals are encouraged to provide pregnant women with information on recommendations for physical activity, particularly in terms of reducing unplanned CSs.


Assessment of unconscious decision aids applied to complex patient-centered medical decisions.
To improve patient health, recent research urges for medical decision aids that are designed to enhance the effectiveness of specific medically related decisions. Many such decisions involve complex information, and decision aids that independently use deliberative (analytical and slower) or intuitive (more affective and automatic) cognitive processes for such decisions result in suboptimal decisions. Unconscious thought can arguably use both intuitive and deliberative (slow and analytic) processes, and this combination may further benefit complex patient (or practitioner) decisions as medical decision aids. Indeed, mounting research demonstrates that individuals render better decisions generally if they are distracted from thinking consciously about complex information after it is presented (but can think unconsciously), relative to thinking about that information consciously or not at all. The current research tested whether the benefits of unconscious thought processes can be replicated using an Internet platform for a patient medical decision involving complex information. This research also explored the possibility that judgments reported after a period of unconscious thought are actually the result of a short period of conscious deliberation occurring during the decision report phase. A total of 173 participants in a Web-based experiment received information about four medical treatments, the best (worst) associated with mostly positive (negative) side-effects/attributes and the others with equal positive-negative ratios. Next, participants were either distracted for 3 minutes (unconscious thought), instructed to think about the information for 3 minutes (conscious thought), or moved directly to the decision task (immediate decision). Finally, participants reported their choice of, and attitudes toward, the treatments while experiencing high, low, or no cognitive load, which varied their ability to think consciously while reporting judgments. Cognitive load was manipulated by having participants memorize semi-random (high), line structured (low), or no dot patterns and recall these intermittently with their decision reports. Overall then, participants were randomly assigned to the conditions of a 3 (thought condition) by 3 (cognitive-load level) between-subjects design. A logistic regression analysis indicated that the odds of participants choosing the best treatment were 2.25 times higher in the unconscious-thought condition compared to the immediate-decision condition (b=.81, Wald=4.32, P=.04, 95% CI 1.048-4.836), and 2.39 times greater compared to the conscious-thought condition (b=.87, Wald=4.87, P=.027, 95% CI 1.103-5.186). No difference was observed between the conscious-thought condition compared to the immediate-decision condition, and cognitive load manipulations did not affect choices or alter the above finding. This research demonstrates a plausible benefit of unconscious thinking as a decision aid for complex medical decisions, and represents the first use of unconscious thought processes as a patient-centered medical decision aid. Further, the quality of decisions reached unconsciously does not appear to be affected by the amount of cognitive load participants experienced.


Most and Least Preferred Colours Differ According to Object Context: New Insights from an Unrestricted Colour Range.
Humans like some colours and dislike others, but which particular colours and why remains to be understood. Empirical studies on colour preferences generally targeted most preferred colours, but rarely least preferred (disliked) colours. In addition, findings are often based on general colour preferences leaving open the question whether results generalise to specific objects. Here, 88 participants selected the colours they preferred most and least for three context conditions (general, interior walls, t-shirt) using a high-precision colour picker. Participants also indicated whether they associated their colour choice to a valenced object or concept. The chosen colours varied widely between individuals and contexts and so did the reasons for their choices. Consistent patterns also emerged, as most preferred colours in general were more chromatic, while for walls they were lighter and for t-shirts they were darker and less chromatic compared to least preferred colours. This meant that general colour preferences could not explain object specific colour preferences. Measures of the selection process further revealed that, compared to most preferred colours, least preferred colours were chosen more quickly and were less often linked to valenced objects or concepts. The high intra- and inter-individual variability in this and previous reports furthers our understanding that colour preferences are determined by subjective experiences and that most and least preferred colours are not processed equally.


Gene polymorphism for microsomal epoxide hydrolase and susceptibility to emphysema in a Japanese population.
Recently, it was reported that gene polymorphism for microsomal epoxide hydrolase (mEPHX), an enzyme involved in the first-pass metabolism of epoxide intermediates, was associated with susceptibility to emphysema. This association was examined in a Japanese population, performing polymerase chain reaction (PCR)-based direct sequencing and restriction fragment length polymorphism assays for variant forms of mEPHX. The subjects consisted of 79 smokers with moderate to severe emphysema diagnosed by lung computed tomography scans, 58 smokers without emphysema, with a comparable smoking history, and 114 consecutive subjects who undertook annual health checkups. The allele frequency of exon 3 Tyr113 to His113, which was reported to confer slow mEPHX activity, was substantially higher in the population control group compared with that of the Caucasian control subjects in a previous study. However, neither the genotype distribution of exon 3, nor that of exon 4 His139 to Arg139, was significantly different between the two groups of smokers. These data indicate that the gene polymorphism for mEPHX is not associated with susceptibility to emphysema in the Japanese population. The discrepancy between the two studies may be explained either by racial difference or by the selection bias of subjects in the previous study, which examined those who had only mild to moderate emphysema with lung cancer or those who were clinically diagnosed as having chronic obstructive pulmonary disease.


Helicobacter pylori in cathartic stools of subjects with and without cimetidine-induced hypochlorhydria.
We previously identified viable Helicobacter pylori in stools from asymptomatic hosts. We now report whether a decrease in gastric acidity enhances faecal shedding. Sixteen asymptomatic H. pylori-positive patients underwent two separate days of phosphosoda-induced diarrhoea, both with normal gastric acidity and under hypochlorhydric conditions induced with the H2-blocker cimetidine. Stool samples were collected for culture to determine the presence of viable H. pylori. Five of the 16 patients gave positive cultures with at least one stool from both normal pH and cimetidine-induced hypochlorhydria. Four were negative for all samples with both. Six gave positive stools only after cimetidine treatments, while one gave positive samples with normal pH but not with cimetidine (two-tailed P value, 0.13; McNemar test). These numbers show a trend suggesting that cimetidine-induced hypochlorhydria increases shedding of viable H. pylori.


Experiences of a National Early Warning Score (NEWS) intervention in care homes during the COVID-19 pandemic: a qualitative interview study.
The COVID-19 pandemic has taken a heavy toll on the care home sector, with residents accounting for up to half of all deaths in Europe. The response to acute illness in care homes plays a particularly important role in the care of residents during a pandemic. Digital recording of a National Early Warning Score (NEWS), which involves the measurement of physical observations, started in care homes in one area of England in 2016. Implementation of a NEWS intervention (including equipment, training and support) was accelerated early in the pandemic, despite limited evidence for its use in the care home setting. To understand how a NEWS intervention has been used in care homes in one area of North-East England during the COVID-19 pandemic, and how it has influenced resident care, from the perspective of stakeholders involved in care delivery and commissioning. A qualitative interview study with care home (n=10) and National Health Service (n=7) staff. Data were analysed using thematic analysis. Use of the NEWS intervention in care homes in this area accelerated during the COVID-19 pandemic. Stakeholders felt that NEWS, and its associated education and support package, improved the response of care homes and healthcare professionals to deterioration in residents' health during the pandemic. Healthcare professionals valued the ability to remotely monitor resident observations, which facilitated triage and treatment decisions. Care home staff felt empowered by NEWS, providing a common clinical language to communicate concerns with external services, acting as an adjunct to staff intuition of resident deterioration. The NEWS intervention formed an important part of the care home response to COVID-19 in the study area. Positive staff perceptions now need to be supplemented with data on the impact on resident health and well-being, workload, and service utilisation, during the pandemic and beyond.


CASSys: an integrated software-system for the interactive analysis of ChIP-seq data.
The mapping of DNA-protein interactions is crucial for a full understanding of transcriptional regulation. Chromatin-immunoprecipitation followed by massively parallel sequencing (ChIP-seq) has become the standard technique for analyzing these interactions on a genome-wide scale. We have developed a software system called CASSys (ChIP-seq data Analysis Software System) spanning all steps of ChIP-seq data analysis. It supersedes the laborious application of several single command line tools. CASSys provides functionality ranging from quality assessment and -control of short reads, over the mapping of reads against a reference genome (readmapping) and the detection of enriched regions (peakdetection) to various follow-up analyses. The latter are accessible via a state-of-the-art web interface and can be performed interactively by the user. The follow-up analyses allow for flexible user defined association of putative interaction sites with genes, visualization of their genomic context with an integrated genome browser, the detection of putative binding motifs, the identification of over-represented Gene Ontology-terms, pathway analysis and the visualization of interaction networks. The system is client-server based, accessible via a web browser and does not require any software installation on the client side. To demonstrate CASSys's functionality we used the system for the complete data analysis of a publicly available Chip-seq study that investigated the role of the transcription factor estrogen receptor-α in breast cancer cells.


Genetic Analysis of Virulence Potential of Escherichia coli O104 Serotypes Isolated From Cattle Feces Using Whole Genome Sequencing.
Escherichia coli O104:H4, a Shiga toxin-producing hybrid pathotype that was implicated in a major foodborne outbreak in Germany in 2011, has not been detected in cattle. However, serotypes of O104, other than O104:H4, have been isolated from cattle feces, with O104:H7 being the most predominant. In this study, we investigated, based on whole genome sequence analyses, the virulence potential of E. coli O104 strains isolated from cattle feces, since cattle are asymptomatic carriers of E. coli O104. The genomes of ten bovine E. coli O104 strains (six O104:H7, one O104:H8, one O104:H12, and two O104:H23) and five O104:H7 isolated from human clinical cases were sequenced. Of all the bovine O104 serotypes (H7, H8, H12, and H23) that were included in the study, only E. coli O104:H7 serotype possessed Shiga toxins. Four of the six bovine O104:H7 strains and one of the five human strains carried stx1c. Three human O104 strains carried stx2, two were of subtype 2a, and one was 2d. Genomes of stx carrying bovine O104:H7 strains were larger than the stx-negative strains of O104:H7 or other serotypes. The genome sizes were proportional to the number of genes carried on the mobile genetic elements (phages, prophages, transposable elements and plasmids). Both bovine and human strains were negative for intimin and other genes associated with the type III secretory system and non-LEE encoded effectors. Plasmid-encoded virulence genes (ehxA, epeA, espP, katP) were also present in bovine and human strains. All O104 strains were negative for antimicrobial resistance genes, except one human strain. Phylogenetic analysis indicated that bovine E. coli O104 strains carrying the same flagellar antigen clustered together and STEC strains clustered separately from non-STEC strains. One of the human O104:H7 strains was phylogenetically closely related to and belonged to the same sequence type (ST-1817) as the bovine O104:H7 STEC strains. This suggests that the bovine feces could be a source of human illness caused by E. coli O104:H7 serotype. Because bovine O104:H7 strains carried virulence genes similar to human clinical strains and one of the human clinical strains was phylogenetically related to bovine strains, the serotype has the potential to be a diarrheagenic pathogen in humans.


Incidence of primary glomerulonephritis in a large North-Eastern Italian area: a 13-year renal biopsy study.
The reported incidence of biopsy-proven primary glomerulonephritis (PGN) varies according to geographical, temporal and environmental factors. Consequently, the development of national/regional registers may help clinicians and researchers to improve knowledge about this important clinical condition. To better define the epidemiology of PGN in our North-Eastern Italian area (∼5 million inhabitants), we evaluated the kidney biopsy records of 2680 adult patients with PGN diagnosis reported from 1998 to 2010 in the 'Triveneto' Register of Renal Biopsies. Statistical analysis showed that the mean age of patients undergoing renal biopsy was gradually increased from 1998 to 2010 (R(2) = 0.82, P < 0.01) with a growing percentage of those aged over 65 years (R(2) = 0.72, P < 0.01). According to the clinical presentation of our PGN patients, we found a significant increase in biopsies performed for acute renal failure (P < 0.01) and a decrement of those for macroscopic haematuria (P < 0.01) and nephritic syndrome (P = 0.04). Moreover, although there has been an unchanged total annual rate of biopsy-proven PGN (P = 0.47), there has been a significant enhancement in the incidence of minimal change disease (MCD, P = 0.04) and extracapillary proliferative glomerulonephritis (ExGN, P = 0.03) over time primarily due to a progressive increase in the mean age of patients affected by both renal diseases. Immunoglobulin A (IgA) nephropathy was the most common glomerulonephritis. Therefore, even if the number of PGN did not diminish during the 13-year study period, we reported considerable changes in the demographical and clinical characteristics of our biopsied patients (older and with acute kidney injury). Additionally, we found a change in the bioptic pattern of our patients over time with a progressive rise of some histological features such as MCD and ExGN. This may reflect not only the progressive ageing of our nephrology patients, but also a change in the biopsy policy of local hospitals.


Grifola frondosa water extract alleviates intestinal inflammation by suppressing TNF-alpha production and its signaling.
TNF-alpha is a major cytokine involved in inflammatory bowel disease (IBD). In this study, water extract of Grifola frondosa (GFW) was evaluated for its protective effects against colon inflammation through the modulation of TNF-alpha action. In coculture of HT-29 human colon cancer cells with U937 human monocytic cells, TNF-alpha-induced monocyte adhesion to HT-29 cells was significantly suppressed by GFW (10, 50, 100 micg/ml). The reduced adhesion by GFW correlated with the suppressed expression of MCP-1 and IL-8, the major IBD-associated chemokines. In addition, treatment with GFW significantly suppressed TNF-alpha-induced reactive oxygen species production and NF-kappaB transcriptional activity in HT-29 cells. In differentiated U937 monocytic cells, LPS-induced TNF-alpha production, which is known to be mediated through NF-kappaB activation, was significantly suppressed by GFW. In an in vivo rat model of IBD, oral administration of GFW for 5 days (1 g/kg per day) significantly inhibited the trinitrobenzene sulfonic acid (TNBS)-induced weight loss, colon ulceration, myeloperoxidase activity, and TNF-alpha expression in the colon tissue. Moreover, the effect of GFW was similar to that of intra-peritoneal injection of 5-aminosalicylic acid (5-ASA), an active metabolite of sulfasalazine, commonly used drug for the treatment of IBD. The results suggest that GFW ameliorates colon inflammation by suppressing production of TNF-alpha as well as its signaling through NF-kappaB leading to the expression of inflammatory chemokines, MCP-1 and IL-8. Taken together, the results strongly suggest GFW is a valuable medicinal food for IBD treatment, and thus may be used as an alternative medicine for IBD.


Response of Commercially Developed Soybean Cultivars and the Ancestral Soybean Lines to Fusarium solani f. sp. glycines.
Sudden death syndrome, caused by Fusarium solani f. sp. glycines, has caused severe damage to soybean production in recent years. One way to control sudden death syndrome is with resistant cultivars. Over a 3-year period, 2,335 publicly and privately developed soybean entries were inoculated and evaluated for their response to F. solani f. sp. glycines under greenhouse conditions. The entries were compared with the susceptible check, Great Lakes 3302 (GL3302), and the moderately resistant checks, plant introductions (PIs) 520733 and 567374. Thirty-eight entries were identified with moderate levels of resistance. Based on foliar ratings, there were no differences (P < 0.05) between the Roundup Ready and conventional cultivars. In all, 90 ancestral lines that represent 99% of the genes in modern U.S. cultivars and 55 lines found in the pedigrees of public cultivars reported to have some resistance were evaluated for their response to F. solani f. sp. glycines. Nine ancestral lines (Aoda, Kim, Jackson, Sioux, Mammoth Yellow, T117, PI 171450, PI 54615-1, and PI 71506) and 12 cultivars or experimental lines (Ina, D83-3349, LN98-4340, LN83-2356, Hartwig, Harosoy, Bedford, Merit, Cutler, Calland, Hill, and Evans) had disease ratings not significantly different (P < 0.05) from PI 520733 or PI 567374. PI 54610, a putative ancestral line, also was found to be moderately resistant.


Chrysoeriol ameliorates TPA-induced acute skin inflammation in mice and inhibits NF-κB and STAT3 pathways.
Chrysoeriol is a flavone found in diverse dietary and medicinal herbs such as Lonicerae Japonicae Flos (the dried flower bud or newly bloomed flower of Lonicera japonica Thunb.). These herbs are commonly used for treating inflammatory diseases. Herbal extracts containing chrysoeriol have been shown to have anti-inflammatory effects and inhibit nuclear factor-kappa B (NF-κB) signaling. Some of these extracts can inhibit signal transducers and activators of transcription 3 (STAT3) signaling in cancer cells. This study aimed to determine whether chrysoeriol has anti-inflammatory effects and whether NF-κB and STAT3 pathways are involved in the effects. A TPA (12-O-tetradecanoylphorbol-13-acetate)-induced ear edema mouse model and LPS-stimulated RAW264.7 cells were used to evaluate the effects of chrysoeriol. Griess reagent was used to measure the production of nitric oxide (NO). Western blot and enzyme-linked immunosorbent assays were employed to detect protein levels. RT-qPCR analyses were used to detect mRNA levels. Haematoxylin and eosin (H&E) staining was employed to examine the pathological conditions in animal tissues. In the mouse model, chrysoeriol ameliorated acute skin inflammation, evidenced by reduced ear thickness, ear weight and number of inflammatory cells in inflamed ear tissues. The compound lowered protein levels of phospho-p65 (Ser536), phospho-STAT3 (Tyr705), inducible nitric oxide synthases (iNOS), cyclooxygenase-2 (COX-2), interleukin 6 (IL-6), IL-1β and tumor necrosis factor α (TNF-α) in mouse swollen ears. In LPS-stimulated RAW264.7 cells, chrysoeriol also lowered levels of these proteins. In addition, chrysoeriol decreased the production of NO and prostaglandin E2; inhibited the phosphorylation of inhibitor of κB (Ser32), p65 (Ser536) and Janus kinase 2 (Tyr1007/1008); decreased nuclear localization of p50, p65 and STAT3; and down-regulated mRNA levels of pro-inflammatory cytokines IL-6, IL-1β and TNF-α that are transcriptionally regulated by NF-κB and STAT3 in the cell model. We for the first time demonstrated that chrysoeriol ameliorates TPA-induced ear edema in mice, and that inhibition of JAK2/STAT3 and IκB/p65 NF-κB pathways are involved in the anti-inflammatory effects of chrysoeriol. This study provides chemical and pharmacological justifications for the use of chrysoeriol-containing herbs in treating inflammatory diseases, and provides pharmacological groundwork for developing chrysoeriol as a novel anti-inflammatory agent.


Renal function in rats with essential fatty acid deficiency.
1. Renal function was studied in 50-, 70-, 90- and 200-day-old rats with essential fatty acid deficiency. The pharmacokinetics of tobramycin was investigated in 90-day-old essential fatty acid-deficient rats. 2. A higher glomerular filtration rate and a higher serum concentration of urea were seen in 50-day-old essential fatty acid-deficient rats compared with age-matched controls. Later, the glomerular filtration rate progressively deteriorated in parallel with a decline in effective renal plasma flow and with a concomittant rise in serum levels of urea and creatinine. The serum concentration of protein was lower in the rats with essential fatty acid deficiency and that of sodium was higher than in the control rats. The non-renal clearance of tobramycin was increased in the rats with essential fatty acid deficiency. 3. The early hyperfiltration in essential fatty acid-deficient rats with the subsequent fall in glomerular filtration rate, which was paralleled by a rise in serum levels of urea and creatinine, as well as the increased non-renal clearance of tobramycin, are in accordance with the clinical manifestations of cystic fibrosis. Rats with essential fatty acid deficiency might be a useful model with which to study the pathophysiological renal changes in cystic fibrosis related to the progressive essential fatty acid deficiency in this disease.


Nurturing Children's Healthy Eating: Position statement.
The relationship between eating a healthy diet and positive health outcomes is well known; nurturing healthy eating among children therefore has the potential to improve public health. A healthy diet occurs when one's usual eating patterns include adequate nutrient intake and sufficient, but not excessive, energy intake to meet the energy needs of the individual. However, many parents struggle to establish healthy eating patterns in their children due to the pressures of modern life. Moreover, healthcare providers often do not have the time or the guidance they need to empower parents to establish healthy eating practices in their children. Based on existing evidence from epidemiologic and intervention research, the Nurturing Children's Healthy Eating collaboration, established by Danone Institute International, has identified four key themes that encourage and support healthy eating practices among children in the modern Western world. The first - positive parental feeding - explores how parenting practices and styles, such as avoiding food restriction, allowing children to make their own food choices, and encouraging children to self-limit their portion sizes, can influence children's dietary intake. The second - eating together - highlights the link between eating socialization through regular family meals and healthful diet among children. The third - a healthy home food environment - explores the impact on eating practices of family resources, food availability/accessibility, parental modeling, and cues for eating. The fourth - the pleasure of eating - associates children's healthy eating with pleasure through repeated exposure to healthful foods, enjoyable social meals, and enhancement of the cognitive qualities (e.g. thoughts or ideas) of healthful foods. This paper reviews the evidence leading to the characterization of these nurturing themes, and ways in which recommendations might be implemented in the home.


The ageing shift worker: a prospective cohort study on need for recovery, disability, and retirement intentions.
This study investigates whether different shift work schedules, compared to day work, are associated with need for recovery (NFR), future disability, and retirement intentions for employees employed within different economic sectors over the course of their careers. Shift work exposure duration and the healthy worker effect are also examined. Data from the prospective Maastricht Cohort Study was used. Subsamples of industry (N=1877, all men) and healthcare (N=818, 624 women and 194 men) workers were separately investigated. GEE and Cox regression analyses were performed to investigate NFR longitudinally. Future disability was investigated using Cox regression, and retirement intentions were investigated using logistic regression analysis. Three-shift industry workers were at risk of becoming a case of elevated NFR during follow-up, compared to industry day workers. Three- and five-shift industry workers were at risk for future disability. In healthcare, irregular shift work was a risk factor for disability among older shift workers. No significant results were found regarding retirement intentions. Findings were probably an underestimation as exposure duration to shift work and the healthy worker effect affected the results. Shift work was associated with higher levels of NFR and a higher risk of disability. However, shift work is a multifaceted concept as different types of shift work schedules are differently associated with these outcomes. Different shift work types exist and shift work schedules allow for optimization, indicating that measures to prevent adverse outcomes should be tailored for different types of shift work and over the course of the work career.


Investigation of continuous changes in the electric-field-induced electronic state in Bi(1-x)Ca(x)FeO(3-δ).
Amongst the most interesting phenomena in correlated oxide systems are the doping-driven competitions between energetically similar ground states found in, e.g., high-Tc superconductors and colossal magnetoresistance manganites. It has recently been reported that doped multiferroics also exhibit this generic concept of phase competition. Here, we employ photoelectron emission microscopy (PEEM) to demonstrate evidence of systematic changes in the electronic structure of Bi(1-x)Ca(x)FeO(3-δ) treated by electrically controlled hole carrier doping, the outcome of which clearly correlates with the local modulation of electronic conductivity observed in the same material.


Associations between community-level disaster exposure and individual-level changes in disability and risk of death for older Americans.
Disasters occur frequently in the United States (US) and their impact on acute morbidity, mortality and short-term increased health needs has been well described. However, barring mental health, little is known about the medium or longer-term health impacts of disasters. This study sought to determine if there is an association between community-level disaster exposure and individual-level changes in disability and/or the risk of death for older Americans. Using the US Federal Emergency Management Agency's database of disaster declarations, 602 disasters occurred between August 1998 and December 2010 and were characterized by their presence, intensity, duration and type. Repeated measurements of a disability score (based on activities of daily living) and dates of death were observed between January 2000 and November 2010 for 18,102 American individuals aged 50-89 years, who were participating in the national longitudinal Health and Retirement Study. Longitudinal (disability) and time-to-event (death) data were modelled simultaneously using a 'joint modelling' approach. There was no evidence of an association between community-level disaster exposure and individual-level changes in disability or the risk of death. Our results suggest that future research should focus on individual-level disaster exposures, moderate to severe disaster events, or higher-risk groups of individuals.


Risks of myeloid malignancies in patients with autoimmune conditions.
Autoimmune conditions are associated with an elevated risk of lymphoproliferative malignancies, but few studies have investigated the risk of myeloid malignancies. From the US Surveillance Epidemiology and End Results (SEER)-Medicare database, 13 486 myeloid malignancy patients (aged 67+ years) and 160 086 population-based controls were selected. Logistic regression models adjusted for gender, age, race, calendar year and number of physician claims were used to estimate odds ratios (ORs) for myeloid malignancies in relation to autoimmune conditions. Multiple comparisons were controlled for using the Bonferroni correction (P<0.0005). Autoimmune conditions, overall, were associated with an increased risk of acute myeloid leukaemia (AML) (OR 1.29) and myelodysplastic syndrome (MDS, OR 1.50). Specifically, AML was associated with rheumatoid arthritis (OR 1.28), systemic lupus erythematosus (OR 1.92), polymyalgia rheumatica (OR 1.73), autoimmune haemolytic anaemia (OR 3.74), systemic vasculitis (OR 6.23), ulcerative colitis (OR 1.72) and pernicious anaemia (OR 1.57). Myelodysplastic syndrome was associated with rheumatoid arthritis (OR1.52) and pernicious anaemia (OR 2.38). Overall, autoimmune conditions were not associated with chronic myeloid leukaemia (OR 1.09) or chronic myeloproliferative disorders (OR 1.15). Medications used to treat autoimmune conditions, shared genetic predisposition and/or direct infiltration of bone marrow by autoimmune conditions, could explain these excess risks of myeloid malignancies.


HP1a recruitment to promoters is independent of H3K9 methylation in Drosophila melanogaster.
Heterochromatin protein 1 (HP1) proteins, recognized readers of the heterochromatin mark methylation of histone H3 lysine 9 (H3K9me), are important regulators of heterochromatin-mediated gene silencing and chromosome structure. In Drosophila melanogaster three histone lysine methyl transferases (HKMTs) are associated with the methylation of H3K9: Su(var)3-9, Setdb1, and G9a. To probe the dependence of HP1a binding on H3K9me, its dependence on these three HKMTs, and the division of labor between the HKMTs, we have examined correlations between HP1a binding and H3K9me patterns in wild type and null mutants of these HKMTs. We show here that Su(var)3-9 controls H3K9me-dependent binding of HP1a in pericentromeric regions, while Setdb1 controls it in cytological region 2L:31 and (together with POF) in chromosome 4. HP1a binds to the promoters and within bodies of active genes in these three regions. More importantly, however, HP1a binding at promoters of active genes is independent of H3K9me and POF. Rather, it is associated with heterochromatin protein 2 (HP2) and open chromatin. Our results support a hypothesis in which HP1a nucleates with high affinity independently of H3K9me in promoters of active genes and then spreads via H3K9 methylation and transient looping contacts with those H3K9me target sites.


Impact of mutations in FLT3, PTPN11 and RAS genes on the overall survival of pediatric B cell precursor acute lymphoblastic leukemia in Brazil.
We analyzed mutations in four genes (FLT3, KRAS/NRAS and PTPN11) that might disrupt the RAS/mitogen activated protein kinase (MAPKinase) signaling pathway, to evaluate their prognostic value in children younger than 16 years old with B-cell precursor acute lymphoblastic leukemia (Bcp-ALL). The overall survival (OS) was determined with the Kaplan-Meier method. MAPKinase genes were mutated in 25.4% and 20.1% of childhood and infant Bcp-ALL, respectively. Children with hyperdiploidy were more prone to harboring a MAPKinase gene mutation (odds ratio [OR] 3.18; 95% confidence interval [CI] 1.07-9.49). The mean OS of all cases was 54.0 months. FLT3 and PTPN11 mutations had no impact on OS. K/NRAS mutations were strongly associated with MLL-AFF1 (OR 5.78; 95% CI 1.00-33.24), and conferred poorer OS (p = 0.034) in univariate analysis.


Effects of variability of practice in music: a pilot study on fast goal-directed movements in pianists.
Variability of Practice (VOP) refers to the acquisition of a particular target movement by practicing a range of varying targets rather than by focusing on fixed repetitions of the target only. VOP has been demonstrated to have beneficial effects on transfer to a novel task and on skill consolidation. This study extends the line of research to musical practice. In a task resembling a barrier-knockdown paradigm, 36 music students trained to perform a wide left-hand interval leap on the piano. Performance at the target distance was tested before and after a 30-min standardized training session. The high-variability group (VAR) practiced four different intervals including the target. Another group (FIX) practiced the target interval only. A third group (SPA) performed spaced practice on the target only, interweaving with periods of not playing. Transfer was tested by introducing an interval novel to either group. After a 24-h period with no further exposure to the instrument, performance was retested. All groups performed at comparable error levels before training, after training, and after the retention (RET) interval. At transfer, however, the FIX group, unlike the other groups, committed significantly more errors than in the target task. After the RET period, the effect was washed out for the FIX group but then was present for VAR. Thus, the results provide only partial support for the VOP hypothesis for the given setting. Additional exploratory observations suggest tentative benefits of VOP regarding execution speed, loudness, and performance confidence. We derive specific hypotheses and specific recommendations regarding sample selection and intervention duration for future investigations. Furthermore, the proposed leap task measurement is shown to be (a) robust enough to serve as a standard framework for studies in the music domain, yet (b) versatile enough to allow for a wide range of designs not previously investigated for music on a standardized basis.


School going adolescents' perception of family climate and mental problems: Results from Kocaeli, Turkey.
To investigate the association between the perception of expressed emotion and psychopathology of school-going adolescents in a non-clinical sample. The cross-sectional field study was conducted in Kocaeli, Turkey, and comprised high-school students during the 2014-15 school years. Data was collected using the Information Collection Form, Strengths and Difficulties Questionnaire and Shortened Level of Expressed Emotion Scale. SPSS 21 was used for data analysis. Of the 487 subjects, 295(60.57%) were girls and 192(39.43%) were boys. The overall median age was 16 (inter-quartile range: 15-17 years). There were significant positive relationships between the scales in terms of emotional problems (p<0.001), behavioural problems (p<0.001), Attention-deficit/hyperactivity disorder (p<0.001), and peer relationship problems (p<0.001). There was also a negative significant relationship between pro-social behaviours and expressed emotion (p<0.001). The concept of perceived expressed emotion in non-clinical adolescents is useful in alarming clinicians and preventing mental disorders.


Counterclockwise rotation of Roux-en-Y limb significantly reduces internal herniation in laparoscopic Roux-en-Y gastric bypass (LRYGB).
Internal hernias continue to be a significant source of morbidity after LRYGB. Literature addressing the technique of Roux limb construction as a predisposing factor is sparse. The objective of this study is to evaluate the impact of Roux limb construction technique on the development of internal hernias. In this study, we included 444 (367 (82.7%) were females and 77 (17.3%) were males, two deaths excluded from the analysis) consecutive patients from our institutional bariatric database who underwent LRYGB. Variables collected include demographics, body mass index (BMI) before and after the procedure, and postoperative small bowel obstruction secondary to internal herniation. Technical details collected include: type of Roux-en-Y limb construction, Peterson's defect closure at initial operation, and reoperative findings. Roux limbs were constructed in 291 patients by a clockwise rotation of the bowel and jejunojejunostomy performed on the right side of the axis of the mesentery (group 1). In 151 patients, the Roux limb was constructed by a counterclockwise rotation of the Roux limb resulting in the jejunojejunostomy on the left side of the axis of the mesentery (group 2). We also analyzed the impact of Peterson's space closure on internal hernias. Fisher's exact test and Chi-square test were used for the analysis. Of a total 442 (mean age, 43.7 ± 10.3 years; mean BMI pre-op was 46.4 ± 5.1; and BMI after median follow-up of 12 months was 34.5 ± 6.98) patients included in the study, 21 (4.7%) internal hernias were identified. Of 21 internal hernias, 17 (81%) were through Peterson's space and four (19%) were through the mesenteric defect. Group 1 patients had significantly higher overall internal hernias (20/291, 6.9% vs. 1/151, 0.7%; P = 0.0018) and Peterson's hernias (16/291, 5.5% vs. 1/151, 0.7%; P = 0.0089) compared with group 2. In addition, no significant difference was noted in the incidence of Peterson's hernia whether the defect was closed or not closed (closed group, 4/117 and 3.4% vs. not closed, 13/325, 4%; P = 1.00). Within the group where Peterson's defect was closed, clockwise rotation and anastomosis on the right side of the axis of the mesentery was associated with significantly higher incidence of Peterson's hernias compared with counterclockwise rotation (4/54 vs. 0/63; P = 0.043). In the group where Peterson's defect was not closed, clockwise rotation was associated with higher incidence of internal hernias that did not reach statistical significance (12/237, 5.1% vs. 1/88, 1.1%; P = 0.12). This study demonstrates that the technique for construction of the Roux limb is a major factor in the development of internal hernias. Construction of the Roux limb with a counterclockwise rotation of the bowel, such that both jejunojejunostomy anastomosis and ligament of Treitz are to the left of the axis of the mesentery significantly reduces the incidence of internal hernias.


An image processing approach to approximating interface textures of microcrystalline silicon layers grown on existing aluminum-doped zinc oxide textures.
We present an algorithm for generating a surface approximation of microcrystalline silicon (μc-Si) layers after plasma enhanced chemical vapor deposition (PECVD) onto surface textured substrates, where data of the textured substrate surface are available as input. We utilize mathematical image processing tools and combine them with an ellipsoid generator approach. The presented algorithm has been tuned for use in thin-film silicon solar cell applications, where textured surfaces are used to improve light trapping. We demonstrate the feasibility of this method by means of optical simulations of generated surface textures, comparing them to simulations of measured atomic force microscopy (AFM) scan data of both Aluminum-doped zinc oxide (AZO, a transparent and conductive material) and μc-Si layers.


Color-tunable organic light-emitting diodes with vertically stacked blue, green, and red colors for lighting and display applications.
We demonstrate independently and simultaneously controlled color-tunable organic light-emitting diodes (OLEDs) with vertically stacked blue, green, and red elements. The blue, green, and red elements were placed at the bottom, middle, and top positions, respectively, forming color-tunable OLEDs. The independently driven blue, green, and red elements in the color-tunable OLEDs exhibited low driving voltages of 5.3 V, 3.0 V, and 4.6 V, as well as high external quantum efficiencies of 11.1%, 10.9%, and 9.6%, respectively, at approximately 1000 cd/m2. Each element in the color-tunable OLEDs showed high-purity blue, green, and red colors with little parasitic emission owing to the delicately designed device structure resultant from optical simulations. The color-tunable OLEDs could produce any colors inside the triangle formed with blue (0.136, 0.261), green (0.246, 0.697), and red (0.614, 0.386) Commission Internationale de l'éclairage (CIE) 1931 color coordinates. In addition, the correlated color temperatures (CCTs) of white colors in the color-tunable OLED can be easily changed from the warm white to the cool white by controlling the red, green, and blue emissions simultaneously. The white colors in the color-tunable OLED have the CIE 1931 color coordinate of (0.304, 0.351), with a CCT of 6289 K and (0.504, 0.440), with a CCT of 2407K at the driving voltage of 5 V (blue), 2.8 V (green), 4.4 V (red), and 4.6 V (blue), 3 V (green), 5 V (red), respectively. Furthermore, the white color in the color-tunable OLED exhibited a high color rendering index (~88.7) due to vertically stacked three color system. Moreover, we successfully fabricated a large-sized, 14 × 12 pixel array of the color-tunable OLEDs to demonstrate lighting and display applications, respectively.


Potential for Paramedic roles in Irish General Practice: A qualitative study of stakeholder's perspectives.
Background: Irish health policy emphasises the role of Primary Care and General Practice however, there is a growing shortage of General Practitioners (GPs) in Ireland. Paramedics have traditionally focused on emergency care in the community. More recently Paramedics have taken on roles in General Practice in international jurisdictions, but not yet in Ireland. This study aimed to explore key stakeholder perceptions of 'the potential for Paramedic roles in Irish General Practice'. Methods: We conducted an exploratory, qualitative stakeholder consultation study incorporating in-depth semi structured telephone interviews followed by thematic analysis. Interviews were conducted with a total of eighteen participants that included six senior Paramedics (Advanced Paramedics), seven General Practitioners (GPs), three Practice Nurses and two Practice Managers. Results: Participants in this study expressed polarised views on the potential for Paramedic roles in Irish General Practice. Paramedics were enthusiastic, highlighting opportunity for professional development and favourable working conditions. GP's, Practice Nurses and Managers were more circumspect and had concerns that Paramedic scope and skillset was not currently aligned to General Practice care. GP's, Practice Nurses and Managers emphasised a greater role for expanded General Practice Nursing. There were varied perceptions on what the potential role of a Paramedic in General Practice might entail, but consensus that Government support would be required to facilitate any potential developments. Conclusions: The findings of this research can inform future development of novel roles in Irish General Practice and suggests that there is appetite from within the Paramedic profession to pursue such roles. A pilot demonstration project, grounded in an action research framework could address data gaps and potential concerns. Any future developments should occur in tandem with and with due consideration for the expansion of General Practice Nursing in Ireland.


Use of latent class models to accommodate inter-laboratory variation in assessing genetic polymorphisms associated with disease risk.
Researchers wanting to study the association of genetic factors with disease may encounter variability in the laboratory methods used to establish genotypes or other traits. Such variability leads to uncertainty in determining the strength of a genotype as a risk factor. This problem is illustrated using data from a case-control study of cervical cancer in which some subjects were independently assessed by different laboratories for the presence of a genetic polymorphism. Inter-laboratory agreement was only moderate, which led to a very wide range of empirical odds ratios (ORs) with the disease, depending on how disagreements were treated. This paper illustrates the use of latent class models (LCMs) and to estimate OR while taking laboratory accuracy into account. Possible LCMs are characterised in terms of the number of laboratory measurements available, and if their error rates are assumed to be differential or non-differential by disease status and/or laboratory. The LCM results give maximum likelihood estimates of laboratory accuracy rates and the OR of the genetic variable and disease, and avoid the ambiguities of the empirical results. Having allowed for possible measurement error in the expure, the LCM estimates of exposure--disease associations are typically stronger than their empirical equivalents. Also the LCM estimates exploit all the available data, and hence have relatively low standard errors. Our approach provides a way to evaluate the association of a polymorphism with disease, while taking laboratory measurement error into account. Ambiguities in the empirical data arising from disagreements between laboratories are avoided, and the estimated polymorphism-disease association is typically enhanced.


High Performance, 3D-Printable Dielectric Nanocomposites for Millimeter Wave Devices.
The creation of millimeter wave, 3D-printable dielectric nanocomposite is demonstrated. Alumina nanoparticles were combined with styrenic block copolymers and solvent to create shear thinning, viscoelastic inks that are printable at room temperature. Particle loadings of up to 41 vol % were achieved. Upon being dried, the highest-performing of these materials has a permittivity of 4.61 and a loss tangent of 0.00298 in the Ka band (26.5-40 GHz), a combination not previously demonstrated for 3D printing. These nanocomposite materials were used to print a simple resonator device with predictable pass-band features.


Open-label 24-week extension study of edaravone (MCI-186) in amyotrophic lateral sclerosis.
We aimed to explore the longer-term efficacy and safety of edaravone in an active-treatment extension period following the double-blind period of the second phase III study. Patients who met all the following criteria (scores ≥2 points on all 12 items of the revised amyotrophic lateral sclerosis functional rating scale [ALSFRS-R], forced vital capacity ≥80%, definite or probable ALS, and disease duration ≤2 years) were randomised to 60 mg intravenous edaravone or placebo for six cycles in the double-blind period, and then offered the opportunity to proceed to this 24-week open-label extension period. One hundred and twenty-three of 137 patients continued to the extension period: 65 edaravone-edaravone (E-E group) and 58 placebo-edaravone (P-E group). Change (mean ± standard deviation; SD) in the ALSFRS-R score from baseline in the double-blind period was -4.1 ± 3.4 and -6.9 ± 5.1 in the E-E group and P-E group, respectively, while it was -8.0 ± 5.6 in the E-E group and -10.9 ± 6.9 in the P-E group over the whole 48-week period. The ALSFRS-R score changed almost linearly throughout Cycles 1-12 in the E-E group. The most commonly reported adverse events were constipation, dysphagia, and contusion. There was no sudden deterioration in the ALSFRS-R score of the E-E group. No safety concerns related to edaravone were detected.


Phosphopantetheinyl transferase CfwA/NpgA is required for Aspergillus nidulans secondary metabolism and asexual development.
Polyketide synthases (PKSs) and/or nonribosomal peptide synthetases (NRPSs) are central components of secondary metabolism in bacteria, plants, and fungi. In filamentous fungi, diverse PKSs and NRPSs participate in the biosynthesis of secondary metabolites such as pigments, antibiotics, siderophores, and mycotoxins. However, many secondary metabolites as well as the enzymes involved in their production are yet to be discovered. Both PKSs and NRPSs require activation by enzyme members of the 4'-phosphopantetheinyl transferase (PPTase) family. Here, we report the isolation and characterization of Aspergillus nidulans strains carrying conditional (cfwA2) and null (DeltacfwA) mutant alleles of the cfwA gene, encoding an essential PPTase. We identify the polyketides shamixanthone, emericellin, and dehydroaustinol as well as the sterols ergosterol, peroxiergosterol, and cerevisterol in extracts from A. nidulans large-scale cultures. The PPTase CfwA/NpgA was required for the production of these polyketide compounds but dispensable for ergosterol and cerevisterol and for fatty acid biosynthesis. The asexual sporulation defects of cfwA, DeltafluG, and DeltatmpA mutants were not rescued by the cfwA-dependent compounds identified here. However, a cfwA2 mutation enhanced the sporulation defects of both DeltatmpA and DeltafluG single mutants, suggesting that unidentified CfwA-dependent PKSs and/or NRPSs are involved in the production of hitherto-unknown compounds required for sporulation. Our results expand the number of known and predicted secondary metabolites requiring CfwA/NpgA for their biosynthesis and, together with the phylogenetic analysis of fungal PPTases, suggest that a single PPTase is responsible for the activation of all PKSs and NRPSs in A. nidulans.


Induction of the epibranchial placodes.
The cranial sensory ganglia, in contrast to those of the trunk, have a dual embryonic origin arising from both neurogenic placodes and neural crest. Neurogenic placodes are focal thickenings of ectoderm, found exclusively in the head of vertebrate embryos. These structures can be split into two groups based on the positions that they occupy within the embryo, dorsolateral and epibranchial. The dorsolateral placodes develop alongside the central nervous system, while the epibranchial placodes are located close to the top of the clefts between the branchial arches. Importantly, previous studies have shown that the neurogenic placodes form under the influence of the surrounding cranial tissues. In this paper, we have analysed the nature of the inductive signal underlying the formation of the epibranchial placodes. We find that epibranchial placodes do not require neural crest for their induction, but rather that it is the pharyngeal endoderm that is the source of the inductive signal. We also find that, while cranial ectoderm is competent to respond to this inductive signal, trunk ectoderm is not. We have further identified the signalling molecule Bmp7 as the mediator of this inductive interaction. This molecule is expressed in a manner consistent with it playing such a role and, when added to ectoderm explants, it will promote the formation of epibranchial neuronal cells. Moreover, the Bmp7 antagonist follstatin will block the ability of pharyngeal endoderm to induce placodal neuronal cells, demonstrating that Bmp7 is required for this inductive interaction. This work answers the long standing question regarding the induction of the epibranchial placodes, and represents the first elucidation of an inductive mechanism, and a molecular effector, underlying the formation of any primary sensory neurons in higher vertebrates.


Modeling denitrification in terrestrial and aquatic ecosystems at regional scales.
Quantifying where, when, and how much denitrification occurs on the basis of measurements alone remains particularly vexing at virtually all spatial scales. As a result, models have become essential tools for integrating current understanding of the processes that control denitrification with measurements of rate-controlling properties so that the permanent losses of N within landscapes can be quantified at watershed and regional scales. In this paper, we describe commonly used approaches for modeling denitrification and N cycling processes in terrestrial and aquatic ecosystems based on selected examples from the literature. We highlight future needs for developing complementary measurements and models of denitrification. Most of the approaches described here do not explicitly simulate microbial dynamics, but make predictions by representing the environmental conditions where denitrification is expected to occur, based on conceptualizations of the N cycle and empirical data from field and laboratory investigations of the dominant process controls. Models of denitrification in terrestrial ecosystems include generally similar rate-controlling variables, but vary in their complexity of the descriptions of natural and human-related properties of the landscape, reflecting a range of scientific and management perspectives. Models of denitrification in aquatic ecosystems range in complexity from highly detailed mechanistic simulations of the N cycle to simpler source-transport models of aggregate N removal processes estimated with empirical functions, though all estimate aquatic N removal using first-order reaction rate or mass-transfer rate expressions. Both the terrestrial and aquatic modeling approaches considered here generally indicate that denitrification is an important and highly substantial component of the N cycle over large spatial scales. However, the uncertainties of model predictions are large. Future progress will be linked to advances in field measurements, spatial databases, and model structures.


MadID, a Versatile Approach to Map Protein-DNA Interactions, Highlights Telomere-Nuclear Envelope Contact Sites in Human Cells.
Mapping the binding sites of DNA- or chromatin-interacting proteins is essential to understanding biological processes. DNA adenine methyltransferase identification (DamID) has emerged as a comprehensive method to map genome-wide occupancy of proteins of interest. A caveat of DamID is the specificity of Dam methyltransferase for GATC motifs that are not homogenously distributed in the genome. Here, we developed an optimized method named MadID, using proximity labeling of DNA by the methyltransferase M.EcoGII. M.EcoGII mediates N6-adenosine methylation in any DNA sequence context, resulting in deeper and unbiased coverage of the genome. We demonstrate, using m6A-specific immunoprecipitation and deep sequencing, that MadID is a robust method to identify protein-DNA interactions at the whole-genome level. Using MadID, we revealed contact sites between human telomeres, repetitive sequences devoid of GATC sites, and the nuclear envelope. Overall, MadID opens the way to identification of binding sites in genomic regions that were largely inaccessible.


Cell surface molecules containing N-acetylgalactosamine are associated with basket cells and neurogliaform cells in cat visual cortex.
In the cerebral cortex, the plant lectin Vicia villosa (VVA) selectively stains the surfaces of nonpyramidal neurons. This lectin binds specifically to alpha- and beta-linked N-acetylgalactosamine (GalNac). VVA-reactive carbohydrate is highly concentrated in layer 4 of the primary visual cortex of the cat, where it is associated exclusively with GABAergic local circuit neurons. We have studied this neuronal subset with intracellular electrophysiological recording and dye marking to identify the particular cell types expressing surface GalNac. Five different types of local circuit neurons were stained intracellularly (N = 45), but only 2 types, the columnar basket and large neurogliaform cells, were also labeled by the lectin (N = 19/45). Lectin negative types included small basket, chandelier, and large bitufted cells (26/45). Spiny stellate and pyramidal neurons were also lectin negative. Electrophysiological recordings revealed differences in the duration of action potentials in smooth versus spiny stellates but no differences between lectin-positive or -negative types. A biochemical analysis of cortical glycoproteins by SDS-PAGE and lectin blotting revealed multiple bands containing GalNac enriched in membrane fractions. These carbohydrate-containing molecules may be part of a biochemical mechanism for linking together cells with common functional properties.


Targeting host store-operated Ca(2+) release to attenuate viral infections.
Viruses coopt host intracellular Ca(2+) signaling pathways to optimize timing and effectiveness of infection stages against barriers to invasion, pathogenesis, replication, and release. Virus-induced changes in free cytosolic Ca(2+) levels facilitate virus adsorption, uncoating, catalysis, toxin production, structural assembly and stabilization, trafficking, and fusion and budding. Ca(2+)-associated alterations in virus status also selectively precipitate host cytopathologies through, among other events, retardation or induction of apoptosis, elevation of metabolic stress and reactive oxygen species production, and promotion of proinflammatory cytokine and chemokine synthesis and release. Viral particles and proteins tune spatiotemporal dynamics of host free cytosolic Ca(2+) concentrations by modulating Ca(2+) entry from the extracellular environment, upstream first or second messengers, ion- and ATP-dependent Ca2+ pumps that sequester or extrude free cytosolic Ca(2+), store-operated Ca(2+) mobilization and leakage, and viral capsid/envelope and downstream host Ca(2+) binding proteins and sensors. Each of these major viral mechanisms, briefly reviewed in this article, presents a suitable drug target capable of mitigating the severity and incidence of viral infections. Given its pivotal role in cellular response regulation, bioenergetics, posttranslational protein and lipid modification and transport, homeostasis, cell motility and morphogenesis, and T lymphocyte proliferation, targeting virally stimulated inositol 1,4,5-trisphoshate (IP3)-mediated store-operated Ca(2+) release especially offers unique, predictable benefits for augmenting immunoprotection in vertebrate clinical populations. We appraise possibilities of modulating this system with experimental proteins that gate activation kinetics of endoplasmic-reticulum-localized Ca(2+)-conducting IP3 receptors via allosteric protein-protein interactions. Such compounds are expected to be valuable in treating primary disease symptoms and sequelae, including virus-associated dementia.


The Salmonella typhi melittin resistance gene pqaB affects intracellular growth in PMA-differentiated U937 cells, polymyxin B resistance and lipopolysaccharide.
Salmonella typhi is the causative agent of typhoid fever in humans. A cell-culture based assay involving the human monocyte macrophage cell line U937 has been developed to examine S. typhi invasion and survival. An S. typhi PhoP- (null) mutant was shown to be restricted in net growth in phorbol myristate acetate (PMA) differentiated U937 (PMA-U937) cells, and an S. typhi PhoPc (constitutive) mutant showed a defect in invasion. Neither of the phoP/Q mutants were growth impaired in HeLa cells, however the PhoPc mutant was impaired in invasion. As opposed to what was found for S. typhi, Salmonella typhimurium wild-type, PhoP- and PhoPc mutants grew equally well in PMA-U937 cells, indicating that the PhoP(-)-mediated net growth restriction in the PMA-U937 cells was S. typhi specific. An S. typhi mutation, pqaB::MudJ, recently shown to be a PhoP-activated locus, was shown to have a net growth defect in PMA-U937 cells. Sequencing of the S. typhipqaB gene revealed it had 98% identity to the fifth gene in a S. typhimurium PmrA/B regulated operon necessary for 4-aminoarabinose lipid A modification and polymyxin B resistance. The pqaB locus was regulated by PmrA/B (whose activity is modulated by PhoP-PhoQ) and the pqaB transposon mutant was sensitive to polymyxin B. The lipopolysaccharides (LPS) of S. typhi and S. typhimurium wild-type, PhoP- and PhoPc mutants, were compared by SDS-PAGE and silver staining. Differences in the LPS profile between the two Salmonella species were observed, and shown to be affected differently by the PhoPc mutation. Additionally, the pqaB::MudJ mutation affected S. typhi LPS. The effects on LPS may have ramifications for the difference between S. typhi and S. typhimurium infection of hosts.


Timing of centrosome separation is important for accurate chromosome segregation.
Spindle assembly, establishment of kinetochore attachment, and sister chromatid separation must occur during mitosis in a highly coordinated fashion to ensure accurate chromosome segregation. In most vertebrate cells, the nuclear envelope must break down to allow interaction between microtubules of the mitotic spindle and the kinetochores. It was previously shown that nuclear envelope breakdown (NEB) is not coordinated with centrosome separation and that centrosome separation can be either complete at the time of NEB or can be completed after NEB. In this study, we investigated whether the timing of centrosome separation affects subsequent mitotic events such as establishment of kinetochore attachment or chromosome segregation. We used a combination of experimental and computational approaches to investigate kinetochore attachment and chromosome segregation in cells with complete versus incomplete spindle pole separation at NEB. We found that cells with incomplete spindle pole separation exhibit higher rates of kinetochore misattachments and chromosome missegregation than cells that complete centrosome separation before NEB. Moreover, our mathematical model showed that two spindle poles in close proximity do not "search" the entire cellular space, leading to formation of large numbers of syntelic attachments, which can be an intermediate stage in the formation of merotelic kinetochores.


Guanine specific binding at a DNA junction formed by d[CG(5-BrU)ACG](2) with a topoisomerase poison in the presence of Co(2+) ions.
The structure of the duplex d[CG(5-BrU)ACG](2) bound to 9-bromophenazine-4-carboxamide has been solved through MAD phasing at 2.0 A resolution. It shows an unexpected and previously unreported intercalation cavity stabilized by the drug and novel binding modes of Co(2+) ions at certain guanine N7 sites. For the intercalation cavity the terminal cytosine is rotated to pair with the guanine of a symmetry-related duplex to create a pseudo-Holliday junction geometry, with two such cavities linked through the minor groove interactions of the N2/N3 guanine sites at an angle of 40 degrees, creating a quadruplex-like structure. The mode of binding of the drug is shown to be disordered, with the major conformations showing the side chain bound to the N7 position of adjacent guanines. The other end of the duplex exhibits a terminal base fraying in the presence of Co(2+) ions linking symmetry-related guanines, causing the helices to intertwine through the minor groove. The stabilization of the structure by the intercalating drug shows that this class of compound may bind to DNA junctions as well as duplex DNA or to strand-nicked DNA ('hemi-intercalated'), as in the cleavable complex. This suggests a structural basis for the dual poisoning of topoisomerase I and II enzymes by this family of drugs.


Cell intrinsic role of COX-2 in pancreatic cancer development.
COX-2 is upregulated in pancreatic ductal adenocarcinomas (PDAC). However, how COX-2 promotes PDAC development is unclear. While previous studies have evaluated the efficacy of COX-2 inhibition via the use of nonsteroidal anti-inflammatory drugs (NSAID) or the COX-2 inhibitor celecoxib in PDAC models, none have addressed the cell intrinsic versus microenvironment roles of COX-2 in modulating PDAC initiation and progression. We tested the cell intrinsic role of COX-2 in PDAC progression using both loss-of-function and gain-of-function approaches. Cox-2 deletion in Pdx1+ pancreatic progenitor cells significantly delays the development of PDAC in mice with K-ras activation and Pten haploinsufficiency. Conversely, COX-2 overexpression promotes early onset and progression of PDAC in the K-ras mouse model. Loss of PTEN function is a critical factor in determining lethal PDAC onset and overall survival. Mechanistically, COX-2 overexpression increases p-AKT levels in the precursor lesions of Pdx1(+); K-ras(G12D)(/+); Pten(lox)(/+) mice in the absence of Pten LOH. In contrast, Cox-2 deletion in the same setting diminishes p-AKT levels and delays cancer progression. These data suggest an important cell intrinsic role for COX-2 in tumor initiation and progression through activation of the PI3K/AKT pathway. PDAC that is independent of intrinsic COX-2 expression eventually develops with decreased FKBP5 and increased GRP78 expression, two alternate pathways leading to AKT activation. Together, these results support a cell intrinsic role for COX-2 in PDAC development and suggest that while anti-COX-2 therapy may delay the development and progression of PDAC, mechanisms known to increase chemoresistance through AKT activation must also be overcome.


Structural and dynamic mechanisms of CBF3-guided centromeric nucleosome formation.
Accurate chromosome segregation relies on the specific centromeric nucleosome-kinetochore interface. In budding yeast, the centromere CBF3 complex guides the deposition of CENP-A, an H3 variant, to form the centromeric nucleosome in a DNA sequence-dependent manner. Here, we determine the structures of the centromeric nucleosome containing the native CEN3 DNA and the CBF3core bound to the canonical nucleosome containing an engineered CEN3 DNA. The centromeric nucleosome core structure contains 115 base pair DNA including a CCG motif. The CBF3core specifically recognizes the nucleosomal CCG motif through the Gal4 domain while allosterically altering the DNA conformation. Cryo-EM, modeling, and mutational studies reveal that the CBF3core forms dynamic interactions with core histones H2B and CENP-A in the CEN3 nucleosome. Our results provide insights into the structure of the budding yeast centromeric nucleosome and the mechanism of its assembly, which have implications for analogous processes of human centromeric nucleosome formation.


Overweight and obesity in relation to cardiovascular disease risk factors among medical students in Crete, Greece.
Recent data indicate increasing rates of adult obesity and mortality from cardiovascular disease (CVD) in Greece. No data, however, are available on prevalence of overweight and obesity in relation to CVD risk factors among young adults in Greece. A total of 989 third-year medical students (527 men, 462 women), aged 22 +/- 2 years, were recruited from the University of Crete during the period 1989-2001. Anthropometric measures and blood chemistries were obtained. The relationships between obesity indices (body mass index [BMI], waist circumference [WC], waist-to-hip ratio [WHpR], waist-to-height ratio [WHtR]) and CVD risk factor variables (blood pressure, glucose, serum lipoproteins) were investigated. Approximately 40% of men and 23% of women had BMI > or = 25.0 kg/m2. Central obesity was found in 33.4% (average percentage corresponding to WC > or = 90 cm, WHpR > or = 0.9 and WHtR > or = 50.0) of male and 21.7% (using WC > or = 80 cm, WHpR > or = 0.8, WHtR > or = 50.0) of female students. Subjects above the obesity indices cut-offs had significantly higher values of CVD risk factor variables. BMI was the strongest predictor of hypertension. WHtR in men and WC in women were the most important indicators of dyslipidaemia. A substantial proportion of Greek medical students were overweight or obese, obesity status being related to the presence of hypertension and dyslipidaemia. Simple anthropometric indices can be used to identify these CVD risk factors. Our results underscore the need to implement health promotion programmes and perform large-scale epidemiological studies within the general Greek young adult population.