The GRAIDS Trial: a cluster randomised controlled trial of computer decision support for the management of familial cancer risk in primary care.
The objective was to evaluate the effect of an assessment strategy using the computer decision support system (the GRAIDS software), on the management of familial cancer risk in British general practice in comparison with best current practice. The design included cluster randomised controlled trial, and involved forty-five general practice teams in East Anglia, UK. Randomised to GRAIDS (Genetic Risk Assessment on the Internet with Decision Support) support (intervention n=23) or comparison (n=22). Training in the new assessment strategy and access to the GRAIDS software (GRAIDS arm) was conducted, compared with an educational session and guidelines about managing familial breast and colorectal cancer risk (comparison) were mailed. Outcomes were measured at practice, practitioner and patient levels. The primary outcome measure, at practice level, was the proportion of referrals made to the Regional Genetics Clinic for familial breast or colorectal cancer that were consistent with referral guidelines. Other measures included practitioner confidence in managing familial cancer (GRAIDS arm only) and, in patients: cancer worry, risk perception and knowledge about familial cancer. There were more referrals to the Regional Genetics Clinic from GRAIDS than comparison practices (mean 6.2 and 3.2 referrals per 10 000 registered patients per year; mean difference 3.0 referrals; 95% confidence interval (CI) 1.2-4.8; P=0.001); referrals from GRAIDS practices were more likely to be consistent with referral guidelines (odds ratio (OR)=5.2; 95% CI 1.7-15.8, P=0.006). Patients referred from GRAIDS practices had lower cancer worry scores at the point of referral (mean difference -1.44 95% CI -2.64 to -0.23, P=0.02). There were no differences in patient knowledge about familial cancer. The intervention increased GPs' confidence in managing familial cancer. Compared with education and mailed guidelines, assessment including computer decision support increased the number and quality of referrals to the Regional Genetics Clinic for familial cancer risk, improved practitioner confidence and had no adverse psychological effects in patients. Trials are registered under N0181144343 in the UK National Research Register.


Making sense of complex phenomena in biology.
The remarkable advances in biotechnology over the past two decades have resulted in the generation of a huge amount of experimental data. It is now recognized that, in many cases, to extract information from this data requires the development of computational models. Models can help gain insight on various mechanisms and can be used to process outcomes of complex biological interactions. To do the latter, models must become increasingly complex and, in many cases, they also become mathematically intractable. With the vast increase in computing power these models can now be numerically solved and can be made more and more sophisticated. A number of models can now successfully reproduce detailed observed biological phenomena and make important testable predictions. This naturally raises the question of what we mean by understanding a phenomenon by modelling it computationally. This paper briefly considers some selected examples of how simple mathematical models have provided deep insights into complicated chemical and biological phenomena and addresses the issue of what role, if any, mathematics has to play in computational biology.


Targeting T-cell malignancies using anti-CD4 CAR NK-92 cells.
Peripheral T-cell lymphomas (PTCLs) are a group of very aggressive non-Hodgkin's lymphomas (NHLs) with poor prognoses and account for a majority of T-cell malignancies. Overall, the standard of care for patients with T-cell malignancies is poorly established, and there is an urgent clinical need for a new approach. As demonstrated in B-cell malignancies, chimeric antigen receptor (CAR) immunotherapy provides great hope as a curative treatment regimen. Because PTCLs develop from mature T-cells, these NHLs are commonly CD4+, and CD4 is highly and uniformly expressed. Therefore, CD4 is an ideal target for PTCL CAR immunotherapy. To that effect, we created a robust third-generation anti-CD4 CAR construct (CD4CAR) and introduced it into clonal NK cells (NK-92). CD4CAR NK-92 cells specifically and robustly eliminated diverse CD4+ human T-cell leukemia and lymphoma cell lines (KARPAS-299, CCRF-CEM, and HL60) and patient samples ex vivo. Furthermore, CD4CAR NK-92 cells effectively targeted KARPAS-299 cells in vivo that modeled difficult-to-access lymphoma nodules, significantly prolonging survival. In our study, we present novel targeting of CD4 using CAR-modified NK cells, and demonstrate efficacy. Combined, our data support CD4CAR NK cell immunotherapy as a potential new avenue for the treatment of PTCLs and CD4+ T-cell malignancies.


Using gene expression data to identify certain gastro-intestinal diseases.
Inflammatory bowel diseases, ulcerative colitis and Crohn's disease are considered to be of autoimmune origin, but the etiology of irritable bowel syndrome remains elusive. Furthermore, classifying patients into irritable bowel syndrome and inflammatory bowel diseases can be difficult without invasive testing and holds important treatment implications. Our aim was to assess the ability of gene expression profiling in blood to differentiate among these subject groups. Transcript levels of a total of 45 genes in blood were determined by quantitative real-time polymerase chain reaction (RT-PCR). We applied three separate analytic approaches; one utilized a scoring system derived from combinations of ratios of expression levels of two genes and two different support vector machines. All methods discriminated different subject cohorts, irritable bowel syndrome from control, inflammatory bowel disease from control, irritable bowel syndrome from inflammatory bowel disease, and ulcerative colitis from Crohn's disease, with high degrees of sensitivity and specificity. These results suggest these approaches may provide clinically useful prediction of the presence of these gastro-intestinal diseases and syndromes.


Out-of-hospital decision making and factors influencing the regional distribution of injured patients in a trauma system.
The decision-making processes used for out-of-hospital trauma triage and hospital selection in regionalized trauma systems remain poorly understood. The objective of this study was to assess the process of field triage decision making in an established trauma system. We used a mixed methods approach, including emergency medical services (EMS) records to quantify triage decisions and reasons for hospital selection in a population-based, injury cohort (2006-2008), plus a focused ethnography to understand EMS cognitive reasoning in making triage decisions. The study included 10 EMS agencies providing service to a four-county regional trauma system with three trauma centers and 13 nontrauma hospitals. For qualitative analyses, we conducted field observation and interviews with 35 EMS field providers and a round table discussion with 40 EMS management personnel to generate an empirical model of out-of-hospital decision making in trauma triage. A total of 64,190 injured patients were evaluated by EMS, of whom 56,444 (88.0%) were transported to acute care hospitals and 9,637 (17.1% of transports) were field trauma activations. For nontrauma activations, patient/family preference and proximity accounted for 78% of destination decisions. EMS provider judgment was cited in 36% of field trauma activations and was the sole criterion in 23% of trauma patients. The empirical model demonstrated that trauma triage is driven primarily by EMS provider "gut feeling" (judgment) and relies heavily on provider experience, mechanism of injury, and early visual cues at the scene. Provider cognitive reasoning for field trauma triage is more heuristic than algorithmic and driven primarily by provider judgment, rather than specific triage criteria.


Current trends and age-based differences of unintentional injury in Japanese children.
Unintentional injury in children is a worldwide public health problem, as it increases the health burden and is a leading cause of death among children. It is important to understand the differences between different age groups of children in regard to unintentional injury, in order to effectively implement child safety education. The present study aimed to determine the current trends of unintentional injury in children, and to identify the differences between different age groups of children with regard to unintentional injury. We identified 1,521 children who attended an 18-month health checkup (18-month group), and 1,368 children who attended a 36-month health checkup (36-month group), between January 1, 2014 and December 31, 2014. The rate of hospital visits associated with unintentional injury was 10.6% (161/1,521) in the 18-month group, and 13.1% (180/1,368) in the 36-month group. In both groups, present/past illness was associated with hospital visits, and in the 36-month group, hospital visits were more common in boys than in girls. The number of unintentional injuries that occurred outdoors was higher in the 36-month group than in the 18-month group. Unintentional injuries resulting from accidental ingestion and falls were more common in the 18-month group, while unintentional injuries resulting from turning over were more common in the 36-month group. In conclusion, the number of hospital visits for unintentional injury might be higher, and the number of preventive actions taken by mothers might be lower, among children attending the 36-month health checkup than among those attending the 18-month health checkup.


Operational sensitivity and acute resetting of aortic baroreceptors in dogs.
Stimulus-response curves of aortic baroreceptors constructed by alternately increasing and decreasing pressure from a normal baseline or set-point differ from curves constructed by varying pressure in one direction only from an abnormally high or low pressure. In anesthetized dogs we recorded impulses from aortic baroreceptors with myelinated fibers, using a pressurized reservoir to control mean aortic blood pressure (MABP). After setting MABP to a baseline of 100 mm Hg (normal MABP in unanesthetized dogs), we constructed baroreceptor response curves by alternately decreasing MABP from 100 to 30 mm Hg, and increasing it from 100 to 180 mm Hg, in each case returning MABP to the baseline to obtain hysteresis loops. All baroreceptors were active at 100 mm Hg, their discharge averaging 15-16 impulses/sec. At all pressures above threshold, baroreceptors fired more when pressure was increasing than when pressure was decreasing. This hysteresis caused the steepest part of the response curve constructed in this manner to span the baseline value, demonstrating that, contrary to previous views, aortic baroreceptors signal decreases in pressure below the normal level, as well as increases above it. We also constructed response curves after holding MABP at a "hypertensive" baseline of 125 mm Hg for 20 minutes. "Hypertensive" curves demonstrated reversible resetting, shifting significantly to the right of "normotensive" curves so that baroreceptor threshold increased on average by 7 mm Hg (P less than 0.01). Both hysteresis and short-term resetting probably result from the viscoelastic behavior of wall elements with which baroreceptors are coupled.


An avirulent ICP34.5 deletion mutant of herpes simplex virus type 1 is capable of in vivo spontaneous reactivation.
The herpes simplex virus type 1 (HSV-1) ICP34.5 gene is a neurovirulence gene in mice. In addition, some ICP34.5 mutants have been reported to have a reduced efficiency of induced reactivation as measured by in vitro explantation of latently infected mouse ganglia. However, since spontaneous reactivation is almost nonexistent in mice, nothing has been reported on the effect of ICP34.5 mutants on spontaneous reactivation in vivo. To examine this, we have deleted both copies of the ICP34.5 neurovirulence gene from a strain of HSV-1 (McKrae) that has a high spontaneous reactivation rate in rabbits and used this mutant to infect rabbit eyes. All rabbits infected with the ICP34.5 mutant virus (d34.5) survived, even at challenge doses greater than 4 x 10(7) PFU per eye. In contrast, a 200-fold-lower challenge dose of 2 x 10(5) PFU per eye was lethal for approximately 50% of rabbits infected with either the wild-type McKrae parental virus or a rescued ICP34.5 mutant in which both copies of the ICP34.5 gene were restored. In mice, the 50% lethal dose of the ICP34.5 mutant was over 10(6) PFU, compared with a value of less than 10 PFU for the rescued virus. The ICP34.5 mutant was restricted for replication in rabbit and mouse eyes and mouse trigeminal ganglia in vivo. The spontaneous reactivation rate in rabbits for the mutant was 1.4% as determined by culturing tear films for the presence of reactivated virus. This was more than 10-fold lower than the spontaneous reactivation rate determined for the rescued virus (19.6%) and was highly significant (P < 0.0001, Fisher exact test). Southern analysis confirmed that the reactivated virus retained both copies of the ICP34.5 deletion. Thus, this report demonstrates that (i) the ICP34.5 gene, known to be a neurovirulence gene in mice, is also important for virulence in rabbits and (ii) in vivo spontaneous reactivation of HSV-1 in the rabbit ocular model, although reduced, can occur in the absence of the ICP34.5 gene.


Liquid Chromatography-Tandem Mass Spectrometry to Define Sortase Cleavage Products.
Sortase enzymes have specific endopeptidase activity, cleaving within a defined pentapeptide sequence at the C-terminal end of their protein substrates. Here, we describe how monitoring sortase cleavage activity can be achieved using peptide substrates. Peptide cleavage can be readily analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS), which allows for the precise definition of cleavage sites. This technique could be used to analyze the peptidase activity of any enzyme, and identify sites of cleavage within any peptide.


Virtual reality and haptics as an assessment device in the postacute phase after stroke.
Virtual reality (VR) technology is altering the health care environment and is changing the options that are available to therapists. This study describes how a haptic device was used as a cinematic assessment utility. Three chronic stroke inpatients at Sahlgrenska University Hospital with left hemisphere damage were assessed. The patients were administered by the box and block manual dexterity test. For comparisons, a reference group was added to the study. Several parameters, including time, speed, and movement of the right upper extremity, were extracted and evaluated. The results indicate that the system shows potential as an assessment device. The feasibility study setup is working well, as is the assessment method. Further research, testing, refinement of the exercises, and use of VR and haptics within neurological rehabilitation are suggested.


Hepatic hepcidin/intestinal HIF-2α axis maintains iron absorption during iron deficiency and overload.
Iron-related disorders are among the most prevalent diseases worldwide. Systemic iron homeostasis requires hepcidin, a liver-derived hormone that controls iron mobilization through its molecular target ferroportin (FPN), the only known mammalian iron exporter. This pathway is perturbed in diseases that cause iron overload. Additionally, intestinal HIF-2α is essential for the local absorptive response to systemic iron deficiency and iron overload. Our data demonstrate a hetero-tissue crosstalk mechanism, whereby hepatic hepcidin regulated intestinal HIF-2α in iron deficiency, anemia, and iron overload. We show that FPN controlled cell-autonomous iron efflux to stabilize and activate HIF-2α by regulating the activity of iron-dependent intestinal prolyl hydroxylase domain enzymes. Pharmacological blockade of HIF-2α using a clinically relevant and highly specific inhibitor successfully treated iron overload in a mouse model. These findings demonstrate a molecular link between hepatic hepcidin and intestinal HIF-2α that controls physiological iron uptake and drives iron hyperabsorption during iron overload.


The impact of oxidative stress on Arabidopsis mitochondria.
Treatment of Arabidopsis cell culture for 16 h with H2O2, menadione or antimycin A induced an oxidative stress decreasing growth rate and increasing DCF fluorescence and lipid peroxidation products. Treated cells remained viable and maintained significant respiratory rates. Mitochondrial integrity was maintained, but accumulation of alternative oxidase and decreased abundance of lipoic acid-containing components during several of the treatments indicated oxidative stress. Analysis of the treatments was undertaken by IEF/SDS-PAGE, comparison of protein spot abundances and tandem mass spectrometry. A set of 25 protein spots increased >3-fold in H2O2/menadione treatments, a subset of these increased in antimycin A-treated samples. A set of 10 protein spots decreased significantly during stress treatments. A specific set of mitochondrial proteins were degraded by stress treatments. These damaged components included subunits of ATP synthase, complex I, succinyl CoA ligase, aconitase, and pyruvate and 2-oxoglutarate dehydrogenase complexes. Nine increased proteins represented products of different genes not found in control mitochondria. One is directly involved in antioxidant defense, a mitochondrial thioredoxin-dependent peroxidase, while another, a thioredoxin reductase-dependent protein disulphide isomerase, is required for protein disulfide redox homeostasis. Several others are generally considered to be extramitochondrial but are clearly present in a highly purified mitochondrial fraction used in this study and are known to play roles in stress response. Using H2O2 as a model stress, further work revealed that this treatment induced a protease activity in isolated mitochondria, putatively responsible for the degradation of oxidatively damaged mitochondrial proteins and that O2 consumption by mitochondria was significantly decreased by H2O2 treatment.


The occurrence and sources of Campylobacter spp., Salmonella enterica and Escherichia coli O157:H7 in the Salmon River, British Columbia, Canada.
In this study, we wished to assess the prevalence and determine the sources of three zoonotic bacterial pathogens (Salmonella, Campylobacter, and Escherichia coli O157:H7) in the Salmon River watershed in southwestern British Columbia. Surface water, sewage, and animal faecal samples were collected from the watershed. Selective bacterial culture and PCR techniques were used to isolate these three pathogens and indicator bacteria from these samples and characterize them. Campylobacter was the most prevalent pathogen in all samples, followed by Salmonella, and E. coli O157:H7. E. coli O157:H7 and Salmonella isolation rates from water, as well as faecal coliform densities correlated positively with precipitation, while Campylobacter isolation rates correlated negatively with precipitation. Analysis of DNA extracted from water samples for the presence of Bacteroides host-species markers, and comparisons of C. jejuni flaA-RFLP types and Salmonella serovars from faecal and water samples provided evidence that human sewage and specific domestic and wild animal species were sources of these pathogens; however, in most cases the source could not be determined or more than one source was possible. The frequent isolation of these zoonotic pathogens in the Salmon River highlights the risks to human health associated with intentional and unintentional consumption of untreated surface waters.


Trajectories of Professional Authority: A Comparative Study of Medical Associations and Professional Authority Claims.
Medical associations not only organize their members' interests but also exercise professional authority within the field of health policy. An important aspect of professional authority is the medical profession's ability to position itself in relation to national health policy and whether its command of professional knowledge enables the profession to claim exclusive authority for reflecting on health policy. This article analyzes and compares how medical associations claim authority over health policy and how they reposition their claims in light of perceived contestations to medical authority in public debates or from the political system. The study is based on a qualitative, descriptive analysis of 975 editorials in the medical associations' lead journals in the United States, the United Kingdom, and Denmark over a period of 60 years. The analysis explores the trajectories of authority claims in the three countries and how professional authority claims may be reconfigured to reflect external changes in health policy institutions. Whereas all the medical associations were highly critical of state-organized health systems in the 1950s and early 1960s, the British and Danish associations seem to shift positions entirely after national health systems are gradually implemented and the associations begin to present themselves as these public institutions' strongest supporters.


Transcriptional profiling of bovine milk using RNA sequencing.
Cow milk is a complex bioactive fluid consumed by humans beyond infancy. Even though the chemical and physical properties of cow milk are well characterized, very limited research has been done on characterizing the milk transcriptome. This study performs a comprehensive expression profiling of genes expressed in milk somatic cells of transition (day 15), peak (day 90) and late (day 250) lactation Holstein cows by RNA sequencing. Milk samples were collected from Holstein cows at 15, 90 and 250 days of lactation, and RNA was extracted from the pelleted milk cells. Gene expression analysis was conducted by Illumina RNA sequencing. Sequence reads were assembled and analyzed in CLC Genomics Workbench. Gene Ontology (GO) and pathway analysis were performed using the Blast2GO program and GeneGo application of MetaCore program. A total of 16,892 genes were expressed in transition lactation, 19,094 genes were expressed in peak lactation and 18,070 genes were expressed in late lactation. Regardless of the lactation stage approximately 9,000 genes showed ubiquitous expression. Genes encoding caseins, whey proteins and enzymes in lactose synthesis pathway showed higher expression in early lactation. The majority of genes in the fat metabolism pathway had high expression in transition and peak lactation milk. Most of the genes encoding for endogenous proteases and enzymes in ubiquitin-proteasome pathway showed higher expression along the course of lactation. This is the first study to describe the comprehensive bovine milk transcriptome in Holstein cows. The results revealed that 69% of NCBI Btau 4.0 annotated genes are expressed in bovine milk somatic cells. Most of the genes were ubiquitously expressed in all three stages of lactation. However, a fraction of the milk transcriptome has genes devoted to specific functions unique to the lactation stage. This indicates the ability of milk somatic cells to adapt to different molecular functions according to the biological need of the animal. This study provides a valuable insight into the biology of lactation in the cow, as well as many avenues for future research on the bovine lactome.


Aperiodic CrSc multilayer mirrors for attosecond water window pulses.
Extending single attosecond pulse technology from currently sub-200 eV to the so called 'water window' spectral range may enable for the first time the unique investigation of ultrafast electronic processes within the core states of bio-molecules as proteins or other organic materials. Aperiodic multilayer mirrors serve as key components to shape these attosecond pulses with a high degree of freedom and enable tailored short pulse pump-probe experiments. Here, we report on chirped CrSc multilayer mirrors, fabricated by ion beam deposition with sub-angstrom precision, designed for attosecond pulse shaping in the 'water window' spectral range.


Associations between respiratory illnesses and secondhand smoke exposure in flight attendants: A cross-sectional analysis of the Flight Attendant Medical Research Institute Survey.
ABSTRACT: BACKGROUND: Secondhand tobacco smoke (SHS) is associated with increased risk of respiratory illness, cancer, and cardiovascular disease. Prior to smoking bans on airlines in the late 1980s, flight attendants were exposed to a significant amount of SHS. In the present study, we examine associations between flight attendant SHS exposure and development of respiratory illnesses and cardiovascular disease. METHODS: Between December 2006 and October 2010, three hundred sixty-two flight attendants completed an online questionnaire with information regarding experience as a flight attendant, medical history, smoking history, and SHS exposure. Rates of illnesses in flight attendants were compared with an age and smoking history matched population sample from NHANES 2005-2006. Logistic regression analysis was used to examine the association of reported medical conditions and pre-ban years of exposure. RESULTS: Compared with the sample from NHANES 2005-2006, flight attendants had increased prevalence of chronic bronchitis (11.7% vs. 7.2%, p < 0.05), emphysema/COPD (3.2% vs. 0.9%, p < 0.03), and sinus problems (31.5% vs. 20.9%, p < 0.002), despite a lower prevalence of medical illnesses including high blood pressure, diabetes, high cholesterol, heart failure, cancer, and thyroid disease. Amongst flight attendants who reported never smoking over their lifetimes, there was not a significant association between years of service as a flight attendant in the pre-smoking ban era and illnesses. However, in this same group, there was a significantly increased risk of daily symptoms (vs. no symptoms) of nasal congestion, throat, or eye irritation per 10-year increase of years of service as a flight attendant prior to the smoking ban (OR 2.14, 95% CI 1.41 - 3.24). CONCLUSIONS: Flight attendants experience increased rates of respiratory illnesses compared to a population sample. The frequency of symptoms of nasal congestion, throat or eye irritation is associated with occupational SHS exposure in the pre-smoking ban era.


Internal state dynamics shape brainwide activity and foraging behaviour.
The brain has persistent internal states that can modulate every aspect of an animal's mental experience1-4. In complex tasks such as foraging, the internal state is dynamic5-8. Caenorhabditis elegans alternate between local search and global dispersal5. Rodents and primates exhibit trade-offs between exploitation and exploration6,7. However, fundamental questions remain about how persistent states are maintained in the brain, which upstream networks drive state transitions and how state-encoding neurons exert neuromodulatory effects on sensory perception and decision-making to govern appropriate behaviour. Here, using tracking microscopy to monitor whole-brain neuronal activity at cellular resolution in freely moving zebrafish larvae9, we show that zebrafish spontaneously alternate between two persistent internal states during foraging for live prey (Paramecia). In the exploitation state, the animal inhibits locomotion and promotes hunting, generating small, localized trajectories. In the exploration state, the animal promotes locomotion and suppresses hunting, generating long-ranging trajectories that enhance spatial dispersion. We uncover a dorsal raphe subpopulation with persistent activity that robustly encodes the exploitation state. The exploitation-state-encoding neurons, together with a multimodal trigger network that is associated with state transitions, form a stochastically activated nonlinear dynamical system. The activity of this oscillatory network correlates with a global retuning of sensorimotor transformations during foraging that leads to marked changes in both the motivation to hunt for prey and the accuracy of motor sequences during hunting. This work reveals an important hidden variable that shapes the temporal structure of motivation and decision-making.


Current perception threshold and the HAVS Stockholm sensorineural scale.
It is important to determine which tests of sensorineural dysfunction identify the neurological damage from hand-arm vibration exposure. To examine the association between the hand-arm vibration syndrome (HAVS) Stockholm sensorineural scale stages and tests of peripheral neurological function including measurement of current perception threshold (CPT) and nerve conduction. All the subjects were men who were assessed for HAVS with a medical and occupational history and physical examination to determine the Stockholm stage, CPT testing at frequencies of 5, 250 and 2000 Hz for the median and ulnar nerves and measurement of nerve conduction carried out in a blinded fashion. A total of 155 of the 157 recruited subjects agreed to take part in the study, a 99% participation rate. CPT was statistically significantly increased (P < 0.001) in both Stockholm sensorineural Stages 1 and > or =2 in comparison to Stage 0 for every frequency and nerve combination. However, CPT could not discriminate well between Stages 1 and > or =2. There was no association between median or ulnar neuropathy measured by nerve conduction and the Stockholm stages. Polychotomous multinomial logistic regression indicated that the CPT measurements at 2000 Hz, corresponding to damage to large myelinated nerve fibres, were most predictive of both Stockholm Stages 1 and > or =2 in comparison to Stage 0. Neuropathy measured by nerve conduction was unrelated to the Stockholm scale stages. CPT was increased above Stage 0 but did not distinguish well between the higher stages of the Stockholm scale.


Ventilators for noninvasive ventilation to treat acute respiratory failure.
The application of noninvasive ventilation (NIV) to treat acute respiratory failure has increased tremendously both inside and outside the intensive care unit. The choice of ventilator is crucial for success of NIV in the acute setting, because poor tolerance and excessive air leaks are significantly correlated with NIV failure. Patient-ventilator asynchrony and discomfort can occur if the physician or respiratory therapist fails to adequately set NIV to respond to the patient's ventilatory demand, so clinicians need to fully understood the ventilator's technical peculiarities (eg, efficiency of trigger and cycle systems, speed of pressurization, air-leak compensation, CO(2) rebreathing, reliability of fraction of inspired oxygen reading, monitoring accuracy). A wide range of ventilators of different complexity have been introduced into clinical practice to noninvasively support patients in acute respiratory failure, but the numerous commercially available ventilators (bi-level, intermediate, and intensive care unit ventilators) have substantial differences that can influence patient comfort, patient-ventilator interaction, and, thus, the chance of NIV clinical success. This report examines the most relevant aspects of the historical evolution, the equipment, and the acute-respiratory-failure clinical application of NIV ventilators.


N-Formylmethionyl peptide receptors on equine leukocytes initiate secretion but not chemotaxis.
The chemotaxis of leukocytes appears to be initiated by the binding of chemotactic factors to the surface of these cells. N-Formylated peptides induce chemotaxis and lysosomal enzyme secretion of leukocytes; because these peptides are available in a purified radiolabeled form, they have been useful in the characterization of receptors for chemotactic factors. Equine polymorphonuclear leukocytes secrete lysosomal enzymes but do not exhibit chemotaxis in respone to the N-formylated peptides, even though they have a high-affinity cell surface receptor for these agents. The specificity of the equine receptor resembles the specificity of the receptor on chemotactically responsive leukocytes from other species. Equine polymorphonuclear leukocytes may thus be an excellent model for the study of the events that lead to a biological response following receptor occupancy.


Conversion of cardiovascular conference abstracts to publications.
The transition of scientific knowledge from discovery into practice is less than ideal. A key step in this translation occurs when presentations from major meetings are published in peer-reviewed literature, yet the completeness and speed of this process are not known. We performed a systematic and automated evaluation of rates, timing, and correlates of publication from scientific abstracts presented at 3 major cardiovascular conferences. Using an automated computer algorithm, we searched the ISI Web of Science to identify peer-reviewed publications of abstracts presented at the American Heart Association (AHA), American College of Cardiology (ACC), and European Society of Cardiology (ESC) scientific sessions from 2006 to 2008. We compared abstract publication rates and journal impact factor between the 3 meetings using multivariable logistic regression modeling. From 2006 to 2008, 11 365, 5005, and 10 838 abstracts were presented at the AHA, ACC, and ESC meetings, respectively. Overall, 30.6% of presented abstracts were published within 2 years of the conference; ranging from 34.5% for AHA to 29.5% for ACC to 27.0% for ESC (P<0.0001). Five years after conference presentation in 2005, these rates had risen slightly to 49.7% for AHA, 42.6% for ACC, and 37.6% for ESC (P<0.0001). After adjustment for abstract characteristics and contributing countries, abstracts presented at the AHA meeting remained more likely for publication relative to the ESC (adjusted odds ratio, 1.24; 95% confidence interval, 1.16-1.34) and the ACC (adjusted odds ratio, 1.20; 95% confidence interval, 1.11-1.29). Median impact factors for subsequent publications varied from 4.8 (interquartile range, 3.8-10.1) for AHA to 4.0 (interquartile range, 3.1-7.5) for ACC and 3.9 (quartile 1-3, 2.5-5.8) for ESC (P for difference between groups <0.01). Clinical science and population science were less likely to be published compared with basic science. One third of abstracts were translated into publications by 2 years after presentation and less than one half by 5 years after presentation. Our findings suggest that efforts to understand the barriers to publication and to facilitate the rapid dissemination of new knowledge are needed to speed up the transition of scientific discovery into clinical practice.


Aerosol features retrieved from solar aureole data: a simulation study concerning a turbid atmosphere.
The characteristics of the solar aureole were evaluated for several cases of a turbid atmosphere in the 3° ≤θ≤30° interval of scattering angles; for each case, the features of the aerosol were retrieved from the simulated aureole data. Computations were carried out with a recently set up radiative transfer code that uses the approximated delta-M method, corrected further for the 1st and 2nd scattering orders. Results showed that the software tested can work out both the direct and the inverse aureole problems with great accuracy and efficiency in several different situations, so it can reliably be used for handling experimental data measured in the field with an aureolemeter. Furthermore, the input parameters of ground albedo, complex refractive index, aerosol radius interval, and measurement angles were varied within a set of values to examine the sensitivity of the retrieval to improperly assumed values of these parameters and to evaluate the most suitable way of determining their correct values. Only data concerning diffuse radiation were elaborated. Results showed that (1) the scanned scattering angles have to be extended up to 40°; (2) the most suitable radius interval for aerosols appears to be from 0.05 to 15 µm; (3) ground albedo A should be independently determined within 15%; and (4) as to the complex refractive index m˜, the real part should be given within 3.5%, and the imaginary part within from 10% to 50%, according to its value. Finally, a procedure through which it is possible to derive A and m˜ by extending the information content of the aureole data is discussed. Improved calibration procedures are also proposed.


Aromatic hydroxylation during the myeloperoxidase-oxidase oxidation of hydrazines.
Benzoic acid was found to be hydroxylated by a mixture of myeloperoxidase (MPO) and the mycobactericidal drug, isoniazid. Aromatic hydroxylation and formation of compound III (oxyperoxidase) were coincident during the MPO-oxidase oxidation of isoniazid which proceeded without augmentation from the reagent hydrogen peroxide. An intermediate of isoniazid reduced ferric MPO to ferrous MPO which associated with dioxygen to form compound III. Aromatic hydroxylation also occurred in a mixture of isoniazid (or phenylhydrazine) and a ferric salt. Hydroxylations in both the enzymatic and nonenzymatic reaction systems were inhibited by the iron chelator, desferal, as well as by the specific hydroxyl radical scavenger, mannitol. To distinguish between the hydroxylating intermediates in the different reaction systems, the unique properties of the natural antioxidant, phytic acid, were exploited. Phytic acid inhibited aromatic hydroxylation in the Fe(3+)-INH system, which is in accordance with its known properties as a powerful inhibitor of iron-driven reactions (.OH formation). By contrast, phytic acid stimulated hydroxylation in the enzymatic system which was accompanied by a concomitant stimulation in the rate of compound III formation. These events were, however, not directly related to each other. Phytic acid had a direct effect on the redox transformation of isoniazid by stimulating superoxide generation during auto-oxidation of the drug. In addition, phytic acid also facilitated compound III decay in the absence of isoniazid, suggesting that it may also regulate the oxygen affinity of MPO, similar to its effect on the oxygenation of haemoglobin. The data on aromatic hydroxylation in the MPO-isoniazid system do not support a role for .OH in the reaction and may fit the model for the P450 mixed oxidase system.


Leadership, staffing and quality of care in nursing homes.
Leadership and staffing are recognised as important factors for quality of care. This study examines the effects of ward leaders' task- and relationship-oriented leadership styles, staffing levels, ratio of registered nurses and ratio of unlicensed staff on three independent measures of quality of care. A cross-sectional survey of forty nursing home wards throughout Norway was used to collect the data. Five sources of data were utilised: self-report questionnaires to 444 employees, interviews with and questionnaires to 13 nursing home directors and 40 ward managers, telephone interviews with 378 relatives and 900 hours of field observations. Separate multi-level analyses were conducted for quality of care assessed by relatives, staff and field observations respectively. Task-oriented leadership style had a significant positive relationship with two of the three quality of care indexes. In contrast, relationship-oriented leadership style was not significantly related to any of the indexes. The lack of significant effect for relationship-oriented leadership style was due to a strong correlation between the two leadership styles (r=0.78). Staffing levels and ratio of registered nurses were not significantly related to any of the quality of care indexes. The ratio of unlicensed staff, however, showed a significant negative relationship to quality as assessed by relatives and field observations, but not to quality as assessed by staff. Leaders in nursing homes should focus on active leadership and particularly task-oriented behaviour like structure, coordination, clarifying of staff roles and monitoring of operations to increase quality of care. Furthermore, nursing homes should minimize use of unlicensed staff and address factors related to high ratios of unlicensed staff, like low staff stability. The study indicates, however, that the relationship between staffing levels, ratio of registered nurses and quality of care is complex. Increasing staffing levels or the ratio of registered nurses alone is not likely sufficient for increasing quality of care.


Defective Interferon Gamma Production by Tumor-Specific CD8+ T Cells Is Associated With 5'Methylcytosine-Guanine Hypermethylation of Interferon Gamma Promoter.
Interferon gamma (IFNγ) supports effector responses of CD8+ cytotoxic T lymphocytes (CTLs) and is a surrogate marker for detection of antigen-specific T cells. Here, we show that tumor-specific CTL clones have impaired IFNγ expression and production upon activation. Assessment of the relationship between IFNγ production and the 5'methylcytosine-guanine (CpG) dinucleotide methylation of the IFNγ promoter using bisulfite treatment has shown that IFNγ- CTL clones accumulates CpG hypermethylation within the promoter at key transcription factor binding sites (-186 and -54), known to be vital for transcription. We confirmed these findings using ex vivo isolated and short-term expanded bulk tumor-specific CTL lines from four cancer patients and demonstrated that IFNγ methylation inversely correlates with transcription, protein level, and cytotoxicity. Altogether, we propose that a sizeable portion of human tumor-specific CTLs are deficient in IFNγ response, contributed by CpG hypermethylation of the IFNγ promoter. Our findings have important implications for immunotherapy strategies and for methods to detect human antigen-specific T cells.


Approaches for establishing the function of regulatory genetic variants involved in disease.
The diversity of regulatory genetic variants and their mechanisms of action reflect the complexity and context-specificity of gene regulation. Regulatory variants are important in human disease and defining such variants and establishing mechanism is crucial to the interpretation of disease-association studies. This review describes approaches for identifying and functionally characterizing regulatory variants, illustrated using examples from common diseases. Insights from recent advances in resolving the functional epigenomic regulatory landscape in which variants act are highlighted, showing how this has enabled functional annotation of variants and the generation of hypotheses about mechanism of action. The utility of quantitative trait mapping at the transcript, protein and metabolite level to define association of specific genes with particular variants and further inform disease associations are reviewed. Establishing mechanism of action is an essential step in resolving functional regulatory variants, and this review describes how this is being facilitated by new methods for analyzing allele-specific expression, mapping chromatin interactions and advances in genome editing. Finally, integrative approaches are discussed together with examples highlighting how defining the mechanism of action of regulatory variants and identifying specific modulated genes can maximize the translational utility of genome-wide association studies to understand the pathogenesis of diseases and discover new drug targets or opportunities to repurpose existing drugs to treat them.


Protective role of melatonin in progesterone production by human luteal cells.
This study investigated whether melatonin protects luteinized granulosa cells from reactive oxygen species (ROS) as an antioxidant to enhance progesterone production in the follicle during ovulation. Follicular fluid was sampled at the time of oocyte retrieval in women undergoing in vitro fertilization and embryo transfer (IVF-ET). Melatonin concentrations in the follicular fluid were positively correlated with progesterone concentrations (r = 0.342, P < 0.05) and negatively correlated with the concentration of 8-hydroxy-2'-deoxyguanosine (8-OHdG), an oxidative stress marker (r = -0.342, P < 0.05). The progesterone and 8-OHdG concentrations were negatively correlated (r = -0.246, P < 0.05). Luteinized granulosa cells were obtained at the time of oocyte retrieval in women undergoing IVF-ET. Cells were incubated with H(2)O(2) (30, 50, 100 μm) in the presence or absence of melatonin (1, 10, 100 μg/mL). Progesterone production by luteinized granulosa cells was significantly inhibited by H(2)O(2). Melatonin treatment overcame the inhibitory effect of H(2) O(2) . Twenty-five patients who had luteal phase defect (serum progesterone concentrations <10 ng/mL during the mid-luteal phase) were divided into two groups during the next treatment cycle: 14 women were given melatonin (3 mg/day at 22:00 hr) throughout the luteal phase and 11 women were given no medication as a control. Melatonin treatment improved serum progesterone concentrations (>10 ng/mL during the mid-luteal phase) in nine of 14 women (64.3%), whereas only two of 11 women (18.1%) showed normal serum progesterone levels in the control group. In conclusion, melatonin protects granulosa cells undergoing luteinization from ROS in the follicle and contributes to luteinization for progesterone production during ovulation.


The effects of forward head posture on expiratory muscle strength in chronic neck pain patients: A cross-sectional study.
This study aims to investigate the relationship between forward head posture (FHP) and respiratory dysfunctions in patients with chronic neck pain. Between June 2014 and November 2016 , a total of 99 patients (11 males, 88 females; mean age 54.1±9 years; range, 38 to 75 years) with chronic neck pain were evaluated for head posture by cervical lateral radiograph measuring the anterior head translation distance (FHPmm) and C7 vertebrae position (C7°). We examined the chest expansion by subtracting chest circumference from the level of xiphoid during maximal inspiration and expiration. Pain severity and neck disability were assessed using the Visual Analog Scale (VAS) and modified Neck Disability Index (MNDI), respectively. The respiratory functions were evaluated using spirometry tests, lung volumes, and maximal inspiratory and expiratory pressures (Pimax and Pemax, respectively). There was a negative correlation between the FHPmm with Pemax% (rho: -0.314; p=0.005). A negative correlation was also observed between C7° and Pemax, Pemax%, forced expiratory volume in one sec (FEV1)/forced vital capacity (FVC)%, forced expiratory flow (FEF)25-75%, and FEF75% (rho:-0.245, -0.349, -0.218, -0.214, and -0.259 respectively; p=0.028, 0.002, 0.035, 0.040, and 0.012, respectively). There was a positive correlation between neck disability and VAS scores (rho: 0.424; p<0.001), while there was a negative correlation between neck disability and chest expansion, maximum voluntary ventilation (rho: -0.201 and -0.217, respectively; p=0.049 and 0.046, respectively). Based on our study results, FHP is associated with expiratory muscle weakness in chronic neck pain patients. To evaluate respiratory dysfunction, chest expansion tests may be useful, although these tests are not specific to muscle weakness. Interventions about FHP and neck pain should focus on the effects of respiratory muscle training.


Embryonic pig pancreatic tissue for the treatment of diabetes in a nonhuman primate model.
Xenotransplantation of pig tissues has great potential to overcome the shortage of organ donors. One approach to address the vigorous immune rejection associated with xenotransplants is the use of embryonic precursor tissue, which induces and utilizes host vasculature upon its growth and development. Recently, we showed in mice that embryonic pig pancreatic tissue from embryonic day 42 (E42) exhibits optimal properties as a beta cell replacement therapy. We now demonstrate the proof of concept in 2 diabetic Cynomolgus monkeys, followed for 393 and 280 days, respectively. A marked reduction of exogenous insulin requirement was noted by the fourth month after transplantation, reaching complete independence from exogenous insulin during the fifth month after transplantation, with full physiological control of blood glucose levels. The porcine origin of insulin was documented by a radioimmunoassay specific for porcine C-peptide. Furthermore, the growing tissue was found to be predominantly vascularized with host blood vessels, thereby evading hyperacute or acute rejection, which could potentially be mediated by preexisting anti-pig antibodies. Durable graft protection was achieved, and most of the late complications could be attributed to the immunosuppressive protocol. While fine tuning of immune suppression, tissue dose, and implantation techniques are still required, our results demonstrate that porcine E-42 embryonic pancreatic tissue can normalize blood glucose levels in primates. Its long-term proliferative capacity, its revascularization by host endothelium, and its reduced immunogenicity, strongly suggest that this approach could offer an attractive replacement therapy for diabetes.


Coronary artery disease and cardiovascular outcomes in patients with non-alcoholic fatty liver disease.
Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome and is associated with cardiovascular risk. The aim of this study was to determine the role of fatty liver in predicting coronary artery disease and clinical outcomes in patients undergoing coronary angiogram. This was a prospective cohort study carried out in a University hospital. Consecutive patients who underwent coronary angiogram had ultrasound screening for fatty liver. Significant cardiovascular disease was defined as ≥50% stenosis in at least one coronary artery. The primary outcome was a composite end point comprising cardiovascular deaths, non-fatal myocardial infarction and the need for further coronary intervention during prospective follow-up. Among 612 recruited patients, 356 (58.2%) had fatty liver by ultrasonography, 318 (52.0%) had elevated serum alanine aminotransferase and 465 (76.0%) had significant coronary artery disease. Coronary artery disease occurred in 84.6% of patients with fatty liver and 64.1% of those without fatty liver (p<0.001). After adjusting for demographic and metabolic factors, fatty liver (adjusted OR 2.31; 95% CI 1.46 to 3.64) and alanine aminotransferase level (adjusted OR 1.01; 95% CI 1.00 to 1.02) remained independently associated with coronary artery disease. At a mean follow-up of 87±22 weeks, 30 (10.0%) patients with fatty liver and 18 (11.0%) patients without fatty liver reached the composite clinical end point (p=0.79). In patients with clinical indications for coronary angiogram, fatty liver is associated with coronary artery disease independently of other metabolic factors. However, fatty liver cannot predict cardiovascular mortality and morbidity in patients with established coronary artery disease.


Combining Fluoxetine and rTMS in Poststroke Motor Recovery: A Placebo-Controlled Double-Blind Randomized Phase 2 Clinical Trial.
Background. Although recent evidence has shown a new role of fluoxetine in motor rehabilitation, results are mixed. We conducted a randomized clinical trial to evaluate whether combining repetitive transcranial magnetic stimulation (rTMS) with fluoxetine increases upper limb motor function in stroke. Methods. Twenty-seven hemiparetic patients within 2 years of ischemic stroke were randomized into 3 groups: Combined (active rTMS + fluoxetine), Fluoxetine (sham rTMS + fluoxetine), or Placebo (sham rTMS + placebo fluoxetine). Participants received 18 sessions of 1-Hz rTMS in the unaffected primary motor cortex and 90 days of fluoxetine (20 mg/d). Motor function was assessed using Jebsen-Taylor Hand Function (JTHF) and Fugl-Meyer Assessment (FMA) scales. Corticospinal excitability was assessed with TMS. Results. After adjusting for time since stroke, there was significantly greater improvement in JTHF in the combined rTMS + fluoxetine group (mean improvement: -214.33 seconds) than in the placebo (-177.98 seconds, P = 0.005) and fluoxetine (-50.16 seconds, P < 0.001) groups. The fluoxetine group had less improvement than placebo on both scales (respectively, JTHF: -50.16 vs -117.98 seconds, P = 0.038; and FMA: 6.72 vs 15.55 points, P = 0.039), suggesting that fluoxetine possibly had detrimental effects. The unaffected hemisphere showed decreased intracortical inhibition in the combined and fluoxetine groups, and increased intracortical facilitation in the fluoxetine group. This facilitation was negatively correlated with motor function improvement (FMA, r2 = -0.398, P = 0.0395). Conclusion. Combined fluoxetine and rTMS treatment leads to better motor function in stroke than fluoxetine alone and placebo. Moreover, fluoxetine leads to smaller improvements than placebo, and fluoxetine's effects on intracortical facilitation suggest a potential diffuse mechanism that may hinder beneficial plasticity on motor recovery.


WE-E-BRB-07: Direct 3D Fluence Calculation from Machine Beam Parameters for VMAT Delivery Verification.
MLC dynamic log files have been clinically used for quality assurance for years. The logged machine parameters and the derived beam 2D fluence maps can be compared to the ones obtained in the treatment plans in order to evaluate the accuracy and consistency of IMRT beam deliveries. In this study, we propose a computationally efficient method, called Direct 3D Fluence Calculation or D3DFC, to extend 2D fluence map derivation to 3D fluence volume computation. The aim is to extend dynalog-based QA from fixed-gantry IMRT to rotational-gantry VMAT. D3DFC calculates the 3D volume of photon fluence distribution directly from the machine parameters (gantry angles, jaw positions, MLC positions, collimator rotation angle, MU) contained in the dynamic log files or DICOM plans, without converting to 2D fluence maps per gantry angle in order to allow higher computation speed and accuracy. For testing, results were verified with film-in-air measurements. 3D fluence volumes computed from VMAT delivery records (with artificially introduced delivery errors) are compared to ones computed from the treatment plans to determine if these delivery errors can be identified. D3DFC is implemented in MATLAB and supports the DICOM plans, Varian MLC dynamic logs and Varian Truebeam machine logs. Computation takes 10 to 20 seconds for a single-arc VMAT plan or delivery records. The results showed that 1 mm MLC errors can be clearly detected using delivery-to-plan fluence volume comparison. Direct computation from machine parameters allows higher computation speed and accuracy. These advantages are useful for beam delivery verification purposes for which (slower) full patient CT based dose computation is less necessary. The calculated 3D photon fluence volume is useful to detect and visually present the VMAT delivery discrepancies.


Analysis of O-glycoforms of the IgA1 hinge region by sequential deglycosylation.
A common renal disease, immunoglobulin A (IgA) nephropathy (IgAN), is associated with glomerular deposition of IgA1-containing immune complexes. IgA1 hinge region (HR) has up to six clustered O-glycans consisting of Ser/Thr-linked N-acetylgalactosamine with β1,3-linked galactose and variable sialylation. IgA1 glycoforms with some galactose-deficient (Gd) HR O-glycans play a key role in IgAN pathogenesis. The clustered and variable O-glycans make the IgA1 glycomic analysis challenging and better approaches are needed. Here, we report a comprehensive analytical workflow for IgA1 HR O-glycoform analysis. We combined an automated quantitative analysis of the HR O-glycopeptide profiles with sequential deglycosylation to remove all but Gd O-glycans from the HR. The workflow was tested using serum IgA1 from healthy subjects. Twelve variants of glycopeptides corresponding to the HR with three to six O-glycans were detected; nine glycopeptides carried up to three Gd O-glycans. Sites with Gd O-glycans were unambiguously identified by electron-transfer/higher-energy collision dissociation tandem mass spectrometry. Extracted ion chromatograms of isomeric glycoforms enabled quantitative assignment of Gd sites. The most frequent Gd site was T236, followed by S230, T233, T228, and S232. The new workflow for quantitative profiling of IgA1 HR O-glycoforms with site-specific resolution will enable identification of pathogenic IgA1 HR O-glycoforms in IgAN.


Pregnancy and pelvic girdle pain: Analysis of pelvic belt on pain.
To analyse pain and functional capacity in women with pelvic girdle pain and to evaluate the effect of pelvic belt on these parameters. Two types of belts were to compare. Pelvic girdle pain is very common during pregnancy. To prevent and relieve pelvic pain, women can use a set of techniques and tools such as a pelvic belt. While scientific evidence is lacking, commercial industries suggest the effectiveness of pelvic belts. Randomised control trial. Forty-six pregnant women with pelvic girdle pain were evaluated. Pain analysis included a quantitative and a qualitative assessment. A daily activities questionnaire was used for functional capacity evaluation. Women were tested at two times during the pregnancy for a longitudinal evaluation, and they used one of the two belt models during their pregnancy. Pelvic pain started between the 14th-21st week of pregnancy. Pain intensity was 60 ± 20 mm. Daily activities could increase pain. The use of belts reduced pain. The intensity of pain decreased by 20 mm on a visual analogue scale. The daily activities were also easier. However, all these conclusions are valid only if pregnant women used belts regularly on short periods. The belts appear to be interesting tools to reduce pelvic pain and improve comfort of pregnant women. This effect might be explained by an analgesic effect with proprioceptive and biomechanical effect. The different types of belts could have differential effects on global, sacroiliac joint and back pain during pregnancy, but this hypothesis requires confirmation. Relevant for patient: to use an easy and validated tool. Relevant for clinical practice: to suggest a tool scientifically validated for patient. Relevant to economic issues: belts decrease pelvic pain and increase comfort of pregnant women. Sick leave could decrease.


Clinical and Histological Findings of Post-Treatment Infection in the Presence of Vertical Root Fracture and Apical Periodontitis: Case Reports.
Apical periodontitis and vertical root fracture (VRF) might coexist in the same root, thus representing a di-agnostic challenge in endodontics. Diagnosis should be based on detailed clinical examination and precise radiographic interpretation in addition to histological examination, if needed. The histological findings of the case presented showed the possible coexistence of a VRF in the coronal third of the root and signs of apical periodontitis caused by intracanal infection in the apical third of the same root. The presentation of this case underlines the importance of histological examination as a valid complementary diagnostic tool with clinical and radiographic examinations in reaching an accurate diagnosis once the root has been extracted.


Empathy is related to clinical competence in medical care.
Most medical schools focus upon aspects of interpersonal functioning such as empathy in the doctor-patient relationship with the aim of training clinically competent doctors. This study investigated the relationship between empathy and clinical competence among medical students. Fifty-seven medical students participated in the study. Clinical competence was assessed in an objective structured clinical examination (OSCE). Empathy was rated by an independent observer of the clinical interaction in OSCE stations. In addition, empathy was self-rated using the Jefferson Scale of Physician Empathy-Student Version. Observed behaviour indicative of empathy, as rated objectively by an independent observer, was strongly associated with clinical competence. The strong association between the behavioural manifestation of empathy and clinical competence was evident across a range of medical conditions and types of consultation. In addition, observable empathy was strongly associated with patients' ratings of the students' performances. Self-rated empathy, however, was not associated with clinical competence. Significant differences in mean total competence scores were noted between students low in observed empathy (mean ± standard deviation [SD]: 165.86 ± 12.92) and students high in observed empathy (mean ± SD: 190.35 ± 14.00) (t[55] = 6.28, p = 0.000, α ≤ 0.01). In medical education, strategies that enhance the behavioural expression of empathy (or at least retard its decay) could help to make medical students appear to be more clinically competent to both examiners and patients. However, if medical students' internal emotions are found to be discrepant with their behaviour, these findings will raise difficult questions regarding the fundamental nature of genuine empathy and alert us to the possibility that medical students may learn that it pays to subscribe to the view that if a person does not feel empathy, he or she can fake it.


Functional characterization and target discovery of glycoside hydrolases from the digestome of the lower termite Coptotermes gestroi.
Lignocellulosic materials have been moved towards the forefront of the biofuel industry as a sustainable resource. However, saccharification and the production of bioproducts derived from plant cell wall biomass are complex and lengthy processes. The understanding of termite gut biology and feeding strategies may improve the current state of biomass conversion technology and bioproduct production. The study herein shows comprehensive functional characterization of crude body extracts from Coptotermes gestroi along with global proteomic analysis of the termite's digestome, targeting the identification of glycoside hydrolases and accessory proteins responsible for plant biomass conversion. The crude protein extract from C. gestroi was enzymatically efficient over a broad pH range on a series of natural polysaccharides, formed by glucose-, xylose-, mannan- and/or arabinose-containing polymers, linked by various types of glycosidic bonds, as well as ramification types. Our proteomic approach successfully identified a large number of relevant polypeptides in the C. gestroi digestome. A total of 55 different proteins were identified and classified into 29 CAZy families. Based on the total number of peptides identified, the majority of components found in the C. gestroi digestome were cellulose-degrading enzymes. Xylanolytic enzymes, mannan- hydrolytic enzymes, pectinases and starch-degrading and debranching enzymes were also identified. Our strategy enabled validation of liquid chromatography with tandem mass spectrometry recognized proteins, by enzymatic functional assays and by following the degradation products of specific 8-amino-1,3,6-pyrenetrisulfonic acid labeled oligosaccharides through capillary zone electrophoresis. Here we describe the first global study on the enzymatic repertoire involved in plant polysaccharide degradation by the lower termite C. gestroi. The biochemical characterization of whole body termite extracts evidenced their ability to cleave all types of glycosidic bonds present in plant polysaccharides. The comprehensive proteomic analysis, revealed a complete collection of hydrolytic enzymes including cellulases (GH1, GH3, GH5, GH7, GH9 and CBM 6), hemicellulases (GH2, GH10, GH11, GH16, GH43 and CBM 27) and pectinases (GH28 and GH29).


Oxidative stress and glomerular filtration barrier injury: role of the renin-angiotensin system in the Ren2 transgenic rat.
TG(mRen2)27 (Ren2) transgenic rats overexpress the mouse renin gene, manifest hypertension, and exhibit increased tissue ANG II levels and oxidative stress. Evidence indicates that elevated tissue ANG II contributes to oxidative stress, increases in glomerular macromolecular permeability, and consequent albuminuria. Furthermore, angiotensin type 1 receptor (AT1R) blockers reduce albuminuria and slow progression of renal disease. However, it is not known whether improvements in glomerular filtration barrier integrity and albuminuria during treatment are related to reductions in oxidative stress and/or kidney renin-angiotensin system (RAS) activity. To investigate the renal protective effects of AT1R blockade, we treated young (6-7 wk old) male Ren2 rats with valsartan (Ren2-V; 30 mg/kg) for 3 wk and measured urine albumin, kidney malondialdehyde (MDA), RAS component mRNAs, and NADPH oxidase subunits (gp91(phox) and Rac1) compared with age-matched untreated Ren2 and Sprague-Dawley (S-D) rats. Basement membrane thickness, slit pore diameter and number, and foot process base width were measured by transmission electron microscopy (TEM). Results indicate that AT1R blockade lowered systolic blood pressure (30%), albuminuria (91%), and kidney MDA (80%) in Ren2-V compared with untreated Ren2 rats. Increased slit pore number and diameter and reductions in basement membrane thickness and podocyte foot process base width were strongly associated with albuminuria and significantly improved following AT1R blockade. AT1R blockade was also associated with increased angiotensin-converting enzyme-2 and neprilysin expression, demonstrating a beneficial shift in balance of renal RAS. Thus reductions in blood pressure, albuminuria, and tissue oxidative stress with AT1R blockade were associated with improved indexes of glomerular filtration barrier integrity and renal RAS in Ren2 rats.


Applying the planned care model to intimate partner violence.
This article presents the steps for organizing a health organization's response to intimate partner violence (IPV) according to the Planned Care Model (PCM). IPV is common and costly and results in poor physical and mental health outcomes for victims and their families. Because most care is not acute, a planned approach that crosses systems may result in more comprehensive and higher quality care. Community collaboration with IPV agencies is especially critical. The health care organization must make IPV a priority and set policies and systems to identify and manage patients, train staff, and measure, monitor, and provide feedback on outcomes. Other key PCM components include: practice design--design systems to identify and track victims, stratify risk, and coordinate care; evidence-based decision support--choose validated IPV screening questions and guidelines for identification, management, and referral and make them available in a systematic way with ongoing assessment and feedback to providers and other members of the health care team; patient self-management--self-man-agement materials should be selected and disseminated to those working with IPV victims; and data information systems--these should support a confidential patient registry and efforts to audit and provide feedback about identification and referral efforts. Process and outcome measures based on the management guidelines and protocols should be developed and monitored, and the results disseminated. Adapting PCM for the management of IPV stretches the traditional acute approach to IPV of screen-identify-refer. It expands the PCM into new realms, including embracing new partners, trying innovative ways to measure return on investment, grappling with ethical dilemmas, and designing a multifactorial evaluation across systems.


Priority setting for health in the context of devolution in Kenya: implications for health equity and community-based primary care.
Devolution changes the locus of power within a country from central to sub-national levels. In 2013, Kenya devolved health and other services from central government to 47 new sub-national governments (known as counties). This transition seeks to strengthen democracy and accountability, increase community participation, improve efficiency and reduce inequities. With changing responsibilities and power following devolution reforms, comes the need for priority-setting at the new county level. Priority-setting arises as a consequence of the needs and demand for healthcare resources exceeding the resources available, resulting in the need for some means of choosing between competing demands. We sought to explore the impact of devolution on priority-setting for health equity and community health services. We conducted key informant and in-depth interviews with health policymakers, health providers and politicians from 10 counties (n = 269 individuals) and 14 focus group discussions with community members based in 2 counties (n = 146 individuals). Qualitative data were analysed using the framework approach. We found Kenya's devolution reforms were driven by the need to demonstrate responsiveness to county contexts, with positive ramifications for health equity in previously neglected counties. The rapidity of the process, however, combined with limited technical capacity and guidance has meant that decision-making and prioritization have been captured and distorted for political and power interests. Less visible community health services that focus on health promotion, disease prevention and referral have been neglected within the prioritization process in favour of more tangible curative health services. The rapid transition in power carries a degree of risk of not meeting stated objectives. As Kenya moves forward, decision-makers need to address the community health gap and lay down institutional structures, processes and norms which promote health equity for all Kenyans.


Characterization of the novel positive allosteric modulator of the metabotropic glutamate receptor 4 ADX88178 in rodent models of neuropsychiatric disorders.
There is growing evidence that activation of metabotropic glutamate receptor 4 (mGlu4) leads to anxiolytic- and antipsychotic-like efficacy in rodent models, yet its relevance to depression-like reactivity remains unclear. Here, we present the pharmacological evaluation of ADX88178 [5-methyl-N-(4-methylpyrimidin-2-yl)-4-(1H-pyrazol-4-yl)thiazol-2-amine], a novel potent, selective, and brain-penetrant positive allosteric modulator of the mGlu4 receptor in rodent models of anxiety, obsessive compulsive disorder (OCD), fear, depression, and psychosis. ADX88178 dose-dependently reduced the number of buried marbles in the marble burying test and increased open-arm exploration in the elevated plus maze (EPM) test, indicative of anxiolytic-like efficacy. Target specificity of the effect in the EPM test was confirmed using male and female mGlu4 receptor knockout mice. In mice, ADX88178 reduced the likelihood of conditioned freezing in the acquisition phase of the fear conditioning test, yet had no carryover effect in the expression phase. Also, ADX88178 dose-dependently reduced duration of immobility in the forced swim test, indicative of antidepressant-like efficacy. ADX88178 reduced DOI (2,5-dimethoxy-4-iodoamphetamine)-mediated head twitches (albeit with no dose-dependency), and MK-801 [(5S,10R)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine]-induced locomotor hyperactivity in mice, but was inactive in the conditioned avoidance response test in rats. The compound showed good specificity as it had no effect on locomotor activity in mice and rats at efficacious doses. Thus, allosteric activation of mGlu4 receptors can be a promising new therapeutic approach for treatment of anxiety, OCD, fear-related disorders, and psychosis.


Interference with the host haemostatic system by schistosomes.
Schistosomes, parasitic flatworms that cause the tropical disease schistosomiasis, are still a threat. They are responsible for 200 million infections worldwide and an estimated 280,000 deaths annually in sub-Saharan Africa alone. The adult parasites reside as pairs in the mesenteric or perivesicular veins of their human host, where they can survive for up to 30 years. The parasite is a potential activator of blood coagulation according to Virchow's triad, because it is expected to alter blood flow and endothelial function, leading to hypercoagulability. In contrast, hepatosplenic schistosomiasis patients are in a hypocoagulable and hyperfibrinolytic state, indicating that schistosomes interfere with the haemostatic system of their host. In this review, the interactions of schistosomes with primary haemostasis, secondary haemostasis, fibrinolysis, and the vascular tone will be discussed to provide insight into the reduction in coagulation observed in schistosomiasis patients. Interference with the haemostatic system by pathogens is a common mechanism and has been described for other parasitic worms, bacteria, and fungi as a mechanism to support survival and spread or enhance virulence. Insight into the mechanisms used by schistosomes to interfere with the haemostatic system will provide important insight into the maintenance of the parasitic life cycle within the host. This knowledge may reveal new potential anti-schistosome drug and vaccine targets. In addition, some of the survival mechanisms employed by schistosomes might be used by other pathogens, and therefore, these mechanisms that interfere with host haemostasis might be a broad target for drug development against blood-dwelling pathogens. Also, schistosome antithrombotic or thrombolytic molecules could form potential new drugs in the treatment of haemostatic disorders.


Phylogenomics Yields New Insight Into Relationships Within Vernonieae (Asteraceae).
Asteraceae, or the sunflower family, is the largest family of flowering plants and is usually considered difficult to work with, not only due to its size, but also because of the abundant cases of polyploidy and ancient whole-genome duplications. Traditional molecular systematics studies were often impaired by the low levels of variation found in chloroplast markers and the high paralogy of traditional nuclear markers like ITS. Next-generation sequencing and novel phylogenomics methods, such as target capture and Hyb-Seq, have provided new ways of studying the phylogeny of the family with great success. While the resolution of the backbone of the family is in progress with some results already published, smaller studies focusing on internal clades of the phylogeny are important to increase sampling and allow morphological, biogeography, and diversification analyses, as well as serving as basis to test the current infrafamilial classification. Vernonieae is one of the largest tribes in the family, accounting for approximately 1,500 species. From the 1970s to the 1990s, the tribe went through several reappraisals, mainly due to the splitting of the mega genus Vernonia into several smaller segregates. Only three phylogenetic studies focusing on the Vernonieae have been published to date, both using a few molecular markers, overall presenting low resolution and support in deepest nodes, and presenting conflicting topologies when compared. In this study, we present the first attempt at studying the phylogeny of Vernonieae using phylogenomics. Even though our sampling includes only around 4% of the diversity of the tribe, we achieved complete resolution of the phylogeny with high support recovering approximately 700 nuclear markers obtained through target capture. We also analyzed the effect of missing data using two different matrices with different number of markers and the difference between concatenated and gene tree analysis.


Postexercise hypotension after maximal short-term incremental exercise depends on exercise modality.
This study investigated postexercise hypotension (PEH) after maximal cardiopulmonary exercise testing (CPET) performed using different exercise modalities. Twenty healthy men (aged 23 ± 3 years) performed 3 maximal CPETs (cycling, walking, and running), separated by 72 h in a randomized, counter-balanced order. Systolic (SBP) and diastolic blood pressure (DBP), heart rate, cardiac output, systemic vascular resistance (SVR), autonomic function (spontaneous baroreflex sensitivity (BRS) and heart rate variability (HRV)), and energy expenditure (EE) were assessed during a 60-min nonexercise control session and for 60 min immediately after each CPET. Total exercise volume (EE during CPET plus 60 min recovery) was significantly higher in running versus cycling and walking CPETs (P ≤ 0.001). Compared with control, only SBP after running CPET was significantly reduced (Δ = -6 ± 8 mm Hg; P < 0.001). Heart rate and cardiac output were significantly increased (P < 0.001) and SVR significantly decreased (P < 0.001) postexercise. BRS and HRV decreased after all CPETs (P < 0.001), whereas sympatho-vagal balance (low- and high-frequency (LF:HF) ratio) increased significantly after all exercise conditions, especially after running CPET (P < 0.001). Changes in SVR, BRS, sympathetic activity (low-frequency component of HRV), and LF:HF ratio were negatively correlated to variations in SBP (range -0.69 to -0.91; P < 0.001) and DBP (range -0.58 to -0.93; P ≤ 0.002). These findings suggest that exercise mode or the total exercise volume are major determinants of PEH magnitude in healthy men. Because of the running CPET, the PEH was primarily related to a decrease in SVR and to an increase in sympatho-vagal balance, which might be a reflex response to peripheral vasodilatation after exercise.


Thiosulfate Mediates Cytoprotective Effects of Hydrogen Sulfide Against Neuronal Ischemia.
Hydrogen sulfide (H2S) exhibits protective effects in various disease models including cerebral ischemia-reperfusion (I/R) injury. Nonetheless, mechanisms and identity of molecules responsible for neuroprotective effects of H2S remain incompletely defined. In the current study, we observed that thiosulfate, an oxidation product of H2S, mediates protective effects of an H2S donor compound sodium sulfide (Na2S) against neuronal I/R injury. We observed that thiosulfate in cell culture medium is not only required but also sufficient to mediate cytoprotective effects of Na2S against oxygen glucose deprivation and reoxygenation of human neuroblastoma cell line (SH-SY5Y) and murine primary cortical neurons. Systemic administration of sodium thiosulfate (STS) improved survival and neurological function of mice subjected to global cerebral I/R injury. Beneficial effects of STS, as well as Na2S, were associated with marked increase of thiosulfate, but not H2S, in plasma and brain tissues. These results suggest that thiosulfate is a circulating "carrier" molecule of beneficial effects of H2S. Protective effects of thiosulfate were associated with inhibition of caspase-3 activity by persulfidation at Cys163 in caspase-3. We discovered that an SLC13 family protein, sodium sulfate cotransporter 2 (SLC13A4, NaS-2), facilitates transport of thiosulfate, but not sulfide, across the cell membrane, regulating intracellular concentrations and thus mediating cytoprotective effects of Na2S and STS. The protective effects of H2S are mediated by thiosulfate that is transported across cell membrane by NaS-2 and exerts antiapoptotic effects via persulfidation of caspase-3. Given the established safety track record, thiosulfate may be therapeutic against ischemic brain injury.


Cosmetic or esthetic dentistry?
This article, through presentation of case studies, defines differences and suggests separate definitions for the terms cosmetic dentistry and esthetic dentistry. Dentistry strives to emulate harmonious form and function for therapy, and modification of appearance is an integral part of dental treatment. Cosmetic dentistry suggests a certain accommodation and is a compromise of current technology. Cosmetic dentistry is commonly selected as an interim procedure that does not necessarily function ideally and does not always emulate the pristine state of a natural dentition. Esthetic dentistry requires less accommodation, incorporates acceptable biologic technology for long-term survival, functions suitably, and mimics the pristine state of the natural dentition. Cosmetic and esthetic dentistry are different in definition, concept, and execution.


Obesity increases risk for pouch-related complications following restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA).
Restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) is the preferred surgical treatment for patients with ulcerative colitis and familial adenomatous polyposis. As obesity is becoming more epidemic in surgical patients, the aim of this study was to investigate if obesity increases complication rates following IPAA. This study was conducted as a retrospective review of patients undergoing IPAA between January 1990 and April 2011. Patients were categorized by body mass index (BMI): BMI < 30 (non-obese) and BMI ≥ 30 (obese). Preoperative patient demographics, operative variables, and postoperative complications were recorded through chart review. The primary outcome studied was cumulative complication rate. A total of 103 non-obese and 75 obese patients were identified who underwent IPAA. Obese patients had an increased rate of overall complications (80 % vs. 64%, p = 0.03), primarily accounted for by increased pouch-related complications (61% vs. 26%, p < 0.01). In particular, obese patients had more anastomotic/pouch strictures (27% vs. 6%, p < 0.01), inflammatory pouch complications (17 % vs. 4%, p < 0.01) and pouch fistulas (12% vs. 3%, p = 0.03). In a regression model, obesity remained a significant risk factor (odds ratio [OR] = 2.86, p = 0.01) for pouch-related complications. Obesity is associated with an increased risk of overall and pouch-related complications following IPAA. Obese patients should be counseled preoperatively about these risks accordingly.


Complex airway disease: an approach to assessment and management.
Research into new treatments for airway disease focuses on severe disease because morbidity, mortality, and health-care costs are substantial and the unmet need is greatest. One reason why outcomes are poor in these patients could be that the clinical expression of disease is heterogeneous and difficult to classify. As a result, guideline-based management algorithms fail. Additionally, difficulties with disease classification and misconceptions about the relation between different aspects of severe airway disease have hindered new drug development. A potential solution is to use a new approach to assess severe airway disease, which moves the diagnostic focus from categorisation of patients to identification and characterisation of the main drivers of disease. This approach will help rather than hinder identification of clinically important phenotypes of disease and will facilitate the development of new phenotype-specific treatment options.


Evi1 is essential for hematopoietic stem cell self-renewal, and its expression marks hematopoietic cells with long-term multilineage repopulating activity.
Ecotropic viral integration site 1 (Evi1), a transcription factor of the SET/PR domain protein family, is essential for the maintenance of hematopoietic stem cells (HSCs) in mice and is overexpressed in several myeloid malignancies. Here, we generate reporter mice in which an internal ribosome entry site (IRES)-GFP cassette is knocked-in to the Evi1 locus. Using these mice, we find that Evi1 is predominantly expressed in long-term HSCs (LT-HSCs) in adult bone marrow, and in the hematopoietic stem/progenitor fraction in the aorta-gonad-mesonephros, placenta, and fetal liver of embryos. In both fetal and adult hematopoietic systems, Evi1 expression marks cells with long-term multilineage repopulating activity. When combined with conventional HSC surface markers, sorting according to Evi1 expression markedly enhances purification of cells with HSC activity. Evi1 heterozygosity leads to marked impairment of the self-renewal capacity of LT-HSCs, whereas overexpression of Evi1 suppresses differentiation and boosts self-renewal activity. Reintroduction of Evi1, but not Mds1-Evi1, rescues the HSC defects caused by Evi1 heterozygosity. Thus, in addition to documenting a specific relationship between Evi1 expression and HSC self-renewal activity, these findings highlight the utility of Evi1-IRES-GFP reporter mice for the identification and sorting of functional HSCs.


Short Is Beautiful: Dimensionality and Measurement Invariance in Two Length of the Basic Psychological Need Satisfaction at Work Scale.
Self-determination theory proposes that all humans have three intrinsic psychological needs: the needs for Autonomy, Competence, and Relatedness. These needs take different forms in different areas of life. The present study examines the psychometric properties of a Swedish version of the Basic Psychological Need Satisfaction at Work (BPNS-W) scale. The fit of 10-factor structures previously suggested for related versions of the scale were compared. Cross-sectional data from 1,200 participants were examined in a confirmatory factor analysis framework. Both the original 21-item version and a reduced 12-item version of the BPNS-W were examined. The General Health Questionnaire was used for validation. The results supported a three-factor solution with correlated error variances for the reversed items. Invariance testing of the long and short scales gave best support to the short scale, for which partial scalar invariance was achieved. The external validity of the short scale was supported by a hierarchical regression analysis in which each need made a unique contribution in predicting psychological well-being. In conclusion, the results corroborate a three-factor structure of BPNS-W. Although not perfect the short scale should, it is argued, be preferred over the long version. Directions for the future development of the scale are discussed.


The importance of neurobiological research to the prevention of psychopathology.
There is both a biological and environmental component to the neural substrates for various forms of psychopathology. Brain dysfunction itself not only constitutes a formidable liability to psychopathology, but also has an impact on environmental and social responses to the individual, compounding the risk for an adverse outcome. Environmental conditions, such as social and physical stimulus deprivation, poverty, traumatic stress, and prenatal drug exposure, can further compromise brain function in the context of existing liabilities. The relationship between genetic and environmental processes is interactive, fluid, and cumulative in their ability to influence an individual's developmental trajectory and alter subsequent behavioral outcomes. Given the codependent relationship between these processes, brain function is now believed to be malleable via manipulations of the environment in ways that may decrease liability for psychopathology. Research that explores these relationships and ways in which interventions can redirect this developmental track may substantially advance both the science and practice of prevention. Studies attempting to isolate the neurobiological effects of socioenvironmental factors are reviewed, implications for intervention strategies are discussed, and a future research agenda is proposed to provide greater insight into specific brain-environment relationships. Armed with this knowledge, prevention scientists may eventually design programs that directly target these effects to reverse or attenuate negative outcomes.


Phosphatidylserine and signal transduction: who needs whom?
The plasma membrane, long considered a simple barrier between the extracellular and intracellular compartments, is now thought to play a pivotal role in many physiological processes that regulate the communication of cells with their environment. On one hand, the plasma membrane directly participates in intracellular signaling; on the other hand, changes in membrane structure contribute to the transcellular transfer of biological information. Among the membrane constituents, phosphatidylserine is a major actor implicated in these effects. Evidence now exists for a role for phosphatidylserine redistribution in modulating the activities of several membrane proteins during signaling in nonapoptotic T lymphocytes.


Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA.
While CRISPR-based gene knock out in mammalian cells has proven to be very efficient, precise insertion of genetic elements via the cellular homology directed repair (HDR) pathway remains a rate-limiting step to seamless genome editing. Under the conditions described here, we achieved up to 56% targeted integration efficiency with up to a six-nucleotide insertion in HEK293 cells. In induced pluripotent stem cells (iPSCs), we achieved precise genome editing rates of up to 45% by co-delivering the Cas9 RNP and donor DNA. In addition, the use of a short double stranded DNA oligonucleotide with 3' overhangs allowed integration of a longer FLAG epitope tag along with a restriction site at rates of up to 50%. We propose a model that favors the design of donor DNAs with the change as close to the cleavage site as possible. For small changes such as SNPs or short insertions, asymmetric single stranded donor molecules with 30 base homology arms 3' to the insertion/repair cassette and greater than 40 bases of homology on the 5' end seems to be favored. For larger insertions such as an epitope tag, a dsDNA donor with protruding 3' homology arms of 30 bases is favored. In both cases, protecting the ends of the donor DNA with phosphorothioate modifications improves the editing efficiency.


Erwinia carotovora has two KdgR-like proteins belonging to the IciR family of transcriptional regulators: identification and characterization of the RexZ activator and the KdgR repressor of pathogenesis.
A novel Erwinia carotovora subsp. carotovora mutant designated RexZ, (regulator of exoenzymes) showed reduced production of the degradative exoenzymes. The rexZ gene product shows similarity of the KdgR regulatory protein from Erwinia chrysanthemi, described as the major repressor of the pectin catabolism pathway genes in the latter species. In vitro DNA-protein interaction experiments demonstrated that the synthesis of the RexZ protein is controlled by the cAMP-CRP (cAMP-receptor protein) complex. Western blot analysis also revealed the presence of a second KdgR homologue (distinct from RexZ) which, like RexZ, was present in all species of the genus Erwinia tested. The corresponding KdgR proteins from both E. carotovora subsp. carotovora and E. carotovora subsp. atroseptica share a high level of sequence identity with the KdgR homologues from E. chrysanthemi and Escherichia coli. Although the E. carotovora subsp. carotovora rexZ regulatory region displayed specific interactions with both the purified E. chrysanthemi KdgR repressor and the partially purified E. carotovora subsp. carotovora KdgR, in vivo quantification revealed that the cellular level of RexZ protein was unaffected by the presence of pectic compounds. This study shows that the complex regulatory network governing virulence in the erwinias involves two totally distinct, but highly conserved, members of the IcIR class of DNA binding proteins: RexZ and KdgR.


Simulation of protein-sugar interactions: a computational model of the complex between ganglioside GM1 and the heat-labile enterotoxin of Escherichia coli.
The cholera toxin from Vibrio cholerae (CT) and the 80% homologous heat-labile toxin of Escherichia coli (LT) are two well-known cases of sugar-binding proteins. The GM1:toxin complexes were chosen as test cases for the elaboration of a computational approach to the modeling of protein-saccharide interactions. The reliability of the method was evaluated on the LT:lactose complex. A model of this complex was built by performing a MC/EM conformational search of the sugar moiety within the binding pocket of LT, using the AMBER* force field and the GB/SA solvation model. The results are a reasonable reproduction of the reported X-ray structure of the complex. The same protocol was then applied to the LT:GM1 complex. The calculations were performed on a substructure that includes the protein shell within 5 A from GM1, three water molecules solvating Glu-51 carboxylate, and two water molecules at crystallographic sites 2 and 3. A satisfactory agreement was found with the recently published X-ray structure of the CT:GM1 complex. All the relevant interactions between the sugar and the residues involved in binding are well reproduced by the calculations. These results suggest that the substructure here identified can be taken as a realistic representation of the toxin binding surface and that the method presented in this paper can be used as a predictive tool in designing artificial LT (CT) binders and thus potential anticholera drugs.


Testing cognitive navigation in unknown territories: homing pigeons choose different targets.
Homing pigeons (Columba livia) are believed to adopt a map-and-compass strategy to find their way home. Surprisingly, to date a clear demonstration of the use of a cognitive map in free-flight experiments is missing. In this study, we investigated whether homing pigeons use a mental map in which - at an unknown release site - their own position, the home loft and a food loft are represented simultaneously. In order to test this, homing pigeons were trained to fly to a 25-30 km distant food loft. A total of 131 hungry and satiated pigeons were then released from an unfamiliar site equidistant from the food loft and the home loft. Their vanishing bearings and homing times were assessed conventionally at four sites, and also their flight tracks from one release site by means of GPS loggers. The vanishing bearings of fed and hungry birds differed significantly at all release sites and a highly significant proportion of hungry birds flew to the food loft, while the fed birds headed home. The GPS experiment revealed a number of pigeons flying very precisely to the food loft, others correcting their flight direction after topography-induced detours. This implies that the pigeons knew their geographical position in relation to the targets, and chose a flight direction according to their locally manipulated needs - clearly the essence of a cognitive navigational map.


Factors associated with physical activity in Australians with hip or knee osteoarthritis.
Physical activity (PA) is recommended for managing osteoarthritis (OA). However, few people with OA are physically active. Understanding the factors associated with PA is necessary to increase PA in this population. This cross-sectional study examined factors associated with leisure-time PA, stretching exercises, and strengthening exercises in people with OA. For a mail survey, 485 individuals, aged 68.0 years (SD = 10.6) with hip or knee OA, were asked about factors that may influence PA participation, including use ofnon-PAOA management strategies and both psychological and physical health-related factors. Associations between factors and each PA outcome were examined in multivariable logistic regression models. Non-PA management strategies were the main factors associated with the outcomes. Information/education courses, heat/cold treatments, and paracetamol were associated with stretching and strengthening exercises (P < .05). Hydrotherapy and magnet therapy were associated with leisure-time PA; using orthotics and massage therapy, with stretching exercises; and occupational therapy, with strengthening exercises (P < .05). Few psychological or health-related factors were associated with the outcomes. Some management strategies may make it easier for people with OA to be physically active, and could be promoted to encourage PA. Providers of strategies are potential avenues for recruiting people with OA into PA programs.


Serotyping dengue virus with isothermal amplification and a portable sequencer.
The recent development of a nanopore-type portable DNA sequencer has changed the way we think about DNA sequencing. We can perform sequencing directly in the field, where we collect the samples. Here, we report the development of a novel method to detect and genotype tropical disease pathogens, using dengue fever as a model. By combining the sequencer with isothermal amplification that only requires a water bath, we were able to amplify and sequence target viral genomes with ease. Starting from a serum sample, the entire procedure could be finished in a single day. The analysis of blood samples collected from 141 Indonesian patients demonstrated that this method enables the clinical identification and serotyping of the dengue virus with high sensitivity and specificity. The overall successful detection rate was 79%, and a total of 58 SNVs were detected. Similar analyses were conducted on 80 Vietnamese and 12 Thai samples with similar performance. Based on the obtained sequence information, we demonstrated that this approach is able to produce indispensable information for etiologically analyzing annual or regional diversifications of the pathogens.


Interaction effect of psychological distress and asthma control on productivity loss?
Little is known about the potential synergistic effect of comorbid psychological distress (PD) and uncontrolled asthma (UA) on productivity loss. We estimated the productivity loss associated with the combination of these two potentially preventable conditions in employed adults with asthma. A population-based random sample of 300 adults with asthma in British Columbia, Canada, was prospectively recruited between Dec 2010 and Aug 2012. PD and productivity loss due to absenteeism and presenteeism was measured using validated instruments, and asthma control was ascertained using 2010 Global Initiative for Asthma management strategy. We used two-part regression models to study the contribution of UA and PD to productivity loss. Compared with reference group (controlled asthma (CA)+noPD), those with UA+noPD had CAD$286 (95%CI $276-297) weekly productivity loss, and those with CA+PD had CAD$465 ($445-485). Those with UA+PD had CAD$449 (437-462) in productivity loss. There was no significant interaction effect of PD with asthma control levels on productivity loss (p=0.22). In patients without PD, uncontrolled asthma was associated with a higher productivity loss than controlled asthma, but this was not the case in patients with PD. This finding can be explained by the fact that the contribution of PD to productivity loss is so large that there is no room for synergy with asthma control. Future studies should assess the impact of interventions that modify PD in patients with asthma.