Epoxyeicosatrienoic acid analog EET-B attenuates post-myocardial infarction remodeling in spontaneously hypertensive rats.
Epoxyeicosatrienoic acids (EETs) and their synthetic analogs have cardiovascular protective effects. Here, we investigated the action of a novel EET analog EET-B on the progression of post-myocardial infarction (MI) heart failure in spontaneously hypertensive rats (SHR). Adult male SHR were divided into vehicle- and EET-B (10 mg/kg/day; p.o., 9 weeks)-treated groups. After 2 weeks of treatment, rats were subjected to 30-min left coronary artery occlusion or sham operation. Systolic blood pressure (SBP) and echocardiography (ECHO) measurements were performed at the beginning of study, 4 days before, and 7 weeks after MI. At the end of the study, tissue samples were collected for histological and biochemical analyses. We demonstrated that EET-B treatment did not affect blood pressure and cardiac parameters in SHR prior to MI. Fractional shortening (FS) was decreased to 18.4 ± 1.0% in vehicle-treated MI rats compared with corresponding sham (30.6 ± 1.0%) 7 weeks following MI induction. In infarcted SHR hearts, EET-B treatment improved FS (23.7 ± 0.7%), markedly increased heme oxygenase-1 (HO-1) immunopositivity in cardiomyocytes and reduced cardiac inflammation and fibrosis (by 13 and 19%, respectively). In conclusion, these findings suggest that EET analog EET-B has beneficial therapeutic actions to reduce cardiac remodeling in SHR subjected to MI.


The PAS study: a randomized controlled trial evaluating the effectiveness of a web-based multiple tailored smoking cessation programme and tailored counselling by practice nurses.
PAS (Personal Advice in Stopping smoking) combines two of the most effective smoking cessation interventions: multiple computer tailoring and tailored counselling by a practice nurse in the general practice. Since May 2009, practice nurses are recruiting smoking patients. Each practice nurse is asked to recruit 15 adult smokers who are willing to quit within 6months and have access to the Internet. Smokers can sign up for PAS through the PAS website and are then randomized into one of three groups receiving multiple tailoring and counselling (MTC), multiple tailoring (MT) or usual care (UC), respectively. All groups receive questionnaires at baseline, 2days after a set quit date and at 6weeks, 6months and 12months follow-up. The MT group receives tailored, iterative feedback letters at the first four measurements points. At 12months follow-up biochemical validation will take place amongst respondents reporting to have quit. The three groups will be compared with regard to quit attempt rate, point prevalence abstinence and continued abstinence by means of logistic multilevel regression analyses. Linear multilevel regression analyses will be used to compare the three groups regarding smoking related beliefs. The present paper provides an extensive description of the development of PAS and of the design of the study towards its effectiveness. This might provide insight into PAS' potentially effective working mechanisms. The results concerning effectiveness may contribute to knowledge about the effectiveness of smoking cessation interventions aimed at smoking adults. Dutch Trial Register NTR1351.


Slowing the Titanic: China's Epic Struggle with Tobacco.
China is home to a third of the world's smokers and, correspondingly, to a third of the world's cases of lung cancer. Beginning in the mid-1990s, a generation or so later than in many Western countries, the Chinese government commenced measures to control tobacco, limiting advertising, banning smoking in many public venues, and increasing taxation. At the time of this review, there are signs that these policies are having some effect, but hundreds of millions of Chinese continue to smoke and rates of diagnosis of lung cancer continue to rise. There is much work to be done and much premature death to be suffered before the epidemic is slowed to the levels reached in Australia or the United States. This article aims to provide, particularly for practicing lung cancer clinicians, a description of patterns of smoking in China, the lung cancer epidemic there, and the stimuli for and barriers to tobacco control imposed by the highly complex and unique regulatory setting of the Chinese tobacco industry. A particular challenge in developing this description has come from the variability of studies published about a huge nation that has enormous diversity in wealth, education, urbanization, and tradition. The studies vary because the data vary. Much information on lung cancer and smoking rates in China comes, for example, from studies of cohorts that may number in the millions yet represent only a small percentage of the population and sometimes only a tiny geographic area of such a vast nation. National registry data on lung cancer in China do not yet cover even a fifth of the national population. Even so, we argue that several major trends can be identified: (1) more than 50% of men smoke regularly; (2) there are more than 300 million smokers in China; (3) almost half a million new cases of lung cancer are diagnosed in China each year; (4) secondhand smoking is a significant problem in China (as elsewhere), accounting for a high proportion of lung cancer cases among women, most of whom do not smoke; (5) corrected for age and sex, lung cancer rates in the Chinese population are as least as high as in Western countries and arguably higher; and (6) a shift over time from squamous cell carcinoma (SCC) to adenocarcinoma is occurring in parallel with that noted in Western populations. Looking to the future, measures to control tobacco in China are strengthening but remain relatively weak. The reinforcement of control is hampered by the circumstance that the tobacco industry remains a powerful state-owned monopoly that participates, through its membership of key government bodies, in the processes that oversee the implementation of tobacco control. The conflict between the government's dual responsibilities for both the production and distribution of tobacco and for the implementation of tobacco control carries inherent contradictions and bias. How long the Chinese government chooses to live with these contradictions will determine how quickly the smoking of tobacco will be reduced and how soon the burden of disease that tobacco inflicts on individuals and society as a whole will be eased.


Core functions for the public health nutrition workforce in Europe: a consensus study.
To assess and develop a consensus among a European panel of public health nutrition workforce stakeholders (academics and employers) regarding core functions required for effective public health nutrition practice. A modified Delphi study involving data from two rounds of questionnaires administered among a panel of public health nutrition workforce stakeholders. Europe. A panel of fifty-three public health nutrition development stakeholders, including thirty-three academics and twenty employers, sampled from eighteen European countries. Panellists rated 50 % (19/38) of the initially listed functions as core (i.e. without which public health capacity is limited), using a majority cut-off (>50 %). Out of the nineteen core functions seven were categorised under the heading Intervention management, emphasising high agreement on the importance of managing interventions in public health nutrition work. Only one of the identified core public health nutrition functions was rated differently between academics and employers, suggesting consistent identification of core functions between stakeholder groups. This consensus on core functions of the public health nutrition workforce in Europe can be used to promote a consistent understanding of the role and value of public health nutritionists as a discrete disciplinary sub-specialty of the public health workforce. The convergence of opinions of academics and employers, as well as comparison with previous international studies, indicates that there is a set of core public health nutrition functions transferable between countries that can be used as a benchmark to guide further development of the public health nutrition workforce in Europe.


Characterization of antibody response in patients with acute and chronic chikungunya virus disease.
Chikungunya virus (CHIKV) is a re-emerging arbovirus capable of causing chronic arthralgia, which can last for months to years. Although neutralizing antibodies have been shown to be important for viral clearance, is it not clear whether the quantitative and qualitative nature of antibodies play a role in progression to chronic disease. To characterize and compare the antibody responses in acute and chronic patients in a prospective observational CHIKV study in Curaçao during the 2014-2015 outbreak. We performed virus neutralization tests and ELISA on plasma samples collected from a prospective observational chikungunya study in Curaçao to compare the complement-dependent and -independent neutralization capacity, as well as the antibody avidity index of acute and chronic patients. We found that there was no significant difference in the virus neutralization titers between patients with acute and chronic chikungunya infection. Furthermore, we found that complement increased the neutralization capacity when large amounts of virus was used. Moreover, we found that patients with acute chikungunya disease had a significantly higher antibody avidity index compared to those with chronic disease. This study suggests that virus neutralization titers in late convalescent sera do not play a role in chronic chikungunya. However, the median antibody avidity was lower in these patients and may therefore suggest a role for antibody avidity in the development of chronic disease.


Microtensile dentin bond strength of self-etching and single-bottle adhesive systems in different cavity configurations.
To evaluate the bond strength of a self-etching (Clearfil SE Bond) and a single-bottle (Excite) adhesive system using two cavity configurations (C-factors 5 and 1). Class I cavities (3 x 4 x 2.5 mm) were prepared in 28 extracted human molars using diamond burs under water cooling. Teeth were divided into 4 groups: G1: Excite, C-factor 5; G2: Excite, C-factor 1; G3: Clearfil, C-factor 5; G4: Clearfil, C-factor 1. To determine C-factor 5, systems were applied to all cavity walls according to the manufacturers' instructions (5 bonded, 1 unbonded). For C-factor 1, lateral walls were isolated using nail varnish, and adhesive systems were only applied to the pulpal floor (5 unbonded, 1 bonded). Cavities were restored using Tetric Ceram composite resin (Ivoclar/Vivadent), and bulk light cured for 40 s (500 mW/cm2). Teeth were stored in distilled water for 24 h at 37 degrees C, and then sectioned using a diamond disk, yielding stick-shaped specimens with a bonded area of approximately 0.8 mm2. Specimens were submitted to the microtensile bond test at a rate of 1 mm/min speed in a universal testing machine. Resin-dentin bond strengths (MPa) were submitted to ANOVA and Tukey's tests (p < 0.05) [number of specimens]: G1 = 35.8b [43]; G2 = 48.9a [55]; G3 = 45.9a [60]; G4 = 49.0a [53]. There was no statistically significant difference between adhesive systems for C-factor 1. For C-factor 5, Clearfil SE Bond produced higher values when compared to the other group. Changes in C-factor only affected the total-etch adhesive system tested. This may be a result of the different filler volume in the self-etching system, and not of the bonding technique itself.


Character evolution and missing (morphological) data across Asteridae.
Our current understanding of flowering plant phylogeny provides an excellent framework for exploring various aspects of character evolution through comparative analyses. However, attempts to synthesize this phylogenetic framework with extensive morphological data sets have been surprisingly rare. Here, we explore character evolution in Asteridae (asterids), a major angiosperm clade, using an extensive morphological data set and a well-resolved phylogeny. We scored 15 phenotypic characters (spanning chemistry, vegetative anatomy, and floral, fruit, and seed features) across 248 species for ancestral state reconstruction using a phylogenetic framework based on 73 plastid genes and the same 248 species. Iridoid production, unitegmic ovules, and cellular endosperm were all reconstructed as synapomorphic for Asteridae. Sympetaly, long associated with asterids, shows complex patterns of evolution, suggesting it arose several times independently within the clade. Stamens equal in number to the petals is likely a synapomorphy for Gentianidae, a major asterid subclade. Members of Lamianae, a major gentianid subclade, are potentially diagnosed by adnate stamens, unilacunar nodes, and simple perforation plates. The analyses presented here provide a greatly improved understanding of character evolution across Asteridae, highlighting multiple characters potentially synapomorphic for major clades. However, several important parts of the asterid tree are poorly known for several key phenotypic features (e.g., degree of petal fusion, integument number, nucellus type, endosperm type, iridoid production). Further morphological, anatomical, developmental, and chemical investigations of these poorly known asterids are critical for a more detailed understanding of early asterid evolution.


Ribosome biogenesis in the yeast Saccharomyces cerevisiae.
Ribosomes are highly conserved ribonucleoprotein nanomachines that translate information in the genome to create the proteome in all cells. In yeast these complex particles contain four RNAs (>5400 nucleotides) and 79 different proteins. During the past 25 years, studies in yeast have led the way to understanding how these molecules are assembled into ribosomes in vivo. Assembly begins with transcription of ribosomal RNA in the nucleolus, where the RNA then undergoes complex pathways of folding, coupled with nucleotide modification, removal of spacer sequences, and binding to ribosomal proteins. More than 200 assembly factors and 76 small nucleolar RNAs transiently associate with assembling ribosomes, to enable their accurate and efficient construction. Following export of preribosomes from the nucleus to the cytoplasm, they undergo final stages of maturation before entering the pool of functioning ribosomes. Elaborate mechanisms exist to monitor the formation of correct structural and functional neighborhoods within ribosomes and to destroy preribosomes that fail to assemble properly. Studies of yeast ribosome biogenesis provide useful models for ribosomopathies, diseases in humans that result from failure to properly assemble ribosomes.


Influence of cycle frequency, muscle strain and muscle length on work and power production of rainbow trout (Oncorhynchus mykiss) ventricular muscle
This study investigates the effects of cycle frequency, strain and length on work and power output of isolated rainbow trout (Oncorhynchus mykiss) ventricular preparations using the work loop technique. These effects are discussed in the context of the whole heart using analogies with heart rate, stroke volume and end-diastolic volume. Power output was dependent on cycle frequency, increasing threefold beween 0.3 and 1.1 Hz. The frequency for maximum power output was approximately 1.1 Hz, corresponding to the frequency for maximum power in perfused heart experiments. The length for maximum work production (Lopt) was found to be the same as the length for maximum isometric force production (Lmax). The decline in net work at lengths greater than Lopt/Lmax was attributed to an increase in passive work (the work done on an unstimulated muscle) or to hysteresis and to a large increase in lengthening work. The strain yielding maximum work decreased with increasing frequency. This is discussed in the context of the decline in stroke volume observed at increased heart rates in vivo. Muscle strain in intact hearts paced at 0.3 Hz was +/-11.9 % (23.8 % peak to peak), a value similar to the optimum strain at 0.3 Hz in vitro (+/-12 %).


Antiepileptic drug effects on subjective and objective cognition.
Cognitive impairment is one of the most common complaints for persons with epilepsy (PWE). These impairments are not only associated with seizures, but are also regularly reported as adverse effects of antiepileptic drugs (AEDs). Previous studies have examined cognitive effects of both AED monotherapy and polytherapy, yet there is limited research on these differences with respect to both subjective and objective cognition. The current study uses data from previous research conducted by the Centers for Disease Control and Prevention (CDC)-sponsored Managing Epilepsy Well (MEW) Network collaborative. We used three distinct archival datasets from the following: (1) the HOBSCOTCH efficacy trial at Dartmouth-Hitchcock Medical Center (HOB-1), (2) the multisite replication trial (HOB-2), and (3) epilepsy self-management research conducted at the NYU School of Medicine. This retrospective analysis combined baseline data from three datasets to determine how the number of AEDs and the type of AEDs were associated with subjective (patient-reported) and objective (examiner-assessed) cognition. Subjective cognition was captured using the cognitive subscale of the Quality of Life in Epilepsy Inventory (QOLIE-31) in all three datasets (n = 224), while objective cognition was measured using the Repeated Battery for the Assessment of Neuropsychological Status (RBANS) in the HOB-1 dataset (n = 65) and the Brief Test of Adult Cognition by Telephone (BTACT) in the HOB-2 dataset (n = 91). Multivariable linear regression was utilized for our initial assessments, followed by propensity score matching to provide stronger control of covariates. Matching was based on significantly different covariates, such as education, depression, and history of prior epilepsy surgery. Nonparametric statistical tests were utilized to compare these matched subjects. Subjective cognitive impairment was significantly worse among individuals on polytherapy (2 + AEDs) compared with those on monotherapy (1 AED) (adjusted p = 0.041). These findings were consistent with our propensity score matched comparison of monotherapy and polytherapy, which indicated that polytherapy was associated with worse overall subjective cognition (adjusted p = 0.01), in addition to impairments on the RBANS (Total score p = 0.05) and specific subdomains of the BTACT (Episodic Verbal Memory p < 0.01, Working Memory p < 0.01, Processing Speed p < 0.01). Interestingly, older generation AEDs were associated with better language performance than newer generation and combined generation AED therapy (RBANS Language p = 0.03). These language-specific findings remained significant after controlling for the effects of topiramate and zonisamide (p = 0.04). A greater number of AEDs is significantly and negatively associated with subjective and objective cognition in PWE, and is in line with previous research. Antiepileptic drug type did not, in itself, appear to be associated with subjective cognition. Our findings suggest that ineffective AEDs should be replaced, rather than introducing additional AEDs to a treatment regimen. Further, while subjective and objective cognition assessments were both sensitive at detecting differences based on AED status, the neuropsychological objective subdomains offer additional and specific insights into how cognition is impaired with AEDs.


Venous Thromboembolism in Hip Fracture Patients: A Subanalysis of the FAITH and HEALTH Trials.
The primary objective of this study was to determine the incidence of symptomatic venous thromboembolism (VTE), including pulmonary embolism (PE) and deep vein thrombosis (DVT), in the hip fracture population. Secondary objectives included determining timing of VTE diagnosis, VTE thromboprophylaxis given, and identifying any factors associated with VTE. Using data from the FAITH and HEALTH trials, the incidence of VTE, including DVT and PE, and the timing of VTE were determined. A multivariable Cox regression analysis was used to determine which factors were associated with increased risk of VTE, including age, treatment for comorbidity, thromboprophylaxis, time to surgery, and method of fracture management. 2520 hip fracture patients were included in the analysis. Sixty-four patients (2.5%) had a VTE [DVT: 36 (1.4%), PE: 28 (1.1%)]. Thirty-five (54.7%) were diagnosed less than 6 weeks postfracture and 29 (45.3%) more than 6 weeks postfracture. One thousand nine hundred ninety-three (79%) patients received thromboprophylaxis preoperatively and 2502 (99%) received thromboprophylaxis postoperatively. The most common method of preoperative (46%) and postoperative (73%) thromboprophylaxis was low molecular weight heparin. Treatment with arthroplasty compared to internal fixation was the only variable associated with increased risk of VTE (hazard ratio 2.67, P = 0.02). The incidence of symptomatic VTE in hip fracture patients recruited to the 2 trials was 2.5%. Although over half of the cases were diagnosed within 6 weeks of fracture, VTE is still prevalent after this period. The majority of patients received thromboprophylaxis. Treatment with arthroplasty rather than fixation was associated with increased incidence of VTE. Therapeutic Level II. See Instructions for Authors for a complete description of levels of evidence.


Single Patient Classifier Assay, Microsatellite Instability, and Epstein-Barr Virus Status Predict Clinical Outcomes in Stage II/III Gastric Cancer: Results from CLASSIC Trial.
Clinical implications of single patient classifier (SPC) and microsatellite instability (MSI) in stage II/III gastric cancer have been reported. We investigated SPC and the status of MSI and Epstein-Barr virus (EBV) as combinatory biomarkers to predict the prognosis and responsiveness of adjuvant chemotherapy for stage II/III gastric cancer. Tumor specimens and clinical information were collected from patients enrolled in CLASSIC trial, a randomized controlled study of capecitabine plus oxaliplatin-based adjuvant chemotherapy. The results of nine-gene based SPC assay were classified as prognostication (SPC-prognosis) and prediction of chemotherapy benefit (SPC-prediction). Five quasimonomorphic mononucleotide markers were used to assess tumor MSI status. EBV-encoded small RNA in situ hybridization was performed to define EBV status. There were positive associations among SPC, MSI, and EBV statuses among 586 patients. In multivariate analysis of disease-free survival, SPC-prognosis [hazard ratio (HR): 1.879 (1.101-3.205), 2.399 (1.415-4.067), p=0.003] and MSI status (HR: 0.363, 95% confidence interval: 0.161-0.820, p=0.015) were independent prognostic factors along with age, Lauren classification, TNM stage, and chemotherapy. Patient survival of SPC-prognosis was well stratified regardless of EBV status and in microsatellite stable (MSS) group, but not in MSI-high group. Significant survival benefit from adjuvant chemotherapy was observed by SPC-Prediction in MSS and EBV-negative gastric cancer. SPC, MSI, and EBV statuses could be used in combination to predict the prognosis and responsiveness of adjuvant chemotherapy for stage II/III gastric cancer.


Effect of Guest Atom Composition on the Structural and Vibrational Properties of the Type II Clathrate-Based Materials AxSi136, AxGe136 and AxSn136 (A = Na, K, Rb, Cs; 0 ≤ x ≤ 24).
Type II clathrates are interesting due to their potential thermoelectric applications. Powdered X-ray diffraction (XRD) data and density functional calculations for NaxSi136 found a lattice contraction as x increases for 0 < x < 8 and an expansion as x increases for x > 8. This is explained by XRD data that shows that as x increases, the Si28 cages are filled first for x < 8 and the Si20 cages are then filled for x > 8. Motivated by this work, here we report the results of first-principles calculations of the structural and vibrational properties of the Type II clathrate compounds AxSi136, AxGe136, and AxSn136. We present results for the variation of the lattice constants, bulk moduli, and other structural parameters with x. These are contrasted for the Si, Ge, and Sn compounds and for guests A = Na, K, Rb, and Cs. We also present calculated results of phonon dispersion relations for Na₄Si136, Na₄Ge136, and Na₄Sn136 and we compare these for the three materials. Finally, we present calculated results for the elastic constants in NaxSi136, NaxGe136, and NaxSn136 for x = 4 and 8. These are compared for the three hosts, as well as for the two compositions.


Liver-specific pRB loss results in ectopic cell cycle entry and aberrant ploidy.
The liver exhibits an exquisitely controlled cell cycle, wherein hepatocytes are maintained in quiescence until stimulated to proliferate. The retinoblastoma tumor suppressor, pRB, plays a central role in proliferative control by inhibiting inappropriate cell cycle entry. In many cases, liver cancer arises due to aberrant cycles of proliferation, and correspondingly, pRB is functionally inactivated in the majority of hepatocellular carcinomas. Therefore, to determine how pRB loss may provide conditions permissive for deregulated hepatocyte proliferation, we investigated the consequence of somatic pRB inactivation in murine liver. We show that liver-specific pRB loss results in E2F target gene deregulation and elevated cell cycle progression during post-natal growth. However, in adult livers, E2F targets are repressed and hepatocytes become quiescent independent of pRB, suggesting that other factors may compensate for pRB loss. Therefore, to probe the consequences of acute pRB inactivation in livers of adult mice, we gave adenoviral-Cre by i.v. injection. We show that acute pRB loss is sufficient to elicit E2F target gene expression and cell cycle entry in adult liver, demonstrating a critical role for pRB in maintaining hepatocyte quiescence. Finally, we show that liver-specific pRB loss results in the development of nuclear pleomorphism associated with elevated ploidy that is evident in adult mice harboring both acute and chronic pRB loss. Together, these results show the crucial role played by pRB in maintaining hepatocyte quiescence and ploidy in adult liver in vivo and underscore the critical importance of delineating the consequences of acute pRB loss in adult animals.


Targeted DNP for biomolecular solid-state NMR.
High-field dynamic nuclear polarization is revolutionizing the scope of solid-state NMR with new applications in surface chemistry, materials science and structural biology. In this perspective article, we focus on a specific DNP approach, called targeted DNP, in which the paramagnets introduced to polarize are not uniformly distributed in the sample but site-specifically located on the biomolecular system. After reviewing the various targeting strategies reported to date, including a bio-orthogonal chemistry-based approach, we discuss the potential of targeted DNP to improve the overall NMR sensitivity while avoiding the use of glass-forming DNP matrix. This is especially relevant to the study of diluted biomolecular systems such as, for instance, membrane proteins within their lipidic environment. We also discuss routes towards extracting structural information from paramagnetic relaxation enhancement (PRE) induced by targeted DNP at cryogenic temperature, and the possibility to recover site-specific information in the vicinity of the paramagnetic moieties using high-resolution selective DNP spectra. Finally, we review the potential of targeted DNP for in-cell NMR studies and how it can be used to extract a given protein NMR signal from a complex cellular background.


Dietary resveratrol prevents the development of food allergy in mice.
Resveratrol is a bioactive polyphenol enriched in red wine that exhibits many beneficial health effects via multiple mechanisms. However, it is unclear whether resveratrol is beneficial for the prevention of food allergy. This study investigated whether resveratrol inhibited the development of food allergy by using a mouse model of the disease. Mice fed standard diet or standard diet plus resveratrol were sensitized by intragastric administration of ovalbumin (OVA) and mucosal adjuvant cholera toxin (CT). Several manifestations of food allergy were then compared between the mice. The effects of resveratrol on T cells or dendritic cells were also examined by using splenocytes from OVA-specific T cell-receptor (TCR) transgenic DO11.10 mice or mouse bone marrow-derived dendritic cells (BMDCs) in vitro. We found that mice fed resveratrol showed reduced OVA-specific serum IgE production, anaphylactic reaction, and OVA-induced IL-13 and IFN-ã production from the mesenteric lymph nodes (MLNs) and spleens in comparison to the control mice, following oral sensitization with OVA plus CT. In addition, resveratrol inhibited OVA plus CT-induced IL-4, IL-13, and IFN-ã production in splenocytes from DO11.10 mice associated with inhibition of GATA-3 and T-bet expression. Furthermore, resveratrol suppressed the OVA plus CT-induced CD25 expression and IL-2 production in DO11.10 mice-splenocytes in association with decreases in CD80 and CD86 expression levels. Finally, resveratrol suppressed CT-induced cAMP elevation in association with decreases in CD80 and CD86 expression levels in BMDCs. Ingestion of resveratrol prevented the development of a food allergy model in mice. Given the in vitro findings, resveratrol might do so by inhibiting DC maturation and subsequent early T cell activation and differentiation via downregulation of CT-induced cAMP activation in mice. These results suggest that resveratrol may have potential for prophylaxis against food allergy.


Digenetic trematodes in South American sea lions from southern Brazilian waters.
The aim of this work was to perform a systematic study to detect and quantify the digenetic trematode infections in South American sea lions from the southern Brazilian coast. Twenty-four South American sea lions, Otaria flavescens (Carnivora: Otaridae), were found dead along the coast of Rio Grande do Sul State, Brazil, between June 2010 and September of 2011. Two trematode species were found in the intestines of O. flavescens, i.e., Stephanoprora uruguayense (Digenea: Echinostomatidae) and Ascocotyle (Phagicola) longa (Digenea: Heterophyidae). Ascocotyle (P.) longa reached a prevalence of 33.3% and mean intensity of 248,500, whereas S. uruguayense showed a prevalence of 4.2% and mean intensity of 202. The 2 trematode species infecting sea lions were likely transmitted by feeding on mullets, Mugil platanus, that commonly harbor heterophyid metacercariae. The present work is the first report of digenetic trematodes infecting O. flavescens in Brazil. The high prevalence and mean intensity values of the 2 trematode species infecting sea lions in the present study suggest caution in human consumption of mullets and other fish, which can be infected with the metacercariae of these trematodes known to have zoonotic potential.


Impact of Mycobacterium tuberculosis RD1-locus on human primary dendritic cell immune functions.
Modern strategies to develop vaccines against Mycobacterium tuberculosis (Mtb) aim to improve the current Bacillus Calmette-Guerin (BCG) vaccine or to attenuate the virulence of Mtb vaccine candidates. In the present study, the impact of wild type or mutated region of difference 1 (RD1) variants on the immunogenicity of Mtb and BCG recombinants was investigated in human primary dendritic cells (DC). A comparative analysis of transcriptome, signalling pathway activation, maturation, apoptosis, cytokine production and capacity to promote Th1 responses demonstrated that DC sense quantitative and qualitative differences in the expression of RD1-encoded factors--ESAT6 and CFP10--within BCG or Mtb backgrounds. Expansion of IFN-γ producing T cells was promoted by BCG::RD1-challenged DC, as compared to their BCG-infected counterparts. Although Mtb recombinants acted as a strong Th-1 promoting stimulus, even with RD1 deletion, the attenuated Mtb strain carrying a C-terminus truncated ESAT-6 elicited a robust Th1 promoting phenotype in DC. Collectively, these studies indicate a necessary but not sufficient role for the RD1 locus in promoting DC immune-regulatory functions. Additional mycobacterial factors are likely required to endow DC with a high Th1 polarizing capacity, a desirable attribute for a successful control of Mtb infection.


Na,K-ATPase polypeptide upregulation responses in lens epithelium.
In a previous study, an increase in Na,K-ATPase alpha 2 expression was detected in the epithelium of porcine lenses exposed to amphotericin B, an ionophore that also increases lens sodium and stimulates active sodium transport. The purpose of the present study was to determine whether an increase of Na,K-ATPase alpha 2 synthesis is a response to an episode of rapid Na-K transport or whether the increase in lens sodium alone can initiate the response. Western blot analyses were conducted to probe for Na,K-ATPase alpha polypeptides in membrane material isolated from porcine lens epithelium. Ouabain-sensitive adenosine triphosphate hydrolysis was used as an index of Na,K-ATPase activity, and lens ion content was determined by atomic absorption spectrophotometry. 86-Rubidium (86Rb) uptake was measured as an indicator for active potassium transport. 86Rb uptake was markedly diminished in lenses exposed to dihydro-ouabain (DHO), signifying inhibition of active sodium-potassium transport. Consistent with this, the sodium content of DHO-treated lenses increased. By western blot analysis, a marked increase of Na,K-ATPase alpha 2 polypeptide could be detected in the epithelium of DHO-treated lenses. To rule out the possibility that apparent stimulation of Na,K-ATPase alpha 2 synthesis stemmed from binding of DHO to Na,K-ATPase sites, experiments were conducted to confirm an increase of Na,K-ATPase alpha 2 polypeptide in the epithelium of lenses exposed to low-potassium medium to inhibit active sodium-potassium transport. Consistent with the apparent increase of Na,K-ATPase polypeptide, Na,K-ATPase activity was detectably increased in epithelial material isolated from lenses pretreated with DHO or low-potassium medium. An increase in Na,K-ATPase alpha 2 polypeptide can occur in the epithelium of lenses subjected to an episode of sodium pump inhibition. This suggests the response could be triggered by an increase in cell sodium and does not necessarily require a period of stimulated active sodium-potassium transport.


Inductive interactions between human dermis and chick chorionic epithelium.
In a study of specificity in mesenchymal-epithelial interactions, human embryonic dermis has been recombined with chick chorionic epithelium and cultured for 7 days on a host chick chorioallantoic membrane. Dermis from the sole of the foot or palm of the hand induces chick chorionic epithelium to form an epidermis that resembles chick rather than human epidermis. Chick epithelium, though it has the capacity to respond to a human dermal stimulus, is limited to forming chick-type tissue. The human dermis was modified in its turn by culture in combination with chick epithelium.


Three-dimensional structures and properties of a transforming and a nontransforming glycine-12 mutant of p21H-ras.
The three-dimensional structures and biochemical properties of two mutants of the G-domain (residues 1-166) of p21H-ras, p21 (G12D) and p21 (G12P), have been determined in the triphosphate-bound form using guanosine 5'-(beta,gamma-imido)triphosphate (GppNHp). They correspond to the most frequent oncogenic and the only nononcogenic mutation of Gly-12, respectively. The G12D mutation is the only mutant analyzed so far that crystallizes in a space group different from wild type, and the atomic model of the protein shows the most drastic changes of structure around the active site as compared to wild-type p21. This is due to the interactions of the aspartic acid side chain with Tyr-32, Gln-61, and the gamma-phosphate, which result in reduced mobility of these structural elements. The interaction between the carboxylate group of Asp-12 and the gamma-phosphate is mediated by a shared proton, which we show by 31P NMR measurements to exist in solution as well. The structure of p21 (G12P) is remarkably similar to that of wild-type p21 in the active site, including the position of the nucleophilic water. The pyrrolidine ring of Pro-12 points outward and seems to be responsible for the weaker affinity toward GAP (GTPase-activating protein) and the failure of GAP to stimulate GTP hydrolysis.


Developmental growth trajectories in understanding of fraction magnitude from fourth through sixth grade.
Development of fraction number line estimation was assessed longitudinally over 5 time points between 4th and 6th grades. Although students showed positive linear growth overall, latent class growth analyses revealed 3 distinct growth trajectory classes: Students who were highly accurate from the start and became even more accurate (n = 154); students who started inaccurate but showed steep growth (n = 121); and students who started inaccurate and showed minimal growth (n = 197). Younger and minimal growth students typically estimated both proper and improper fractions as being less than 1, failing to base estimates on the relation between the numerator and denominator. Class membership was highly predictive of performance on a statewide-standardized mathematics test as well as on a general fraction knowledge measure at the end of 6th grade, even after controlling for mathematic-specific abilities, domain-general cognitive abilities, and demographic variables. Multiplication fluency, classroom attention, and whole number line estimation acuity at the start of the study predicted class membership. The findings reveal that fraction magnitude understanding is central to mathematical development. (PsycINFO Database Record


Revising Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, criteria for the bipolar disorders: Phase I of the AREDOC project.
To derive new criteria sets for defining manic and hypomanic episodes (and thus for defining the bipolar I and II disorders), an international Task Force was assembled and termed AREDOC reflecting its role of Assessment, Revision and Evaluation of DSM and other Operational Criteria. This paper reports on the first phase of its deliberations and interim criteria recommendations. The first stage of the process consisted of reviewing Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, and recent International Classification of Diseases criteria, identifying their limitations and generating modified criteria sets for further in-depth consideration. Task Force members responded to recommendations for modifying criteria and from these the most problematic issues were identified. Principal issues focussed on by Task Force members were how best to differentiate mania and hypomania, how to judge 'impairment' (both in and of itself and allowing that functioning may sometimes improve during hypomanic episodes) and concern that rejecting some criteria (e.g. an imposed duration period) might risk false-positive diagnoses of the bipolar disorders. This first-stage report summarises the clinical opinions of international experts in the diagnosis and management of the bipolar disorders, allowing readers to contemplate diagnostic parameters that may influence their clinical decisions. The findings meaningfully inform subsequent Task Force stages (involving a further commentary stage followed by an empirical study) that are expected to generate improved symptom criteria for diagnosing the bipolar I and II disorders with greater precision and to clarify whether they differ dimensionally or categorically.


Plasmodium falciparum: a family of sulphated glycoconjugates disrupts erythrocyte rosettes.
The ability of Plasmodium falciparum-infected erythrocytes to form spontaneous rosettes with uninfected red cells is a parasite adhesion property which has been associated with severe malaria. The mechanism of rosetting remains unknown, but the ability of heparin to disrupt rosettes has been recognised previously. In this paper we show that a group of sulphated glycoconjugates including sulphatide, dextran sulphate, and fucoidan are more effective rosette reversing agents than heparin and are active against both laboratory strains and wild isolates. Other related anionic glycosaminoglycans such as the chondroitin sulphates A, B, and C and hyaluronic acid have no effect on rosette formation. This family of sulphated glycoconjugates which are active against rosettes is also known to inhibit sporozoite invasion of hepatocytes and merozoite reinvasion of erythrocytes, suggesting that sulphated glycoconjugate interaction may be an important process in cell adhesion at different stages in the plasmodial life cycle.


Downregulation of hepatic multi-drug resistance protein 1 (MDR1) after copper exposure.
Copper homeostasis is strictly regulated in mammalian cells. We investigated the adaptation of hepatocytes after long-term copper exposure. Copper-resistant hepatoma HepG2 cell lines lacking ATP7B were generated. Growth, copper accumulation, gene expression, and transport were determined. Hepatocyte-like cells derived from a Wilson disease (WD) patient and the liver of a WD animal model were also studied. The rapidly gained copper resistance was found to be stable, as subculturing of cells in the absence of added copper (weaning) did not restore copper sensitivity. Intracellular copper levels and the expression of MT1 and HSP70 were increased, whereas the expression of CTR1 was reduced. However, the values normalized after weaning. In contrast, downregulation of multi-drug resistance protein 1 (MDR1), encoding P-glycoprotein (P-gp), was shown to be permanent. Calcein assays confirmed the downregulation of MDR1 in the resistant cell lines. MDR1 knockdown by siRNA resulted in increased copper resistance and decreased intracellular copper. Treatment of the resistant cells with verapamil, a known inducer of MDR1, was followed by increased copper-induced toxicity. Downregulation of MDR1 was also observed in hepatocyte-like cells derived from a WD patient after copper exposure. In addition, MDR1 was downregulated in Long-Evans Cinnamon rats when the liver copper was elevated. The results indicate that downregulation of MDR1 is an adaptation of hepatic cells after sustained copper exposure when ATP7B is non-functional. Our data add to the versatile functions of MDR1 in the hepatocyte and may have an impact on the treatment of copper-related diseases, prominently WD.


Temperature, temperature extremes, and mortality: a study of acclimatisation and effect modification in 50 US cities.
The authors examined the increase in mortality associated with hot and cold temperature in different locations, the determinants of the variability in effect estimates, and its implications for adaptation. The authors conducted a case-crossover study in 50 US cities. They used daily mortality and weather data for 6 513 330 deaths occurring during 1989-2000. Exposure was assessed using two approaches. First, the authors determined exposure to extreme temperatures using city-specific indicator variables based on the local temperature distribution. Secondly, they used piecewise linear variables to assess exposure to temperature on a continuous scale above/below a threshold. Effects of hot and cold temperature were examined in season-specific models. In a meta-analysis of the city-specific results, the authors examined several city characteristics as effect modifiers. Mortality increases associated with both extreme cold (2-day cumulative increase 1.59% (95% CI 0.56 to 2.63)) and extreme heat (5.74% (95% CI 3.38 to 8.15)) were found, the former being especially marked for myocardial infarction and cardiac arrest deaths. The increase in mortality was less marked at less extreme temperatures. The effect of extreme cold (defined as a percentile) was homogeneous across cities with different climates, suggesting that only the unusualness of the cold temperature (and not its absolute value) had a substantial impact on mortality (that is, acclimatisation to cold). Conversely, heat effects were quite heterogeneous, with the largest effects observed in cities with milder summers, less air conditioning and higher population density. Adjustment for ozone led to similar results, but some residual confounding could be present due to other uncontrolled pollutants. The authors confirmed in a large sample of cities that both cold and hot temperatures increase mortality risk. These findings suggest that increases in heat-related mortality due to global warming are unlikely to be compensated for by decreases in cold-related mortality and that population acclimatisation to heat is still incomplete.


Comparison of outlier identification methods in hospital surgical quality improvement programs.
Surgeons and hospitals are being increasingly assessed by third parties regarding surgical quality and outcomes, and much of this information is reported publicly. Our objective was to compare various methods used to classify hospitals as outliers in established surgical quality assessment programs by applying each approach to a single data set. Using American College of Surgeons National Surgical Quality Improvement Program data (7/2008-6/2009), hospital risk-adjusted 30-day morbidity and mortality were assessed for general surgery at 231 hospitals (cases = 217,630) and for colorectal surgery at 109 hospitals (cases = 17,251). The number of outliers (poor performers) identified using different methods and criteria were compared. The overall morbidity was 10.3% for general surgery and 25.3% for colorectal surgery. The mortality was 1.6% for general surgery and 4.0% for colorectal surgery. Programs used different methods (logistic regression, hierarchical modeling, partitioning) and criteria (P < 0.01, P < 0.05, P < 0.10) to identify outliers. Depending on outlier identification methods and criteria employed, when each approach was applied to this single dataset, the number of outliers ranged from 7 to 57 hospitals for general surgery morbidity, 1 to 57 hospitals for general surgery mortality, 4 to 27 hospitals for colorectal morbidity, and 0 to 27 hospitals for colorectal mortality. There was considerable variation in the number of outliers identified using different detection approaches. Quality programs seem to be utilizing outlier identification methods contrary to what might be expected, thus they should justify their methodology based on the intent of the program (i.e., quality improvement vs. reimbursement). Surgeons and hospitals should be aware of variability in methods used to assess their performance as these outlier designations will likely have referral and reimbursement consequences.


Optimal vascular delay in cardiomyoplasty following latissimus dorsi muscle isolation in dogs.
Distal ischemia of the latissimus dorsi muscle (LDM) during cardiomyoplasty is a recognized complication that can reduce the muscle's function and mechanical effects. A 2-week vascular recovery period is recommended to allow revascularization and adhesion to the heart. It is not clear, however, that a 2-week vascular delay is optimal after LDM isolation. This study was designed to evaluate both the regional blood flow (RBF) in canine LDM flaps immediately after perforators were ligated, and the effect of 1-, 2-, and 3-week vascular delays on the regional perfusion of LDM flaps without electrical stimulation. A catheter-access device connected to the left atrium was implanted in the left back subcutaneous layer in each of six adult mongrel dogs when the LDM was dissected. Five different colored microspheres were injected at five different time points: pre- and post-dissection, and after 1-, 2-, and 3-week vascular delays. At each time point, reference blood was withdrawn from the femoral artery at a fixed speed. The LDMs were removed post mortem for RBF determination. Both tissue and reference blood samples were spectrophotometrically processed to quantify the amount of dye. Proximal RBF did not decrease immediately after dissection when compared to the control (0.28 +/- 0.10 vs 0.26 +/- 0.05 mL.g-1.min-1, p > 0.05), but it decreased after a 1-week vascular delay (0.11 +/- 0.02 vs 0.26 +/- 0.05 mL.g-1.min-1, p < 0.01), and then returned to normal after a 3-week vascular delay (0.21 +/- 0.06 vs 0.26 +/- 0.05 mL.g-1.min-1, p > 0.05). RBF decreased immediately after dissection in the middle and distal segments, and did not return to the control value even after a 3-week vascular delay. RBF in the LDM was depressed by surgical dissection in this canine model. Without electrical stimulation, the middle and distal portions of the LDM remained compromised even after a 3-week vascular delay. The clinical effects of progressive programmed electrical stimulation of the LDM at different periods of vascular delay remain to be studied.


What is new in electronic-cigarettes research?
Electronic cigarettes (e-cigarettes) and other vaping devices have seen extraordinary growth in use in the past 10 years, and companies are accelerating their development of new products and marketing efforts. In turn, researchers have increased their efforts to determine how e-cigarettes affect health, how marketing these products impacts adolescents and how the use of e-cigarettes may affect adolescents' use of other tobacco products. Products like Juul were not on the market 2 years ago; thus, frequent updates on the topic are essential. Studies have begun to demonstrate that users of the newer pod systems are exposed to high levels of nicotine, as well as other chemicals. These products are highly marketed, with a strong emphasis on how adolescents can use them surreptitiously. This is concerning to researchers, and other studies have continued to demonstrate that teens who use e-cigarettes are more likely to go on to use combusted tobacco. Further research has also failed to demonstrate that e-cigarettes are useful for people wishing to quit smoking combusted tobacco. E-cigarettes and vaping systems are not safe products and should not be used by adolescents.


Ethnic/racial disparities in hospital procedure volume for lung resection for lung cancer.
Ethnic/racial minorities experience poorer outcomes from lung cancer than non-Hispanic whites. Higher hospital procedure volume is associated with better survival from lung resection for lung cancer. We examined whether (1) ethnic/racial minorities are more likely to obtain lung resections at lower volume hospitals, (2) ethnicity/race is associated with inpatient mortality, (3) hospital volume mediates this association, and (4) hospital selection is mediated by racial/ethnic segregation, differences in insurance coverage, or limited hospital choice. Six years of data from the Nationwide Inpatient Sample (NIS 1998-2003, unweighted n = 50,245, weighted n = 129,506) were used in multivariate models controlling for sociodemographic factors, case complexity, and hospital characteristics. Additional analyses were conducted using the Area Resource File, which provided data on ethnic density and number of surgical hospitals in the hospital region. Blacks/African Americans (odds ratio [OR] = 0.45; 0.34-0.58) and Latinos (OR = 0.44; 0.32-0.63) had lower odds of obtaining lung resection at a high-volume hospital than non-Hispanic whites. Blacks/African Americans (OR = 1.30; 1.01-1.67), Latinos (OR = 1.41; 1.02-1.94), and other racial/ethnic minorities (OR = 1.46; 1.04-2.06) also had higher odds of dying in hospital, but this association was statistically nonsignificant after controlling for hospital volume. Hospital location was not associated with lung resection procedure volume, nor did location mediate the association between ethnicity/race and hospital volume. Ethnic/racial minorities are obtaining lung resection in lower volume hospitals and are more likely to die in hospital. Hospital volume is associated with higher mortality, but health insurance, segregation, and number of surgical hospitals within a county do not account for observed disparities.


Effects of IL-4 and IL-13 on total and allergen specific IgE production by cultured PBMC from allergic patients determined with recombinant pollen allergens.
Interleukin (IL)-4 and IL-13 have been shown to be potent switch factors for IgE synthesis in human B cells. In this study we investigated the effects of recombinant human IL-4 and IL-13 on total and allergen specific IgE synthesis by peripheral blood mononuclear cells (PBMC) from pollen allergic patients and healthy control individuals. Peripheral blood mononuclear cells (PBMC) from allergic patients were investigated for their capacity to produce allergen specific IgE in vitro. Total protein extracts from birch pollen and timothy grass pollen as well as purified recombinant birch pollen allergens, Bet v I, birch profilin (Bet v II) and recombinant timothy grass pollen allergens, Phl p I, Phl p II, and Phl p V were used to measure specific IgE-antibody synthesis in cell culture supernatants by IgE-immunoblot and ELISA. PBMC obtained from allergic patients spontaneously secreted allergen specific IgE in the culture supernatants. Addition of Interleukin 4, Interleukin 13 and anti-CD40 antibody to the cultures alone or in combinations significantly induced total IgE production whereas allergen specific IgE production was not affected. Our results indicate that the peripheral blood of allergic individuals contains long lived allergen specific B cells which have already switched to IgE production and which are not sensitive to IL-4 and IL-13 treatment. These results may have implications on attempts to use cytokines or cytokine antagonists in therapy of Type I allergy.


The Rpd3 core complex is a chromatin stabilization module.
The S. cerevisiae Rpd3 large (Rpd3L) and small (Rpd3S) histone deacetylase (HDAC) complexes are prototypes for understanding transcriptional repression in eukaryotes [1]. The current view is that they function by deacetylating chromatin, thereby limiting accessibility of transcriptional factors to the underlying DNA. However, an Rpd3 catalytic mutant retains substantial repression capability when targeted to a promoter as a LexA fusion protein [2]. We investigated the HDAC-independent properties of the Rpd3 complexes biochemically and discovered a chaperone function, which promotes histone deposition onto DNA, and a novel activity, which prevents nucleosome eviction but not remodeling mediated by the ATP-dependent RSC complex. These HDAC-independent activities inhibit Pol II transcription on a nucleosomal template. The functions of the endogenous Rpd3 complexes can be recapitulated with recombinant Rpd3 core complex comprising Sin3, Rpd3, and Ume1. To test the hypothesis that Rpd3 contributes to chromatin stabilization in vivo, we measured histone H3 density genomewide and found that it was reduced at promoters in an Rpd3 deletion mutant but partially restored in a catalytic mutant. Importantly, the effects on H3 density are most apparent on RSC-enriched genes [3]. Our data suggest that the Rpd3 core complex could contribute to repression via a novel nucleosome stabilization function.


Lipocalin-2 Promotes Pancreatic Ductal Adenocarcinoma by Regulating Inflammation in the Tumor Microenvironment.
Lipocalin-2 (LCN2) promotes malignant development in many cancer types. LCN2 is upregulated in patients with pancreatic ductal adenocarcinoma (PDAC) and in obese individuals, but whether it contributes to PDAC development is unclear. In this study, we investigated the effects of Lcn2 depletion on diet-induced obesity, inflammation, and PDAC development. Mice with acinar cell-specific expression of KrasG12D were crossed with Lcn2-depleted animals and fed isocaloric diets with varying amounts of fat content. Pancreas were collected and analyzed for inflammation, pancreatic intraepithelial neoplasia (PanIN), and PDAC. We also used a syngeneic orthotopic PDAC mouse model to study tumor growth in the presence or absence of Lcn2 expression. In addition, to understand the mechanistic role of how LCN2 could be mediating PDAC, we studied LCN2 and its specific receptor solute carrier family 22 member 17 (SLC22A17) in human pancreatic cancer stellate cells (PSC), key mediators of the PDAC stroma. Depletion of Lcn2 diminished extracellular matrix deposition, immune cell infiltration, PanIN formation, and tumor growth. Notably, it also increased survival in both obesity-driven and syngeneic orthotopic PDAC mouse models. LCN2 modulated the secretion of proinflammatory cytokines in PSC of the PDAC tumor microenvironment, whereas downregulation of LCN2-specific receptor SLC22A17 blocked these effects. Our results reveal how LCN2 acts in the tumor microenvironment links obesity, inflammation, and PDAC development. Cancer Res; 77(10); 2647-60. ©2017 AACR.


Regularized multi-structural shape modeling of the knee complex based on deep functional maps.
The incorporation of a-priori knowledge on the shape of anatomical structures and their variation through Statistical Shape Models (SSMs) has shown to be very effective in guiding highly uncertain image segmentation problems. In this paper, we construct multiple-structure SSMs of purely geometric nature, that describe the relationship between adjacent anatomical components through Canonical Correlation Analysis. Shape inference is then conducted based on a regularization term on the shape likelihood providing more reliable structure representations. A fundamental prerequisite for performing statistical shape analysis on a set of objects is the identification of corresponding points on their associated surfaces. We address the correspondence problem using the recently proposed Functional Maps framework, which is a generalization of point-to-point correspondence to manifolds. Additionally, we show that, by incorporating techniques from the deep learning theory into this framework, we can further enhance the ability of SSMs to better capture the shape variation in a given dataset. The efficiency of our approach is illustrated through the creation of 3D models of the human knee complex in two application scenarios: incomplete or noisy shape reconstruction and missing structure estimation.


The effect of adjunctive armodafinil on cognitive performance and psychopathology in antipsychotic-treated patients with schizophrenia/schizoaffective disorder: a randomized, double-blind, placebo-controlled trial.
The efficacy, safety and tolerability of adjunctive armodafinil for cognitive performance, and negative and affective symptoms, were examined in 60 patients with schizophrenia or schizoaffective disorder. This was a 6-week, double-blind, placebo-controlled, fixed dose trial of armodafinil (150 mg/d) augmentation in patients with clinically stable schizophrenia or schizoaffective disorder. Cognition, psychopathology, alertness/wakefulness and adverse effects were assessed with standardized rating instruments. The primary endpoint was performance on measures of attention/vigilance. Patients were randomly allocated to adjunctive armodafinil or placebo. There was a significant Drug×Time interaction effect for attention/vigilance, due to modest non-significant worsening in the armodafinil group and improvement in the armodafinil group [CPT-Pairs d', F(1,40)=6.2, p=0.017]. However, it became non-significant after correction for multiple comparisons. There were no differences between armodafinil and placebo in other cognitive domains or psychopathology measures. However, armodafinil was associated with significant improvement in the Scale for the Assessment of Negative Symptoms (SANS) anhedonia-asociality [F(1,41)=4.1, p=0.05]. There were no significant differences in neurocognitive measures between adjunctive armodafinil and placebo in this 6-week study. Armodafinil improved anhedonia-asociality, but not other negative symptom domains.


Linking mass media campaigns to pictorial warning labels on cigarette packages: a cross-sectional study to evaluate effects among Mexican smokers.
This study assessed the effects of pictorial health warning labels (HWLs) and a linked media campaign in Mexico. Cross-sectional data were collected from a population-based sample of 1756 adult smokers, aged 18-55 years, during the initial implementation of pictorial HWLs, which some smokers had seen on cigarette packages while others had seen only the text-based HWLs. Exposure to the campaign and pictorial HWLs was assessed with aided recall methods, and other questions addressed attention and cognitive impact of HWLs, knowledge related to HWL and campaign content, and quit-related thoughts and behaviours. Logistic and linear regression models were estimated to determine associations between key outcomes and intervention exposure. In bivariate and multivariate adjusted models, recall of pictorial HWLs and of the campaign were positively associated with greater attention to and cognitive impact of HWLs, whereas only pictorial HWL exposure was associated with having refrained from smoking due to HWLs. Both recall of pictorial HWLs and of the campaign were independently associated with greater knowledge of secondhand smoke harms and toxic tobacco constituents. Smokers who recalled only the pictorial HWLs were more likely to try to quit than smokers who recalled neither the pictorial HWLs nor the campaign (17% vs 6%, p<0.001). Consistent with other studies, adult smokers' exposure to new pictorial HWLs in Mexico was associated with psychosocial and behavioural responses related to quit behaviour. Exposure to the complementary media campaign was associated with independent additive effects on campaign-related knowledge, and it enhanced psychosocial responses to pictorial HWLs.


Aortic pulse wave velocity measurement in systemic sclerosis patients.
Systemic sclerosis (SSc) is characterized by endothelial dysfunction and widespread microangiopathy. However, a macrovascular damage could be also associated. Aortic pulse wave velocity (aPWV) is known to be a reliable indicator of arterial stiffness and a useful prognostic predictor of cardiovascular events. Moreover, aPWV may be easily measured by non-invasive, user-friendly tool. Aim of our study was to evaluate aPWV alterations in a series of SSc patients. The aPWV was evaluated in 35 consecutive female SSc patients and 26 sex- and age-matched healthy controls. aPWV alterations were correlated with cardiopulmonary involvement. A significant increase of aPWV was observed in SSc patients compared to controls (9.4 ± 3.2 m/s vs 7.3 ± 1 m/s; P = 0.002). In particular, 14/35 (40%) SSc patients and only 1/26 (4%) controls (P=0.0009) showed increased aPWV (>9 m/s cut-off value). Moreover, echocardiography evaluation showed an increased prevalence of right atrial and ventricular dilatation (atrial volume: 23.6 ± 6.2 mL vs 20.3 ± 4.3 mL, P=0.026; ventricular diameter 19.5 ± 4.9 mm vs 15.9 ± 1.6 mm; P=0.001) associated to higher values of pulmonary arterial systolic pressure (PAPs) in SSc patients (31.5 ± 10.4 mmHg vs 21.6 ± 2.9 mmHg; P<0.0001; 40% of SSc patients showed an abnormal PAPs). Clinically, SSc patients presented a reduction of six-minute walking test (413 ± 96 m vs 491 ± 49 m; P=0.001), not correlated with pulmonary function tests. Increased aPWV values were evidenced only in SSc patients >50 years old. Furthermore, altered aPWV was more frequently associated with limited cutaneous pattern, longer disease duration (≥ 5 years), and/or presence of anticentromere antibody (ACA). A significantly higher prevalence of abnormally increased aPWV was evidenced in SSc patients compared to healthy controls. The possibility of more pronounced and diffuse vascular damage in a particular SSc subset (ACA-positive subjects with limited cutaneous scleroderma and longer disease duration) might be raised.


Crystal structure combined with genetic analysis of the Thermus thermophilus ribosome recycling factor shows that a flexible hinge may act as a functional switch.
Ribosome recycling factor (RRF), in concert with elongation factor EF-G, is required for disassembly of the posttermination complex of the ribosome after release of polypeptides. The crystal structure of Thermus thermophilus RRF was determined at 2.6 A resolution. It is a tRNA-like L-shaped molecule consisting of two domains: a long three-helix bundle (domain 1) and a three-layer beta/alpha/beta sandwich (domain 2). Although the individual domain structures are similar to those of Thermotoga maritima RRF (Selmer et al., Science, 1999, 286:2349-2352), the interdomain angle differs by 33 degrees in two molecules, suggesting that the hinge between two domains is potentially flexible and responsive to different conditions of crystal packing. The hinge connects hydrophobic junctions of domains 1 and 2. The structure-based genetic analysis revealed the strong correlation between the hinge flexibility and the in vivo function of RRF. First, altering the hinge flexibility by making alanine or serine substitutions for large-size residues conserved at the hinge loop and nearby in domain 1 frequently gave rise to gain of function except a Pro residue conserved at the hinge loop. Second, the hinge defect resulting from a too relaxed hinge structure can be compensated for by secondary alterations in domain 1 that seem to increase the hydrophobic contact between domain 1 and the hinge loop. These results show that the hinge flexibility is vital for the function of RRF and that the steric interaction between the hinge loop and domains 1 and 2 restricts the interdomain angle and/or the hinge flexibility. These results indicate that RRF possesses an architectural difference from tRNA regardless of a resemblance to tRNA shape: RRF has a "gooseneck" elbow, whereas the tRNA elbow is rigid, and the direction of flex of RRF and tRNA is at a nearly right angle to each other. Moreover, surface electrostatic potentials of the two RRF proteins are dissimilar and do not mimic the surface potential of tRNA or EF-G. These properties will add a new insight into RRF, suggesting that RRF is more than a simple tRNA mimic.


26Postoperative diagnosis and outcome in patients with revision arthroplasty for aseptic loosening.
The most common cause of implant failure is aseptic loosening (AL), followed by prosthetic joint infection (PJI). This study evaluates the incidence of PJI among patients operated with suspected AL and whether the diagnosis of PJI was predictive of subsequent implant failure including re-infection, at 2 years of follow up. Patients undergoing revision hip or knee arthroplasty due to presumed AL from February 2009 to September 2011 were prospectively evaluated. A sonication fluid of prosthesis and tissue samples for microbiology and histopathology at the time of the surgery were collected. Implant failure include recurrent or persistent infection, reoperation for any reason or need for chronic antibiotic suppression. Of 198 patients with pre-and intraoperative diagnosis of AL, 24 (12.1 %) had postoperative diagnosis of PJI. After a follow up of 31 months (IQR: 21 to 38 months), 9 (37.5 %) of 24 patients in the PJI group had implant failure compared to only 1 (1.1 %) in the 198 of AL group (p < 0.0001). Sensitivity of sonicate fluid culture (>20 CFU) and peri-prosthetic tissue culture were 87.5 % vs 66.7 %, respectively. Specificities were 100 % for both techniques (95 % CI, 97.9-100 %). A greater number of patients with PJI (79.1 %) had previous partial arthroplasty revisions than those patients in the AL group (56.9 %) (p = 0.04). In addition, 5 (55.5 %) patients with PJI and implant failure had more revision arthroplasties during the first year after the last implant placement than those patients with PJI without implant failure (1 patient; 6.7 %) (RR 3.8; 95 % CI 1.4-10.1; p = 0.015). On the other hand, 6 (25 %) patients finally diagnosed of PJI were initially diagnosed of AL in the first year after primary arthroplasty, whereas it was only 16 (9.2 %) patients in the group of true AL (RR 2.7; 95 % CI 1.2-6.1; p = 0.03). More than one tenth of patients with suspected AL are misdiagnosed PJI. Positive histology and positive peri-implant tissue and sonicate fluid cultures are highly predictive of implant failure in patients with PJI. Patients with greater number of partial hip revisions for a presumed AL had more risk of PJI. Early loosening is more often caused by hidden PJI than late loosening.


Rab10 is involved in basolateral transport in polarized Madin-Darby canine kidney cells.
The sorting of newly synthesized membrane proteins to the cell surface is an important mechanism of cell polarity. To identify more of the molecular machinery involved, we investigated the function of the small GTPase Rab10 in polarized epithelial Madin-Darby canine kidney cells. We find that GFP-tagged Rab10 localizes primarily to the Golgi during early cell polarization. Expression of an activated Rab10 mutant inhibits biosynthetic transport from the Golgi and missorts basolateral cargo to the apical membrane. Depletion of Rab10 by RNA interference has only mild effects on biosynthetic transport and epithelial polarization, but simultaneous inhibition of Rab10 and Rab8a more strongly impairs basolateral sorting. These results indicate that Rab10 functions in trafficking from the Golgi at early stages of epithelial polarization, is involved in biosynthetic transport to the basolateral membrane and may co-operate with Rab8.


"It's a funny old game". Football as an educational metaphor within induction to practice-based interprofessional learning.
The Common Learning Programme in the North East of England (CLPNE) sought to introduce interprofessional education into the practice setting for pre-registration health and social care students. Students, clinical educators/mentors, and facilitators met within groups over a period of 3 - 6 weeks to explore interprofessional working and learning together. This paper evaluates the use of a game, the Football Stadium, to stimulate participants' exploration of practice-based interprofessional working and learning at CLPNE induction sessions. Data consisting of verbal and written feedback from students and clinical educators/mentors, and field notes from facilitators covering 22 CLPNE pilot sites (February 2003 - July 2005) was supplemented by researcher observation at 12 sites. Two themes emerged from the data: the use of the Football Stadium as an "ice-breaker" at team induction and, the use of the Football Stadium as a vehicle to facilitate reflective learning. Key issues included personal identity and role within a novice--expert continuum, creating and developing the team environment and, enhancing and developing learning communities. Although recognized as requiring careful, sensitive facilitation, the Football Stadium is a simple means to present learning opportunities for interprofessional education within a non-threatening learning environment that facilitates active participation.


Epitope convergence of broadly HIV-1 neutralizing IgA and IgG antibody lineages in a viremic controller.
Decrypting the B cell ontogeny of HIV-1 broadly neutralizing antibodies (bNAbs) is paramount for vaccine design. Here, we characterized IgA and IgG bNAbs of three distinct B cell lineages in a viremic controller, two of which comprised only IgG+ or IgA+ blood memory B cells; the third combined both IgG and IgA clonal variants. 7-269 bNAb in the IgA-only lineage displayed the highest neutralizing capacity despite limited somatic mutation, and delayed viral rebound in humanized mice. bNAbs in all three lineages targeted the N332 glycan supersite. The 2.8-Å resolution cryo-EM structure of 7-269-BG505 SOSIP.664 complex showed a similar pose as 2G12, on an epitope mainly composed of sugar residues comprising the N332 and N295 glycans. Binding and cryo-EM structural analyses showed that antibodies from the two other lineages interact mostly with glycans N332 and N386. Hence, multiple B cell lineages of IgG and IgA bNAbs focused on a unique HIV-1 site of vulnerability can codevelop in HIV-1 viremic controllers.


Human invariant V alpha 24-J alpha Q TCR supports the development of CD1d-dependent NK1.1+ and NK1.1- T cells in transgenic mice.
A sizable fraction of T cells expressing the NK cell marker NK1.1 (NKT cells) bear a very conserved TCR, characterized by homologous invariant (inv.) TCR V alpha 24-J alpha Q and V alpha 14-J alpha 18 rearrangements in humans and mice, respectively, and are thus defined as inv. NKT cells. Because human inv. NKT cells recognize mouse CD1d in vitro, we wondered whether a human inv. V alpha 24 TCR could be selected in vivo by mouse ligands presented by CD1d, thereby supporting the development of inv. NKT cells in mice. Therefore, we generated transgenic (Tg) mice expressing the human inv. V alpha 24-J alpha Q TCR chain in all T cells. The expression of the human inv. V alpha 24 TCR in TCR C alpha(-/-) mice indeed rescues the development of inv. NKT cells, which home preferentially to the liver and respond to the CD1d-restricted ligand alpha-galactosylceramide (alpha-GalCer). However, unlike inv. NKT cells from non-Tg mice, the majority of NKT cells in V alpha 24 Tg mice display a double-negative phenotype, as well as a significant increase in TCR V beta 7 and a corresponding decrease in TCR V beta 8.2 use. Despite the forced expression of the human CD1d-restricted TCR in C alpha(-/-) mice, staining with mCD1d-alpha-GalCer tetramers reveals that the absolute numbers of peripheral CD1d-dependent T lymphocytes increase at most by 2-fold. This increase is accounted for mainly by an increased fraction of NK1.1(-) T cells that bind CD1d-alpha-GalCer tetramers. These findings indicate that human inv. V alpha 24 TCR supports the development of CD1d-dependent lymphocytes in mice, and argue for a tight homeostatic control on the total number of inv. NKT cells. Thus, human inv. V alpha 24 TCR-expressing mice are a valuable model to study different aspects of the inv. NKT cell subset.


Patterns and predictors of non-use of family planning methods in Ethiopian women: a panel survey.
Family planning (FP) is a key global health issue for achieving millennium development goals by 2015. The aim of this study was to identify predictors of changes in FP practice before and after interventions among Ethiopian women of reproductive age in a rural area. This study used a panel survey to identify predictors that placed women at greater risk for non-use of FP methods. The sample consisted of 407 women aged 19-49 years old living in a rural community of Ethiopia who completed both a baseline and follow-up survey. Trained local enumerators conducted face-to-face interviews during home visits. Patterns of FP practice were categorized into four groups: FP use at both baseline and follow-up; use at baseline but non-use at follow-up; non-use at baseline but use at follow-up; non-use at both baseline and follow-up. Logistic regression and classification and regression tree analyses were used. In a binary logistic regression, women over 35 years old with a negative attitude towards FP and resided in the Sibu area were less likely to start FP practice. Women over 35 years who were in poor health and resided in Sibu were less likely to continue FP practice. For the decision tree, age was the primary node for FP non-users at baseline while area of residence was the primary node for FP users at baseline. Age, health status, attitude towards FP, and area of residence are the primary considerations when developing FP interventions for Ethiopian women in a resource-limited rural area.


Pathologic interpretation of transbronchial biopsy for acute rejection of lung allograft is highly variable.
Despite the standardization of pathologic grading of acute rejection in transbronchial lung biopsies following lung transplantation, the reproducibility of pathologic diagnosis has not been adequately evaluated. To determine the interobserver variability for pathologic grading of acute rejection, 1566 biopsies from 845 subjects in the Lung Allograft Rejection Gene Expression Observational study were regraded by a pathology panel blinded to the original diagnosis and compared to the grade of acute rejection assigned by individual center pathologists. The study panel confirmed 49.1% of center pathologists' A0 grades, but upgraded 5.7% to A1 and 2.7% to grade ≥ A2 rejection; 42.5% were regraded as AX. Of 268 grade A1 samples, 21.2% were confirmed by the pathology panel; 18.7% were upgraded to ≥ A2 and 35.8% were downgraded to A0 with 24.3% being regraded as AX. Lastly, 53.5% of ≥ A2 cases were confirmed, but 15.7% were downgraded to grade A0 and 18.4% cases to A1, while 12.4% were regraded as AX. The kappa value for interobserver agreement was 0.183 (95%CI 0.147-0.220, p < 0.001). The results for B grade interpretation were similar. Suboptimal sampling is common and a high degree of variability exists in the pathologic interpretation of acute rejection in transbronchial biopsies.


A novel representation of the conformational structure of transfer RNAs. Correlation of the folding patterns of the polynucleotide chain with the base sequence and the nucleotide backbone torsions.
A novel description of the conformational characteristics of all the individual nucleotides and the phosphodiesters in tRNAs is presented in the form of a circular plot. This representation furnishes information of the base sequence with the folding patterns of the polynucleotide chain as one traverses along the circumference and with the individual nucleotide and phosphodiester linkage torsions along the radii. The circular plot obtained for yeast tRNAPhe strikingly distinguishes the helical and the loop regions. The variation of the different nucleotide torsions along the entire chain length and their effect on the secondary helical and tertiary loop regions become readily apparent.


Genetic and systems level analysis of Drosophila sticky/citron kinase and dFmr1 mutants reveals common regulation of genetic networks.
In Drosophila, the genes sticky and dFmr1 have both been shown to regulate cytoskeletal dynamics and chromatin structure. These genes also genetically interact with Argonaute family microRNA regulators. Furthermore, in mammalian systems, both genes have been implicated in neuronal development. Given these genetic and functional similarities, we tested Drosophila sticky and dFmr1 for a genetic interaction and measured whole genome expression in both mutants to assess similarities in gene regulation. We found that sticky mutations can dominantly suppress a dFmr1 gain-of-function phenotype in the developing eye, while phenotypes produced by RNAi knock-down of sticky were enhanced by dFmr1 RNAi and a dFmr1 loss-of-function mutation. We also identified a large number of transcripts that were misexpressed in both mutants suggesting that sticky and dFmr1 gene products similarly regulate gene expression. By integrating gene expression data with a protein-protein interaction network, we found that mutations in sticky and dFmr1 resulted in misexpression of common gene networks, and consequently predicted additional specific phenotypes previously not known to be associated with either gene. Further phenotypic analyses validated these predictions. These findings establish a functional link between two previously unrelated genes. Microarray analysis indicates that sticky and dFmr1 are both required for regulation of many developmental genes in a variety of cell types. The diversity of transcripts regulated by these two genes suggests a clear cause of the pleiotropy that sticky and dFmr1 mutants display and provides many novel, testable hypotheses about the functions of these genes. As both of these genes are implicated in the development and function of the mammalian brain, these results have relevance to human health as well as to understanding more general biological processes.


Predictors of PFOA levels in a community surrounding a chemical plant.
Perfluorooctanoic acid (PFOA) is considered a probable human carcinogen by the U.S. Environmental Protection Agency. It does not exist in nature but has been used widely since World War II. It is present in the serum of most Americans at about 4-5 ng/mL, although the routes of exposure remain unknown. We examined predictors of PFOA in mid-Ohio Valley residents living near a chemical plant that until recently released large quantities of PFOA into the environment, contaminating drinking water. We studied 69,030 residents in six contaminated water districts who participated in a 2005-2006 survey involving a questionnaire and blood tests. Of these, 64,251 had complete data on PFOA and covariates. We also analyzed a subset (71%) for whom we had occupational history. We ran linear regression models to determine serum PFOA predictors. Mean PFOA serum level was 83.0 ng/mL (median, 28.2). The most important predictors were current (median for all districts, 38.4; highest district, 224.1) and past (median, 18.6) residence in contaminated water districts, and current (median, 147.8) and past (median, 74.9) employment at the chemical plant (R(2) model = 0.55). PFOA was higher for males, those consuming local vegetables, and those using well water rather than public water, and lower for those using bottled water. PFOA was higher at younger and older ages. PFOA levels in this population varied with distance of residence from the plant and employment at the plant. Effects of age and sex reflected prior findings. Effects of other demographic and lifestyle covariates were relatively weak.


Physical properties of alginate hydrogels and their effects on in vitro follicle development.
The mechanical properties and density of natural and synthetic extracellular matrices are known to affect cellular processes and regulate tissue formation. In this report, these factors were independently investigated for their role in ovarian follicle development. The matrix composition was controlled through decreasing the solids concentration or the molar mass of the encapsulating biomaterial, alginate. Decreasing matrix stiffness and solids concentration enhanced follicle growth and coordinated differentiation of the follicle cell types, as evidenced by antral cavity formation, theca cell differentiation, oocyte maturation, and relative hormone production levels. While a stiff environment favored high progesterone and androgen secretion, decreasing alginate stiffness resulted in estrogen production which exceeded progesterone and androgen accumulation. These studies reveal, for the first time, a direct link between the biomechanical environment and follicle function, and suggest a novel non-hormonal mechanism regulating follicle development.


Co-ordinate single-cell expression of LEE4- and LEE5-encoded proteins of Escherichia coli O157:H7.
Escherichia coli O157:H7 is a zoonotic pathogen that can express a type III secretion system (TTSS) considered important for colonization and persistence in ruminants. E. coli O157:H7 strains have been shown to vary markedly in levels of protein secreted using the TTSS and this study has confirmed that a high secretion phenotype is more prevalent among isolates associated with human disease than isolates shed by healthy cattle. The variation in secretion levels is a consequence of heterogeneous expression, being dependent on the proportion of bacteria in a population that are actively engaged in protein secretion. This was demonstrated by indirect immunofluorescence and eGFP fusions that examined the expression of locus of enterocyte effacement (LEE)-encoded factors in individual bacteria. In liquid media, the expression of EspA, tir::egfp, intimin, but not map::egfp were co-ordinated in a subpopulation of bacteria. In contrast to E. coli O157:H7, expression of tir::egfp in EPEC E2348/69 was equivalent in all bacteria although the same fusion exhibited variable expression when transformed into an E. coli O157:H7 background. An E. coli O157:H7 strain deleted for the LEE demonstrated weak but variable expression of tir::egfp indicating that the elements controlling the heterogeneous expression lie outside the LEE. The research also demonstrated the rapid induction of tir::egfp and map::egfp on contact with bovine epithelial cells. This control in E. coli O157:H7 may be required to limit exposure of key surface antigens, EspA, Tir and intimin during colonization of cattle but allow their rapid production on contact with bovine gastrointestinal epithelium at the terminal rectum.


Local Sphingosine Kinase 1 Activity Improves Islet Transplantation.
Pancreatic islet transplantation is a promising clinical treatment for type 1 diabetes, but success is limited by extensive β-cell death in the immediate posttransplant period and impaired islet function in the longer term. Following transplantation, appropriate vascular remodeling is crucial to ensure the survival and function of engrafted islets. The sphingosine kinase (SK) pathway is an important regulator of vascular beds, but its role in the survival and function of transplanted islets is unknown. We observed that donor islets from mice deficient in SK1 (Sphk1 knockout) contain a reduced number of resident intraislet vascular endothelial cells. Furthermore, we demonstrate that the main product of SK1, sphingosine-1-phosphate, controls the migration of intraislet endothelial cells in vitro. We reveal in vivo that Sphk1 knockout islets have an impaired ability to cure diabetes compared with wild-type controls. Thus, SK1-deficient islets not only contain fewer resident vascular cells that participate in revascularization, but likely also a reduced ability to recruit new vessels into the transplanted islet. Together, our data suggest that SK1 is important for islet revascularization following transplantation and represents a novel clinical target for improving transplant outcomes.


Pharmacokinetics of vancomycin in critically ill infants undergoing extracorporeal membrane oxygenation.
Extracorporeal membrane oxygenation (ECMO) is a widely used therapy for neonates with respiratory failure. Because of sepsis, many of these infants require antibiotics like vancomycin during ECMO treatment. ECMO transiently alters renal function and increases the circulating blood volume by 75%. Initial vancomycin pharmacokinetics were determined in 12 infants undergoing ECMO to determine an adequate drug administration regimen. Vancomycin dosage was based on current recommendations for weight and gestational age. Pharmacokinetic parameters were determined by fitting the data to a two compartment model. This study yielded a mean steady-state volume of distribution of 1.1 +/- 0.5 (range, 0.6 to 2.1) liters/kg and a mean vancomycin clearance of 0.78 +/- 0.19 (range, 0.49 to 1.07) ml/min/kg. The mean vancomycin half-life was 16.9 +/- 9.5 (range, 8.8 to 42.9) h. Nomogram-calculated creatinine clearance was a significant predictor of vancomycin terminal rate constant and clearance. These data suggest alterations in the pharmacokinetics of vancomycin in infants on ECMO. With the goal of achieving vancomycin concentrations in serum above the MIC for the offending pathogen while using the least amount of the drug necessary, new administration guidelines for term infants without renal impairment undergoing ECMO should be 20 mg of vancomycin per kg at an interval of 24 h. With significant renal impairment, the interval should be extended on the basis of concentrations in serum. In comparison with previously published data, the neonates undergoing ECMO in our study demonstrated a much larger volume of distribution, a lower clearance, and consequently a longer vancomycin half-life.


Clinician-researcher's perspectives on clinical research during the COVID19 pandemic.
The outcome of well-performed clinical research is essential for evidence-based patient management during pandemics. However, conducting clinical research amidst a pandemic requires researchers to balance clinical and research demands. We seek to understand the values, experiences, and beliefs of physicians working at the onset of the COVID-19 pandemic in order to inform clinical research planning. We aim to understand whether pandemic settings affect physician comfort with research practices, and how physician experiences shape their understanding of research in a pandemic setting. A survey tool was adapted to evaluate familiarity and comfort with research during a pandemic. A cross-sectional, online questionnaire was distributed across Canadian research networks early in the COVID-19 outbreak. The survey was administered between March 11th and 17th, 2020, during a time of local transmission but prior to the surge of cases. We aimed to recruit into the survey physicians in infectious disease and critical care research networks across Canada. Of the 133 physician respondents, 131 (98%) considered it important to conduct clinical research during the COVID-19 pandemic. Respondents were more accepting of adaptations to the research process in during a pandemic compared to in a non-pandemic setting, including conducting research with deferred consent (χ2 = 8.941, 95% CI: -0.264, -0.085, p = 0.003), using non-identifiable observational data with a waiver of consent with a median score of 97 out of 100 (IQR: 79.25-100) vs median 87 out of 100 (IQR: 63-79) (95% CI: -12.43, 0.054, p = 0.052). The majority felt that research quality is not compromised during pandemics. Physicians consider it important to conduct research during a pandemic, highlighting the need to expedite research activities in pandemic settings. Respondents were more accepting of adaptations to the research process for research conducted during a pandemic, compared to that conducted in its absence of a pandemic.


Single port training in Latin-America-first uniportal video-assisted thoracoscopic surgery masterclass in Santiago, Chile.
Video-assisted thoracoscopic surgery (VATS) surgery has become the standard technique in Thoracic Surgery since its introduction 20 years ago. Single port VATS appeared as the next step in its evolution, with rapid development since the first uniportal video-assisted thoracoscopic surgery lobectomy (VATS lobectomy) by Dr. Diego Gonzalez-Rivas. During the last 5 years, the approach has been simplified, standardized and taught in many countries, courses, live surgery and dedicated programs, with reproducible results. Hands-on courses represent the best way to learn a new surgical technique, as it shortens learning curves and decreases complications. We present the first training course in single port VATS in our country, which became the first hands course in Thoracic Surgery in Chile.


Inhibition of human platelet aggregation by S-nitrosothiols. Heme-dependent activation of soluble guanylate cyclase and stimulation of cyclic GMP accumulation.
Recent studies have suggested that cyclic GMP accumulation in platelets mediates the antiaggregatory effects of certain nitrogen oxide-containing agents such as sodium nitroprusside, nitric oxide, nitrosoguanidines, and related agents. The vasodilator effect of these agents may involve the formation of S-nitrosothiol intermediates which relax vascular smooth muscle, elevate tissue levels of cyclic GMP, and activate guanylate cyclase. The purpose of this study was to investigate the effects of various synthetic S-nitrosothiols on human platelet aggregation. The S-nitroso derivatives of N-acetylpenicillamine, cysteine, and beta-D-thioglucose inhibited human platelet aggregation in a concentration-dependent fashion when ADP, collagen, U46619, or sodium arachidonate was employed as the aggregating agent. The antiaggregatory effects of the S-nitrosothiols were associated with a rapid and marked increase in intracellular platelet cyclic GMP levels, whereas cyclic AMP levels remained unchanged. Additionally, S-nitrosothiols disaggregated platelets which had been aggregated while concomitantly elevating platelet cyclic GMP levels. Moreover, guanylate cyclase, partially purified from the soluble fraction of human platelets, was markedly activated by S-nitrosothiols in a heme-dependent manner. Methemoglobin, a hemoprotein with a high affinity for nitric oxide, partially reversed the antiaggregatory effects, attenuated the accumulation of cyclic GMP, and inhibited the activation of guanylate cyclase by S-nitrosothiols. These data are consistent with the hypothesis that S-nitrosothiols could serve as active intermediates in the inhibitory action of sodium nitroprusside, nitric oxide, and related nitrogen oxides on platelet aggregation.


Diversity in fine specificity and T cell receptor usage of the human CD4+ cytotoxic T cell response specific for the immunodominant myelin basic protein peptide 87-106.
Multiple sclerosis (MS), a human demyelinating disease, is thought to be caused by an autoimmunologic process, and myelin basic protein (MBP) is considered a likely autoantigen. Studies of T cell lines (TCL) responding to different parts of the MBP molecule have indicated that amino acids 87 through 106 contain an immunodominant epitope of MBP. We have demonstrated previously that amino acids 89 through 99 represent the core of this 87-106 peptide epitope. Importantly, this epitope is not only encephalitogenic in SJL/J mice and Lewis rats but also has been shown to be recognized by human cytotoxic TCL in the context of four HLA-DR molecules that are associated with MS in different geographic areas. If the immune response to MBP peptide 87-106 was homogeneous with respect to epitope specificity and TCR usage, specific immunotherapies targeting the interaction of peptide, MHC, and TCR might be possible. In this study, the fine specificity of 29 CD4+ cytotoxic, long term, and limiting dilution TCL that had been generated against whole MBP and were derived from four MS patients and two healthy relatives was dissected using truncated and alanine-substituted peptides for the 87-106 peptide. In addition, the TCR alpha and beta chain usage of 15 CD4+ TCL was determined. Using truncated peptides, the presence of several nested immunogenic epitopes within amino acids 87 to 106 was demonstrated. TCL with identical restriction elements and similar responses to truncated peptides could be differentiated further using alanine-substituted peptides. Finally, heterogeneity of TCR usage was shown not only for those lines that differed in their peptide specificity but also for some that showed identical responses and were restricted by the same HLA-DR antigen. In conclusion, the CD4+ cytotoxic T cell response to the immunodominant MBP peptide 87-106 demonstrates a high degree of heterogeneity at the level of fine specificity and TCR usage. These findings indicate that specific immunotherapies aimed at TCR in MS will probably be more complicated than previously anticipated.


Efficacy and safety of artemether-lumefantrine dispersible tablets compared with crushed commercial tablets in African infants and children with uncomplicated malaria: a randomised, single-blind, multicentre trial.
Combination treatments, preferably containing an artemisinin derivative, are recommended to improve efficacy and prevent Plasmodium falciparum drug resistance. Our aim was to show non-inferiority of a new dispersible formulation of artemether-lumefantrine to the conventional crushed tablet in the treatment of young children with uncomplicated malaria. We did a randomised non-inferiority study on children weighing 5-35 kg with uncomplicated P falciparum malaria in Benin, Kenya, Mali, Mozambique, and Tanzania. The primary outcome measure was PCR-corrected 28-day parasitological cure rate. We aimed to show non-inferiority (with a margin of -5%) of dispersible versus crushed tablet. We constructed an asymptotic one-sided 97.5% CI on the difference in cure rates. A computer-generated randomisation list was kept centrally and investigators were unaware of the study medication administered. We used a modified intention-to-treat analysis. This trial is registered with ClinicalTrials.gov, number NCT00386763. 899 children aged 12 years or younger were randomly assigned to either dispersible (n=447) or crushed tablets (n=452). More than 85% of patients in each treatment group completed the study. 812 children qualified for the modified intention-to-treat analysis (n=403 vs n=409). The PCR-corrected day-28 cure rate was 97.8% (95% CI 96.3-99.2) in the group on dispersible formulation and 98.5% (97.4-99.7) in the group on crushed formulation. The lower bound of the one-sided 97.5% CI was -2.7%. The most common drug-related adverse event was vomiting (n=33 [7%] and n=42 [9%], respectively). No signs of ototoxicity or relevant cardiotoxicity were seen. A six-dose regimen of artemether-lumefantrine with the new dispersible formulation is as efficacious as the currently used crushed tablet in infants and children, and has a similar safety profile.


Mycobacterium branderi sp. nov., a new potential human pathogen.
A number of mycobacterial strains with similar growth characteristics, metabolic properties, and lipid compositions, which were previously placed in the Helsinki group (E. Brander, E. Jantzen, R. Huttunen, A. Juntunen, and M.-L. Katila, J. Clin. Microbiol. 30:1972-1975, 1992), were characterized by performing 16S rRNA gene sequencing. Of the 14 strains studied, 9 had a unique, previously undescribed sequence in the variable region of 16S rRNA. These nine strains, all of which were isolated from respiratory tract specimens, were nonpigmented and grew at 25 degrees C to 45 degrees C, reaching full colony size after 2 to 3 weeks. They produced arylsulfatase, nicotinamidase, and pyrazinamidase and were negative for Tween 80 hydrolysis, catalase, urease, and nitrate reductase activities, and niacin. Their glycolipid patterns were identical. A mycolic acid analysis performed by using thin-layer chromatography showed that these organisms contained alpha-mycolates, ketomycolates, and carboxy mycolates. Gas-liquid chromatography revealed that 2-eicosanol was the major alcohol and hexacosanoic acid was the major mycolic acid cleavage product. On the basis of their growth, biochemical, and lipid characteristics and their unique 16S rRNA sequence, we propose that these organisms should be assigned to a new species, Mycobacterium branderi. Comparative 16S rRNA sequencing revealed that this new species is closely related to Mycobacterium celatum, Mycobacterium cookii, and Mycobacterium xenopi. Strains 52157T (T = type strain) and 43548 have been deposited in the American Type Culture Collection as strains ATCC 51789 and ATCC 51788, respectively.