Structural variation in the chicken genome identified by paired-end next-generation DNA sequencing of reduced representation libraries.
Variation within individual genomes ranges from single nucleotide polymorphisms (SNPs) to kilobase, and even megabase, sized structural variants (SVs), such as deletions, insertions, inversions, and more complex rearrangements. Although much is known about the extent of SVs in humans and mice, species in which they exert significant effects on phenotypes, very little is known about the extent of SVs in the 2.5-times smaller and less repetitive genome of the chicken. We identified hundreds of shared and divergent SVs in four commercial chicken lines relative to the reference chicken genome. The majority of SVs were found in intronic and intergenic regions, and we also found SVs in the coding regions. To identify the SVs, we combined high-throughput short read paired-end sequencing of genomic reduced representation libraries (RRLs) of pooled samples from 25 individuals and computational mapping of DNA sequences from a reference genome. We provide a first glimpse of the high abundance of small structural genomic variations in the chicken. Extrapolating our results, we estimate that there are thousands of rearrangements in the chicken genome, the majority of which are located in non-coding regions. We observed that structural variation contributes to genetic differentiation among current domesticated chicken breeds and the Red Jungle Fowl. We expect that, because of their high abundance, SVs might explain phenotypic differences and play a role in the evolution of the chicken genome. Finally, our study exemplifies an efficient and cost-effective approach for identifying structural variation in sequenced genomes.


Risk of adverse outcomes associated with concomitant use of clopidogrel and proton pump inhibitors following acute coronary syndrome.
Prior mechanistic studies reported that omeprazole decreases the platelet inhibitory effects of clopidogrel, yet the clinical significance of these findings is not clear. To assess outcomes of patients taking clopidogrel with or without a proton pump inhibitor (PPI) after hospitalization for acute coronary syndrome (ACS). Retrospective cohort study of 8205 patients with ACS taking clopidogrel after discharge from 127 Veterans Affairs hospitals between October 1, 2003, and January 31, 2006. Vital status information was available for all patients through September 30, 2006. All-cause mortality or rehospitalization for ACS. Of 8205 patients taking clopidogrel after discharge, 63.9% (n = 5244) were prescribed PPI at discharge, during follow-up, or both and 36.1% (n = 2961) were not prescribed PPI. Death or rehospitalization for ACS occurred in 20.8% (n = 615) of patients taking clopidogrel without PPI and 29.8% (n = 1561) of patients taking clopidogrel plus PPI. In multivariable analyses, use of clopidogrel plus PPI was associated with an increased risk of death or rehospitalization for ACS compared with use of clopidogrel without PPI (adjusted odds ratio [AOR], 1.25; 95% confidence interval [CI], 1.11-1.41). Among patients taking clopidogrel after hospital discharge and prescribed PPI at any point during follow-up (n = 5244), periods of use of clopidogrel plus PPI (compared with periods of use of clopidogrel without PPI) were associated with a higher risk of death or rehospitalization for ACS (adjusted hazard ratio, 1.27; 95% CI, 1.10-1.46). In analyses of secondary outcomes, patients taking clopidogrel plus PPI had a higher risk of hospitalizations for recurrent ACS compared with patients taking clopidogrel without PPI (14.6% vs 6.9%; AOR, 1.86 [95% CI, 1.57-2.20]) and revascularization procedures (15.5% vs 11.9%; AOR, 1.49 [95% CI, 1.30-1.71]), but not for all-cause mortality (19.9% vs 16.6%; AOR, 0.91 [95% CI, 0.80-1.05]). The association between use of clopidogrel plus PPI and increased risk of adverse outcomes also was consistent using a nested case-control study design (AOR, 1.32; 95% CI, 1.14-1.54). In addition, use of PPI without clopidogrel was not associated with death or rehospitalization for ACS among patients not taking clopidogrel after hospital discharge (n = 6450) (AOR, 0.98; 95% CI, 0.85-1.13). Concomitant use of clopidogrel and PPI after hospital discharge for ACS was associated with an increased risk of adverse outcomes than use of clopidogrel without PPI, suggesting that use of PPI may be associated with attenuation of benefits of clopidogrel after ACS.


Adaptive vibration suppression system: an iterative control law for a piezoelectric actuator shunted by a negative capacitor.
An adaptive system for the suppression of vibration transmission using a single piezoelectric actuator shunted by a negative capacitance circuit is presented. It is known that by using a negative-capacitance shunt, the spring constant of a piezoelectric actuator can be controlled to extreme values of zero or infinity. Because the value of spring constant controls a force transmitted through an elastic element, it is possible to achieve a reduction of transmissibility of vibrations through the use of a piezoelectric actuator by reducing its effective spring constant. Narrow frequency range and broad frequency range vibration isolation systems are analyzed, modeled, and experimentally investigated. The problem of high sensitivity of the vibration control system to varying operational conditions is resolved by applying an adaptive control to the circuit parameters of the negative capacitor. A control law that is based on the estimation of the value of the effective spring constant of a shunted piezoelectric actuator is presented. An adaptive system which achieves a self-adjustment of the negative capacitor parameters is presented. It is shown that such an arrangement allows the design of a simple electronic system which offers a great vibration isolation efficiency under variable vibration conditions.


Variational region-based segmentation using multiple texture statistics.
This paper investigates variational region-level criterion for supervised and unsupervised texture-based image segmentation. The focus is given to the demonstration of the effectiveness and robustness of this region-based formulation compared to most common variational approaches. The main contributions of this global criterion are twofold. First, the proposed methods circumvent a major problem related to classical texture based segmentation approaches. Existing methods, even if they use different and various texture features, are mainly stated as the optimization of a criterion evaluating punctual pixel likelihoods or similarity measure computed within a local neighborhood. These approaches require sufficient dissimilarity between the considered texture features. An additional limitation is the choice of the neighborhood size and shape. These two parameters and especially the neighborhood size significantly influence the classification performances: the neighborhood must be large enough to capture texture structures and small enough to guarantee segmentation accuracy. These parameters are often set experimentally. These limitations are mitigated with the proposed variational methods stated at the region-level. It resorts to an energy criterion defined on image where regions are characterized by nonparametric distributions of their responses to a set of filters. In the supervised case, the segmentation algorithm consists in the minimization of a similarity measure between region-level statistics and texture prototypes and a boundary based functional that imposes smoothness and regularity on region boundaries. In the unsupervised case, the data-driven term involves the maximization of the dissimilarity between regions. The proposed similarity measure is generic and permits optimally fusing various types of texture features. It is defined as a weighted sum of Kullback-Leibler divergences between feature distributions. The optimization of the proposed variational criteria is carried out using a level-set formulation. The effectiveness and the robustness of this formulation at region-level, compared to classical active contour methods, are evaluated for various Brodatz and natural images.


RNA-seq implicates deregulation of the immune system in the pathogenesis of diverticulitis.
Individuals with diverticula or outpouchings of the colonic mucosa and submucosa through the colonic wall have diverticulosis, which is usually asymptomatic. In 10-25% of individuals, the diverticula become inflamed, resulting in diverticulitis. Very little is known about the pathophysiology or gene regulatory pathways involved in the development of diverticulitis. To identify these pathways, we deep sequenced RNAs isolated from full-thickness sections of sigmoid colon from diverticulitis patients and control individuals. Specifically for diverticulitis cases, we analyzed tissue adjacent to areas affected by chronic disease. Since the tissue was collected during elective sigmoid resection, the disease was in a quiescent state. A comparison of differentially expressed genes found that gene ontology (GO) pathways associated with the immune response were upregulated in diverticulitis patients compared with nondiverticulosis controls. Next, weighted gene coexpression network analysis was performed to identify the interaction among coexpressed genes. This analysis revealed RASAL3, SASH3, PTPRC, and INPP5D as hub genes within the brown module eigengene, which highly correlated (r = 0.67, P = 0.0004) with diverticulitis. Additionally, we identified elevated expression of downstream interacting genes. In summary, transcripts associated with the immune response were upregulated in adjacent tissue from the sigmoid colons of chronic, recurrent diverticulitis patients. Further elucidating the genetic or epigenetic mechanisms associated with these alterations can help identify those at risk for chronic disease and may assist in clinical decision management.NEW & NOTEWORTHY By using an unbiased approach to analyze transcripts expressed in unaffected colonic tissues adjacent to those affected by chronic diverticulitis, our study implicates that a defect in the immune response may be involved in the development of the disease. This finding expands on the current data that suggest the pathophysiology of diverticulitis is mediated by dietary, age, and obesity-related factors. Further characterizing the immunologic differences in diverticulitis may better inform clinical decision-making.


Embryo selection versus natural selection: how do outcomes of comprehensive chromosome screening of blastocysts compare with the analysis of products of conception from early pregnancy loss (dilation and curettage) among an assisted reproductive technology population?
To compare the incidence of numerical chromosomal abnormalities (NCAs) reported after preimplantation genetic screening (PGS) analysis compared with that reported after cytogenetic analysis of products of conception after spontaneous abortion. Retrospective study. Private academic in vitro fertilization center. Cytogenetic reports of patients who underwent an IVF cycle with PGS of at least one biopsied embryo were compared with cytogenetic analysis reported from patients who had dilation and curettage (D&C) for the treatment of a spontaneous abortion after assisted reproductive technology (ART) treatment. None. Frequencies for each numerical chromosomal abnormality from both groups were compared. A total of 1,069 NCAs were reported after PGS (trisomy 54.3%, monosomy 45.7%, no polyploidies), resulting in a trisomy/monosomy ratio of 0.82. A total of 447 NCAs was reported after D&C (trisomy 83%, polyploidy 10.7%, monosomy 6.3%). The aneuploidies most frequently identified were similar in both groups and included 15, 16, 18, 21, and 22. Monosomies (n = 28, 6.3%) were rarely observed in the group that underwent D&C after ART. This review provides an analysis of the most commonly identified NCAs after PGS and in first-trimester D&C samples in an infertile population utilizing ART. Although monosomies comprised >50% of all cytogenetic anomalies identified after PGS, there were very few identified in the post-D&C samples. This suggests that although monosomies occur frequently in the IVF population, they commonly do not implant. Despite this difference, this study demonstrated that the specific NCAs observed after PGS analysis and D&C were comparable.


Effect of garlic supplementation on serum C-reactive protein level: A systematic review and meta-analysis of randomized controlled trials.
The present study was designed to systematically review randomized controlled trials (RCTs) that report on the effects of garlic supplementation on serum C-reactive protein (CRP) levels. We conducted a literature search of Scopus, PubMed, Cochrane Library, and Google Scholar up to January 2018. Weighted mean differences (WMD) were estimated for net change in serum CRP. Subgroup analyses were also performed by duration of study, dose of supplementation, baseline CRP level, and the quality of studies. From 438 articles found and screened in our initial search, nine RCTs with the sum of total sample size of 363 were included in the meta-analysis. Compared with the controls, garlic intake significantly reduced the concentrations of serum CRP by 0.8 mg/L (95% CI [-1.5, -0.1], p = 0.02) with the evidence of heterogeneity among studies. Subgroup analyses showed that garlic significantly lowered CRP by 0.82 mg/L (95% CI [-1.02, -0.62], p < 0.001) among studies with a daily garlic dose ≥1,200 mg/day and by 2.44 mg/L (95% CI [-4.02, -0.87], p = 0.002) among studies with baseline CRP ≥2 mg/L. Current data confirmed that garlic supplementation would reduce serum CRP levels. However, the changes were related to the supplemental doses and baseline levels of serum CRP.


Stromal cell-derived factor-1 (chemokine C-X-C motif ligand 12) and chemokine C-X-C motif receptor 4 are required for migration of gonadotropin-releasing hormone neurons to the forebrain.
Gonadotropin-releasing hormone (GnRH) neurons migrate from the vomeronasal organ (VNO) in the nasal compartment to the basal forebrain in mice, beginning on embryonic day 11 (E11). These neurons use vomeronasal axons as guides to migrate through the nasal mesenchyme. Most GnRH neurons then migrate along the caudal branch of the vomeronasal nerve to reach the hypothalamus. We show here that stromal cell-derived factor-1 [SDF-1, also known as chemokine C-X-C motif ligand 12 (CXCL12)] is expressed in the embryonic nasal mesenchyme from as early as E10 in an increasing rostral to caudal gradient that is most intense at the border of the nasal mesenchyme and the telencephalon. Chemokine C-X-C motif receptor 4 (CXCR4), the receptor for SDF-1, is expressed by neurons in the olfactory epithelium and VNO. Cells derived from these sensory epithelia, including migrating GnRH neurons and ensheathing glial precursors of the migrating mass (MM), also express CXCR4, suggesting that they may use SDF-1 as a chemokine. In support of this, most GnRH neurons of CXCR4-/- mice fail to exit the VNO at E13, and comparatively few GnRH neurons reach the forebrain. There is also a significant decrease in the total number of GnRH neurons in CXCR4-/- mice and an increase in cell death within the VNO relative to controls. The MM is smaller in CXCR4-/- mice, suggesting that some MM cells also require SDF-1/CXCR4 function for migration and survival.


The effect of selection and referral biases for the treatment of localised prostate cancer with surgery or radiation.
Consultation with radiation oncologists, in addition to urologists, is advocated for patients diagnosed with prostate cancer. Treatment patterns for patients receiving consultation from radiation oncologists in addition to urologists have not previously been described. We conducted a matched cohort study of men with newly diagnosed non-metastatic prostate cancer in Ontario, Canada. Patients who underwent consultation with a radiation oncologist prior to treatment were matched 1:1 with patients managed by a urologist alone based on tumour and patient characteristics. We examined rates of active treatment (surgery or radiotherapy) within one year following diagnosis. Among 5708 matched pairs (11,416 patients), those who received radiation oncology consultation were more likely to undergo active treatments whether they had intermediate or high-risk disease (88.6% vs. 65.9%, p < 0.0001; adjusted odds ratio 4.0, 95% CI: 3.6-4.4) or low-risk disease (56.1% vs. 13.3%, p < 0.0001; adjusted odds ratio 8.4, 95% CI: 6.7-10.6). This effect persisted after considering age, comorbidity, tumour volume and year of diagnosis. Patients newly diagnosed with prostate cancer who receive radiation oncology consultation are associated with a higher rate of active treatment, compared to patients managed by urologists only. Selection and referral biases, and unmeasured confounding such as patient preference must be considered as important factors attributing this association.


Clinical and Radiological Characterization of Progressive Multifocal Leukoencephalopathy in HIV-Infected Patients: A Retrospective Analysis and Review of the Literature.
Progressive multifocal leukoencephalopathy is a demyelinating disease of the central nervous system caused by John Cunningham virus, mostly associated with immunodeficiency conditions, such as the human immunodeficiency virus infection. Progressive multifocal leukoencephalopathy can have multiple clinical features and usually presents a typical lesion pattern on brain magnetic resonance imaging. Its course may be rapidly progressive, although immunological responsiveness can be associated with an improved prognosis. We performed a retrospective analysis of the clinical and radiological data from patients admitted in our institution between January 2005 and April 2014 with the diagnosis of definitive progressive multifocal leukoencephalopathy (ICD10:A81.2) in the setting of human immunodeficiency virus infection. Twenty-one patients were included in our study, mostly men (n = 20, 95.2%). Mean age at diagnosis was 39 years. Motor deficits were the most common clinical finding. John Cunningham virus-DNA was detected in the cerebral spinal fluid in 20 patients (95.2%). Brain imaging studies most commonly disclosed bilateral supratentorial, asymmetric lesions. Four (19%) patients developed immune reconstitution inflammatory syndrome in the follow-up. Therapeutic approach included initiation and continuation/optimization of antiretroviral therapy, with adjunctive therapy with corticosteroids in four patients. Seventeen (81%) patients died during the study period; median survival time following progressive multifocal leukoencephalopathy diagnosis was 3 months (range 1 - 13). The results of our study are in accordance with the data previously published on progressive multifocal leukoencephalopathy in human immunodeficiency virus patients. Progressive multifocal leukoencephalopathy is predominantly associated with severe immunosuppression, particularly in patients who are not under anti-retroviral therapy, and usually presents with motor and cognitive symptoms and signs. A typical bilateral asymmetric pattern in conventional magnetic resonance imaging is present in the majority of the patients. There is no specific therapy for progressive multifocal leukoencephalopathy and it is usually fatal, although outcomes can improve with highly active anti-retroviral therapy. Immune reconstitution inflammatory syndrome is also an important complication related with progressive multifocal leukoencephalopathy, usually associated with anti-retroviral therapy. Progressive multifocal leukoencephalopathy-immune reconstitution inflammatory syndrome presents with different imaging characteristics from progressive multifocal leukoencephalopathy and treatment with steroids can improve survival. The mortality rate and long-term neurological morbidity associated with progressive multifocal leukoencephalopathy are quite high. These data should increase clinician awareness to the occurrence of progressive multifocal leukoencephalopathy among human immunodeficiency virus patients and highlight the important role of magnetic resonance imaging, as early diagnosis may beassociated with better outcome.


Hesperidin potentiates the neuroprotective effects of diazepam and gabapentin against pentylenetetrazole-induced convulsions in mice: Possible behavioral, biochemical and mitochondrial alterations.
Epilepsy is a chronic neurological disorder with complex pathophysiology. Several evidences suggest a role of oxidative stress and mitochondrial dysfunction in pathophysiology of epilepsy. Hesperidin (Hesp) acts as a powerful anti-oxidant agent against superoxide, singlet oxygen, and hydroxyl radicals. Thus, this study was undertaken to evaluate the possible neuroprotective mechanism of Hesp against pentylenetetrazole (PTZ)-induced convulsions in mice. Sixty males Laca mice (20-25 g) were randomly divided into 10 treatment groups (n = 6). Seven days pretreatment of Hesp (100, 200 mg/kg, p.o.) was carried out before PTZ (80 mg/kg, intraperitoneal [i.p.]) challenge, whereas diazepam (DZP) (0.2, 0.5 mg/kg) and gabapentin (Gbp) (10, 20 mg/kg) were administered i.p. 30 min before PTZ administration, that is, on 7(th) day. Following PTZ challenge, severity of convulsions (onset of jerks, myoclonic seizures, extensor phase and death), brain anti-oxidant enzyme levels and mitochondrial complex enzymes activities were estimated. Single i.p. PTZ (80 mg/kg) challenge demonstrated severe convulsions, oxidative damage (raised lipid peroxidation [LPO], nitrite concentration as well as depleted reduced glutathione, superoxide dismutase and catalase levels), and depletion of mitochondrial enzyme Complex (I, II, IV) activities. Hesp (200 mg/kg), DZP (0.5 mg/kg) and Gbp (20 mg/kg) pretreatments attenuated PTZ induced behavioral, biochemical and mitochondrial alterations. However, administration of Hesp (100 mg/kg) in combination with DZP (0.2 mg/kg) or Gbp (10 mg/kg) potentiated their neuroprotective effect, which was significant as compared to their effects in PTZ treated animals. Hesp possesses potent anticonvulsant activity which might be mediated through modulation of gamma-amino butyric acid/benzodiazepine receptor action.


Growth factor requirements of human colorectal tumour cells: relations to cellular differentiation.
Human colorectal tumour lines that exhibit different degrees of differentiation were studied to define their growth requirements. The poorly differentiated cell lines SW620, SW480, SW48 and SW837 proliferated in Dulbecco's modified Eagle's medium without exogenously added growth factors. In contrast, the moderately differentiated cell lines SW1222, HT29, PC/JW and LS174T proliferated only in medium supplemented with growth factor. SW1222 and HT29 cells required transferrin for growth, which was improved by other growth-promoting factors including epidermal growth factor (SW1222) and sodium selenite (HT29). PC/JW and LS174T required both insulin and transferrin for optimal growth. The tumour cell lines could be passaged continuously in serum-free medium supplemented with growth factor and in some cases they grew better than in serum-supplemented medium. The serum-free growth conditions should prove useful for studies on differentiation in colorectal cell lines and their interactions with growth factors.


Subcellular fractionation method to study endosomal trafficking of Kaposi's sarcoma-associated herpesvirus.
Virus entry involves multiple steps and is a highly orchestrated process on which successful infection collectively depends. Entry processes are commonly analyzed by monitoring internalized virus particles via Western blotting, polymerase chain reaction, and imaging techniques that allow scientist to track the intracellular location of the pathogen. Such studies have provided abundant direct evidence on how viruses interact with receptor molecules on the cell surface, induce cell signaling at the point of initial contact with the cell to facilitate internalization, and exploit existing endocytic mechanisms of the cell for their ultimate infectious agenda. However, there is dearth of knowledge in regards to trafficking of a virus via endosomes. Herein, we describe an optimized laboratory procedure to isolate individual organelles during different stages of endocytosis by performing subcellular fractionation. This methodology is established using Kaposi's sarcoma-associated herpesvirus (KSHV) infection of human foreskin fibroblast (HFF) cells as a model. With KSHV and other herpesviruses alike, envelope glycoproteins have been widely reported to physically engage target cell surface receptors, such as integrins, in interactions leading to entry and subsequent infection. Subcellular fractionation was used to isolate early and late endosomes (EEs and LEs) by performing a series of centrifugations steps. Specifically, a centrifugation step post-homogenization was utilized to obtain the post-nuclear supernatant containing intact intracellular organelles in suspension. Successive fractionation via sucrose density gradient centrifugation was performed to isolate specific organelles including EEs and LEs. Intracellular KSHV trafficking was directly traced in the isolated endosomal fractions. Additionally, the subcellular fractionation approach demonstrates a key role for integrins in the endosomal trafficking of KSHV. The results obtained from fractionation studies corroborated those obtained by traditional imaging studies. This study is the first of its kind to employ a sucrose flotation gradient assay to map intracellular KSHV trafficking in HFF cells. We are confident that such an approach will serve as a powerful tool to directly study intracellular trafficking of a virus, signaling events occurring on endosomal membranes, and dynamics of molecular events within endosomes that are crucial for uncoating and virus escape into the cytosol.


Metacognition Modules: A Scaffolded Series of Online Assignments Designed to Improve Students' Study Skills.
Many first-year biology students begin college with high aspirations but limited skills in terms of those needed for their success. Teachers are increasingly focused on students' lack of metacognitive awareness combined with students' inability to self-regulate learning behaviors. To address this need, we have designed a series of out-of-class assignments to provide explicit instruction on memory and learning. Our metacognition modules consist of six video assignments with reflective journaling prompts, allowing students to explore the relationship between the learning cycle, neuroplasticity, memory function, expert and novice thinking, and effective study strategies. By setting lessons on improving study behavior within a biological context, we help students grasp the reason for changing their behavior based on an understanding of biological functions and their application to learning. Students who complete these scaffolded journaling assignments show a shift toward a growth mindset and a consistent ability to evaluate the efficacy of their own study behaviors. In this article, we discuss the modules and student assignments, as well as provide in depth support for faculty who wish to adopt the modules for their own courses.


Are opinions based on science: modelling social response to scientific facts.
As scientists we like to think that modern societies and their members base their views, opinions and behaviour on scientific facts. This is not necessarily the case, even though we are all (over-) exposed to information flow through various channels of media, i.e. newspapers, television, radio, internet, and web. It is thought that this is mainly due to the conflicting information on the mass media and to the individual attitude (formed by cultural, educational and environmental factors), that is, one external factor and another personal factor. In this paper we will investigate the dynamical development of opinion in a small population of agents by means of a computational model of opinion formation in a co-evolving network of socially linked agents. The personal and external factors are taken into account by assigning an individual attitude parameter to each agent, and by subjecting all to an external but homogeneous field to simulate the effect of the media. We then adjust the field strength in the model by using actual data on scientific perception surveys carried out in two different populations, which allow us to compare two different societies. We interpret the model findings with the aid of simple mean field calculations. Our results suggest that scientifically sound concepts are more difficult to acquire than concepts not validated by science, since opposing individuals organize themselves in close communities that prevent opinion consensus.


The influence of esomeprazole and cisapride on gastroesophageal reflux during anesthesia in dogs.
Gastroesophageal reflux (GER) is common in anesthetized dogs and can cause esophagitis, esophageal stricture, and aspiration pneumonia. To determine whether preanesthetic IV administration of esomeprazole alone or esomeprazole and cisapride increases esophageal pH and decreases the frequency of GER in anesthetized dogs using combined multichannel impedance and pH monitoring. Sixty-one healthy dogs undergoing elective orthopedic surgery procedures. Prospective, randomized, placebo-controlled study. Dogs were randomized to receive IV saline (0.9% NaCl), esomeprazole (1 mg/kg) alone, or a combination of esomeprazole (1 mg/kg) and cisapride (1 mg/kg) 12-18 hours and 1-1.5 hours before anesthetic induction. An esophageal pH/impedance probe was utilized to measure esophageal pH and detect GER. Eight of 21 dogs in the placebo group (38.1%), 8 of 22 dogs in the esomeprazole group (36%), and 2 of 18 dogs in the combined esomeprazole and cisapride group (11%) had ≥ 1 episode of GER on impedance testing during anesthesia (P < .05). Esomeprazole was associated with a significant increase in gastric and esophageal pH (P = .001), but the drug did not significantly decrease the frequency of GER (P = .955). Concurrent administration of cisapride was associated with a significant decrease in the number of reflux events (RE) compared to the placebo and esomeprazole groups (P < .05). Preanesthetic administration of cisapride and esomeprazole decreases the number of RE in anesthetized dogs, but administration of esomeprazole alone was associated with nonacid and weakly acidic reflux in all but 1 dog.


The activity of human aquaporin 1 as a cGMP-gated cation channel is regulated by tyrosine phosphorylation in the carboxyl-terminal domain.
In addition to a constitutive water channel activity, several studies suggest Aquaporin-1 (AQP1) functions as a nonselective monovalent cation channel activated by intracellular cGMP, although variability in responsiveness between preparations has led to controversy in the field. Data here support the hypothesis that responsiveness of the AQP1 ionic conductance to cGMP is governed by tyrosine phosphorylation. Wild-type and mutant human AQP1 channels expressed in Xenopus laevis oocytes were characterized by two-electrode voltage clamp and optical osmotic swelling analyses. Quadruple mutation by site-directed mutagenesis of barrier hydrophobic residues (Val50, Leu54, Leu170, Leu174) to alanines in the central pore induced inward rectification of the ionic current and shifted reversal potential by approximately +10 mV, indicating increased permeability of tetraethylammonium ion. Introduction of cysteine at lysine 51 in the central pore (K51C) in a cysteine-less template created new sensitivity to block of the conductance by mercuric ion. Mutations of candidate consensus sites and pharmacological manipulation of serine and threonine phosphorylation did not alter cGMP-dependent responses; however, mutation of tyrosine Y253C or pharmacological dephosphorylation prevented ion channel activation. Modification of Y253C by covalent addition of a negatively charged group [2-sulfonatoethyl methanethiosulfonate sodium salt (MTSES)] rescued the cGMP-activated conductance response, an effect reversed by dithiothreitol. Results support the proposal that phosphorylation of tyrosine Tyr253 in the carboxyl terminal domain, confirmed by Western blot, acts as a master switch regulating responsiveness of AQP1 ion channels to cGMP, and the tetrameric central pore is the ion permeation pathway. These findings advance resolution of a standing controversy and expand our understanding of AQP1 as a multifunctional regulated channel.


Control of the hierarchical assembly of π-conjugated optoelectronic peptides by pH and flow.
Self-assembled nanoaggregates of π-conjugated peptides possess optoelectronic properties due to electron delocalization over the conjugated peptide groups that make them attractive candidates for the fabrication of bioelectronic materials. We present a computational and theoretical study to resolve the microscopic effects of pH and flow on the non-equilibrium morphology and kinetics of early-stage assembly of an experimentally-realizable optoelectronic peptide that displays pH triggerable assembly. Employing coarse-grained molecular dynamics simulations, we probe the effects of pH on growth kinetics and aggregate morphology to show that control of the peptide protonation state by pH can be used to modulate the assembly rates, degree of molecular alignment, and resulting morphologies within the self-assembling nanoaggregates. We also quantify the time and length scales at which convective flows employed in directed assembly compete with microscopic diffusion to show that flow influences cluster alignment and assembly rate during early-stage assembly only at extremely high shear rates. This suggests that observed improvements in optoelectronic properties at experimentally-accessible shear rates are due to the alignment of large aggregates of hundreds of monomers on time scales in excess of hundreds of nanoseconds. Our work provides new fundamental understanding of the effects of pH and flow to control the morphology and kinetics of early-stage assembly of π-conjugated peptides and lays the groundwork for the rational manipulation of environmental conditions to direct assembly and the attendant emergent optoelectronic properties.


A2BCD: a concise guide for asthma management.
The management of asthma has changed fundamentally during the past two decades. Precise assessment and phenotyping are now required to establish individually targeted treatment with disease-modifying anti-asthmatic drugs (DMAADs). Patients with asthma are often managed by primary care doctors or non-respiratory specialists in secondary care. However, the implementation of complex asthma guidelines in non-specialised care remains a challenge. There is a need for easy-to-understand, concise guides for general practice. In this Viewpoint, we propose a one-page practical guide for asthma management, titled A2BCD, with four components: dual assessment (A2) of asthma (ie, diagnosis and phenotype, plus asthma control and future risks); basic measures (B; eg, education, self-management skills, regular physical activity, and avoidance of asthma triggers); identification and treatment of comorbidities (C) of asthma (eg, chronic rhinosinusitis, obesity, or sleep apnoea); and phenotype-specific, individually targeted treatment with DMAADs (D), including individual inhalation schemes based on inhaled corticosteroids, leukotriene modifiers, biologics, and allergen immunotherapy.


Anti-inflammatory activity of polyphenolics from açai (Euterpe oleracea Martius) in intestinal myofibroblasts CCD-18Co cells.
The demand for tropical fruits high in polyphenolics including açai (Euterpe oleracea Mart.) has been increasing based on ascribed health benefits and antioxidant properties. This study evaluated the anti-inflammatory activities of açai polyphenolics in human colon myofibroblastic CCD-18Co cells to investigate the suppression of reactive oxygen species (ROS), and mRNA and protein expression of inflammatory proteins. Non-cytotoxic concentrations of açai extract, 1-5 mg gallic acid equivalent L(-1), were selected. The generation of ROS was induced by lipopolysaccharide (LPS) and açai extract partially reversed this effect to 0.53-fold of the LPS-control. Açai extract (5 mg GAE L(-1)) down-regulated LPS-induced mRNA-expression of tumor necrosis factor alpha, TNF-α (to 0.42-fold), cyclooxygenase 2, COX-2 (to 0.61-fold), toll-like receptor-4, TLR-4 (to 0.52-fold), TNF receptor-associated factor 6, TRAF-6 (to 0.64-fold), nuclear factor kappa-B, NF-κB (to 0.76-fold), vascular cell adhesion molecule 1, VCAM-1 (to 0.71-fold) and intercellular adhesion molecule 1, ICAM-1 (to 0.68-fold). The protein levels of COX-2, TLR-4, p-NF-κB and ICAM-1 were induced by LPS and the açai extract partially reversed this effect in a dose-dependent manner. These results suggest the anti-inflammatory effect of açai polyphenolic extract in intestinal cells are at least in part mediated through the inhibition of ROS and the expression of TLR-4 and NF-κB. Results indicate the potential for açai polyphenolics in the prevention of intestinal inflammation.


GABAA receptor subunit immunoreactivity in primate visual cortex: distribution in macaques and humans and regulation by visual input in adulthood.
Subunit proteins that make up functional GABAA receptors were localized immunocytochemistry in the primary visual cortex (area 17) of adult monkeys and humans. Immunoreactivity for the alpha 1, beta 2/3, and gamma 2 subunits is greatest in layers (II-III, IVA and IVC) of monkey area 17 that contain the highest density of GABA neurons and terminals. Immunostaining for each subunit is unevenly distributed in layers II and III, where patches of immunoreactivity correspond to regions of intense cytochrome oxidase (CO) staining, and in layer IVA, where intense immunoreactivity forms a honeycomb pattern identical to the CO staining pattern. Immunoreactivity for the subunits is localized principally within the neuropil, which, by simultaneous comparison with the distribution of microtubule-associated protein immunostaining, was found to include bundles of thin dendrites and zones of numerous dendritic segments. In addition, gamma 2 immunostaining surrounds the somata of a subpopulation of GABAergic neurons, immunoreactive for the calcium-binding protein parvalbumin. All three subunits are present in the somata and processes of neurons that occupy the white matter subjacent to monkey area 17. In human visual cortex, the alpha 1, beta 2/3, and gamma 2 subunits are distributed in a manner similar to that found in monkeys, with relatively intense immunostaining in layers IVC and IVA. In layer IVC, vertical stripes of intense receptor immunostaining (20-30 microns wide) alternate with wider stripes of pale immunostaining (30-60 microns wide). In the upper and lower halves of IVC beta, these stripes form lattices similar to those in layers IVC and IVA of monkeys. Following monocular deprivation by intravitreal injections of TTX in adult monkeys, immunoreactivity for each subunit in layer IVC consists of alternating intensely and lightly stained stripes. Comparison with the pattern of CO staining indicates that intense immunostaining for alpha 1, beta 2/3, and gamma 2 occurs in normal-eye stripes while abnormally light immunostaining is present in deprived-eye stripes. For all three subunits, immunoreactivity in deprived-eye stripes is reduced within 5 d of monocular deprivation and remains abnormally low for deprivations that extend to at least 30 d. These findings indicate that each of several GABAA receptor subunits adopt similar laminar and compartmental distributions in monkey and human area 17 and are likely to be expressed by the same neurons. The deprivation-dependent reduction in immunoreactivity for alpha 1, beta 2/3, and gamma 2 subunits suggests that all are regulated by visually driven activity.(ABSTRACT TRUNCATED AT 400 WORDS)


Effect of Microplastic on the Gills of the Shore Crab Carcinus maenas.
Microscopic plastic debris (microplastics, <5 mm in diameter) is ubiquitous in the marine environment. Previous work has shown that microplastics may be ingested and inhaled by the shore crab Carcinus maenas, although the biological consequences are unknown. Here, we show that acute aqueous exposure to polystyrene microspheres (8 μm) with different surface coatings had significant but transient effects on branchial function. Microspheres inhaled into the gill chamber had a small but significant dose-dependent effect on oxygen consumption after 1 h of exposure, returning to normal levels after 16 h. Ion exchange was also affected, with a small but significant decrease in hemolymph sodium ions and an increase in calcium ions after 24 h post-exposure. To further asses the effects on osmoregulation, we challenged crabs with reduced salinity after microplastic exposure. Neither microspheres nor natural sediments altered the crab's response to osmotic stress regardless of plastic concentration added. Carboxylated (COOH) and aminated (NH2) polystyrene microspheres were distributed differently across the gill surface, although neither had a significant adverse impact on gill function. These results illustrate the extent of the physiological effects of microplastics compared to the physiological resilience of shore crabs in maintaining osmoregulatory and respiratory function after acute exposure to both anthropogenic plastics and natural particles.


The Hemodynamic Response of Spreading Depolarization Observed by Near Infrared Spectroscopy After Aneurysmal Subarachnoid Hemorrhage.
Electrocorticography (ECoG) in brain-injured patients allows to detect spreading depolarization, a potential mechanism of secondary ischemia. Here, we describe the relationship of spreading depolarization with changes in cerebral hemodynamics using a brain tissue probe applying near infrared spectroscopy (NIRS). Simultaneous ECoG and NIRS monitoring was performed in a patient with severe aneurysmal subarachnoid hemorrhage. Changes in cerebral blood oxygenation and regional cerebral blood volume were studied before and after the occurrence of spreading depolarization. Cerebral blood flow measurements were performed daily using an indocyanine green dye dilution mode. Single events of spreading depolarizations demonstrated with transient hyperoxic responses and increase in cerebral blood volume. On the other hand, temporal clusters of recurrent spreading depolarizations were associated with prolonged hypoxic responses and decrease in cerebral blood volume. Cerebral blood flow measurements showed higher values before compared to after onset of spreading depolarization (33.7 ± 8.4 vs. 24.2 ± 4.5 ml/100 g/min). The findings suggest that NIRS monitoring in the cerebral white matter might reflect the hemodynamic signature of spreading depolarization detected by ECoG recordings. This is of potential interest for the further development of both neuromonitoring methods.


Early feeding practices and associated factors in Sudan: a cross-sectional analysis from multiple Indicator cluster survey.
In efforts to reduce neonatal mortality, the World Health Organization (WHO) has included breastfeeding among its recommended packages of interventions. Early initiation of breastfeeding and avoidance of prelacteal feeding are key contributors to optimal feeding practices. This study aims to assess the prevalence and associated factors of early breastfeeding practices in Sudan. This study utilises the cross-sectional nationally-representative Sudan Multiple Indicator Cluster Survey (MICS) conducted in 2014. The sample includes women who had a live birth in the two years before the survey and their self-report on early breastfeeding practices, namely early initiation and prelacteal feeding. Percentages of these early breastfeeding practices indicators were estimated accounting for the complex survey design. Multivariable logistic regression analyses were used to examine the factors associated with these outcomes. Of 5622 mothers, 69% initiated breastfeeding within one hour of birth, 72% avoided prelacteal feeding in the first three days after birth, and 51% met the criteria for both (i.e. practised optimal early feeding practice). Optimal early feeding varied across regions of Sudan. Birth by caesarean section (Adjusted Odds Ratio [AOR] 0.34; 95% CI 0.25, 0.47) and at a health facility (AOR 0.75; 95% CI 0.60, 0.94) were negatively associated with optimal early feeding practice. Mothers with secondary education (AOR 1.62; 95% CI 1.30, 2.02), those who desired their pregnancy at the time (AOR 1.31; 95% CI 1.08, 1.60), those who were assisted by a skilled birth attendant at birth (AOR 1.48; 95% CI 1.19, 1.83), and those who gave birth to female infants (AOR 1.16; 95% CI 1.02, 1.33) had higher odds of use optimal early feeding practice. Similarly, the odds of optimal early feeding increased with parity and maternal age. Only half of Sudanese mothers practised optimal early feeding practice, with important differences between regions in the country. Early feeding practices in Sudan are associated with various maternal, child and community level factors. The findings suggest the need to develop breastfeeding promotion programs with consideration of regional variations and healthcare system interventions.


Disease dynamics of Porites bleaching with tissue loss: prevalence, virulence, transmission, and environmental drivers.
The prevalence, number of species affected, and geographical extent of coral diseases have been increasing worldwide. We present ecological data on the coral disease Porites bleaching with tissue loss (PBTL) from Kaneohe Bay, Oahu (Hawaii, USA), affecting P. compressa. This disease is prevalent throughout the year, although it shows spatio-temporal variability with peak prevalence during the warmer summer months. Temporal variability in disease prevalence showed a strong positive relationship with elevated water temperature. Spatially, PBTL prevalence peaked in clearer waters (lower turbidity) with higher water flow and higher densities of parrotfish, together explaining approximately 26% of the spatial variability in PBTL prevalence. However, the relatively poor performance of the spatial model suggests that other, unmeasured factors may be more important in driving spatial prevalence. PBTL was not transmissible through direct contact or the water column in controlled aquaria experiments, suggesting that this disease may not be caused by a pathogen, is not highly infectious, or perhaps requires a vector for transmission. In general, PBTL results in partial tissue mortality of affected colonies; on average, one-third of the tissue is lost. This disease can affect the same colonies repeatedly, suggesting a potential for progressive damage which could cause increased tissue loss over time. P. compressa is the main framework-building species in Kaneohe Bay; PBTL therefore has the potential to negatively impact the structure of the reefs at this location.


The mammalian single-minded (SIM) gene: mouse cDNA structure and diencephalic expression indicate a candidate gene for Down syndrome.
We have recently isolated a human homolog (hSIM) of the Drosophila single-minded (sim) gene from the Down syndrome critical region of chromosome 21 using the exon trapping method. The Drosophila sim gene encodes a transcription factor that regulates the development of the central nervous system midline cell lineage. To elucidate the structure of the mammalian SIM protein, we have isolated cDNA clones from a mouse embryo cDNA library. The cDNA clones encode a polypeptide of 657 amino acids with a bHLH (basic-helix-loop-helix) domain, characteristic of a large family of transcription factors, and a PAS (Per-Arnt-Sim) domain in the amino-terminal half region. Both of these domains have striking sequence homology with human SIM and Drosophila SIM proteins. In contrast, the carboxy-terminal half of the mouse SIM protein consists of a proline-rich region with no sequence homology to the Drosophila SIM protein. A similar proline-rich domain is known for the activator domain of a number of transcription factors. Whole-mount embryo in situ hybridization experiments revealed that the SIM mRNA is expressed prominently in the diencephalon of mouse embryos at 8-9.5 days postcoitum. The structural characteristics of the mouse SIM protein and its expression in the diencephalon during embryogenesis strongly suggest that the newly isolated mammalian SIM homolog may play a critical role in the development of the mammalian central nervous system. We propose that the human SIM gene may be one of the pathogenic genes of Down syndrome.


Cemented versus cementless hemiarthroplasty for intracapsular neck of femur fracture--a comparison of 60,848 matched patients using national data.
National guidelines recommend cemented hemiarthroplasty for intracapsular fractured neck of femur (NOF), based on evidence of less pain, better mobility and lower costs. We aimed to compare complications following cemented and cementless implants, using the national hospital episode statistics (HES) database in England. Dislocation, revision, return to theatre and medical complications were extracted for all patients with NOF fracture who underwent hemiarthroplasty between January 2005 and December 2008. To make a 'like for like' comparison all 30,424 patients with a cementless implant were matched to 30,424 cemented implants (from a total of 42,838) in terms of age, sex and Charlson co-morbidity score. In the cementless group, 18-month revision (1.62% versus 0.57% (OR 2.90, p<0.001)), 4-year revision (2.45% versus 1.11% (OR 2.28, p<0.001)) and 30-day chest infection (8.14% versus 7.23% (OR 1.14, p=0.028)) were significantly higher. Four-year dislocation rate was higher in cemented implants (0.60% versus 0.26% (OR 0.45, p<0.001)). No significant differences were seen in return to theatre or other medical complications. In this national analysis of matched patients mid-term revision and perioperative chest infection was significantly higher in the cementless group. This supports the published evidence and national guidelines recommending cement fixation of hemiarthroplasty.


Mirizzi syndrome.
Mirizzi syndrome is an important complication of gallstone disease. If not recognized preoperatively, it can result in significant morbidity and mortality. Preoperative diagnosis may be difficult despite the availability of multiple imaging modalities. Ultrasonography (US), CT, and magnetic resonance cholangiopancreatography (MRCP) are common initial tests for suspected Mirizzi syndrome. Typical findings on US suggestive of Mirizzi syndrome are a shrunken gallbladder, impacted stone(s) in the cystic duct, a dilated intrahepatic tree, and common hepatic duct with a normal-sized common bile duct. The main role of CT is to differentiate Mirizzi syndrome from a malignancy in the area of porta hepatis or in the liver. MRI and MRCP are increasingly playing an important role and have the additional advantage of showing the extent of inflammation around the gallbladder that can help in the differentiation of Mirizzi syndrome from other gallbladder pathologies such as gallbladder malignancy. Endoscopic retrograde cholangiopancreatography (ERCP) is the gold standard in the diagnosis of Mirizzi syndrome. It delineates the cause, level, and extent of biliary obstruction, as well as ductal abnormalities, including fistula. ERCP also offers a variety of therapeutic options, such as stone extraction and biliary stent placement. Percutaneous cholangiogram can provide information similar to ERCP; however, ERCP has an additional advantage of identifying a low-lying cystic duct that may be missed on percutaneous cholangiogram. Wire-guided intraductal US can provide high-resolution images of the biliary tract and adjacent structures. Treatment is primarily surgical. Open surgery is the current standard for managing patients with Mirizzi syndrome. Good short- and long-term results with low mortality and morbidity have been reported with open surgical management. Laparoscopic management is contraindicated in many patients because of the increased risk of morbidity and mortality associated with this approach. Endoscopic treatment may serve as an alternative in patients who are poor surgical candidates, such as elderly patients or those with multiple comorbidities. Endoscopic treatment also can serve as a temporizing measure to provide biliary drainage in preparation for an elective surgery.


Ligand-Protein Affinity Studies Using Long-Lived States of Fluorine-19 Nuclei.
The lifetimes TLLS of long-lived states or TLLC of long-lived coherences can be used for the accurate determination of dissociation constants of weak protein-ligand complexes. The remarkable contrast between signals derived from LLS or LLC in free and bound ligands can be exploited to search for weak binders with large dissociation constants KD > 1 mM that are important for fragment-based drug discovery but may escape detection by other screening techniques. Alternatively, the high sensitivity of the proposed method can be exploited to work with large ligand-to-protein ratios, with an evident advantage of reduced consumption of precious proteins. The detection of (19)F-(19)F long-lived states in suitably designed fluorinated spy molecules allows one to perform competition binding experiments with high sensitivity while avoiding signal overlap that tends to hamper the interpretation of proton spectra of mixtures.


The important but weakening maize yield benefit of grain filling prolongation in the US Midwest.
A better understanding of recent crop yield trends is necessary for improving the yield and maintaining food security. Several possible mechanisms have been investigated recently in order to explain the steady growth in maize yield over the US Corn-Belt, but a substantial fraction of the increasing trend remains elusive. In this study, trends in grain filling period (GFP) were identified and their relations with maize yield increase were further analyzed. Using satellite data from 2000 to 2015, an average lengthening of GFP of 0.37 days per year was found over the region, which probably results from variety renewal. Statistical analysis suggests that longer GFP accounted for roughly one-quarter (23%) of the yield increase trend by promoting kernel dry matter accumulation, yet had less yield benefit in hotter counties. Both official survey data and crop model simulations estimated a similar contribution of GFP trend to yield. If growing degree days that determines the GFP continues to prolong at the current rate for the next 50 years, yield reduction will be lessened with 25% and 18% longer GFP under Representative Concentration Pathway 2.6 (RCP 2.6) and RCP 6.0, respectively. However, this level of progress is insufficient to offset yield losses in future climates, because drought and heat stress during the GFP will become more prevalent and severe. This study highlights the need to devise multiple effective adaptation strategies to withstand the upcoming challenges in food security.


ACCELERATE and European Medicines Agency Paediatric Strategy Forum for medicinal product development of checkpoint inhibitors for use in combination therapy in paediatric patients.
The third multistakeholder Paediatric Strategy Forum organised by ACCELERATE and the European Medicines Agency focused on immune checkpoint inhibitors for use in combination therapy in children and adolescents. As immune checkpoint inhibitors, both as monotherapy and in combinations have shown impressive success in some adult malignancies and early phase trials in children of single agent checkpoint inhibitors have now been completed, it seemed an appropriate time to consider opportunities for paediatric studies of checkpoint inhibitors used in combination. Among paediatric patients, early clinical studies of checkpoint inhibitors used as monotherapy have demonstrated a high rate of activity, including complete responses, in Hodgkin lymphoma and hypermutant paediatric tumours. Activity has been very limited, however, in more common malignancies of childhood and adolescence. Furthermore, apart from tumour mutational burden, no other predictive biomarker for monotherapy activity in paediatric tumours has been identified. Based on these observations, there is collective agreement that there is no scientific rationale for children to be enrolled in new monotherapy trials of additional checkpoint inhibitors with the same mechanism of action of agents already studied (e.g. anti-PD1, anti-PDL1 anti-CTLA-4) unless additional scientific knowledge supporting a different approach becomes available. This shared perspective, based on scientific evidence and supported by paediatric oncology cooperative groups, should inform companies on whether a paediatric development plan is justified. This could then be proposed to regulators through the available regulatory tools. Generally, an academic-industry consensus on the scientific merits of a proposal before submission of a paediatric investigational plan would be of great benefit to determine which studies have the highest probability of generating new insights. There is already a rationale for the evaluation of combinations of checkpoint inhibitors with other agents in paediatric Hodgkin lymphoma and hypermutated tumours in view of the activity shown as single agents. In paediatric tumours where no single agent activity has been observed in multiple clinical trials of anti-PD1, anti-PDL1 and anti-CTLA-4 agents as monotherapy, combinations of checkpoint inhibitors with other treatment modalities should be explored when a scientific rationale indicates that they could be efficacious in paediatric cancers and not because these combinations are being evaluated in adults. Immunotherapy in the form of engineered proteins (e.g. monoclonal antibodies and T cell engaging agents) and cellular products (e.g. CAR T cells) has great therapeutic potential for benefit in paediatric cancer. The major challenge for developing checkpoint inhibitors for paediatric cancers is the lack of neoantigens (based on mutations) and corresponding antigen-specific T cells. Progress critically depends on understanding the immune macroenvironment and microenvironment and the ability of the adaptive immune system to recognise paediatric cancers in the absence of high neoantigen burden. Future clinical studies of checkpoint inhibitors in children need to build upon strong biological hypotheses that take into account the distinctive immunobiology of childhood cancers in comparison to that of checkpoint inhibitor responsive adult cancers.


Within- and between-culture variation: individual differences and the cultural logics of honor, face, and dignity cultures.
The CuPS (Culture × Person × Situation) approach attempts to jointly consider culture and individual differences, without treating either as noise and without reducing one to the other. Culture is important because it helps define psychological situations and create meaningful clusters of behavior according to particular logics. Individual differences are important because individuals vary in the extent to which they endorse or reject a culture's ideals. Further, because different cultures are organized by different logics, individual differences mean something different in each. Central to these studies are concepts of honor-related violence and individual worth as being inalienable versus socially conferred. We illustrate our argument with 2 experiments involving participants from honor, face, and dignity cultures. The studies showed that the same "type" of person who was most helpful, honest, and likely to behave with integrity in one culture was the "type" of person least likely to do so in another culture. We discuss how CuPS can provide a rudimentary but integrated approach to understanding both within- and between-culture variation.


The effects of super-flux (high performance) dialyzer on plasma glycosylated pro-B-type natriuretic peptide (proBNP) and glycosylated N-Terminal proBNP in end-stage renal disease patients on dialysis.
Plasma BNP levels are predictive of prognosis in hemodialysis patients. However, recent studies showed that the current BNP immunoassay cross-reacts with glycosylated proBNP, and the NT-proBNP assay underestimates glycosylated NT-proBNP. In addition, the recently developed high performance dialyzer removes medium-sized molecular solutes such as β2-microgloburin. We therefore investigated the effects of high performance dialysis on measured levels of glycosylated proBNP, glycosylated NT-proBNP and other BNP-related peptides in end-stage renal disease (ESRD) patients on hemodialysis. The relationships between clinical parameters and BNP-related molecule were also investigated. We used our newly developed immunoassay to measure plasma total BNP and proBNP in 105 normal subjects and 36 ESRD patients before and after hemodialysis. Plasma NT-proBNP was measured using Elecsys II after treatment with or without deglycosylating enzymes. We also measured plasma ANP and cGMP using radioimmunoassays. All the measured BNP-related peptides were significantly higher in ESRD patients than healthy subjects. Total BNP (-38.9%), proBNP (-29.7%), glycoNT-proBNP (-45.5%), nonglycoNT-proBNP (-53.4%), ANP (-50.4%) and cGMP (-72.1%) were all significantly reduced after hemodialysis, and the magnitude of the reduction appeared molecular weight- dependent. Both the proBNP/total BNP and glycoNT-proBNP/nonglycoNT-proBNP ratios were increased after hemodialysis. The former correlated positively with hemodialysis vintage and negatively with systolic blood pressure, while the latter correlated positively with parathyroid hormone levels. These results suggest that hemodialysis using super-flux dialyzer removes BNP-related peptides in a nearly molecular weight-dependent manner. The ProBNP/total BNP and glycoNT-proBNP/nonglycoNT-proBNP ratios appear to be influenced by hemodialysis-related parameters in ESRD patients on hemodialysis.


Collective Pulsing in Xeniid Corals: Part I-Using Computer Vision and Information Theory to Search for Coordination.
Xeniid corals (Cnidaria: Alcyonacea), a family of soft corals, include species displaying a characteristic pulsing behavior. This behavior has been shown to increase oxygen diffusion away from the coral tissue, resulting in higher photosynthetic rates from mutualistic symbionts. Maintaining such a pulsing behavior comes at a high energetic cost, and it has been proposed that coordinating the pulse of individual polyps within a colony might enhance the efficiency of fluid transport. In this paper, we test whether patterns of collective pulsing emerge in coral colonies and investigate possible interactions between polyps within a colony. We video recorded different colonies of Heteroxenia sp. in a laboratory environment. Our methodology is based on the systematic integration of a computer vision algorithm (ISOMAP) and an information-theoretic approach (transfer entropy), offering a vantage point to assess coordination in collective pulsing. Perhaps surprisingly, we did not detect any form of collective pulsing behavior in the colonies. Using artificial data sets, however, we do demonstrate that our methodology is capable of detecting even weak information transfer. The lack of a coordination is consistent with previous work on many cnidarians where coordination between actively pulsing polyps and medusa has not been observed. In our companion paper, we show that there is no fluid dynamic benefit of coordinated pulsing, supporting this result. The lack of coordination coupled with no obvious fluid dynamic benefit to grouping suggests that there may be non-fluid mechanical advantages to forming colonies, such as predator avoidance and defense.


Transboundary spread of highly pathogenic avian influenza through poultry commodities and wild birds: a review.
The extensive circulation of the H5N1 highly pathogenic avian influenza (HPAI) virus in animals and the human health implications which it poses have led to extensive research in unexplored fields and thus a re-assessment of our understanding of this infection. Moreover, widespread infection of poultry has raised concerns about the food safety and trade implications of this infection, necessitating revised international trade regulations. The role of wild birds has been much debated and resources have been invested to clarify the role that they may play in the spread of infection. It is now clear that wild birds may be responsible for primary introduction in a previously free area. To date it is still unclear whether HPAI infection may be maintained in wild bird populations for extended periods of time. This paper reviews existing knowledge on the transboundary spread of HPAI through poultry and poultry commodities and summarises evidence of spread through wild birds.


Noninvasive detection of human cardiac transplant rejection with indium-111 antimyosin (Fab) imaging.
Diagnosis of rejection after cardiac transplantation is currently made by right ventricular endomyocardial biopsy. To evaluate antimyosin imaging as a noninvasive means of detecting human cardiac rejection, the Fab fragment of murine monoclonal antimyosin antibodies was labeled with indium-111 and given intravenously to 18 patients (age 45 +/- 12 years) in 20 studies 7 days to 9 years after transplantation. Endomyocardial biopsy specimens were obtained at the time of each imaging study. Eight patients had positive scans confirmed by biopsy as rejection, and eight patients had negative scans and no evidence of rejection on biopsy. Discordance was observed in four studies, two with positive scans and no rejection on biopsy and two with negative scans and positive biopsy. The sensitivity, specificity, and overall accuracy of the technique were each 80%. Imaging with radiolabeled antimyosin antibody Fab fragments may be of value in the noninvasive identification of rejection in the cardiac transplant recipient.


Retroauricular Endoscope-Assisted Approach to the Neck: Early Experience in Latin America.
Introduction There has been a significant increase in concern towards improving aesthetic and functional outcomes without compromising the oncologic effectiveness in head and neck surgery. In this subset, endoscope-assisted and robotic procedures allowed the development of new approaches to the neck, including the retroauricular access, which is now routinely used, especially in Korea. Objectives This study aims to provide a descriptive analysis of our initial experience with retroauricular endoscope-assisted approach assessing feasibility, safety, and aesthetic results. Methods Prospective analysis of the first 11 eligible patients submitted to retroauricular endoscope-assisted approach for neck procedures in the Head and Neck Surgery Department at AC Camargo Cancer Center. Results A total of 18 patients were included in this study, comprising 7 supraomohyoid neck dissections, 8 submandibular gland excisions, 3 thyroid lobectomies, and one paraganglioma excision. There was no significant local complications, surgical accident, or need for conversion into conventional open procedure in this series. Conclusion Our initial experience has shown us that this approach is feasible, safe, oncologically efficient, and applicable to selected cases, with a clear cosmetic benefit.


β-Cell-specific protein kinase A activation enhances the efficiency of glucose control by increasing acute-phase insulin secretion.
Acute insulin secretion determines the efficiency of glucose clearance. Moreover, impaired acute insulin release is characteristic of reduced glucose control in the prediabetic state. Incretin hormones, which increase β-cell cAMP, restore acute-phase insulin secretion and improve glucose control. To determine the physiological role of the cAMP-dependent protein kinase (PKA), a mouse model was developed to increase PKA activity specifically in the pancreatic β-cells. In response to sustained hyperglycemia, PKA activity potentiated both acute and sustained insulin release. In contrast, a glucose bolus enhanced acute-phase insulin secretion alone. Acute-phase insulin secretion was increased 3.5-fold, reducing circulating glucose to 58% of levels in controls. Exendin-4 increased acute-phase insulin release to a similar degree as PKA activation. However, incretins did not augment the effects of PKA on acute-phase insulin secretion, consistent with incretins acting primarily via PKA to potentiate acute-phase insulin secretion. Intracellular calcium signaling was unaffected by PKA activation, suggesting that the effects of PKA on acute-phase insulin secretion are mediated by the phosphorylation of proteins involved in β-cell exocytosis. Thus, β-cell PKA activity transduces the cAMP signal to dramatically increase acute-phase insulin secretion, thereby enhancing the efficiency of insulin to control circulating glucose.


Response to the COVID-19 epidemic: a report from Shenzhen, China.
The whole world is now facing an unprecedented pandemic with over 1.8 million confirmed cases and more than one hundred thousand deaths. To counter the pandemic, Shenzhen created a central command and control structure based on the only designated hospital- Shenzhen Third People's Hospital which is a large general hospital specialized on infectious diseases in the bay area. The hospital has taken many decisive and effective actions to respond to the epidemic. Here, we will describe and share healthcare experiences from Shenzhen and call for international cooperation and collaboration.


Clinical management with efalizumab of a patient with psoriasis and comorbid vitiligo.
Although the precise pathogenesis is not known, vitiligo appears to be an autoimmune disease involving T cell-mediated melanocyte destruction. Efalizumab, a recombinant, humanized monoclonal antibody, targets T cells, the key mediators of the immunopathogenesis of psoriasis. Although a concomitant presentation of vitiligo with psoriasis is uncommon, several cases have been reported previously in the literature. A case of a patient with vitiligo and psoriasis who was treated with efalizumab to alleviate the symptoms of psoriasis is described. Over the course of the diseases, the patient had been treated unsuccessfully with numerous therapies. The patient initiated efalizumab with a 0.7 mg/kg conditioning dose and then continued on 1 mg/kg weekly. After 2 months of efalizumab therapy, the psoriasis symptoms were reduced, and the vitiligo had visibly improved in some areas. The patient has remained on efalizumab therapy with no evidence of an exacerbation of vitiligo. The management of acute flares is also discussed. This case is illustrative of a patient with psoriasis and vitiligo who was treated successfully with efalizumab.


Electropolymerized molecularly imprinted polypyrrole film for sensing of clofibric acid.
Piezoelectric quartz crystals and analogous gold substrates were electrochemically coated with molecularly imprinted polypyrrole films for pulsed amperometric detection (PAD) of clofibric acid, a metabolite of clofibrate. Cyclic voltammetry data obtained during polymerization and deposited weight estimations revealed a decrease of the polymerization rate with increasing clofibric acid concentration. XPS measurements indicated that clofibric acid could be removed after imprinting with an aqueous ethanol solution, which was further optimized by using PAD. Zeta potential and contact angle measurements revealed differences between molecularly imprinted (MIP) and non-imprinted polymer (NIP) layers. Binding experiments with clofibric acid and other substances showed a pronounced selectivity of the MIP for clofibric acid vs. carbamazepine, but the response of MIP and NIP to 2,4-dichlorophenoxyacetic acid was higher than that for clofibric acid. A smooth surface, revealed by AFM measurements, with roughness of 6-8 nm for imprinted and non-imprinted layers, might be a reason for an excessively low density of specific binding sites for clofibric acid. Furthermore, the decreased polymerization rate in the presence of clofibric acid might not result in well-defined polymer structures, which could be the reason for the lower sensitivity.


Comparison of Object-Based Image Analysis Approaches to Mapping New Buildings in Accra, Ghana Using Multi-Temporal QuickBird Satellite Imagery.
The goal of this study was to map and quantify the number of newly constructed buildings in Accra, Ghana between 2002 and 2010 based on high spatial resolution satellite image data. Two semi-automated feature detection approaches for detecting and mapping newly constructed buildings based on QuickBird very high spatial resolution satellite imagery were analyzed: (1) post-classification comparison; and (2) bi-temporal layerstack classification. Feature Analyst software based on a spatial contextual classifier and ENVI Feature Extraction that uses a true object-based image analysis approach of image segmentation and segment classification were evaluated. Final map products representing new building objects were compared and assessed for accuracy using two object-based accuracy measures, completeness and correctness. The bi-temporal layerstack method generated more accurate results compared to the post-classification comparison method due to less confusion with background objects. The spectral/spatial contextual approach (Feature Analyst) outperformed the true object-based feature delineation approach (ENVI Feature Extraction) due to its ability to more reliably delineate individual buildings of various sizes. Semi-automated, object-based detection followed by manual editing appears to be a reliable and efficient approach for detecting and enumerating new building objects. A bivariate regression analysis was performed using neighborhood-level estimates of new building density regressed on a census-derived measure of socio-economic status, yielding an inverse relationship with R2 = 0.31 (n = 27; p = 0.00). The primary utility of the new building delineation results is to support spatial analyses of land cover and land use and demographic change.


Mortality, length of stay, and inpatient charges for heart failure patients at public versus private hospitals in South Korea.
This study compared in-hospital mortality within 30 days of admission, lengths of stay, and inpatient charges among patients with heart failure admitted to public and private hospitals in South Korea. We obtained health insurance claims data for all heart failure inpatients nationwide between November 1, 2011 and May 31, 2012. These data were then matched with hospital-level data, and multi-level regression models were examined. A total of 8406 patients from 253 hospitals, including 31 public hospitals, were analyzed. The in-hospital mortality rate within 30 days of admission was 0.92% greater and the mean length of stay was 1.94 days longer at public hospitals than at private hospitals (mortality: 5.18% and 4.26%, respectively; LOS: 12.08 and 10.14 days, respectively). The inpatient charges were 11.4% lower per case and 24.5% lower per day at public hospitals than at private hospitals. After adjusting for patient- and hospital-level confounders, public hospitals had a 1.62-fold higher in-hospital mortality rate, a 16.5% longer length of stay, and an 11.7% higher inpatient charge per case than private hospitals, although the charges of private hospitals were greater in univariate analysis. We recommend that government agencies and policy makers continue to monitor quality of care, lengths of stay in the hospital, and expenditures according to type of hospital ownership to improve healthcare outcomes and reduce spending.


Barriers to and enablers of uptake of and adherence to antiretroviral therapy in the context of integrated HIV and tuberculosis treatment among adults in sub-Saharan Africa: a protocol for a systematic literature review.
The scale-up of integrated Human Immunodeficiency Virus (HIV) and tuberculosis (TB) treatment has been an important intervention to curb the burden of HIV and TB co-infection worldwide. Uptake of and adherence to antiretroviral therapy (ART) are key determinants of the quality and therapeutic endpoints of this intervention. This study aims to conduct an up-to-date collection and synthesis of evidence on barriers to and facilitators of uptake of and adherence to ART in HIV/TB integrated treatment programs in sub-Saharan Africa (SSA). A systematic review of peer-reviewed literature on the uptake of and adherence to ART in the context of integrated therapy for HIV and TB in SSA will be performed. We will review qualitative and quantitative studies reporting on the uptake of and adherence to ART during integrated treatment for TB and HIV among adults. These will include studies that involve HIV-infected TB patients initiating ART and studies involving PLWHA already on ART who are newly diagnosed with TB. Qualitative studies, quantitative studies, randomised trials and observational studies will be included. Six databases including Medline and Embase will be searched for relevant studies published from March 2004 to July 2019. Two authors will independently screen the search output and retrieve full texts of eligible studies. Disagreements between the two authors will be resolved by arbitration by a third author. Data will be abstracted from the eligible studies and synthesis will be done through descriptive synthesis for qualitative data and meta-analysis for quantitative data. This study will be a review of the literature and will not involve primary collection of individuals' data. Amendments to the protocol will be documented in the final review. The final study will be published in a peer-reviewed journal and presented at conferences. The review is expected to contribute to improving strategies to enhance uptake of and adherence to ART in integrated care. CRD42019131933.


Reversible middle cerebral artery embolization in dogs without intracranial surgery.
Using dogs, we developed an intravascular model for reversible middle cerebral artery occlusion that does not involve intracranial surgery or enucleation. Using silicone plastic plugs with a suture embedded within them, we embolized the middle cerebral artery in 19 dogs via the cervical carotid artery. The free end of the suture remained accessible in the neck, and after variable dwell times traction was placed on the suture and the plug was withdrawn. Placement of the plug in the middle cerebral artery produced ischemia in the basal ganglia. The degree and distribution of cortical ischemia were variable as evidence by the pathologically documented scattered nature of infarcts that resulted when the plug was left permanently in the middle cerebral artery and when it was removed after 1 or 2 hours. Angiography demonstrated occlusion of the middle cerebral artery with the plug in place as well as reperfusion when the plug was withdrawn. This modification of a previously described model of middle cerebral artery occlusion provides an opportunity to study structural, physiologic, and biochemical events occurring in acutely hypoperfused cerebral tissue as well as critical changes leading to irreversible injury without the disadvantages of surgical manipulation required by all previous models of reversible cerebral ischemia.


Major adverse cardiovascular events in non-valvular atrial fibrillation with chronic obstructive pulmonary disease: the ARAPACIS study.
Chronic obstructive pulmonary disease (COPD) increases the risk of mortality in non-valvular atrial fibrillation (NVAF) patients. Data on the relationship of COPD to major cardiovascular events (MACE) in AF have not been defined. The aim of the study is to assess the predictive value of COPD on incident MACE in NVAF patients over a 3-year follow-up. In the Atrial Fibrillation Registry for Ankle-Brachial Index Prevalence Assessment-Collaborative Italian Study (ARAPACIS) cohort, we evaluate the impact of COPD on the following clinical endpoints: MACE (including vascular death, fatal/non-fatal MI and stroke/TIA), cardiovascular (CV) death and all-cause mortality. Among 2027 NVAF patients, patients with COPD (9%) are more commonly male, elderly and at higher thromboembolic risk. During a median 36.0 months follow-up, 186 patients experienced MACE: vascular death (n = 72), MI (n = 57), stroke/TIA (n = 57). All major outcomes (including stroke/TIA, MI, vascular death, and all-cause death) are centrally adjudicated. Kaplan-Meier curves show that NVAF patients with COPD are at higher risk for MACE (p < 0.001), CV death (p < 0.001) and all-cause death (p < 0.001). On Cox proportional hazard analysis, COPD is an independent predictor of MACE (Hazard ratio [HR] 1.77, 95% Confidence Intervals [CI] 1.20-2.61; p = 0.004), CV death (HR 2.73, 95% CI 1.76-4.23; p < 0.0001) and all-cause death (HR 2.16, 95% CI 1.48-3.16; p < 0.0001). COPD is an independent predictor of MACE, CV death and all-cause death during a long-term follow-up of NVAF patients.


Albumin versus crystalloid solutions in patients with the acute respiratory distress syndrome: a systematic review and meta-analysis.
In patients with acute respiratory distress syndrome (ARDS) fluid therapy might be necessary. The aim of this systematic review and meta-analysis is to determine the effects of colloid therapy compared to crystalloids on mortality and oxygenation in adults with ARDS. Randomized controlled trials (RCTs) were identified through a systematic literature search of MEDLINE, EMBASE, CENTRAL and LILACS. Articles published up to 15th February 2013 were independently screened, abstracted, and assessed (Cochrane Risk of Bias Tool) to provide evidence-based therapy recommendations. RCTs were eligible if they compared colloid versus crystalloid therapy on lung function, inflammation, damage or mortality in adults with ARDS. Primary outcome parameters were respiratory mechanics, gas exchange lung inflammation and damage as well as hospital mortality. Kidney function, need for renal replacement therapy, hemodynamic stabilization and intensive care unit (ICU) length of stay served as secondary outcomes. A total of 3 RCTs out of 4130 potential trials found in the databases were selected for qualitative and quantitative analysis totaling 206 patients who received either albumin or saline. Overall risk of bias was unclear to high in the identified trials. Calculated pooled risk of death was not statistically significant (albumin 34 of 100 (34.0%) versus 40 of 104 (38.5%), relative risk (RR) = 0.89, 95% confidence interval (CI) 0.62 to 1.28, P = 0.539). Weighted mean difference (WMD) in PaO2/FiO2 (mmHg) improved in the first 48 hours (WMD = 62, 95% CI 47 to 77, P <0.001, I2 = 0%) after therapy start and remained stable after 7 days (WMD = 20, 95% CI 4 to 36, P = 0.017, I2 = 0%). There is a high need for RCTs investigating the effects of colloids in ARDS patients. Based on the findings of this review, colloid therapy with albumin improved oxygenation but did not affect mortality.


Characterization of major histocompatibility complex (MHC) DRB exon 2 and DRA exon 3 fragments in a primary terrestrial rabies vector (Procyon lotor).
The major histocompatibility complex (MHC) presents a unique system to explore links between genetic diversity and pathogens, as diversity within MHC is maintained in part by pathogen driven selection. While the majority of wildlife MHC studies have investigated species that are of conservation concern, here we characterize MHC variation in a common and broadly distributed species, the North American raccoon (Procyon lotor). Raccoons host an array of broadly distributed wildlife diseases (e.g., canine distemper, parvovirus and raccoon rabies virus) and present important human health risks as they persist in high densities and in close proximity to humans and livestock. To further explore how genetic variation influences the spread and maintenance of disease in raccoons we characterized a fragment of MHC class II DRA exon 3 (250 bp) and DRB exon 2 (228 bp). MHC DRA was found to be functionally monomorphic in the 32 individuals screened; whereas DRB exon 2 revealed 66 unique alleles among the 246 individuals screened. Between two and four alleles were observed in each individual suggesting we were amplifying a duplicated DRB locus. Nucleotide differences between DRB alleles ranged from 1 to 36 bp (0.4-15.8% divergence) and translated into 1 to 21 (1.3-27.6% divergence) amino acid differences. We detected a significant excess of nonsynonymous substitutions at the peptide binding region (P = 0.005), indicating that DRB exon 2 in raccoons has been influenced by positive selection. These data will form the basis of continued analyses into the spatial and temporal relationship of the raccoon rabies virus and the immunogenetic response in its primary host.


Effect of Interscalene Brachial Plexus Block on the Pulmonary Function of Obese Patients: A Prospective, Observational Cohort Study.
The effect of interscalene block (ISB) on pulmonary function of obese participants has not been investigated. The goal of this study is to assess the association of obesity (body mass index [BMI] >29 kg/m vs BMI <25 kg/m) and change in forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1) after ISB in participants undergoing outpatient shoulder surgery. This prospective, observational cohort study compared obese (BMI >29 kg/m) and normal-weight (BMI <25 kg/m) groups undergoing ISB for ambulatory shoulder surgery, on preblock and postblock FVC and FEV1, at 30 minutes postblock and in the postanesthesia care unit (PACU). The primary outcome in this study was FVC% change (percentage change from preblock to postblock values of FVC) at 30 minutes postblock in the supine position. Secondary outcomes included FVC% change at PACU and in the sitting position, FEV1% change (percentage change from preblock to postblock values of FEV1), FVC, FEV1, incidence of diaphragmatic paresis, modified Borg scale for perceived dyspnea, Richmond Agitation-Sedation Scale scores for sedation, and intraoperative airway events. Fourteen participants were recruited to each group. The mean (standard deviation) BMI in the normal-weight and obese groups was 23 (1.7) and 33 (3.1) kg/m, respectively. ISB success rate was 100%. All participants demonstrated hemidiaphragmatic paresis after ISB. Compared to the normal-weight group, in the sitting position, the obese group had a significant decrease in FVC% change at 30 minutes (-30 [10.5] vs -23 [7.2], P = .046) and an FEV1% change in the PACU (-40 [12.6] vs -27 [13.9], P = .02). No difference was found for measurements taken in the supine position. A repeated-measures analysis demonstrated that, adjusted for position, there is no significant group effect on FVC% change or FEV1% change from 30 minutes to PACU. The 2 groups were not different in terms of breathlessness and sedation at 30 minutes (P = .67, P = .48, respectively) and in the PACU (P = .69, P > .99, respectively) nor in the occurrence of intraoperative airway events (P > .99). ISB is associated with greater FVC and FEV1 reductions in obese participants undergoing shoulder surgery compared to normal-weight participants. Neither time (30 minutes versus PACU) nor position (sitting versus supine) affected this relationship. Despite these changes, obesity was not associated with increased clinical respiratory symptoms or events.


Deciphering Direct and Indirect Effects of Neurokinin B and GnRH in the Brain-Pituitary Axis of Tilapia.
Neurokinin B (NKB) and its cognate receptor (NK3R) are emerging as important components of the neuroendocrine regulation of reproduction. Unlike mammalian tac3, which encodes only one mature peptide (namely NKB), two mature peptides are predicted for each tac3 gene in fish and frogs. Therefore, it was designated as Neurokinin F (NKF). Hormone analogs with high and long-lasting biological activity are important tools for physiological and biological research; however, the availability of piscine-specific analogs is very limited. Therefore, we have developed specific NKB and NKF analogs based on the structure of the mammalian NKB analog-senktide. These analogs, specifically designed for longer half-lives by methylation of proteolysis sites, exhibited activity equal to those of the native NKB and NKF in short-term signal-transduction assays of tilapia NKB receptors. However, the analogs were found to be able to significantly increase the release of luteinizing hormone (LH), follicle stimulating hormone (FSH) and growth hormone (GH) in tilapia, as fast as 1 h after intraperitoneal (IP) injection. The impact of the analogs on LH and FSH secretion lasted longer compared to the effect of native peptides and salmon GnRH analog (sGnRHa). In addition, we harvested pituitaries 24 h post injection and measured LH, FSH and GH mRNA synthesis. Both analogs elevated mRNA levels of LH and GH, but only NKB analog increased FSH mRNA levels in the pituitary and all GnRH forms in the brain. NKB receptors were co-localized with all three types the GnRH neurons in tilapia brain in situ. We previously showed a direct effect of NKB at the pituitary level, and these new results suggest that the stronger impact of the NKB analog on GTH release is also due to an indirect effect through the activation of GnRH neurons. These results suggest that novel synthetic NKB analogs may serve as a tool for both research and agricultural purposes. Finally, the biological activity and regulatory role of NKB in tilapia brain and pituitary suggest that the NKB/NKBR system in fish is an important reproductive regulator in a similar way to the kisspeptin system in mammals.


Bordetella parapertussis isolates not expressing pertactin circulating in France.
Surprisingly, most Bordetella parapertussis isolates collected in France since 2007 do not express pertactin, owing to mutations in the structural gene encoding this protein. We used a 454 pyrosequencing strategy to study and compare the genetics of two B. parapertussis isolates (one expressing pertactin and one not expressing pertactin) and the reference strain. No region of difference was detected between the genomes of the two isolates and the genome of the reference strain. No increase in repeated sequences between both isolates was found, and there were very few sequence differences. Using cellular and animal models, we found no substantial difference between the pathogenicity of these B. parapertussis isolates, which is consistent with clinical data. The emergence of these isolates, indicating that pertactin expression is not essential for virulence for B. parapertussis, is discussed.


Evidence-based Risk Stratification for Sport Medicine Procedures During the COVID-19 Pandemic.
Orthopaedic practices have been markedly affected by the emergence of the COVID-19 pandemic. Despite the ban on elective procedures, it is impossible to define the medical urgency of a case solely on whether a case is on an elective surgery schedule. Orthopaedic surgical procedures should consider COVID-19-associated risks and an assimilation of all available disease dependent, disease independent, and logistical information that is tailored to each patient, institution, and region. Using an evidence-based risk stratification of clinical urgency, we provide a framework for prioritization of orthopaedic sport medicine procedures that encompasses such factors. This can be used to facilitate the risk-benefit assessment of the timing and setting of a procedure during the COVID-19 pandemic.


Inhibition of nitric oxide production aggravates myocardial hypoperfusion during exercise in the presence of a coronary artery stenosis.
Regulation of coronary vasomotor tone during myocardial hypoperfusion is incompletely understood. The present study was performed to test the hypothesis that endogenous production of nitric oxide contributes to resistance vessel dilation distal to a coronary artery stenosis that results in myocardial ischemia during exercise. Seven dogs instrumented with a Doppler velocity probe, hydraulic occluder, and indwelling microcatheter in the left anterior descending coronary artery (LAD) were studied during treadmill exercise in the presence of a coronary artery stenosis before and after intracoronary infusion of NG-nitro-L-arginine (LNNA, 20 mg/kg). This dose of LNNA inhibited the maximal increase in LAD flow produced by intracoronary acetylcholine by 82 +/- 5% but did not alter the response to intracoronary nitroprusside. Coronary pressure distal to the stenosis was maintained constant during the control period and after administration of LNNA. LNNA increased aortic and left ventricular systolic and end-diastolic pressures at rest and during exercise. During control in the absence of a stenosis, LNNA had no effect on coronary blood flow. In the presence of a stenosis that decreased distal coronary pressure to 55 +/- 2 mm Hg, mean myocardial blood flow measured with microspheres was 1.09 +/- 0.13 mL.min-1.g-1 in the LAD-dependent and 2.57 +/- 0.50 mL.min-1.g-1 in the posterior control region, respectively. With no change in distal coronary pressure, LNNA decreased mean myocardial blood flow in the LAD region to 0.68 +/- 0.11 mL.min-1.g-1 (P < .01). To avoid systemic hemodynamic effects, LNNA was administered in a dose of 1.5 mg/kg IC to four additional dogs. This low dose inhibited the coronary blood flow increases produced by acetylcholine by 61 +/- 5% but was devoid of systemic hemodynamic effects. During exercise in the presence of a coronary stenosis that decreased coronary pressure to 52 +/- 1 mm Hg, this dose of LNNA decreased mean myocardial blood flow from 0.89 +/- 0.23 to 0.66 +/- 0.21 mL.min-1.g-1 (P < .02). These data demonstrate that nitric oxide contributes to the maintenance of myocardial perfusion distal to a flow-limiting coronary artery stenosis during exercise.


Antiphospholipid antibodies and antiphospholipid syndrome in patients presenting with immune thrombocytopenic purpura: a prospective cohort study.
The pathogenetic role and the clinical importance of the presence of antiphospholipid antibodies (APAs) in patients with immune thrombocytopenic purpura (ITP) are not clear. In this study, the prevalence and clinical significance of APAs were investigated in patients with ITP. Eighty-two newly diagnosed ITP patients were prospectively studied. They were evaluated for the presence of lupus anticoagulant (LA) and immunoglobulin G/M anticardiolipin antibodies (ACAs). Thirty-one patients (37.8%) were APA positive at diagnosis. No statistically significant differences were found between the APA-positive and APA-negative groups regarding gender, initial platelet counts, or response to methylprednisolone therapy. After 5 years of follow-up, cumulative thrombosis-free survival of APA-positive (n = 31) and APA-negative (n = 51) ITP patients was 39% and 97.7%, respectively. A significant difference was found between these groups by log-rank test (P =.0004). In addition, LA was an important risk marker for the development of thrombosis in ITP patients. After a median follow-up of 38 months, 14 ITP patients (45%) who had APA positivity developed clinical features (thrombosis or fetal losses) of antiphospholipid syndrome (APS). There were no differences between the APA-positive patients with and without APS regarding the initial platelet counts, response to the therapy, or ACA positivity. The positivity rate for LA was significantly higher in those patients with ITP who developed APS (chi(2): P =.0036; relative risk 7.15; 95% confidence interval, 1.7-47). In conclusion, this study indicates that a significant proportion of patients initially presenting with ITP and APA positivity developed APS. In patients with ITP, the persistent presence of APAs is an important risk factor for the development of APS.


SOD1 in cerebral spinal fluid as a pharmacodynamic marker for antisense oligonucleotide therapy.
Therapies designed to decrease the level of SOD1 are currently in a clinical trial for patients with superoxide dismutase (SOD1)-linked familial amyotrophic lateral sclerosis (ALS). To determine whether the SOD1 protein in cerebral spinal fluid (CSF) may be a pharmacodynamic marker for antisense oligonucleotide therapy and a disease marker for ALS. Antisense oligonucleotides targeting human SOD1 were administered to rats expressing SOD1G93A. The human SOD1 protein levels were measured in the rats' brain and CSF samples. In human CSF samples, the following proteins were measured: SOD1, tau, phosphorylated tau, VILIP-1, and YKL-40. Ninety-three participants with ALS, 88 healthy controls, and 89 controls with a neurological disease (55 with dementia of the Alzheimer type, 19 with multiple sclerosis, and 15 with peripheral neuropathy). Antisense oligonucleotide-treated SOD1G93A rats had decreased human SOD1 messenger RNA levels (mean [SD] decrease of 69% [4%]) and decreased protein levels (mean [SD] decrease of 48% [14%]) in the brain. The rats' CSF samples showed a similar decrease in hSOD1 levels (mean [SD] decrease of 42% [14%]). In human CSF samples, the SOD1 levels varied a mean (SD) 7.1% (5.7%) after additional measurements, separated by months, were performed. The CSF SOD1 levels were higher in the participants with ALS (mean [SE] level, 172 [8] ng/mL; P<.05) and the controls with a neurological disease (mean [SE] level, 172 [6] ng/mL; P<.05) than in the healthy controls (mean [SE] level, 134 [4] ng/mL). Elevated CSF SOD1 levels did not correlate with disease characteristics in participants with ALS or controls with dementia of the Alzheimer type, but they did correlate with tau, phosphorylated tau, VILIP-1 and YKL-40 levels in controls with dementia of the Alzheimer type. SOD1 in CSF may be an excellent pharmacodynamic marker for SOD1-lowering therapies because antisense oligonucleotide therapy lowers protein levels in the rat brain and rat CSF samples and because SOD1 levels in CSF samples from humans are stable over time.