Co-localization and confinement of ecto-nucleotidases modulate extracellular adenosine nucleotide distributions.
Nucleotides comprise small molecules that perform critical signaling roles in biological systems. Adenosine-based nucleotides, including adenosine tri-, di-, and mono-phosphate, are controlled through their rapid degradation by diphosphohydrolases and ecto-nucleotidases (NDAs). The interplay between nucleotide signaling and degradation is especially important in synapses formed between cells, which create signaling 'nanodomains'. Within these 'nanodomains', charged nucleotides interact with densely-packed membranes and biomolecules. While the contributions of electrostatic and steric interactions within such nanodomains are known to shape diffusion-limited reaction rates, less is understood about how these factors control the kinetics of nucleotidase activity. To quantify these factors, we utilized reaction-diffusion numerical simulations of 1) adenosine triphosphate (ATP) hydrolysis into adenosine monophosphate (AMP) and 2) AMP into adenosine (Ado) via two representative nucleotidases, CD39 and CD73. We evaluate these sequentially-coupled reactions in nanodomain geometries representative of extracellular synapses, within which we localize the nucleotidases. With this model, we find that 1) nucleotidase confinement reduces reaction rates relative to an open (bulk) system, 2) the rates of AMP and ADO formation are accelerated by restricting the diffusion of substrates away from the enzymes, and 3) nucleotidase co-localization and the presence of complementary (positive) charges to ATP enhance reaction rates, though the impact of these contributions on nucleotide pools depends on the degree to which the membrane competes for substrates. As a result, these contributions integratively control the relative concentrations and distributions of ATP and its metabolites within the junctional space. Altogether, our studies suggest that CD39 and CD73 nucleotidase activity within junctional spaces can exploit their confinement and favorable electrostatic interactions to finely control nucleotide signaling.


Rh-Catalyzed aminative dearomatization of 2-naphthols.
A direct aminative dearomatization of 2-naphthols was achieved. In the presence of 1 mol% Rh2(esp)2 and 3 equivalents of O-(2,4-dinitrophenyl)hydroxylamine (DPH) as readily available aminating reagents, the reactions of 2-naphthols afforded unprotected α-amino-β-naphthalenones in good yields under mild reaction conditions. The conditions were compatible with gram-scale reaction, and the product could undergo diverse transformations.


Identification of autophagy-related genes in osteoarthritis articular cartilage and their roles in immune infiltration.
Autophagy plays a critical role in the progression of osteoarthritis (OA), mainly by regulating inflammatory and immune responses. However, the underlying mechanisms remain unclear. This study aimed to investigate the potential relevance of autophagy-related genes (ARGs) associated with infiltrating immune cells in OA. GSE114007, GSE169077, and ARGs were obtained from the Gene Expression Omnibus (GEO) database and the Human Autophagy database. R software was used to identify the differentially expressed autophagy-related genes (DEARGs) in OA. Functional enrichment and protein-protein interaction (PPI) analyses were performed to explore the role of DEARGs in OA cartilage, and then Cytoscape was utilized to screen hub ARGs. Single-sample gene set enrichment analysis (ssGSEA) was used to conduct immune infiltration analysis and evaluate the potential correlation of key ARGs and immune cell infiltration. Then, the expression levels of hub ARGs in OA were further verified by the GSE169077 and qRT-PCR. Finally, Western blotting and immunohistochemistry were used to validate the final hub ARGs. A total of 24 downregulated genes and five upregulated genes were identified, and these genes were enriched in autophagy, mitophagy, and inflammation-related pathways. The intersection results identified nine hub genes, namely, CDKN1A, DDIT3, FOS, VEGFA, RELA, MAP1LC3B, MYC, HSPA5, and HSPA8. GSE169077 and qRT-PCR validation results showed that only four genes, CDKN1A, DDT3, MAP1LC3B, and MYC, were consistent with the bioinformatics analysis results. Western blotting and immunohistochemical (IHC) showed that the expression of these four genes was significantly downregulated in the OA group, which is consistent with the qPCR results. Immune infiltration correlation analysis indicated that DDIT3 was negatively correlated with immature dendritic cells in OA, and FOS was positively correlated with eosinophils. CDKN1A, DDIT3, MAP1LC3B, and MYC were identified as ARGs that were closely associated with immune infiltration in OA cartilage. Among them, DDIT3 showed a strong negative correlation with immature dendritic cells. This study found that the interaction between ARGs and immune cell infiltration may play a crucial role in the pathogenesis of OA; however, the specific interaction mechanism needs further research to be clarified. This study provides new insights to further understand the molecular mechanisms of immunity involved in the process of OA by autophagy.


Predictors of postpartum sexual activity and function in a diverse population of women.
The purpose of this study was to identify predictors of postpartum sexual activity and functioning in a diverse population of women using the Sexual Health Outcomes in Women Questionnaire (SHOW-Q). This was a prospective study of 160 postpartum women assessing relationships between demographic factors, mode of birth, depression, breastfeeding, and sexual activity and function. Questionnaires were administered over the telephone 8 to 10 weeks postpartum and in person 6 to 8 months postpartum. Primary outcomes were sexual activity at 8 to 10 weeks postpartum and global and subscale SHOW-Q scores at 6 to 8 months postpartum; the primary predictor was mode of birth. Associations were assessed using multiple linear and logistic regression analyses. Seventy-five percent of this population (n = 140 at 8-10 weeks, n = 129 at 6-8 months) gave birth vaginally, and 60.7% resumed sexual activity by 8 to 10 weeks postpartum. Only multiparity was associated with increased odds of having resumed sexual activity by 8 to 10 weeks postpartum (adjusted odds ratio [aOR], 2.44; P = .03), whereas older age was associated with decreased odds (aOR, 0.92; P = .02) of having resumed sexual activity. Women who were depressed (effect estimate, -13.3; P = .01), older (-1.1, P = .01), or exclusively breastfeeding (-16.5, P < .001) had significantly poorer sexual satisfaction, whereas multiparous women reported better sexual satisfaction (11.1, P = .03). A significant relationship between mode of birth and SHOW-Q scores did not emerge, although we did observe a trend toward lower SHOW-Q scores among women who underwent cesarean compared with those giving birth vaginally. Multiparity and younger age predict early resumption of sexual activity, whereas depression and breastfeeding are associated with poorer postpartum sexual functioning. The relationship between mode of birth and resumed sexual activity or postpartum sexual function remains uncertain.


Biomartr: genomic data retrieval with R.
Retrieval and reproducible functional annotation of genomic data are crucial in biology. However, the current poor usability and transparency of retrieval methods hinders reproducibility. Here we present an open source R package, biomartr , which provides a comprehensive easy-to-use framework for automating data retrieval and functional annotation for meta-genomic approaches. The functions of biomartr achieve a high degree of clarity, transparency and reproducibility of analyses. The biomartr package implements straightforward functions for bulk retrieval of all genomic data or data for selected genomes, proteomes, coding sequences and annotation files present in databases hosted by the National Center for Biotechnology Information (NCBI) and European Bioinformatics Institute (EMBL-EBI). In addition, biomartr communicates with the BioMart database for functional annotation of retrieved sequences. Comprehensive documentation of biomartr functions and five tutorial vignettes provide step-by-step instructions on how to use the package in a reproducible manner. The open source biomartr package is available at https://github.com/HajkD/biomartr and https://cran.r-project.org/web/packages/biomartr/index.html . email@example.com. Supplementary data are available at Bioinformatics online.


LZTS1 downregulation confers paclitaxel resistance and is associated with worse prognosis in breast cancer.
The Leucine Zipper Tumor Suppressor 1 (LZTS1) is a tumor suppressor gene, located at chromosome 8p22, which is frequently altered in human cancer. In normal tissue, its ubiquitous expression regulates cell mitosis by the stabilization of microtubule networks. LZTS1-deficient mouse embryonic fibroblasts have been shown to have an accelerated mitotic progression, and a higher resistance to taxanes, microtubule-stabilizing drugs. We investigate the role of Lzts1 in paclitaxel-resistance in breast cancer cells. Downregulation of Lzts1 expression significantly decreases sensitivity to paclitaxel in vitro. We further analyzed Lzts1 expression by immunohistochemistry in 270 primary breast cancer samples and 16 normal breast specimens. Lzts1 was significantly downregulated in breast cancer samples and its deregulation was associated with a higher incidence of tumor recurrence, and to a worse overall survival. Moreover, Lzts1-negative tumors were associated with unfavorable outcome after taxanes-based therapy. Thus our data suggest that Lzts1 deregulation is involved in breast cancer and its immunohistochemical evaluation may serve as a prognostic factor for breast cancer therapy.


A domino effect in drug action: from metabolic assault towards parasite differentiation.
Awareness is growing that drug target validation should involve systems analysis of cellular networks. There is less appreciation, though, that the composition of networks may change in response to drugs. If the response is homeostatic (e.g. through upregulation of the target protein), this may neutralize the inhibitory effect. In this scenario the effect on cell growth and survival would be less than anticipated based on affinity of the drug for its target. Glycolysis is the sole free-energy source for the deadly parasite Trypanosoma brucei and is therefore a possible target pathway for anti-trypanosomal drugs. Plasma-membrane glucose transport exerts high control over trypanosome glycolysis and hence the transporter is a promising drug target. Here we show that at high inhibitor concentrations, inhibition of trypanosome glucose transport causes cell death. Most interestingly, sublethal concentrations initiate a domino effect in which network adaptations enhance inhibition. This happens via (i) metabolic control exerted by the target protein, (ii) decreases in mRNAs encoding the target protein and other proteins in the same pathway, and (iii) partial differentiation of the cells leading to (low) expression of immunogenic insect-stage coat proteins. We discuss how these 'anti-homeostatic' responses together may facilitate killing of parasites at an acceptable drug dosage.


Therapy Dogs' and Handlers' Behavior and Salivary Cortisol During Initial Visits in a Complex Medical Institution: A Pilot Study.
Therapy dogs provide health benefits for individuals who suffer from illnesses, such as dementia, depression, loneliness, and aggression. Therapy dogs' impact on human health has been thoroughly studied; however, studies on dog welfare have been limited. Additionally, as dogs have evolved with humans, they have learned to read non-verbal social cues. Dogs can read humans' non-verbal body language and can react to their emotions. However, the body language of dogs is poorly understood and can lead to dog owner-directed aggression. Communication plays a vital role to be a cohesive therapy team. The purpose of this study was to assess perceived stress and cortisol concentrations in therapy dogs and their handlers during the first three visits in a hospital setting. Moreover, the study aimed to investigate whether, while in an overstimulating environment, a therapy dog handler can observe his or her dog's body language and correlate such observations to the dog's stress. Nine therapy dog teams from Mayo Clinic's Caring Canine Program participated in this study. A baseline salivary cortisol was collected from the handler and therapy dog each day of the visits. Once the team arrived, a pre-visit salivary cortisol was collected from the handler and therapy dog and, afterward, a post-visit salivary cortisol. Handlers were also asked to fill out a perceived stress survey on their own stress and that of their therapy dogs'. Behavior was documented by a staff member and the handler. For each visit, the therapy dogs were at the hospital on average 47 min and visited with nine people. There was significant correlation (P = 0.02) between the owner's perceived stress of his or her therapy dog and the dog's salivary cortisol concentrations. The handlers noted medium to high stress, and those dogs had higher cortisol concentrations post-visit. There was no significant difference in salivary cortisol for the handler and therapy dog over the course of the three visits and comparing pre- and post-visit. Overall, the dogs displayed mixed behaviors, with the three most reported being panting, lip licking, and yawning. However, salivary cortisol results suggest that the handlers and therapy dogs maintained their welfare state throughout the visits.


Microchip-based immunomagnetic detection of circulating tumor cells.
Screening for circulating tumor cells (CTCs) in blood has been an object of interest for evidence of progressive disease, status of disease activity, recognition of clonal evolution of molecular changes and for possible early diagnosis of cancer. We describe a new method of microchip-based immunomagnetic CTC detection, in which the benefits of both immunomagnetic assay and the microfluidic device are combined. As the blood sample flows through the microchannel closely above arrayed magnets, cancer cells labeled with magnetic nanoparticles are separated from blood flow and deposited at the bottom wall of the glass coverslip, which allows direct observation of captured cells with a fluorescence microscope. A polydimethylsiloxane (PDMS)-based microchannel fixed on a glass coverslip was used to screen blood samples. The thin, flat dimensions of the microchannel, combined with the sharp magnetic field gradient in the vicinity of arrayed magnets with alternate polarities, lead to an effective capture of labeled cells. Compared to the commercially available CellSearch™ system, fewer (25%) magnetic particles are required to achieve a comparable capture rate, while the screening speed (at an optimal blood flow rate of 10 mL h(-1)) is more than five times faster than those reported previously with a microchannel-based assay. For the screening experiment, blood drawn from healthy subjects into CellSave™ tubes was spiked with cultured cancer cell lines of COLO205 and SKBR3. The blood was then kept at room temperature for 48 hours before the screening, emulating the actual clinical cases of blood screening. Customized Fe(3)O(4) magnetic nanoparticles (Veridex Ferrofluid™) conjugated to anti-epithelial cell adhesion molecule (EpCAM) antibodies were introduced into the blood samples to label cancer cells, and the blood was then run through the microchip device to capture the labelled cells. After capture, the cells were stained with fluorescent labelled anti-cytokeratin, DAPI and anti-CD45. Subsequent immunofluorescence images were taken for the captured cells, followed by comprehensive computer aided analysis based on fluorescence intensities and cell morphology. Rare cancer cells (from ∼1000 cells down to ∼5 cells per mL) with very low tumor cell to blood cell ratios (about 1 : 10(7) to 10(9), including red blood cells) were successfully detected. Cancer cell capture rates of 90% and 86% were demonstrated for COLO205 and SKBR3 cells, respectively.


Dislocations Accelerate Oxygen Ion Diffusion in La0.8Sr0.2MnO3 Epitaxial Thin Films.
Revealing whether dislocations accelerate oxygen ion transport is important for providing abilities in tuning the ionic conductivity of ceramic materials. In this study, we report how dislocations affect oxygen ion diffusion in Sr-doped LaMnO3 (LSM), a model perovskite oxide that serves in energy conversion technologies. LSM epitaxial thin films with thicknesses ranging from 10 nm to more than 100 nm were prepared by pulsed laser deposition on single-crystal LaAlO3 and SrTiO3 substrates. The lattice mismatch between the film and substrates induces compressive or tensile in-plane strain in the LSM layers. This lattice strain is partially reduced by dislocations, especially in the LSM films on LaAlO3. Oxygen isotope exchange measured by secondary ion mass spectrometry revealed the existence of at least two very different diffusion coefficients in the LSM films on LaAlO3. The diffusion profiles can be quantitatively explained by the existence of fast oxygen ion diffusion along threading dislocations that is faster by up to 3 orders of magnitude compared to that in LSM bulk.


Evaluation of Tobacco Control Policies in San Francisco Homeless Housing Programs.
The 2014 Surgeon General's Report noted that high smoking rates in vulnerable populations such as the homeless have been a persistent public health problem; smoking prevalence among individuals experiencing homelessness exceeds 70%. Historically, service providers for the homeless have not enacted comprehensive tobacco control policies. We conducted a qualitative study of homeless housing programs in San Francisco. Administrators representing 9 of the city's 11 homeless service agencies were interviewed to assess institutional smoking-related policies and cessation programs and perceived barriers and receptivity to instituting tobacco control interventions. Respondents indicated that although most programs had adopted smoke-free grounds and some had eliminated evidence of staff smoking, the smoking status of clients was assessed only when required by funders. None of the programs offered smoking cessation interventions. Most administrators were receptive to adopting policies that would promote a tobacco-free culture; however, they noted that their clients had unique challenges that made traditional smoking cessation programs unfeasible. Homeless housing programs in San Francisco have not yet adopted a tobacco-free culture. Existing policies were created in response to external mandates, and smoking cessation programs may need to be modified in order to effectively reach clients.


Evidence for alpha-ketoglutarate dehydrogenase monomer activity with the aid of enzyme immobilization.
Immobilization of pigeon breast muscle alpha-ketoglutarate dehydrogenase on CNBr-activated Sepharose 4B was carried out. Conditions for dissociation of the dimeric enzyme bound to the carrier via a single subunit were determined. Immobilized enzyme monomers with a specific activity higher than that of the dimer were obtained. The immobilized subunits are capable of reassociating with the soluble ones; this is accompanied by the restoration of the initial amount of the matrix-bound protein and the reconstitution of the activity typical of the immobilized enzyme original preparations.


Cost-Effectiveness of Early Treatment with Originator Biologics or Their Biosimilars After Methotrexate Failure in Patients with Established Rheumatoid Arthritis.
Evidence supports the clinical benefits of early aggressive biologic treatment in patients with rheumatoid arthritis (RA) who have an inadequate response to conventional synthetic disease-modifying antirheumatic drugs (csDMARDs), but the cost-effectiveness of early intervention with originator biologics such as tumor necrosis factor inhibitors (TNFis) or their biosimilars has not been well studied. We developed a Markov model to estimate lifetime costs and utilities for patients with established RA who do not respond to methotrexate (MTX) therapy. A cost-effectiveness analysis was conducted comparing a standard intervention pathway (addition of originator biologic TNFis to MTX monotherapy at 12 months) and two early intervention pathways (either addition of originator biologic TNFis or addition of biosimilar TNFis to MTX monotherapy at 6 months). Early intervention with an originator biologic TNFi at 6 months was associated with increases in total lifetime costs of £1692 and utilities of 0.10 quality-adjusted life-years (QALYs) per patient compared with standard intervention at 12 months, resulting in an incremental cost-effectiveness ratio (ICER) of £17,335/QALY. Early intervention with a biosimilar TNFi increased costs by £70 and utilities by 0.10 QALYs per patient and was associated with an ICER of £713/QALY. Switching from MTX monotherapy to combination therapy with either an originator biologic or biosimilar TNFis at 6 months after csDMARD failure in patients with RA was cost-effective at a threshold of £30,000/QALY. Pfizer Inc.


Reliability assessment of a rod photoreceptor outer segment grading system.
The purpose of this study was to assess the reliability of a rod photoreceptor outer segment (PR-OS) grading system based on the analysis of 1 microm thick retinal sections obtained from Xenopus laevis whole-eye organ cultures. Digitally captured images, representative of the entire spectrum of rod PR-OS organization levels, were selected and coded numerically. A total of 102 individual rod PR-OS profiles were graded according to a six-step classification scheme based on the percentage of rod PR-OS membrane organization. Unweighted (exact agreement) and weighted kappa (kappa) coefficients (for use with ordered categorical rating scales) were calculated. Differences between kappa coefficients were tested for by chi-square analysis. To investigate the intra- and inter-rater variability and the possible presence of an interaction of the measurements with time, a repeated-measures analysis of variance was performed. The overall unweighted and weighted intra-rater kappa coefficients were 0.78 and 0.92, respectively. The overall unweighted and weighted inter-rater kappa coefficients were 0.73 and 0.90, respectively. There was no significant difference between raters or between first and second reading, nor was interaction between raters and time of rating documented. Individual kappa coefficients were equivalent both between raters and between sessions. Intra- and inter-rater agreement was within one step in 100% of cases. The estimated values of the kappa coefficients are consistent with a good to excellent degree of reliability and reproducibility of this rod PR-OS grading system. This system will be useful in the assessment of rod PR-OS morphology in studies of photoreceptor physiology and pathology.


Glutathione S-transferase polymorphisms and outcome of chemotherapy in childhood acute myeloid leukemia.
Glutathione S-transferase theta (GSTT1) and mu (GSTM1) genes are polymorphic, the genes being absent in approximately 15% and 50% of the population, respectively. Because glutathione S-transferases may be involved in the metabolism of chemotherapy drugs, we hypothesized that presence or absence of the genes may influence the outcome of treatment for childhood acute myeloid leukemia (AML). We genotyped GSTT1 and GSTM1 in 306 children with AML receiving chemotherapy on Children's Cancer Group therapeutic studies. Outcomes were compared in those with and without GSTT1 and GSTM1 genes. Patients with the GSTT1-negative genotype had reduced survival compared with those with at least one GSTT1 allele (GSTT1 positive) (52% v 40% at 5 years; log-rank P =.05). A multivariate model of survival adjusted for age group, sex, WBC count, chloroma, CNS involvement, and French-American-British group confirmed the increased risk of death in the GSTT1-null cases (relative risk, AQ 1.6; P =.02). The frequency of death in remission was increased in GSTT1-negative cases compared with GSTT1-positive cases (24% v 12%, log-rank P =.05). The frequency of relapse from end of induction was similar in GSTT1-negative and GSTT1-positive cases (38% v 35%, log-rank P =.5). Children who lacked GSTT1 had greater toxicity and reduced survival after chemotherapy for AML compared with children with at least one GSTT1 allele. If confirmed in further studies, GSTT1 genotype might be useful in selecting appropriate chemotherapy regimens for children with AML.


New Carboxytriazolyl Amphiphilic Derivatives of Calix[4]arenes: Aggregation and Use in CuAAC Catalysis.
This work focuses on the synthesis of a new series of amphiphilic derivatives of calix[4]arenes for the copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The aggregation properties of synthesized calix[4]arenes were studied using various techniques (fluorescence spectroscopy, nanoparticle tracking analysis, and dynamic light scattering). Increasing the length of the alkyl substituent led to stronger hydrophobic interactions, which increased polydispersity in solution. The zwitterionic nature of the synthesized calix[4]arenes was established using different types of dyes (Eosin Y for anionic structures and Rhodamine 6G for cationic structures). The synthesized calix[4]arenes were used as organic stabilizers for CuI. The catalytic efficiency of CuI-calix[4]arene was compared with that of the phase transfer catalyst tetrabutylammonium bromide (TBAB) and the surfactant sodium dodecyl sulfate (SDS). For all calixarenes, the selectivity in the CuAAC reaction was higher than that observed when TBAB and SDS were estimated.


Thin film solar cell design based on photonic crystal and diffractive grating structures.
In this paper we present novel light trapping designs applied to multiple junction thin film solar cells. The new designs incorporate one dimensional photonic crystals as band pass filters that reflect short light wavelengths (400 - 867 nm) and transmit longer wavelengths(867 -1800 nm) at the interface between two adjacent cells. In addition, nano structured diffractive gratings that cut into the photonic crystal layers are incorporated to redirect incoming waves and hence increase the optical path length of light within the solar cells. Two designs based on the nano structured gratings that have been realized using the scattering matrix and particle swarm optimization methods are presented. We also show preliminary fabrication results of the proposed devices.


Platelet Glycoprotein-Ib (GPIb) May Serve as a Bridge between Type 2 Diabetes Mellitus (T2DM) and Atherosclerosis, Making It a Potential Target for Antiplatelet Agents in T2DM Patients.
Type 2 diabetes mellitus (T2DM) is a persistent metabolic condition that contributes to the development of cardiovascular diseases. Numerous studies have provided evidence that individuals with T2DM are at a greater risk of developing cardiovascular diseases, typically two to four times more likely than those without T2DM, mainly due to an increased risk of atherosclerosis. The rupture of an atherosclerotic plaque leading to pathological thrombosis is commonly recognized as a significant factor in advancing cardiovascular diseases caused by TD2M, with platelets inducing the impact of plaque rupture in established atherosclerosis and predisposing to the primary expansion of atherosclerosis. Studies suggest that individuals with T2DM have platelets that display higher baseline activation and reactivity than those without the condition. The expression enhancement of several platelet receptors is known to regulate platelet activation signaling, including platelet glycoprotein-Ib (GPIb). Furthermore, the high expression of platelet GP1b has been reported to increase the risk of platelet adhesion, platelet-leucocyte interaction, and thrombo-inflammatory pathology. However, the study exploring the role of GP1b in promoting platelet activation-induced cardiovascular diseases in T2DM patients is still limited. Therefore, we summarize the important findings regarding pathophysiological continuity between T2DM, platelet GPIb, and atherosclerosis and highlight the potential therapy targeting GPIb as a novel antiplatelet agent for preventing further cardiovascular incidents in TD2M patients.


Gut microbiota of elite female football players is not altered during an official international tournament.
The current study aimed to investigate if the gut microbiota composition of elite female football players changes during an official international tournament. The study was conducted throughout ten consecutive days, encompassing seven training sessions, and three official matches. The matches were separated by 48-72 h. Seventeen elite female football players from the Portuguese women's national football team participated in the study. Fecal samples were collected at two time points: at the beginning and end of the tournament. Fecal microbiota was analyzed by sequencing the 16S rRNA gene. Throughout the study, the duration and rating of perceived exertion (RPE) were recorded after training sessions and matches. The internal load was determined by the session RPE. The gut microbiota of players was predominantly composed of bacteria from the phyla Firmicutes (50% of relative abundance) and Bacteroidetes (20%); the genera Faecalibacterium (29%) and Collinsella (16%); the species Faecalibacterium prausnitzii (30%) and Collinsella aerofaciens (17%). Overall, no significant changes were observed between time points (p ≥ 0.05). Also, no relationship was found between any exercise parameter and the gut microbiota composition (p ≥ 0.05). These findings demonstrate that the physical and physiological demands of training and matches of an official international tournament did not change the gut microbiota composition of elite female football players. Furthermore, it supports that the gut microbiota of athletes appears resilient to the physical and physiological demands of training and match play.


Conditional inactivation of Foxc1 and Foxc2 in neural crest cells leads to cardiac abnormalities.
Cardiac neural crest cells (cNCCs) are required for normal heart development. cNCCs are a multipotent and migratory cell lineage that differentiates into multiple cell types. cNCCs migrate into the developing heart to contribute to the septation of the cardiac outflow tract (OFT). Foxc1 and Foxc2 are closely related members of the FOX (Forkhead box) transcription factor family and are expressed in cNCC during heart development. However, the precise role of Foxc1 and Foxc2 in cNCCs has yet to be fully described. We found that compound NCC-specific Foxc1;Foxc2 mutant embryos exhibited persistent truncus arteriosus (PTA), ventricular septal defects (VSDs), and thinning of the ventricular myocardium. Loss of Foxc1/c2 expression in cNCCs resulted in abnormal patterns of cNCC migration into the OFT without the formation of the aorticopulmonary septum. Further, loss of Foxc1 expression in cNCCs resulted in normal OFT development but abnormal ventricular septal formation. In contrast, loss of Foxc2 expression in NCCs led to no obvious cardiac abnormalities. Together, we provide evidence that Foxc1 and Foxc2 in cNCCs are cooperatively required for proper cNCC migration, the formation of the OFT septation, and the development of the ventricles. Our data also suggests that Foxc1 expression may play a larger role in ventricular development compared to Foxc2.


Searching online for health-related information by people suffering from respiratory allergy and asthma: the results of a survey study.
The objective of the study was to compare online behaviour in groups of persons suffering from respiratory allergy and/or asthma and in the reference group. Comparative analysis of two independent questionnaire studies. Study A was carried out in the Masovian Voivodeship, Poland in 2013. As many as 1,728 questionnaires were collected and 51 respondents were selected for analysis (study A, reference group). Study B was carried out in 2014 in a group of 59 patients of the outpatient allergy clinic. More than 92% of the general population (study A) declared that they used the Internet to search for health-related information, health protection, while in the group of patients (study B) the same was declared by 97%. Both studies showed that more than 74% of respondents looked for online information on a specific disease (p > 0.05). Information on how other persons cope with similar health issues was searched by 20% (study A) and 33% (study B) of respondents. Advice on different methods of treatment of a given disease was sought by 40% and 51% of respondents, respectively. Nearly 51% of respondents in study A and 56% of subjects in study B searched online for health information when they did not feel well or were sick. Both in the group of chronically ill persons and in the reference group, it is common to search online for health- -related information. Persons suffering from respiratory allergy and/or asthma verified the physician's diagnosis on the Internet much more often than did the reference group.


Comparative Study of Single-stranded Oligonucleotides Secondary Structure Prediction Tools.
Single-stranded nucleic acids (ssNAs) have important biological roles and a high biotechnological potential linked to their ability to bind to numerous molecular targets. This depends on the different spatial conformations they can assume. The first level of ssNAs spatial organisation corresponds to their base pairs pattern, i.e. their secondary structure. Many computational tools have been developed to predict the ssNAs secondary structures, making the choice of the appropriate tool difficult, and an up-to-date guide on the limits and applicability of current secondary structure prediction tools is missing. Therefore, we performed a comparative study of the performances of 9 freely available tools (mfold, RNAfold, CentroidFold, CONTRAfold, MC-Fold, LinearFold, UFold, SPOT-RNA, and MXfold2) on a dataset of 538 ssNAs with known experimental secondary structure. The minimum free energy-based tools, namely mfold and RNAfold, and some tools based on artificial intelligence, namely CONTRAfold and MXfold2, provided the best results, with [Formula: see text] of exact predictions, whilst MC-fold seemed to be the worst performing tool, with only [Formula: see text] of exact predictions. In addition, UFold and SPOT-RNA are the only options for pseudoknots prediction. Including in the analysis of mfold and RNAfold results 5-10 suboptimal solutions further improved the performances of these tools. Nevertheless, we could observe issues in predicting particular motifs, such as multiple-ways junctions and mini-dumbbells, or the ssNAs whose structure has been determined in complex with a protein. In addition, our benchmark shows that some effort has to be paid for ssDNA secondary structure predictions. In general, Mfold, RNAfold, and MXfold2 seem to currently be the best choice for the ssNAs secondary structure prediction, although they still show some limits linked to specific structural motifs. Nevertheless, actual trends suggest that artificial intelligence has a high potential to overcome these remaining issues, for example the recently developed UFold and SPOT-RNA have a high success rate in predicting pseudoknots.


Functional performance in children with Down syndrome.
The purpose of this study was to describe home and community functional performance in 5-year-old children with Down syndrome. In a cross-sectional study of 5-year-old children with Down syndrome in Norway (N=43), functional performance was measured with the Norwegian translation of the Pediatric Evaluation of Disability Inventory (PEDI). Additional descriptive information related to health, disabilities, and function was also gathered. The children showed a wide range of functional performance. Performance of self-care activities appeared most delayed on activities that required fine motor skills. Children appeared less affected in basic functional mobility skills. Parents' identified their main concerns as language functioning and, for the children not yet toilet trained, the management of bladder and bowel control in relation to starting school. The results provide baseline information regarding typical levels of functional performance in children with Down syndrome at 5 years of age. However, the broad range of functional performance across children indicates a need for caution in generalizing the results to an individual child.


Diaphragmatic hernia in bovines.
Three cases of diaphragmatic hernia in bovines were encountered over a period of 5 years. The successful surgical repair of one case is reported.


Addressing Burnout among Healthcare Technology Management Professionals.
This article suggests ways in which healthcare delivery organizations (HDOs) may address and mitigate burnout among healthcare technology management (HTM) professionals. Burnout is defined as a state of emotional and physical exhaustion resulting from workplace stressors. Although not intended to be a fully comprehensive empirical analysis of causes of burnout, this article discusses four likely causes for burnout that are particularly relevant to HTM professionals in HDOs: (1) gaps in developing a trained and sustainable workforce, (2) growing demand for HTM expertise across knowledge domains, (3) increasing occupational stressors related to patient safety, and (4) perceived lack of understanding of employee needs. Next, based on existing literature, the article proposes four possible solutions in response to each of these issues: (1) developing human resource planning and mentoring to ensure adequate transfer of skills and knowledge, (2) implementing ways to accomplish knowledge filtering, (3) integrating HTM workers in quality improvement initiatives, and (4) implementing employee voice practices. The article concludes by discussing the possible benefits that may be derived from implementing these solutions.


The relationship between childhood aerobic fitness and brain functional connectivity.
Several studies have indicated that higher levels of childhood aerobic fitness is associated with superior cognitive function, and this association is disproportionately observed in tasks requiring greater top-down control. We designed the current study to clarify the relationship between childhood fitness and top-down control in terms of functional connectivity among brain regions, by evaluating phase-locking values (PLVs), which is a measure of frequency-specific phase synchrony between electroencephalographic signals during a visual search task. Lower-fit and higher-fit children performed a visual search task that included feature search and conjunction search conditions. The conjunction search condition required greater top-down control to reduce interference from task-irrelevant distractors that shared a basic feature with the target. Results indicated that higher-fit children exhibited higher response accuracy relative to lower-fit children across search conditions. The results of PLVs showed that higher-fit children had greater functional connectivity for the conjunction relative to the feature search condition, whereas lower-fit children showed no difference in functional connectivity between search conditions. Furthermore, PLVs showed different time courses between groups; that is, higher-fit children sustained upregulation of top-down control throughout the task period, whereas lower-fit children transiently upregulated top-down control after stimulus onset and could not sustain the upregulation. These findings suggest that higher levels of childhood aerobic fitness is related to brain functional connectivity involved in the sustained upregulation of top-down control.


Pyridinylimidazole p38 mitogen-activated protein kinase inhibitors block intracellular Toxoplasma gondii replication.
Toxoplasma gondii is a medically important, obligate intracellular parasite. Little is known regarding factors that regulate its replication within cells. Such knowledge would further understanding of T. gondii pathogenesis, and might lead to novel therapeutic strategies. Mitogen-activated protein kinases (MAPKs) govern diverse cellular processes including proliferation and differentiation. We now show that treatment of T. gondii-infected cells with SB203580 or SB202190, substituted pyridinylimidazoles that are potent inhibitors of human p38 MAPK, inhibits intracellular T. gondii replication. Several independent experimental approaches suggest that the anti-proliferative effects of pyridinylimidazoles depend on direct action on tachyzoites, not the host cell: (i) selective inhibition of host p38 MAPK using recombinant adenoviruses had little effect on tachyzoite replication, (ii) pyridinylimidazole-treated tachyzoites developed abnormal morphology suggesting defective parasite division, and (iii) pyridinylimidazole-resistant mutant tachyzoites were developed through culture in progressively higher drug concentrations. We hypothesise that pyridinylimidazoles target a human p38 MAPK homologue in tachyzoites that regulates their replication. Phylogenetic data suggest that T. gondii likely encodes a p38 MAPK homologue, but such a homologue is absent from the incomplete Toxoplasma genomic data base. As all eukaryotic pathogens, including agents of malaria, leishmaniasis and trypanosomiasis encode endogenous MAPKs, drugs inhibiting endogenous MAPK activation may represent a novel, potentially broadly-acting class of anti-parasitic agents. Pyridinylimidazoles also represent tools to elucidate factors governing intracellular tachyzoite replication.


Imaging Findings of an Intraluminal Duodenal Diverticulum Associated with Adult Duodeno-Duodenal Intussusception and Recurrent Pancreatitis: A Case Report.
Intraluminal duodenal diverticulum (IDD) is a rare congenital abnormality, consisting of a sac-like mucosal lesion in the duodenum. Cases of IDD can present with gastrointestinal bleeding, duodenal obstruction, or pancreatitis. Here, we report a rare case of a 25-year-old female presenting with IDD complicated by duodeno-duodenal intussusception and recurrent pancreatitis. The diagnosis was based on findings from radiologic examinations (CT and MRI), upper gastrointestinal series (barium swallow), and gastroduodenofiberscopy. Laparoscopic excision of the presumed duodenal duplication was performed. The subsequent histopathologic evaluation of the excised sac revealed normal mucosa on both sides, but the absence of a proper muscle layer confirmed the diagnosis of IDD. Radiologic detection of a saccular structure in the second portion of the duodenum can indicate IDD with duodeno-duodenal intussusception as the lead point.


Structural brain mutant of Drosophila melanogaster with reduced cell number in the medulla cortex and with normal optomotor yaw response.
KS58, one out of six known alleles of the small optic lobes (sol) gene in Drosophila melanogaster, reduces the cell number in the medulla cortex by degeneration of ganglion cells in the pupae to about 50%. Also, about half the volume of the medulla and lobula complex neuropils is missing. Many Golgistained cells in the mutant optic lobes resemble their homologues in wild type. However, special classes of transmedullary columnar neurons projecting to the lobula or to both lobula and lobula plate are not seen in the mutant. Some neurons linking the lobula complex to the central brain send branches to the medulla (the branches do not exist in wild type); some other types seem to be missing. The fate mapping of the KS58 focus reveals a location ventral to the head bristles and in sine oculis (so) flies the mutation further reduces the rudiments of the optic lobes normally seen. Therefore the sol phenotype is not induced by mutant eyes and the primary gene action seems to be on nervous tissue. The structural alterations of the small optic lobes are reflected in visual orientation behavior. The optomotor yaw response, however, is almost quantitatively preserved. The respective neural network should still be present in the mutant optic lobes.


Studies of the denaturation and partial renaturation of ovalbumin.
1. The denaturation of ovalbumin by the reagents sodium dodecyl sulphate and guanidinium chloride was investigated, by following the changes in sedimentation velocity, optical rotatory dispersion and viscosity as a function of denaturant concentration. 2. With sodium dodecyl sulphate both the optical-rotatory-dispersion parameters a(0) and b(0) become more negative, the sedimentation coefficient decreases and the viscosity increases; significant differences in the denaturation profiles are observed. The change in each parameter is indicative of only limited denaturation. 3. With guanidinium chloride the transition occurs over the concentration range 1-4m: more extensive changes occur in all the physical parameters than with sodium dodecyl sulphate. The values of a(0) and b(0) are indicative of complete denaturation. Reduction by mercaptoethanol produces only minor further changes. 4. Renaturation was attempted from both denaturants, the removal of reagent being accomplished reversibly by controlled slow dialysis. Partial renaturation was observed, but aggregated or insoluble material was produced in both cases at relatively low concentrations of denaturant. Similar behaviour was observed with fully reduced protein in guanidinium chloride-mercaptoethanol; complete renaturation could not be brought about even at very low protein concentrations.


The use of clinical practice guidelines in primary care: professional mindlines and control mechanisms.
To identify the relevant barriers and enablers perceived by primary care professionals in implementing the recommendations of clinical practice guidelines (CPG). Two focus groups were conducted with primary care physicians and nurses in Catalonia (Spain) between October and December 2012. Thirty-nine health professionals were selected based on their knowledge and daily use of CPG. Finally, eight general practitioners and eight nurses were included in the discussion groups. Participants were asked to share their views and beliefs on the accessibility of CPG, their knowledge and use of these documents, the content and format of CPG, dissemination strategy, training, professional-patient relationship, and the use of CPG by the management structure. We recorded and transcribed the content verbatim and analysed the data using qualitative analysis techniques. Physicians believed that, overall, CPG were of little practical use and frequently referred to them as a largely bureaucratic management control instrument that threatened their professional autonomy. In contrast, nurses believed that CPG were rather helpful tools in their day-to-day practice, although they would like them to be more sensitive to the current role of nurses. Both groups believed that CPG did not provide a response to most of the decisions they faced in the primary care setting. Compliance with CPG recommendations would be improved if these documents were brief, non-compulsory, not cost-containment oriented, more based on nursing care models, sensitive to the specific needs of primary care patients, and integrated into the computer workstation.


Novel 2019 coronavirus structure, mechanism of action, antiviral drug promises and rule out against its treatment.
In the past two decades, the world has faced several infectious disease outbreaks. Ebola, Influenza A (H1N1), SARS, MERS, and Zika virus have had a massive global impact in terms of economic disruption, the strain on local and global public health. Most recently, the global outbreak of novel coronavirus 2019 (SARS-CoV-2) that causes COVID-19 is a newly discovered virus from the coronavirus family in Wuhan city, China, known to be a great threat to the public health systems. As of 15 April 2020, The Johns Hopkins University estimated that the COVID-19 affected more than two million people, resulting in a death toll above 130,000 around the world. Infected people in Europe and America correspond about 40% and 30% of the total reported cases respectively. At this moment only few Asian countries have controlled the disease, but a second wave of new infections is expected. Predicting inhibitor and target to the COVID-19 is an urgent need to protect human from the disease. Therefore, a protocol to identify anti-COVID-19 candidate based on computer-aided drug design is urgently needed. Thousands of compounds including approved drugs and drugs in the clinical trial are available in the literature. In practice, experimental techniques can measure the time and space average properties but they cannot be captured the structural variation of the COVID-19 during the interaction of inhibitor. Computer simulation is particularly suitable to complement experiments to elucidate conformational changes at the molecular level which are related to inhibition process of the COVID-19. Therefore, computational simulation is essential tool to elucidate the phenomenon. The structure-based virtual screening computational approach will be used to filter the best drugs from the literature, the investigate the structural variation of COVID-19 with the interaction of the best inhibitor is a fundamental step to design new drugs and vaccines which can combat the coronavirus. This mini-review will address novel coronavirus structure, mechanism of action, and trial test of antiviral drugs in the lab and patients with COVID-19.


Initial stages of CO2 adsorption on CaO: a combined experimental and computational study.
Room temperature adsorption of carbon dioxide (CO2) on monocrystalline CaO(001) thin films grown on a Mo(001) substrate was studied by infrared reflection-absorption spectroscopy (IRAS) and quantum chemical calculations. For comparison, CO2 adsorption was examined on poorly ordered, nanoparticulate CaO films prepared on Ru(0001). For both systems, CO2 readily adsorbs on the clean CaO surface. However, additional bands were observable on the CaO/Ru(0001) films compared with CaO/Mo(001), because the stricter IRAS surface selection rules do not apply to adsorption on the disordered thin films grown on Ru(0001). Spectral evolution with increasing exposure of the IRA bands suggested the presence of several adsorption sites which are consecutively populated by CO2. Density functional calculations showed that CO2 adsorption occurs as monodentate surface carbonate (CO32-) species at monatomic step sites and other low-coordinated sites, followed by formation of carbonates on terraces, which dominate at increasing CO2 exposure. To explain the coverage-dependent IRAS results, we propose CO2 surface islanding from the onset, most likely in the form of pairs and other chain-like species, which were calculated as thermodynamically favorable. The calculated adsorption energy for isolated CO2 on the terrace sites (184 ± 10 kJ mol-1) is larger than the adsorption energy obtained by temperature programmed desorption (∼120-140 kJ mol-1) and heat of adsorption taken from microcalorimetry measurements at low coverage (∼125 kJ mol-1). However, the calculated adsorption energies become less favorable when carbonate chains intersect on CaO terraces, forming kinks. Furthermore, our assignments of the initial stages of CO2 adsorption are consistent with the observed coverage effect on the CO2 adsorption energy measured by microcalorimetry and the IRAS results.


Treat-to-target study for improved outcome in polyarticular juvenile idiopathic arthritis.
Juvenile idiopathic arthritis is one of the most prevalent chronic inflammatory diseases in children. Evidence suggests that early effective treatment minimises the burden of disease during childhood and in further life. We hypothesise that a guided treat-to-target (T2T) approach is superior to routine care in polyarticular juvenile idiopathic arthritis (pJIA) in terms of reaching a clinical remission after 12 months of treatment. Patients with early and active pJIA were enrolled. Targets for treatment were the following: Recognisable Juvenile Arthritis Disease Activity Score (JADAS) improvement after 3 months, acceptable disease at 6 months, minimal disease activity at 9 months and as primary endpoint remission after 12 months. Initially, patients received methotrexate. Failure to meet a defined target required treatment modification at the specified intervals. The choice of biologics was not influenced by the protocol. Finally, T2T patients were compared with a cohort of matched controls of patients with pJIA with unguided therapy documented by BIKER. Sixty-three patients were enrolled. Treatment targets after 3/6/9 and 12 months were reached by 73%/75%/77% and 48% of patients. Fifty-four patients completed the protocol. Compared with matched controls, on T2T guidance significantly more patients reached JADAS remission (48% vs 32%; OR 1.96 (1.1-3.7); p=0.033) and JADAS minimal disease activity (JADAS-MDA) (76% vs 59%; OR 2.2 (1.1-4.4); p=0.028). Patients from the T2T cohort received a biologic significantly more frequent (50% vs 9% after 12 months; OR 9.8 (4.6-20.8); p<0.0001). The T2T concept was feasible and superior to unguided treatment. High rates of patients reached JADAS-MDA and JADA remission after 12 months. Approximately half of the patients achieved their therapy goals without a biologic.


Overexpression of a Cinnamyl Alcohol Dehydrogenase-Coding Gene, GsCAD1, from Wild Soybean Enhances Resistance to Soybean Mosaic Virus.
Soybean mosaic virus (SMV) is the most prevalent soybean viral disease in the world. As a critical enzyme in the secondary metabolism of plants, especially in lignin synthesis, cinnamyl alcohol dehydrogenase (CAD) is widely involved in plant growth and development, and in defense against pathogen infestation. Here, we performed RNAseq-based transcriptome analyses of a highly SMV-resistant accession (BYO-15) of wild soybean (Glycine soja) and a SMV-susceptible soybean cultivar (Williams 82), also sequenced together with a resistant plant and a susceptible plant of their hybrid descendants at the F3 generation at 7 and 14 days post-inoculation with SMV. We found that the expression of GsCAD1 (from G. soja) was significantly up-regulated in the wild soybean and the resistant F3 plant, while the GmCAD1 from the cultivated soybean (G. max) did not show a significant and persistent induction in the soybean cultivar and the susceptible F3 plant, suggesting that GsCAD1 might play an important role in SMV resistance. We cloned GsCAD1 and overexpressed it in the SMV-susceptible cultivar Williams 82, and we found that two independent GsCAD1-overexpression (OE) lines showed significantly enhanced SMV resistance compared with the non-transformed wild-type (WT) control. Intriguingly, the lignin contents in both OE lines were higher than the WT control. Further liquid chromatography (HPLC) analysis showed that the contents of salicylic acid (SA) were significantly more improved in the OE lines than that of the wild-type (WT), coinciding with the up-regulated expression of an SA marker gene. Finally, we observed that GsCAD1-overexpression affected the accumulation of SMV in leaves. Collectively, our results suggest that GsCAD1 enhances resistance to SMV in soybeans, most likely by affecting the contents of lignin and SA.


[Status of resistance to insecticides in field strains of the Blatella germanica species (Dictyoptera: Blattellidae) from Pinar del Río municipality].
A study of the levels of resistance to 10 insecticides: 4 organophosphate compounds (malathion, clorpirifos, methylpyrimifos and diazinon), 2 carbamates (propoxur and bendiocarb) and 4 pyrethroids (cypermethrin, deltamethrin, lamdacyhalothrin and cyfluthrin) was conducted in 5 strains of Blatella germanica (Linnaeus, 1767) collected in the field of Pinar del Rio. High levels of resistance to bediocarb, cypermethrin and deltamethrin insecticides; low level of resistance to diazinon; from moderate to high resistance to methyl-pyrimifos, as well as susceptibility to one insecticide in each study group: clorpirifos (organophosphate), propoxur (carbamate) and cyfluthrin (pyrethroid); were detected. Only a strain presented low resistance to malathion (Inicio Carlos Manuel) and to lambda-cyhalothrin (Consejo Celso Maragoto). Cypermethrin-deltamethrin cross resistance was evidenced. It did not affect the susceptibility to lambda-cyalothrin and cyfluthrin.


Computed tomographic, magnetic resonance imaging, and ultrasonographic characteristics of retrobulbar meningiomas in 15 dogs.
Meningioma is the most common tumor of the cranium in dogs and an important differential diagnosis for a potentially treatable disease that can be found in the periorbital tissues. The objective of this retrospective, case series study was to describe the CT, MRI, and US characteristics of confirmed retrobulbar meningiomas in a group of dogs. Medical records from multiple institutions were searched for canine patients with CT, MRI, and/or US imaging of a cytologically or histologically confirmed retrobulbar meningioma. Fifteen dogs met the inclusion criteria. Retrobulbar meningiomas typically appeared as a relatively well-defined conical to ovoid mass within the retrobulbar space, most often along the optic nerve and expanding the extraocular muscle cone. On CT, masses were predominantly soft tissue attenuating and variably heterogeneously contrast enhancing. While MRI features were variable, moderate to marked contrast enhancement was seen in all cases. Many of the tumors had evidence of partial mineralization, best appreciated on CT in nine patients, but also suspected based on susceptibility artifacts in three MRI cases, one of which was confirmed on CT. Regional osteolysis was a rare finding, noted in three cases, but was often accompanied by cranial cavity extension (2/3). Cranial cavity extension was also seen in the absence of regional osteolysis, identified in a total of six patients. On US, masses were echogenic and compressed the globe. The findings were consistent with previous gross and histologic descriptions and supported prioritizing retrobulbar meningioma as a differential diagnosis for dogs with the described imaging characteristics.


A back-to-back zig-zag model for higher order chromatin structure.
The evidence for the next level of chromatin structure above the 'zig-zag' is examined and a system of large, uncoiled, 'hair-pin' loops, folded until the zig-zags are back to back, is proposed as being more compatible with the evidence than coiling models. The loop model suggests a simple way for the base sequences on the DNA to control the higher order folding of chromatin, and is also strongly suggestive of a simple mechanism ensuring homologous alignment of nucleofilaments at synapse.


Future therapeutic directions for factor Xa inhibition in the prophylaxis and treatment of thrombotic disorders.
The targeted mechanism of factor Xa inhibition has been studied extensively, initially as prophylaxis for venous thromboembolism (VTE) in the orthopedic surgical setting. Future therapeutic directions for selective factor Xa inhibition in the management of other thrombotic diseases are discussed. Thromboembolic diseases can occur in the venous or arterial sides of the circulatory system. Factor Xa inhibition is a targeted approach to anticoagulation that resulted from significant advances in our understanding of the coagulation cascade. The factor Xa inhibitor fondaparinux has been studied extensively in the orthopedic surgical setting for the prophylaxis of VTE. Current investigations that are under way or completed evaluate the efficacy and safety of fondaparinux for the management of various thrombotic diseases. The future development of fondaparinux resides primarily in three therapeutic areas: prevention of VTE, treatment of VTE, and treatment of acute coronary syndromes. For the prevention of VTE, fondaparinux has been studied as extended prophylaxis following hip fracture surgery (PENTHIFRA Plus), for use in high-risk abdominal surgical patients (PEGASUS and APOLLO), and for use in medical patients (ARTEMIS). Studies evaluating fondaparinux for the treatment of VTE are part of the large MATISSE clinical program (MATISSE DVT and MATISSE PE). Fondaparinux was investigated in phase 2 studies for the treatment of acute coronary syndromes, including acute ST-segment myocardial infarction (PENTALYSE) and unstable angina (PENTUA). Encouraging data from these trials are the basis for phase 3 programs in this area (MICHELANGELO). The orthopedic prophylactic and nonorthopedic clinical programs for fondaparinux in the management of thrombosis support the concept that targeted inhibition of coagulation is an effective advance in antithrombotic therapy.


Symptom Adverse Events and Quality of Life of Children With Advanced Cancer: Results From a Longitudinal Study Using the Pediatric Patient-Reported Outcomes-Common Terminology Criteria for Adverse Events.
Background: The presence of poorly controlled symptoms negatively impacts the quality of life (QoL) throughout cancer treatment. The purpose of this multisite study was to explore the relationship between QoL and symptom adverse events (AEs) in children with advanced cancer over 6 months. Method: A prospective and longitudinal descriptive study design was used to collect QoL and symptom AE data from children aged 2 to 18 with advanced cancer. QoL was measured using the Pediatric Quality of Life Inventory (PedsQLTM) Cancer Module 3.0 and symptom AEs were measured using the Pediatric Patient-Reported Outcome-Common Terminology Criteria for AEs (PRO-CTCAEs®). Descriptive statistics were used to describe QoL and symptom AE data. Correlational analyses and generalized linear mixed models were used to examine the relationship between symptom AEs and QoL. Results: Forty-nine children participated in the study. The mean total PedsQLTM score was 73.86 for the sample across all time points. Children diagnosed with a central nervous system (CNS) tumor reported poorer QoL compared to children diagnosed with a hematologic malignancy or non-CNS solid tumor. Symptom frequency AEs of anxiety, pain, nausea, insomnia, hot flashes, and fatigue severity demonstrated the strongest and most significant negative correlation with total QoL scores. Analyses of the relationship between QoL and symptom AEs over time revealed time-specific significant differences with children who experienced frequency AEs of nausea, and anxiety reporting poorer QoL at time point 4 (week 8). Discussion: The Ped PRO-CTCAE® and PedsQLTM can be used to evaluate the relationship between symptom AEs and QoL in practice and in future research.


Synthesis of low-temperature, fast, single-firing body for porcelain stoneware tiles with coal gangue.
Coal gangue is a major industrial solid waste in China, causing great environment pollution. According to phase diagram theory, a low-temperature, fast, single-firing body mix for porcelain stoneware tiles was designed in the quaternary system CaO--MgO--Al₂O₃--SiO₂, using coal gangue as the main raw material. The coal gangue was from Baishan city, Jilin province and mainly composed of kaolinite and quartz. Mineralogical compositions and microstructures of some selected samples sintered at different temperatures were identified with X-ray diffraction (XRD) and scanning electron microscopy (SEM), respectively. The results indicated that the optimal body mix was the one containing 34 wt% coal gangue sintered at 1170°C for about 1 h, with rupture strength of 43 MPa and water absorption of 0.22%. The main crystalline phases of the sintered body were quartz, anorthite and mullite.


Language in Behavioral Variant Frontotemporal Dementia: Another Stone to Be Turned in Latin America.
Beyond canonical deficits in social cognition and interpersonal conduct, behavioral variant frontotemporal dementia (bvFTD) involves language difficulties in a substantial proportion of cases. However, since most evidence comes from high-income countries, the scope and relevance of language deficits in Latin American bvFTD samples remain poorly understood. As a first step toward reversing this scenario, we review studies reporting language measures in Latin American bvFTD cohorts relative to other groups. We identified 24 papers meeting systematic criteria, mainly targeting phonemic and semantic fluency, naming, semantic processing, and comprehension skills. The evidence shows widespread impairments in these domains, often related to overall cognitive disturbances. Some of these deficits may be as severe as in other diseases where they are more widely acknowledged, such as Alzheimer's disease. Considering the prevalence and informativeness of language deficits in bvFTD patients from other world regions, the need arises for more systematic research in Latin America, ideally spanning multiple domains, in diverse languages and dialects, with validated batteries. We outline key challenges and pathways of progress in this direction, laying the ground for a new regional research agenda on the disorder.


Affinity chromatography of the anterior pituitary D2-dopamine receptor.
The D2-dopamine receptor from bovine anterior pituitary has been solubilized with digitonin and purified approximately 1000-fold by affinity chromatography on a new affinity support. This support consists of a (carboxymethylene)oximino derivative of the D2-selective antagonist spiperone (CMOS) covalently attached to Sepharose 4B through a long side chain. The interaction of the solubilized receptor activity with the affinity gel was biospecific. Dopaminergic drugs blocked adsorption of solubilized receptor activity to the CMOS-Sepharose with the appropriate D2-dopaminergic potency and stereoselectivity. For agonists, (-)-N-n-propylnorapomorphine greater than 2-amino-6,7-dihydroxytetrahydronaphthalene approximately equal to apomorphine greater than dopamine, whereas for antagonists (+)-butaclamol much greater than (-)-butaclamol. The same D2-dopaminergic specificity was observed for elution of receptor activity from the gel. To observe eluted receptor binding activity, reconstitution of the eluted material into phospholipid vesicles was necessary. Typically, 70-80% of the solubilized receptor was adsorbed by CMOS-Sepharose, and 40-50% of the adsorbed activity could be recovered after reconstitution of the eluted material. The overall recovery of D2-receptor activity from bovine anterior pituitary membranes was 12-15% with specific binding activity of approximately 150 pmol/mg. The reconstituted affinity-purified receptor bound ligands with the expected D2-dopaminergic specificity, stereoselectivity, and rank order of potency.


A non-cationic nucleic acid nanogel for the delivery of the CRISPR/Cas9 gene editing tool.
Herein, we report a non-cationic DNA-crosslinked nanogel for intracellular delivery of a Cas9 and single guide RNA (Cas9/sgRNA) complex. A DNA-grafted polycaprolactone brush (DNA-g-PCL) is first loaded with the Cas9/sgRNA complex and then crosslinked by DNA linkers via nucleic acid hybridization to form a nanosized hydrogel, in which the gene editing tools are embedded and protected inside. With compact architecture, the Cas9/sgRNA complex-containing nanogel exhibited excellent physiological stability against nuclease digestion and enhanced cellular uptake efficiency, making the delivery system a promising tool for target genome editing.


Effect on plasma lipids and lipoproteins of replacing partially hydrogenated fish oil with vegetable fat in margarine.
We have compared the effects on lipoproteins and haemostatic variables of two hard margarines with similar functional properties, one traditional margarine containing partially hydrogenated fish oil (PHFO), and one experimental margarine based on vegetable oil (VO). Both were all-purpose cooking margarines with nearly identical functional properties. Trans fatty acids from PHFO in the traditional margarine were replaced mostly by saturated, monounsaturated and trans fatty acids of vegetable origin in the new formulation. Both test margarines contained approximately the same amount of cis polyunsaturated fatty acids. Sixteen female normolipidaemic students consumed each diet with the two test margarines for 14 d in random order (crossover design). The amount of fat was 31% energy in the PHFO diet and 32% energy in the VO diet. The test margarines provided approximately 26% energy in both diets. In the PHFO diet 7.8% of the energy was derived from trans fatty acids and 9.2% from saturated fatty acids (12:0, 14:0 and 16:0) while in the VO diet, 1.1% energy was derived from trans fatty acids and 13.3% from saturated fatty acids (12:0, 14:0 and 16:0). The natural content of cholesterol in PHFO was deliberately not balanced by addition of cholesterol to the VO diet, thus the PHFO diet contained 215 mg and the VO diet 86 mg cholesterol per 8.5 MJ. LDL-cholesterol concentration was 19% higher in subjects on the PHFO diet compared with the VO diet (P < 0.01). The ratio LDL-cholesterol:HDL-cholesterol was 12.6% higher in subjects on the PHFO diet compared with the VO diet (P < 0.01). The level of apolipoprotein (apo)A-I was 6% lower in subjects on the PHFO diet compared with the VO diet (P < 0.01). The ratio apoB:apoA-I was 10.4% higher in subjects on the PHFO diet than on the VO diet (P < 0.01). There were no significant differences in total cholesterol, HDL-cholesterol, triacylglycerols, apoB, lipoprotein(a) and haemostatic variables between the diets. Our results demonstrate that PHFO, with its unfavourable effects on plasma lipids, can be replaced by vegetable oils in margarine without appreciable loss of functional properties but with significant improvement in the effects on plasma lipoproteins.


The anterior olfactory nucleus: quantitative study of dendritic morphology.
The anterior olfactory nucleus (AON) occupies a crucial position within the olfactory circuit, as it is able to influence function in nearly every major synaptic processing stage of both the ipsilateral and the contralateral pathways. Nevertheless, very little is known about the region's internal organization and circuitry. The present study provides basic quantitative and qualitative data on the morphology of several cell types within the two major regions of the AON, pars externa and pars principalis. In pars externa two types of cells are analyzed, the "classical" cell (type I), containing only apically directed dendrites with large spines, and a previously unreported cell with basilar dendrites and complex, spiny apical processes (type II). In pars principalis the characteristic pyramidal cell is described both on the basis of the depth of the cell bodies in the cell layer comprising the structure and on the basis of their radial location. Several other nonpyramidal neurons are also described. The findings provide useful basic information necessary for understanding and modeling the circuitry of the AON.


Nociceptin/orphanin FQ in inflammation and sepsis.
Nociceptin/orphanin FQ, N/OFQ, and its receptor NOP represent a non-opioid branch of the opioid superfamily that were first studied for their effects on pain responses. Both N/OFQ and NOP are involved in a wide range of 'non-pain' responses including immunomodulation and cardiovascular control. There is now growing interest in this system in inflammation and sepsis, which is the focus of this review article. The N/OFQ-NOP system is present in immune cells and N/OFQ modifies immunocyte function. On the basis of various in vitro and in vivo studies, N/OFQ increases the inflammatory response in healthy anaesthetized animals and in those with a septic or inflammatory process. It affects tissue perfusion, increases capillary leakage and inflammatory markers, and leads to immune cell chemotaxis. Moreover, NOP activation produces bradycardia and hypotension. Systemic N/OFQ administration also increased mortality in an animal model of sepsis, and there is limited evidence for increased plasma N/OFQ concentrations in patients with sepsis who died compared with those who survived. There is a need for further observational and mechanistic studies in patients with established inflammatory processes or sepsis. These studies may facilitate the design of appropriate clinical studies to evaluate NOP ligands as modifiers of the inflammatory response.


From sea squirts to squirrelfish: facultative trace element hyperaccumulation in animals.
The hyperaccumulation of trace elements is a widely characterized phenomenon in plants, bacteria, and fungi, but has received little attention in animals. However, there are numerous examples of animals that specifically and facultatively accumulate trace elements in the absence of elevated environmental concentrations. Metal hyperaccumulating animals are usually marine invertebrates, likely owing to environmental (e.g. constant exposure via the water) and physiological (e.g. osmoconforming and reduced integument permeability) factors. However, there are examples of terrestrial animals (insect larvae) and marine vertebrates (e.g. squirrelfish) that accumulate high body and/or tissue metal burdens. This review examines examples of animal hyperaccumulation of the elements arsenic, copper, iron, titanium, vanadium and zinc, describing mechanisms by which accumulation occurs and, where possible, hypothesizing functional roles. Groups such as the ascidians (sea squirts), molluscs (gastropods, bivalves and cephalopods) and polychaete annelids feature prominently as animals with hyperaccumulating capacity. Many of these species are potential model organisms offering insight into fundamental processes underlying metal handling, with relevance to human disease and aquatic metal toxicity, and some offer promise in applied fields such as bioremediation.


[A false historic representation of the martyrdom of St. Apollonia: decapitation].
The author quotes the only known historical witness on the life and death of Santa Apollonia: the description given by the historian Eusebius. According to his description of the martyrdom of the Saint, her teeth were extracted and her jawbones broken. An extensive iconography exists in museums, churches and private collections. According to statues and images of the Saint, the martyrdom took place by stoning or else by the use of instruments of torture such as pincers, gouge and gammer, and dentists' forceps. Although it is most probable that recourse was taken to lapidation, it is also likely that the above instruments were used, because it agrees with the technic then in use in dental surgery and does not disagree in any way with the historical facts. Such is not the case regarding the beheading. However, the author quotes three groups of iconographic documents displaying that kind of torture. One of them, shown for the first time, appears in a Book of hours of French origin, dating from the 15th century, which exists in the Calouste Gulbenkian Museum, in Lisbon, Portugal.


Cohabitation, Marijuana Use, and Heavy Alcohol Use in Young Adulthood.
Research on the association between cohabitation and substance use has been inconsistent, with some studies indicating an elevated risk among cohabiters and others finding either no difference in risk or a reduced risk of substance use. However, studies of this association have not utilized a causal modeling empirical framework. The purpose of this research was to assess whether cohabitation has a causal effect on two forms of substance use among young adults: marijuana and heavy alcohol use. Three waves of data from the National Survey of Youth and Religion (n = 2,202; 2002-2008), a representative sample of young adults in the United States, and an augmented inverse probability weighting (AIPW) model designed for multivalued treatment effects estimation, were used to assess the association between cohabitation and substance use. The findings indicated that cohabitation was associated with more frequent marijuana use only among females. Much of the effect of cohabitation, though, was due to previous factors, including substance use, that affected whether young adults cohabit or not. Moreover, there was no evidence that cohabitation had a causal impact on heavy alcohol use. Conclusions/Importance: There is little evidence that cohabitation has a causal impact on substance use in general. However, among young women, those who cohabited reported higher levels of marijuana use than those who remained single. Future research should address why this group is at particular risk of substance use.


Serum Amyloid A Is a Biomarker of Disease Activity and Health-Related Quality-of-Life in Patients with Antineutrophil Cytoplasmic Antibody-Associated Vasculitis.
Serum amyloid A (SAA) is one of the acute phase proteins synthesized in hepatocytes and secreted by various inflammation or infectious stimuli. We investigated the clinical implication of measuring SAA in patients with antineutrophil cytoplasmic antibody- (ANCA-) associated vasculitis (AAV). Seventy-five patients who had been classified as AAV and enrolled in our prospective observational cohort for AAV patients were included. Clinical and laboratory data were obtained on the day of blood sampling, and SAA was measured by ELISA kits. Birmingham Vasculitis Activity Score (BVAS) and Short-Form 36-Item Health Survey (SF-36) were assessed for disease activity and health-related quality-of-life (HRQoL) measures. We stratified patients into having high BVAS when the BVAS was over the median values, and those with either low SF-36 PCS or low SF-36 MCS were defined as having poor HRQoL. Multivariate logistic regression analysis was conducted to estimate independent predictors of high BVAS. The relative risk (RR) was analyzed using the contingency tables and the chi-squared test. SAA was positively correlated with BVAS (r = 0.642) and FFS (r = 0.367) and was inversely correlated with both the SF-36 physical component summary (r = -0.456) and mental component summary scores (r = -0.394). Furthermore, SAA was significantly correlated with acute phase reactants ESR (r = 0.611) and CRP (r = 0.629). Patients with high BVAS exhibited significantly higher SAA than those with low BVAS (1317.1 ng/mL vs. 423.1 ng/mL). In multivariable logistic regression analysis, serum albumin (odds ratio (OR) 0.132) and SAA > 1173.6 ng/mL (OR 15.132) were independently associated with high BVAS. The risk of having high BVAS and poor HRQoL in patients with SAA > 1173.6 ng/mL was higher than in those with SAA ≤ 1173.6 ng/mL (RR 3.419 and 1.493). Our results suggest that SAA might be a useful biomarker in assessing disease activity and HRQoL in AAV.


Validation of the Chinese Handwriting Analysis System (CHAS) for primary school students in Hong Kong.
There are more children diagnosed with specific learning difficulties in recent years as people are more aware of these conditions. Diagnostic tool has been validated to screen out this condition from the population (SpLD test for Hong Kong children). However, for specific assessment on handwriting problem, there seems a lack of standardized and objective evaluation tool to look into the problems. The objective of this study was to validate the Chinese Handwriting Analysis System (CHAS), which is designed to measure both the process and production of handwriting. The construct validity, convergent validity, internal consistency and test-retest reliability of CHAS was analyzed using the data from 734 grade 1-6 students from 6 primary schools in Hong Kong. Principal Component Analysis revealed that measurements of CHAS loaded into 4 components which accounted for 77.73% of the variance. The correlation between the handwriting accuracy obtained from HAS and eyeballing was r=.73. Cronbach's alpha of all measurement items was .65. Except SD of writing time per character, all the measurement items regarding handwriting speed, handwriting accuracy and pen pressure showed good to excellent test-retest reliability (r=.72-.96), while measurement on the numbers of characters which exceeded grid showed moderate reliability (r=.48). Although there are still ergonomic, biomechanical or unspecified aspects which may not be determined by the system, the CHAS can definitely assist therapists in identifying primary school students with handwriting problems and implement interventions accordingly.


The Impact of a Shortage of Pharmacy-Prepared Ephedrine Syringes on Intraoperative Medication Use.
Anesthesia-related medication shortages have become increasingly common in the United States. We tested whether a local shortage of pharmacy-prepared ephedrine syringes, replaced by provider-prepared ephedrine, was associated with provider-level changes in ephedrine and phenylephrine use and patient-level changes in intraoperative hemodynamics. Consecutive patients undergoing general and orthopedic surgery at a tertiary care center were included 1 month before and 1 month after the start of the pharmacy-prepared ephedrine syringe shortage. Lowest mean arterial blood pressure and slowest heart rate were obtained as measures of hemodynamics. Adjusted associations were tested using mixed-effects regression with repeated measures. Three hundred four patients before the shortage and 298 patients after the shortage began were included. The administration of at least 1 bolus of ephedrine was significantly more common before versus during the shortage (148/304 [48.7]% vs 117/298 [39.3]%; P = 0.0199). After adjusting for age, sex, ASA physical status, surgery type, anesthesia provider, and operative duration, patients were significantly less likely to receive ephedrine during the shortage (relative risk [RR] = 0.78 [95% confidence interval {CI}, 0.61-0.96]; P = 0.0198) and more likely to receive a phenylephrine bolus (RR = 1.27 [95% CI, 1.02-1.51]; P = 0.0357). Patient hemodynamics assessed by slowest heart rate or lowest mean arterial blood pressure did not differ significantly during the shortage. There was an alteration in medication administration patterns during a shortage of pharmacy-prepared syringes. Changes in ephedrine and phenylephrine use were noted; however, patient hemodynamics remained comparable. Provider use patterns were sensitive even to a relative and not absolute medication shortage as observed in this study.


Processing of natural time series of intensities by the visual system of the blowfly.
A major problem that a visual system faces is how to fit the large intensity variation of natural image streams into the limited dynamic range of its neurons. One of the means to accomplish this is through the use of gain control. In order to investigate this, natural time series of intensities were measured, as well as the responses of blowfly photoreceptors and Large Monopolar Cells (LMCs) to these time series. Time series representative of what each photoreceptor of a real visual system would normally receive were measured with an optical system measuring the light intensity of a spot comparable with the field of view of single human foveal cones. This system was worn on a headband by a freely walking person. Resulting time series have rms-contrasts ranging from an average of 0.45 for 1-sec segments to 1.39 for 100-sec segments (both when limited to frequencies up to 100 Hz). Power spectra behave approximately as 1/f (f: temporal frequency). Measured time series were subsequently presented to fly photoreceptors and LMCs by playing them back on an LED. The results show that fast gain controls indeed keep the response within the dynamic range of the cells and that a large part of this range is actually used for packing the information in natural time series.


[Sulphonylurea derivatives and the cardiovascular system].
Sulphonylurea derivatives (SUD) are a mainstay of treatment of type 2 diabetes but questions have been raised about the potential adverse effects of these drugs as far as cardiovascular functions are concerned. An early prospective study which examined the effects of glycaemic control with various agents on coronary heart disease was stopped in the 70-ies due to excess cardiovascular mortality in the group receiving SUD of first generation: tolbutamide. The discovery of ATP-sensitive potassium channels in the heart, their role in ischaemic heart disease and mechanisms of endogenous cardiac cell protection--preconditioning have brought back concerns of SUD safety. Recently, a new SUD--glimepiride--claimed as the first representant of III generation of these agents--has been introduced into clinical practice. Glimepiride appears to be devoid of vascular ATP-sensitive potassium channels binding properties. Postulated and confirmed in animal experimental studies cardioprotective features of glimepiride were evaluated in a randomised, placebo-controlled study with glimepiride and glibenclamide comparing effects of these drugs on ischaemic preconditioning during angioplasty of high grade coronary artery stenoses in patients with stable angina. Myocardial ischaemia was quantified by intracoronary electrocardiography and time to occurrence of angina during vessel occlusion was measured. The results of the study confirmed glimepiride effects of maintaining myocardial preconditioning. The article summarises current knowledge of SUD influence on cardiovascular system and discusses some differences in pharmacodynamics of glimepiride which appear to provide this agent with clinical advantages over conventional SUD at least in cardiovascular aspects.


Gastric cancers emerging after H. pylori eradication arise exclusively from non-acid-secreting areas.
Although Helicobacter pylori (H. pylori) eradication has some inhibitory effects on the subsequent development of gastric cancer, there are sporadic cases of gastric cancer even after successful eradication. The pathogenesis of gastric cancer emerging after H. pylori eradication remains to be clarified. In this study, employing Congo-red chromoendoscopy, which is capable of visualizing the acid-secreting fundic mucosa, we investigated the topographic relationship of the acid secretion pattern to the occurrence site of gastric cancers emerging after eradication. Fourteen consecutive patients who suffered from new gastric cancer after eradication, defined as lesions that were discovered at least 2 years after the eradication, were prospectively enrolled. Whether the neoplasias arose from acid-secreting or non-acid-secreting areas was evaluated with Congo-red chromoendoscopy. Biopsy specimens taken from the two areas were subjected to histologic evaluation and immunohistochemistry for Ki-67 and p53. The mean period from the eradication to the subsequent occurrence of gastric cancer was 74 (44) months. There were two cancer lesions in 5 cases, and thus there was a total 19 lesions from 14 cases. Congo-red chromoendoscopy revealed that all 19 lesions arose exclusively from non-acid-secreting areas. Histological examination revealed sustained hyperproliferation and accumulation of p53 protein was frequently detectable in non-acid-secreting areas. Genetic alteration such as p53 mutation seems to be already present in the residual non-acid-secreting areas after eradication, areas that could be the origin of gastric carcinogenesis after eradication. Identification of such high-risk areas should be a promising approach for estimating the individual cancer risk after eradication.


Fractionation of hemp hurds by organosolv pretreatment and its effect on production of lignin and sugars.
Fractionation of hemp hurds into its three main components, cellulose, hemicellulose, and lignin, was carried out using organosolv pretreatment. The effect of processing parameters, such as temperature, catalyst concentration, reaction time, and methanol (MeOH) concentration, on the dissolution and recovery of hemicellulose and lignin was determined. More than 75% of total hemicellulose and 75% of total lignin was removed in a single step with low amounts of degradation products under the following conditions: 165 °C, 3% H2 SO4 , 20 min reaction time, and 45% MeOH. Enzymatic hydrolysis of the residual pretreated biomass yielded up to 60% of cellulose-to-glucose conversion. The maximum recovery of the main components was obtained at a combined severity factor value of around one. Characterization of pretreated biomass and isolated lignin was carried out with FTIR and 2D (13) C-(1) H correlation HSQC NMR spectroscopy, the latter technique providing detailed structural information about the obtained methanol organosolv lignin (MOSL). Results suggested that xylopyranoside is the major carbohydrate associated with hemp lignin. The chemical properties of MOSL samples in terms of their phenolic group content and antioxidant capacity were also investigated. The results showed that MOSL samples have a high phenolic group content and antioxidant capacity relative to Klason lignin.


Coaching Intensity, Adherence to Essential Birth Practices, and Health Outcomes in the BetterBirth Trial in Uttar Pradesh, India.
Coaching can improve the quality of care in primary-level birth facilities and promote birth attendant adherence to essential birth practices (EBPs) that reduce maternal and perinatal mortality. The intensity of coaching needed to promote and sustain behavior change is unknown. We investigated the relationship between coaching intensity, EBP adherence, and maternal and perinatal health outcomes using data from the BetterBirth Trial, which assessed the impact of a complex, coaching-based implementation of the World Health Organization's Safe Childbirth Checklist in Uttar Pradesh, India. For each birth, we defined multiple coaching intensity metrics, including coaching frequency (coaching visits per month), cumulative coaching (total coaching visits accrued during the intervention), and scheduling adherence (coaching delivered as scheduled). We considered coaching delivered at both facility and birth attendant levels. We assessed the association between coaching intensity and birth attendant adherence to 18 EBPs and with maternal and perinatal health outcomes using regression models. Coaching frequency was associated with modestly increased EBP adherence. Delivering 6 coaching visits per month to facilities was associated with adherence to 1.3 additional EBPs (95% confidence interval [CI]=0.6, 1.9). High-frequency coaching delivered with high coverage among birth attendants was associated with greater improvements: providing 70% of birth attendants at a facility with at least 1 visit per month was associated with adherence to 2.0 additional EBPs (95% CI=1.0, 2.9). Neither cumulative coaching nor scheduling adherence was associated with EBP adherence. Coaching was generally not associated with health outcomes, possibly due to the small magnitude of association between coaching and EBP adherence. Frequent coaching may promote behavior change, especially if delivered with high coverage among birth attendants. However, the effects of coaching were modest and did not persist over time, suggesting that future coaching-based interventions should explore providing frequent coaching for longer periods.


Cardiovascular risk factors burden in Saudi Arabia: The Africa Middle East Cardiovascular Epidemiological (ACE) study.
Limited data exist on the epidemiology of cardiovascular risk factors in Saudi Arabia, particularly in relation to the differences between Saudi nationals and expatriates in Saudi Arabia. The aim of this analysis was to describe the current prevalence of cardiovascular risk factors among patients attending general practice clinics across Saudi Arabia. In this cross-sectional epidemiological analysis of the Africa Middle East Cardiovascular Epidemiological (ACE) study, the prevalence of cardiovascular risk factors (hypertension, diabetes, dyslipidemia, obesity, smoking, abdominal obesity) was evaluated in adults attending primary care clinics in Saudi Arabia. Group comparisons were made between patients of Saudi ethnicity (SA nationals) and patients who were not of Saudi ethnicity (expatriates). A total of 550 participants were enrolled from different clinics across Saudi Arabia [aged (mean ± standard deviation) 43 ± 11 years; 71% male]. Nearly half of the study cohort (49.8%) had more than three cardiovascular risk factors. Dyslipidemia was the most prevalent risk factor (68.6%). The prevalence of hypertension (47.5%) and dyslipidemia (75.5%) was higher among expatriates when compared with SA nationals (31.4% vs. 55.1%, p = 0.0003 vs. p < 0.0001, respectively). Conversely, obesity (52.6% vs. 41.0%; p = 0.008) and abdominal obesity (65.5% vs. 52.2%; p = 0.0028) were higher among SA nationals vs. expatriates. Modifiable cardiovascular risk factors are highly prevalent in SA nationals and expatriates. Programmed community-based screening is needed for all cardiovascular risk factors in Saudi Arabia. Improving primary care services to focus on risk factor control may ultimately decrease the incidence of coronary artery disease and improve overall quality of life. The ACE trial is registered under NCT01243138.


Cryptococcus neoformans. I. Nonencapsulated mutants.
Seven nonencapsulated mutants of Cryptococcus neoformans were isolated from an encapsulated strain of human origin. Initially, the mutants were avirulent for mice. After several months of subculturing, six of the seven isolates reverted to the encapsulated state and possessed varying degrees of virulence. The results of these experiments suggest that a strong correlation exists between the presence of a capsule and the virulence of C. neoformans.


SPRING - an image processing package for single-particle based helical reconstruction from electron cryomicrographs.
Helical reconstruction from electron cryomicrographs has become a routine technique for macromolecular structure determination of helical assemblies since the first days of Fourier-based three-dimensional image reconstruction. In the past decade, the single-particle technique has had an important impact on the advancement of helical reconstruction. Here, we present the software package SPRING that combines Fourier based symmetry analysis and real-space helical processing into a single workflow. One of the most time-consuming steps in helical reconstruction is the determination of the initial symmetry parameters. First, we propose a class-based helical reconstruction approach that enables the simultaneous exploration and evaluation of many symmetry combinations at low resolution. Second, multiple symmetry solutions can be further assessed and refined by single-particle based helical reconstruction using the correlation of simulated and experimental power spectra. Finally, the 3D structure can be determined to high resolution. In order to validate the procedure, we use the reference specimen Tobacco Mosaic Virus (TMV). After refinement of the helical symmetry, a total of 50,000 asymmetric units from two micrographs are sufficient to reconstruct a subnanometer 3D structure of TMV at 6.4Å resolution. Furthermore, we introduce the individual programs of the software and discuss enhancements of the helical reconstruction workflow. Thanks to its user-friendly interface and documentation, SPRING can be utilized by the novice as well as the expert user. In addition to the study of well-ordered helical structures, the development of a streamlined workflow for single-particle based helical reconstruction opens new possibilities to analyze specimens that are heterogeneous in symmetries.


Soluble forms of Toll-like receptor (TLR)2 capable of modulating TLR2 signaling are present in human plasma and breast milk.
Dysregulation of the initial, innate immune response to bacterial infection may lead to septic shock and death. Toll-like receptors (TLRs) play a crucial role in this innate immune response, and yet the regulatory mechanisms controlling microbial-induced TLR triggering are still to be fully understood. We have therefore sought specific regulatory mechanisms that may modulate TLR signaling. In this study, we tested for the possible existence of a functionally active soluble form of TLR2. We demonstrated the existence of natural soluble forms of TLR2 (sTLR2), which we show to be capable of modulating cell activation. We found that blood monocytes released sTLR2 constitutively and that the kinetics of sTLR2 release increased upon cell activation. Analysis of cells expressing the human TLR2 cDNA or its c-myc-tagged version indicated that sTLR2 resulted from the posttranslational modification of the TLR2 protein in an intracellular compartment. Moreover, an intracellular pool of sTLR2 is maintained. sTLR2 was found naturally expressed in breast milk and plasma. Milk sTLR2 levels mirrored those of the TLR coreceptor soluble CD14. Depletion of sTLR2 from serum resulted in an increased cellular response to bacterial lipopeptide. Notably, serum sTLR2 was lower in tuberculosis patients. Coimmunoprecipitation experiments and computational molecular docking studies showed an interaction between sTLR2 and soluble CD14 in plasma and milk. These findings suggest the existence of a novel and specific innate immune mechanism regulating microbial-induced TLR triggering, and may lead to new therapeutics for the prevention and/or treatment of severe infectious diseases.


The N-terminal Arg residue is essential for autocatalytic activation of a lipopolysaccharide-responsive protease zymogen.
Factor C, a serine protease zymogen involved in innate immune responses in horseshoe crabs, is known to be autocatalytically activated on the surface of bacterial lipopolysaccharides, but the molecular mechanism of this activation remains unknown. In this study, we show that wild-type factor C expressed in HEK293S cells exhibits a lipopolysaccharide-induced activity equivalent to that of native factor C. Analysis of the N-terminal addition, deletion, or substitution mutants shows that the N-terminal Arg residue and the distance between the N terminus and the tripartite of lipopolysaccharide-binding site are essential factors for autocatalytic activation, and that the positive charge of the N terminus may interact with an acidic amino acid(s) of the molecule to convert the zymogen into an active form. Chemical cross-linking experiments indicate that the N terminus is required to form a complex of the factor C molecules in a sufficiently close vicinity to be chemically cross-linked on the surface of lipopolysaccharides. We propose a molecular mechanism of the autocatalytic activation of the protease zymogen on lipopolysaccharides functioning as a platform to induce specific protein-protein interaction between the factor C molecules.