Perceived discrimination and medication adherence in black hypertensive patients: the role of stress and depression.
To examine the relationship between perceived discrimination and medication adherence among black people with hypertension and the role of stress and depressive symptoms in this relationship. Perceived racial discrimination has been associated with poor health outcomes in blacks; its relationship to medication adherence among hypertensive patients remains untested. We measured perceived racial discrimination at baseline, stress and depressive symptoms at 6 months, and medication adherence at 12 months among patients enrolled in a 30-site cluster-randomized controlled trial testing a patient and physician-targeted intervention to improve blood pressure. A mediational method with bootstrapping (stratified by site) confidence intervals was used to estimate the indirect association between perceived discrimination and medication adherence through stress and depression. Of 1056 patients from 30 sites enrolled in the trial, 463 had complete data on all four measures at 6 and 12 months and were included in the analyses. Adjusting for clustering, perceived discrimination was associated with poor medication adherence (B = 0.138, p = .011) at 12 months, and with stress (B = 2.24, p = .001) and depression (B = 1.47, p = .001) at 6 months. When stress and depression were included in the model, there was a 65% reduction in the total association of perceived discrimination with medication adherence, and the relationship was no longer significant (B = 0.049, p = .35). Perceived discrimination is associated with poor medication adherence among hypertensive blacks, and stress and depressive symptoms may account for this relationship. clinicaltrials.gov Identifier: NCT00233220.


Acidification of phagosomes containing Salmonella typhimurium in murine macrophages.
Salmonella species are facultative intracellular pathogens. Following entry into mammalian host cells, they reside in membrane-bound vacuoles, resist killing, and replicate. In this work, we investigated the importance of phagosomal pH in the ability of Salmonella typhimurium to survive and replicate within macrophages. Intraphagosomal pH was measured in situ by recording the fluorescence intensity of a pH-sensitive probe, DM-NERF dextran. The majority of vacuoles containing S. typhimurium (live, heat killed, or formalin fixed) acidified from pH > or = 6.0 to between pH 4.0 and 5.0 within 60 min after formation. In contrast, Mycobacterium avium-containing vacuoles failed to acidify even at later time points. Acidification of S. typhimurium-containing vacuoles was completely blocked by treatment of host cells with bafilomycin A, a specific inhibitor of vacuolar proton-ATPases. Bafilomycin inhibition of vacuolar acidification from the onset of infection significantly decreased the survival of S. typhimurium in macrophages. Furthermore, bafilomycin treatment at 2, 4, 8, or even 12 h postinfection decreased the percentage of recoverable bacteria by up to 20-fold. Loss of bacterial viability was seen with several other reagents which, like bafilomycin, raise the pH of phagosomal compartments but are not directly lethal to the bacteria or host cells. Thus, we conclude that Salmonella-containing phagosomes acidify soon after formation and hypothesize that an acidic environment is necessary for survival and replication of the bacteria within the macrophage.


Progressive encephalomyelitis with rigidity: A Taiwanese case and review of literature.
Progressive encephalomyelitis with rigidity and myoclonus (PERM) is a rare disorder. However, the outcome is still variable with different serological and tumor associations, and the elements to good response with less relapse is yet to be elucidated. We present a case and obtain a literature review of patients with PERM and make comparisons based on different serological groups. We also analyze patients with idiopathic PERM that had detailed medical records. 81 patients were collected and analyzed. The largest group were glycine receptor-antibody (GlyR-Ab)-positive (70%), and the seropositive-GlyR-Ab-negative group had better response to immunotherapy. Malignancy can occur up to 2 years from the presentation of PERM. Among the 18 cases with detailed records, the patients who had good outcome initiate immunotherapy within 2 months from presentation. 9 of the 12 patients who experienced no relapse had non-steroid immunotherapy. The maximal interval time of relapse was 24 months. We recommend tumor surveillance up to 2 years in patients with PERM and early administration of immunotherapies and maintain with non-steroid immunotherapy with or without oral corticosteroid for a minimum of 2 years to reduce the risk of relapse in GlyR-Ab-positive patients.


Molecular phenotype of inflammatory bowel disease-associated neoplasms with microsatellite instability.
Patients with inflammatory bowel disease (IBD) are at increased risk of developing colorectal cancer (CRC). We sought to determine the frequency of high-level microsatellite instability (MSI-H) and the mutational and methylation profile of MSI-H IBD-related neoplasms (IBDNs). A total of 124 IBDNs (81 cancers, 43 dysplasias) from 78 patients were studied for the frequency of MSI-H and hypermethylation of 3 target genes: MLH1 , HPP1 , and RAB-32 . Fifteen MSI-H IBDNs were characterized according to their profile of frameshift mutations in 28 mononucleotide repeats and compared with 46 sporadic MSI-H CRCs. Nineteen of 124 IBDNs were MSI-H. The frequency of frameshift mutations in coding mononucleotide repeats was significantly lower in MSI-H IBDNs than in sporadic MSI-H CRCs for TGFBR2 (7 of 14 vs 34 of 43 samples; P = .047) and ACVR2 (3 of 14 vs 25 of 43 samples; P = .029). In contrast, ICA1 was mutated in 3 of 9 MSI-H IBDNs vs 2 of 54 sporadic MSI-H CRCs ( P = .028). HPP1 and RAB32 methylation was independent of MSI status and was observed in 4 of 59 and 0 of 64 nondysplastic mucosae, 20 of 38 and 1 of 25 dysplasias, and 28 of 61 and 20 of 60 carcinomas, respectively. The profiles of coding microsatellite mutations (instabilotypes) differ significantly between MSI-H IBDNs and MSI-H sporadic CRCs. Specifically, TGFBR2 and ACVR2 mutations are significantly rarer in MSI-H IBDNs than in MSI-H sporadic CRCs. Furthermore, HPP1 methylation occurs early, in 7% of nondysplastic and approximately half of dysplastic mucosae, whereas RAB32 methylation occurs at the transition to invasive growth, being rarer in dysplasias.


Rejuvenation in Deep Thermal Cycling of a Generic Model Glass: A Study of Per-Particle Energy Distribution.
We investigate the effect of low temperature (cryogenic) thermal cycling on a generic model glass and observe signature of rejuvenation in terms of per-particle potential energy distributions. Most importantly, these distributions become broader and its average values successively increase when applying consecutive thermal cycles. We show that linear dimension plays a key role for these effects to become visible, since we do only observe a weak effect for a cubic system of roughly one hundred particle diameter but observe strong changes for a rule-type geometry with the longest length being two thousand particle diameters. A consistent interpretation of this new finding is provided in terms of a competition between relaxation processes, which are inherent to glassy systems, and excitation due to thermal treatment. In line with our previous report (Bruns et al., PRR 3, 013234 (2021)), it is shown that, depending on the parameters of thermal cycling, rejuvenation can be either too weak to be detected or strong enough for a clear observation.


A mouse model of pathological small intestinal epithelial cell apoptosis and shedding induced by systemic administration of lipopolysaccharide.
The gut barrier, composed of a single layer of intestinal epithelial cells (IECs) held together by tight junctions, prevents the entrance of harmful microorganisms, antigens and toxins from the gut lumen into the blood. Small intestinal homeostasis is normally maintained by the rate of shedding of senescent enterocytes from the villus tip exactly matching the rate of generation of new cells in the crypt. However, in various localized and systemic inflammatory conditions, intestinal homeostasis can be disturbed as a result of increased IEC shedding. Such pathological IEC shedding can cause transient gaps to develop in the epithelial barrier and result in increased intestinal permeability. Although pathological IEC shedding has been implicated in the pathogenesis of conditions such as inflammatory bowel disease, our understanding of the underlying mechanisms remains limited. We have therefore developed a murine model to study this phenomenon, because IEC shedding in this species is morphologically analogous to humans. IEC shedding was induced by systemic lipopolysaccharide (LPS) administration in wild-type C57BL/6 mice, and in mice deficient in TNF-receptor 1 (Tnfr1(-/-)), Tnfr2 (Tnfr2(-/-)), nuclear factor kappa B1 (Nfκb1(-/-)) or Nfĸb2 (Nfĸb2(-/-)). Apoptosis and cell shedding was quantified using immunohistochemistry for active caspase-3, and gut-to-circulation permeability was assessed by measuring plasma fluorescence following fluorescein-isothiocyanate-dextran gavage. LPS, at doses ≥0.125 mg/kg body weight, induced rapid villus IEC apoptosis, with peak cell shedding occurring at 1.5 hours after treatment. This coincided with significant villus shortening, fluid exudation into the gut lumen and diarrhea. A significant increase in gut-to-circulation permeability was observed at 5 hours. TNFR1 was essential for LPS-induced IEC apoptosis and shedding, and the fate of the IECs was also dependent on NFκB, with signaling via NFκB1 favoring cell survival and via NFκB2 favoring apoptosis. This model will enable investigation of the importance and regulation of pathological IEC apoptosis and cell shedding in various diseases.


Cognitive Effects of Social Media Use: A Case of Older Adults.
Research on the effects of social media use at older ages has largely focused on social benefits. Yet, participation in these new media forms may result in other favorable outcomes, such as improved cognitive functioning. Using a wait list-control design, this study examines the effects of social media engagement among novice adult social media users, aged 65 and older, in four cognitive domains: attention, processing speed, working memory, and inhibitory control. Baseline and multiple post-tests indicate improvement of intervention participants in inhibitory control. These findings demonstrate that the benefits of social media use at older ages extend beyond mere social engagement, and into other domains of everyday well-being.


Solvent-induced reversed stereoselectivity in reciprocal resolutions of mandelic acid and erythro-2-amino-1,2-diphenylethanol.
Solvent-induced chirality switching in reciprocal optical resolution between mandelic acid (1) and erythro-2-amino-1,2-diphenylethanol (2) has been demonstrated. The stereochemistry of the deposited salts was controlled by changing the crystallization solvent from 1-PrOH or 1-BuOH to 1,4-dioxane. It was revealed from (1)H NMR spectra, thermogravimetric analysis, and X-ray crystallography of the salts that an equimolar amount of the crystallization solvent was incorporated in each diastereomeric salt. On the basis of the crystal structures, it was found that both the hydrogen-bonding ability and the size of the solvent molecule played an important role. Differences in the formed hydrogen-bonding networks (columnar or sheetlike structure) and their packing manner were found to be crucial for the reversed stereoselectivity. Furthermore, pseudopolymorphic salt crystals that incorporated 1,4-dioxane were obtained during the enantioseparation of racemic 2, and their solid-state properties were examined by measurement of their IR spectra. This solvent-induced dual stereocontrol technique was successfully applied to the successive resolution process, eliminating the need to change the resolving agent for access to both enantiomers of 1 and 2.


On the accuracy of dose prediction near metal fixation devices for spine SBRT.
The metallic fixations used in surgical procedures to support the spine mechani-cally usually consist of high-density materials. Radiation therapy to palliate spinal cord compression can include prophylactic inclusion of potential tumor around the site of such fixation devices. Determination of the correct density and shape of the spine fixation device has a direct effect on the dose calculation of the radiation field. Even with the application of modern computed tomography (CT), under- or overestimation of dose, both immediately next to the device and in the surround-ing tissues, can occur due to inaccuracies in the dose prediction algorithm. In this study, two commercially available dose prediction algorithms (Eclipse AAA and ACUROS), EGSnrc Monte Carlo, and GAFchromic film measurements were com-pared for a clinical spine SBRT case to determine their accuracy. An open six-field plan and a clinical nine-field IMRT plan were applied to a phantom containing a metal spine fixation device. Dose difference and gamma analysis were performed in and around the tumor region adjacent to the fixation device. Dose calculation inconsistency was observed in the open field plan. However, in the IMRT plan, the dose perturbation effect was not observed beyond 5 mm. Our results suggest that the dose effect of the metal fixation device to the spinal cord and the tumor volume is not observable, and all dose calculation algorithms evaluated can provide clinically acceptable accuracy in the case of spinal SBRT, with the tolerance of 95% for gamma criteria of 3%/3 mm.


Hemodynamics in internal carotid artery occlusion examined by positron emission tomography.
Using positron emission tomography in nine patients with minor strokes, unilateral internal carotid artery occlusion, and good collateral circulation through the anterior portion of the circle of Willis, we analyzed regional cerebral blood flow, cerebral metabolic rate of oxygen, oxygen extraction fraction, and cerebral blood volume. These studies allowed quantification of the regional hemodynamic status, especially in relation to watershed areas. Compared with eight normal controls, the patients had significantly (p less than 0.01) decreased regional cerebral blood flow in the middle cerebral artery territory and the surrounding watershed areas of the occluded hemisphere. The oxygen extraction fraction rose with the distance from the anterior portion of the circle of Willis, attaining the highest value in the superior parietal and posterior temporo-occipital watershed area. A concomitant decrease in the cerebral blood flow/cerebral blood volume ratio suggested reduction in the mean blood flow velocity, whereby elevated blood viscosity would be more liable to reduce cerebral blood flow. These findings suggest hemodynamic vulnerability of the watershed areas after internal carotid artery occlusion in persons with good collateral circulation through the anterior portion of the circle of Willis. Our results also emphasize the importance of systemic hemodynamic factors such as blood pressure and circulating blood volume in the genesis of watershed infarction.


Next-of-kin perceptions of physician responsiveness to symptoms of hospitalized patients near death.
Many different medical providers visit critically ill patients during a hospitalization, and patients and family members may not feel any physician is truly in charge of care. This study explores whether perceiving that a physician was clearly in charge is associated with reports by surviving next of kin about the responsiveness of physicians to symptoms in hospitalized patients near the end of life. We conducted telephone interviews with surviving next of kin of adult patients (n = 1107) who died in one of five New York City teaching hospitals between April 1998 and June 1999 after a minimum 3-day inpatient stay. Next-of-kin ratings of whether physicians did "all they could" all or most of the time in response to patient pain, dyspnea, and affective distress (confusion, depression or emotional distress) were compared by whether the next of kin reported one or more physicians "clearly in charge" of care, adjusting for patient and next-of-kin characteristics. More than 80% of patients were reported to have experienced often serious pain, dyspnea, or affective distress. Physicians were rated as responsive to pain by 79.1% of respondents, to dyspnea by 84.9%, and to affective distress by 66.6%. Ratings of physician responsiveness to pain (p = 0.001) and affective distress (p = 0.001) were significantly lower among patients for whom no physician was seen as clearly in charge of care. This finding is consistent with the view that ensuring that a physician coordinates the care of seriously ill, hospitalized patients may improve symptom management. Further research is warranted to establish causality and identify optimal models of care.


Long-Range Communication Network in the Type 1B Bone Morphogenetic Protein Receptor.
Protein-protein interactions are recognized as a fundamental phenomenon that is intimately associated with biological functions and thus are ideal targets for developing modulators for regulating biological functions. A challenge is to identify a site that is situated away from but functionally connected to the protein-protein interface. We employed bone morphogenetic proteins (BMPs) and their receptors as a model system to develop a strategy for identifying such a network of communication. Accordingly, using computational analyses with the COREX/BEST algorithm, we uncovered an overall pattern connecting various regions of BMPR-1B ectodomain, including the four conserved residues in the protein-protein interface. In preparation for testing the long-range effects of mutations of distal residues for future studies, we examined the extent of measurable perturbation of the four conserved residues by determination of the conformation and relative affinities of these BMPR-1B mutants for ligands BMP-2, -6, and -7 and GDF-5. Results suggest no significant structural changes in the receptor but do suggest that the four residues play different roles in defining ligand affinity and both intra- and intermolecular interactions play a role in defining ligand affinity. Thus, these results established two primary but necessary goals: (1) the baseline knowledge of perturbation of conserved interfacial residues for future reference and (2) the ability of the computational approach to identify the distal residues connecting to the interfacial residues. The data presented here provide the foundation for future experiments to identify the effects of distal residues that affect the specificity and affinity of BMP recognition. Protein-protein interactions are integral reactions in essentially all biological activities such as gene regulation and age-related development. Often, diseases are consequences of the alteration of these intermacromolecular interactions, which are thus recognized as a legitimate target for developing modulators for regulating biological functions. One approach is to design ligands that bind to the protein-protein interface. Another is to identify an allosteric site, an advantage of which is bypassing the potential challenge in competing for high-affinity interfacial interactions or a specific interface in a superassembly of multiple macromolecules. However, a challenge of this approach is identifying a site that is situated away from but functionally connected to the protein-protein interface.


The 11-item workplace organizational policies and practices questionnaire (OPP-11): examination of its construct validity, factor structure, and predictive validity in injured workers with upper-limb disorders.
Reliable and valid measurement of workplace organizational policies and practices (OPPs) is needed to evaluate their influences on the prevention and management of occupational injuries. Injured workers (n = 614) attending an upper-limb specialty clinic operated by the Workplace Safety & Insurance Board of Ontario were recruited for a 1-year study. The OPP-11, completed at baseline, was examined for scaling properties (floor/ceiling effects, internal consistency, and item-convergent/discriminant validity), construct validity, and factor structure. Predictive validity for longitudinal work disability was examined using multivariable logistic regressions. Strong scaling properties were observed for the OPP-11. High internal consistency (Cronbach's alpha = 0.80-0.90) and expected associations with comparator constructs were evident. A 4-domain structure (fit statistics: CFI = 0.98, TLI = 0.97, and RMSEA = 0.06) was supported. Higher OPP-11 predicted more favorable 12-month work disability outcomes, after adjusting for demographic and health attributes. The OPP-11 is valid for use in upper-limb disorders. Expanding the ergonomic practices domain could be considered.


Cloning and genetic diversity analysis of a new P5CS gene from common bean (Phaseolus vulgaris L.).
Delta(1)-pyrroline-5-carboxylate synthetase (P5CS) is the rate-limiting enzyme involved in the biosynthesis of proline in plants. By the 3' rapid amplification of cDNA ends (3'-RACE) approach, a 2,246-bp cDNA sequence was obtained from common bean (Phaseolus vulgaris L.), denominated PvP5CS2 differing from another P5CS gene that we cloned previously from common bean (PvP5CS). The predicted amino acid sequence of PvP5CS2 has an overall 93.2% identity GmP5CS (Glycine max L. P5CS). However, PvP5CS2 shows only 83.7% identity in amino acid sequence to PvP5CS, suggesting PvP5CS2 represents a homolog of the soybean P5CS gene. Abundant indel (insertion and deletion events) and SNP (single nucleotide polymorphisms) were found in the cloned PvP5CS2 genome sequence when comparing 24 cultivated and 3 wild common bean accessions and these in turn reflected aspects of common bean evolution. Sequence alignment showed that genotypes from the same gene pool had similar nucleotide variation, while genotypes from different gene pools had distinctly different nucleotide variation for PvP5CS2. Furthermore, diversity along the gene sequence was not evenly distributed, being low in the glutamic-g-semialdehyde dehydrogenase catalyzing region, moderate in the Glu-5-kinase catalyzing region and high in the intervening region. Neutrality tests showed that PvP5CS2 was a conserved gene undergoing negative selection. A new marker (Pv97) was developed for genetic mapping of PvP5CS2 based on an indel between DOR364 and G19833 sequences and the gene was located between markers Bng126 and BMd045 on chromosome b01. The relationship of PvP5CS2 and a previously cloned pyrroline-5-carboxylate synthetase gene as well as the implications of this work on selecting for drought tolerance in common bean are discussed.


Phonon hydrodynamics in two-dimensional materials.
The conduction of heat in two dimensions displays a wealth of fascinating phenomena of key relevance to the scientific understanding and technological applications of graphene and related materials. Here, we use density-functional perturbation theory and an exact, variational solution of the Boltzmann transport equation to study fully from first-principles phonon transport and heat conductivity in graphene, boron nitride, molybdenum disulphide and the functionalized derivatives graphane and fluorographene. In all these materials, and at variance with typical three-dimensional solids, normal processes keep dominating over Umklapp scattering well-above cryogenic conditions, extending to room temperature and more. As a result, novel regimes emerge, with Poiseuille and Ziman hydrodynamics, hitherto typically confined to ultra-low temperatures, characterizing transport at ordinary conditions. Most remarkably, several of these two-dimensional materials admit wave-like heat diffusion, with second sound present at room temperature and above in graphene, boron nitride and graphane.


Maternal and perinatal outcomes of twin pregnancy in 23 low- and middle-income countries.
Twin pregnancies in low- and middle-income countries (LMICs) pose a high risk to mothers and newborns due to inherent biological risks and scarcity of health resources. We conducted a secondary analysis of the WHO Global Survey dataset to analyze maternal and perinatal outcomes in twin pregnancies and factors associated with perinatal morbidity and mortality in twins. We examined maternal and neonatal characteristics in twin deliveries in 23 LMICs and conducted multi-level logistic regression to determine the association between twins and adverse maternal and perinatal outcomes. 279,425 mothers gave birth to 276,187 (98.8%) singletons and 6,476 (1.2%) twins. Odds of severe adverse maternal outcomes (death, blood transfusion, ICU admission or hysterectomy) (AOR 1.85, 95% CI 1.60-2.14) and perinatal mortality (AOR 2.46, 95% CI 1.40-4.35) in twin pregnancies were higher, however early neonatal death (AOR 2.50, 95% CI 0.95-6.62) and stillbirth (AOR 1.22, 95% CI 0.58-2.57) did not reach significance. Amongst twins alone, maternal age <18, poor education and antenatal care, nulliparity, vaginal bleeding, non-cephalic presentations, birth weight discordance >15%, born second, preterm birth and low birthweight were associated with perinatal mortality. Marriage and caesarean section were protective. Twin pregnancy is a significant risk factor for maternal and perinatal morbidity and mortality in low-resource settings; maternal risk and access to safe caesarean section may determine safest mode of delivery in LMICs. Improving obstetric care in twin pregnancies, particularly timely access to safe caesarean section, is required to reduce risk to mother and baby.


ACE inhibitors: upper respiratory symptoms.
Cough and angioedema are well-known adverse reactions of ACE inhibitors. However, other adverse effects of upper airways such as postnasal drainage, rhinitis and nasal blockage, are less frequently recognised. These might share the same pathophysiological mechanism: bradykinin accumulation. We present two patients with ACE inhibitor-induced upper respiratory symptoms that improved after the discontinuation of ACE-inhibitors and substitution with angiotensin II receptor blockers. The incidence of these adverse events is not accurately known, since these are not required to be reported, but it is estimated to be low. This presents challenges to the physician and demonstrates the importance of keeping it as a differential diagnosis. Most physicians are aware of ACE inhibitor-induced cough but not of ACE inhibitor-induced nasal blockage, rhinitis or postnasal drainage. Identifying these can avoid unnecessary diagnostic tests and inappropriate treatment.


Allelic variation in the effector genes of the tomato pathogen Cladosporium fulvum reveals different modes of adaptive evolution.
The allelic variation in four avirulence (Avr) and four extracellular protein (Ecp)-encoding genes of the tomato pathogen Cladosporium fulvum was analyzed for a worldwide collection of strains. The majority of polymorphisms observed in the Avr genes are deletions, point mutations, or insertions of transposon-like elements that are associated with transitions from avirulence to virulence, indicating adaptive evolution of the Avr genes to the cognate C. fulvum resistance genes that are deployed in commercial tomato lines. Large differences in types of polymorphisms between the Avr genes were observed, especially between Avr2 (indels) and Avr4 (amino-acid substitutions), indicating that selection pressure favors different types of adaptation. In contrast, only a limited number of polymorphisms were observed in the Ecp genes, which mostly involved synonymous modifications. A haplotype network based on the polymorphisms observed in the effector genes revealed a complex pattern of evolution marked by reticulations that suggests the occurrence of genetic recombination in this presumed asexual fungus. This, as well as the identification of strains with identical polymorphisms in Avr and Ecp genes but with opposite mating-type genes, suggests that development of complex races can be the combined result of positive selection and genetic recombination.


The assessment and appraisal of regenerative medicines and cell therapy products: an exploration of methods for review, economic evaluation and appraisal.
The National Institute for Health and Care Excellence (NICE) commissioned a 'mock technology appraisal' to assess whether changes to its methods and processes are needed. This report presents the findings of independent research commissioned to inform this appraisal and the deliberations of a panel convened by NICE to evaluate the mock appraisal. Our research included reviews to identify issues, analysis methods and conceptual differences and the relevance of alternative decision frameworks, alongside the development of an exemplar case study of chimeric antigen receptor (CAR) T-cell therapy for treating acute lymphoblastic leukaemia. An assessment of previous evaluations of regenerative medicines found that, although there were a number of evidential challenges, none was unique to regenerative medicines or was beyond the scope of existing methods used to conceptualise decision uncertainty. Regarding the clinical evidence for regenerative medicines, the issues were those associated with a limited evidence base but were not unique to regenerative medicines: small non-randomised studies, high variation in response and the intervention subject to continuing development. The relative treatment effects generated from single-arm trials are likely to be optimistic unless it is certain that the historical data have accurately estimated the efficacy of the control agent. Pivotal trials may use surrogate end points, which, on average, overestimate treatment effects. To reduce overall uncertainty, multivariate meta-analysis of all available data should be considered. Incorporating indirectly relevant but more reliable (more mature) data into the analysis can also be considered; such data may become available as a result of the evolving regulatory pathways being developed by the European Medicines Agency. For the exemplar case of CAR T-cell therapy, target product profiles (TPPs) were developed, which considered the 'curative' and 'bridging to stem-cell transplantation' treatment approaches separately. Within each TPP, three 'hypothetical' evidence sets (minimum, intermediate and mature) were generated to simulate the impact of alternative levels of precision and maturity in the clinical evidence. Subsequent assessments of cost-effectiveness were undertaken, employing the existing NICE reference case alongside additional analyses suggested within alternative frameworks. The additional exploratory analyses were undertaken to demonstrate how assessments of cost-effectiveness and uncertainty could be impacted by alternative managed entry agreements (MEAs), including price discounts, performance-related schemes and technology leasing. The panel deliberated on the range of TPPs, evidence sets and MEAs, commenting on the likely recommendations for each scenario. The panel discussed the challenges associated with the exemplar and regenerative medicines more broadly, focusing on the need for a robust quantification of the level of uncertainty in the cost-effective estimates and the potential value of MEAs in limiting the exposure of the NHS to high upfront costs and loss associated with a wrong decision. It is to be expected that there will be a significant level of uncertainty in determining the clinical effectiveness of regenerative medicines and their long-term costs and benefits, but the existing methods available to estimate the implications of this uncertainty are sufficient. The use of risk sharing and MEAs between the NHS and manufacturers of regenerative medicines should be investigated further. The National Institute for Health Research Health Technology Assessment programme.


Indicators of accuracy of consumer health information on the Internet: a study of indicators relating to information for managing fever in children in the home.
To identify indicators of accuracy for consumer health information on the Internet. The results will help lay people distinguish accurate from inaccurate health information on the Internet. Several popular search engines (Yahoo, AltaVista, and Google) were used to find Web pages on the treatment of fever in children. The accuracy and completeness of these Web pages was determined by comparing their content with that of an instrument developed from authoritative sources on treating fever in children. The presence on these Web pages of a number of proposed indicators of accuracy, taken from published guidelines for evaluating the quality of health information on the Internet, was noted. Correlation between the accuracy of Web pages on treating fever in children and the presence of proposed indicators of accuracy on these pages. Likelihood ratios for the presence (and absence) of these proposed indicators. One hundred Web pages were identified and characterized as "more accurate" or "less accurate." Three indicators correlated with accuracy: displaying the HONcode logo, having an organization domain, and displaying a copyright. Many proposed indicators taken from published guidelines did not correlate with accuracy (e.g., the author being identified and the author having medical credentials) or inaccuracy (e.g., lack of currency and advertising). This method provides a systematic way of identifying indicators that are correlated with the accuracy (or inaccuracy) of health information on the Internet. Three such indicators have been identified in this study. Identifying such indicators and informing the providers and consumers of health information about them would be valuable for public health care.


Hemiascomycetous yeasts at the forefront of comparative genomics.
With more than a dozen species fully sequenced, as many as this partially sequenced, and more in development, yeasts are now used to explore the frontlines of comparative genomics of eukaryotes. Innovative procedures have been developed to compare and annotate genomes at various evolutionary distances, to identify short cis-acting regulatory elements, to map duplications, or to align syntenic blocks. Human and plant pathogens, in addition to yeasts that show a variety of interesting physiological properties, are included in this multidimensional comparative survey, which encompasses a very broad evolutionary range. As major steps of the evolutionary history of hemiascomycetous genomes emerge, precise questions on the general mechanisms of their evolution can be addressed, using both experimental and in silico methods.


Light-induced dark states of organic fluochromes enable 30 nm resolution imaging in standard media.
We show that high quantum efficiency fluorophores can exhibit reversible photobleaching. This observation provides the basis for an imaging technique we call reversible photobleaching microscopy. We demonstrate applicability of this technique using antibody labeled biological samples in standard aqueous (or glycerol based) media to produce far-field images at approximately 30 nm resolution. Our novel method relies on intense illumination to reversibly induce a very long-lived (>10 s) dark state from which single fluorochromes slowly return stochastically. As in other localization microscopy methods, reversible photobleaching microscopy localizes single fluorochromes, but has the advantage that specialized photoactivatible and photoswitchable molecules or special immersion/embedding media are not required.


Modelling Metabolic Shifts during Cardiomyocyte Differentiation, Iron Deficiency and Transferrin Rescue Using Human Pluripotent Stem Cells.
Cardiomyocytes rely on specialised metabolism to meet the high energy demand of the heart. During heart development, metabolism matures and shifts from the predominant utilisation of glycolysis and glutamine oxidation towards lactate and fatty acid oxidation. Iron deficiency (ID) leads to cellular metabolism perturbations. However, the exact alterations in substrate metabolism during ID are poorly defined. Using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM), the present study investigated changes in major metabolic substrate utilisation in the context of ID or upon transferrin rescue. Typically, during hiPSC-CM differentiation, the greatest increase in total metabolic output and rate was seen in fatty acid metabolism. When ID was induced, hiPSC-CMs displayed increased reliance on glycolytic metabolism, and six TCA cycle, five amino acid, and four fatty acid substrates were significantly impaired. Transferrin rescue was able to improve TCA cycle substrate metabolism, but the amino acid and fatty acid metabolism remained perturbed. Replenishing iron stores partially reverses the adverse metabolic changes that occur during ID. Understanding the changes in metabolic substrate utilisation and their modification may provide potential for discovery of new biomarkers and therapeutic targets in cardiovascular diseases.


A new population and habitat for Neotricula aperta in the Mekong river of northeastern Thailand: a DNA sequence-based phylogenetic assessment confirms identifications and interpopulation relationships.
Neotricula aperta (Gastropoda: Pomatiopsidae), the snail intermediate host of Schistosoma mekongi, is found in Cambodia, Laos, and Thailand. We update information on the distribution of this species in the Mekong River and its tributary, the Mun River, in Thailand. DNA sequences of a portion of the mitochondrial cytochrome c oxidase subunit 1 were obtained from N. aperta collected from different locations and used to confirm species and strain identities. Specimens of the β-strain were found in the Mun River, whereas specimens of the γ-strain were found in the Mekong River. The γ-strain (with molecular confirmation of identity) is newly reported from Nong Khai Province, where it occurred in a habitat novel for this species: under paving slabs instead of under natural bed rocks, where agal aufwuchs is extensively located on the islet in the middle of the Mekong River. The new location is approximate 400 km upstream from the nearest previously known site for this species.


Identification of Pseudomonas mosselii BS011 gene clusters required for suppression of Rice Blast Fungus Magnaporthe oryzae.
Pseudomonas is a Gram-negative, rod-shaped bacteria. Many members of this genus displayed remarkable physiological and metabolic activity against different plant pathogens. However, Pseudomonas mosselii has not yet been characterized in biocontrol against plant disease. Here we isolated a strain of P. mosselii BS011 from the rhizosphere soil of rice plants, and the isolate showed strong inhibitory activity against the rice blast fungus Magnaporthe oryzae. Further we sequenced the complete genome of BS011, which consist of 5.75 Mb with a circular chromosome, 5,170 protein-coding genes, 23 rRNA and 78 tRNA operons. Bioinformatic analysis revealed that seven gene clusters may be involved in the biosynthesis of metabolites. Gene deletion experiments demonstrated that the gene cluster c-xtl is required for inhibitory activity against M. oryzae. Bioassay showed that the crude extract from BS011 fermentation sample significantly inhibited the development of M. oryzae at a concentration of 10 μg/ml. Besides, we illustrated that the crude extract of BS011 impaired the appressorial formation in a dose dependent manner. Collectively our results revealed that P. mosselii BS011 is a promising biocontrol agent and the gene cluster c-xtl is essential for inhibiting the development of M. oryzae.


Anxiety and depression have no influence on the cancellation and pregnancy rates of a first IVF or ICSI treatment.
After many years of research, the impact of psychological distress on the IVF treatment outcome is still unclear. This study aimed to determine the influences of anxiety and depression before and during IVF or ICSI treatment on the cancellation and pregnancy rates of inductees. In a multicentre prospective cohort study, we assessed anxiety and depression at baseline and the procedural anxiety level one day before oocyte retrieval, with the short versions of the State Anxiety Inventory (STAI) and the Beck Depression Inventory-Primary Care (BDI-PC). The effect of baseline anxiety and depression on the cancellation and pregnancy rates of 783 women in their first IVF or ICSI treatment was evaluated. We also determined if a change in anxiety from the start of treatment until just before oocyte retrieval affects the pregnancy rate. The predictive value of distress was assessed while controlling for several factors in subfertility treatment. Neither baseline nor procedural anxiety, nor depression affected the ongoing pregnancy rates, with odds ratios (ORs) of 1.04 (95% CI 0.82-1.33), 0.96 (95% CI 0.77-1.20) and 0.85 (95% CI 0.65-1.10), respectively. Neither did the anxiety gain score affect the pregnancy rate, OR 1.08 (95% CI 0.83-1.41). A cancellation of treatment could not be predicted by either anxiety or depression, OR 1.16 (95% CI 0.83-1.63) and 0.85 (95% CI 0.59-1.22), respectively. Inductees in IVF treatment can be reassured that anxiety and depression levels before and during treatment have no significant influence on the cancellation and pregnancy rates.


Purine-scaffold Hsp90 inhibitors.
Hsp90 is a molecular chaperone with important roles in regulating the function of several proteins with potential pathogenic activity. Because many of these proteins are involved in cancer and neurodegenerative promoting pathways, Hsp90 has emerged as an attractive therapeutic target in these diseases. Molecules that bind to the N-terminal nucleotide pocket of Hsp90 inhibit its activity, and consequently, disrupt client protein function. A number of these inhibitors from several chemical classes are now known, and some are already in clinical trials. This review focuses on the purine class of Hsp90 inhibitors, their discovery through rational design, and on efforts aimed towards their optimization and development into clinically viable drugs for the treatment of cancer. Their potential towards neurodegenerative diseases will also be touched upon.


Numerical demonstration of the validity of the Rayleigh hypothesis.
The Rayleigh hypothesis and the related method of diffraction analysis are revisited. It is shown that the Rayleigh method can be applied to deep grating modeling without numerical problems and that it gives any desired accuracy whatever the groove depth. This proves the validity of the Rayleigh hypothesis and rehabilitates the Rayleigh method.


Lung simulation to support non-invasive pulmonary blood flow measurement in Acute Respiratory Distress Syndrome in animals.
Patients undergoing mechanical lung ventilation are at risk of lung injury. A noninvasive bedside lung monitor may benefit these patients. The Inspired Sinewave Test (IST) can measure cardio-pulmonary parameters noninvasively. We propose a lung simulation to improve the measurement of pulmonary blood flow using IST. The new method was applied to 12 pigs' data before lung injury (control) and after lung injury (ARDS model). Results using the lung simulation shown improvements in correlation in both simulated data (R2 increased from 0.98 to 1) and pigs' data (R2 increased from <0.001 to 0.26). Paired blood flow measurements were performed by both the IST (noninvasive) and thermodilution (invasive). In the control group, the bias of the two methods was negligible (0.02L/min), and the limit of agreement was from -1.20 to 1.18 L/min. The bias was -0.68 L/min in the ARDS group and with a broader limit of agreement (-2.49 to 1.13 L/min).Clinical Relevance- the inspired sinewave test can be used to measure cardiac output noninvasively in mechanically ventilated subjects with and without acute respiratory distress syndrome.


Identification of central projections from amylin-activated neurons to the lateral hypothalamus.
The ability of the pancreatic hormone amylin to inhibit food intake relies on a direct activation of the area postrema (AP). This activation is synaptically transmitted to the nucleus of the solitary tract (NTS), the lateral parabrachial nucleus (LPB), the central amygdaloid nucleus (Ce) and the lateral bed nucleus of stria terminalis (BSTL). Interestingly, neurons of the rostro-dorsal lateral hypothalamic area (dLHA), which are activated during fasting, are inhibited by peripheral amylin, although they lack amylin receptors. Using the retrograde tracer cholera toxin-B (Ctb) we analyzed whether the dLHA receives neuronal projections from amylin-activated brain areas. The anterograde tracer biotinylated dextran-amine (BDA) was used to confirm the projections and to identify further neuronal pathways potentially involved in amylin signaling. We identified dense projections from the amylin activated neurons in the LPB and sparse projections from the NTS to the dLHA. LPB fiber efferents were found in close proximity to dLHA nuclei activated by 24h of fasting. The AP and the Ce showed no projections to the dLHA. Dense efferents were also observed from the LPB to other hypothalamic areas, namely to the ventromedial, dorsomedial, paraventricular and arcuate nuclei. This study provides neuroanatomical evidence that among the amylin activated areas, the LPB provides the strongest input to the dLHA, thus it may mediate the amylin-induced inhibition of the dLHA.


Exenatide decreases liver fat content and epicardial adipose tissue in patients with obesity and type 2 diabetes: a prospective randomized clinical trial using magnetic resonance imaging and spectroscopy.
To conduct a prospective randomized trial to investigate the effect of glucagon-like peptide-1 (GLP-1) analogues on ectopic fat stores. A total of 44 obese subjects with type 2 diabetes uncontrolled on oral antidiabetic drugs were randomly assigned to receive exenatide or reference treatment according to French guidelines. Epicardial adipose tissue (EAT), myocardial triglyceride content (MTGC), hepatic triglyceride content (HTGC) and pancreatic triglyceride content (PTGC) were assessed 45 min after a standardized meal with 3T magnetic resonance imaging and proton magnetic resonance spectroscopy before and after 26 weeks of treatment. The study population had a mean glycated haemoglobin (HbA1c) level of 7.5 ± 0.2% and a mean body mass index of 36.1 ± 1.1 kg/m(2) . Ninety five percent had hepatic steatosis at baseline (HTGC ≥ 5.6%). Exenatide and reference treatment led to a similar improvement in HbA1c (-0.7 ± 0.3% vs. -0.7 ± 0.4%; p = 0.29), whereas significant weight loss was observed only in the exenatide group (-5.5 ± 1.2 kg vs. -0.2 ± 0.8 kg; p = 0.001 for the difference between groups). Exenatide induced a significant reduction in EAT (-8.8 ± 2.1%) and HTGC (-23.8 ± 9.5%), compared with the reference treatment (EAT: -1.2 ± 1.6%, p = 0.003; HTGC: +12.5 ± 9.6%, p = 0.007). No significant difference was observed in other ectopic fat stores, PTGC or MTGC. In the group treated with exenatide, reductions in liver fat and EAT were not associated with homeostatic model assessment of insulin resistance index, adiponectin, HbA1c or fructosamin change, but were significantly related to weight loss (r = 0.47, p = 0.03, and r = 0.50, p = 0.018, respectively). Our data indicate that exenatide is an effective treatment to reduce liver fat content and epicardial fat in obese patients with type 2 diabetes, and these effects are mainly weight loss dependent.


Protein footprinting by pyrite shrink-wrap laminate.
The structure of macromolecules and their complexes dictate their biological function. In "footprinting", the solvent accessibility of the residues that constitute proteins, DNA and RNA can be determined from their reactivity to an exogenous reagent such as the hydroxyl radical (·OH). While ·OH generation for protein footprinting is achieved by radiolysis, photolysis and electrochemistry, we present a simpler solution. A thin film of pyrite (cubic FeS2) nanocrystals deposited onto a shape memory polymer (commodity shrink-wrap film) generates sufficient ·OH via Fenton chemistry for oxidative footprinting analysis of proteins. We demonstrate that varying either time or H2O2 concentration yields the required ·OH dose-oxidation response relationship. A simple and scalable sample handling protocol is enabled by thermoforming the "pyrite shrink-wrap laminate" into a standard microtiter plate format. The low cost and malleability of the laminate facilitates its integration into high throughput screening and microfluidic devices.


PKA-induced stimulation of ROMK1 channel activity is governed by both tethering and non-tethering domains of an A kinase anchor protein.
We have used the patch-clamp technique to explore the role of A Kinase Anchor Proteins (AKAP) in mediating the effect of cAMP on ROMK1 channels expressed in the Xenopus oocytes. Addition of membrane permeant cAMP analogs increased channel activity only in oocytes injected with ROMK1 and AKAP79 cRNA but had no effect on channel activity in oocytes injected with ROMK1 alone. Using the two-electrode voltage clamp technique, we determined that application of H89, a potent inhibitor of protein kinase A (PKA), abolished the stimulatory effect of cAMP/forskolin. To investigate the role of AKAP specificity in conferring cAMP responses to ROMK1 channels, we examined channel activity in oocytes expressing ROMK1 and either AKAP18, AKAP-KL or AKAP75. Addition of cAMP failed to increase channel current in oocytes expressing ROMK1 and either AKAP18 or AKAP-KL. In contrast, cAMP increased ROMK1 channel activity by 33% in oocytes coexpressing AKAP75, the bovine homologue of AKAP79. The effect of cAMP on ROMK1 in oocytes coexpressing AKAP75 is inhibited by H89. Since all three AKAPs bind PKAII, the results suggest that a unique structural domain in AKAP75/79 collaborates with the PKAII binding site and enables a productive association of PKA with ROMK1 channels. Deletion of either the membrane targeting region of AKAP75 (AKAP45) or PKAII binding domain of AKAP75 (AKAP75DeltaC) abolished the effects of forskolin on ROMK1 channels. This suggests that the membrane targeting and the PKA binding domains of AKAP75 are essential for the effect of cAMP. However, the nature of the AKAP, that interacts with ROMK1 in the native tissue, remains to be determined because AKAP75/79 are not expressed in the kidney. We conclude that the regulation of ROMK1 channels by PKA requires the involvement of the cell membrane-directed AKAPs that are able to specifically link PKA to the target channel protein.


Interaction of phospholipid transfer protein with human tear fluid mucins.
In addition to circulation, where it transfers phospholipids between lipoprotein particles, phospholipid transfer protein (PLTP) was also identified as a component of normal tear fluid. The purpose of this study was to clarify the secretion route of tear fluid PLTP and elucidate possible interactions between PLTP and other tear fluid proteins. Human lacrimal gland samples were stained with monoclonal antibodies against PLTP. Heparin-Sepharose (H-S) affinity chromatography was used for specific PLTP binding, and coeluted proteins were identified with MALDI-TOF mass spectrometry or Western blot analysis. Immunoprecipitation assay and blotting with specific antibodies helped to identify and characterize PLTP-mucin interaction in tear fluid. Human tear fluid PLTP is secreted from the lacrimal gland. MALDI-TOF analysis of H-S fractions identified several candidate proteins, but protein-protein interaction assays revealed only ocular mucins as PLTP interaction partners. We suggest a dual role for PLTP in human tear fluid: (1) to scavenge lipophilic substances from ocular mucins and (2) to maintain the stability of the anterior tear lipid film. PLTP may also play a role in the development of ocular surface disease.


The obesity and inflammatory marker haptoglobin attracts monocytes via interaction with chemokine (C-C motif) receptor 2 (CCR2).
Obesity is a chronic low inflammatory state. In the obesity condition the white adipose tissue (WAT) is massively infiltrated with monocytes/macrophages, and the nature of the signals recruiting these inflammatory cells has yet to be fully elucidated. Haptoglobin (Hp) is an inflammatory marker and its expression is induced in the WAT of obese subjects. In an effort to elucidate the biological significance of Hp presence in the WAT and of its upregulation in obesity we formulated the hypothesis that Hp may serve as a macrophage chemoattractant. We demonstrated by chemotaxis assay that Hp is able to attract chemokine (C-C motif) receptor 2 (CCR2)-transfected pre-B lymphocytes and monocytes in a dose-dependent manner. Moreover, Hp-mediated migration of monocytes is impaired by CCR2-specific inhibition or previous cell exposure to monocyte chemoattractant protein 1 (MCP1) (also known as CCR2 ligand or chemokine (C-C motif) ligand 2 (CCL2)). Downstream effects of Hp/CCR2 interaction were also investigated: flow cytometry proved that monocytes treated with Hp show reduced CCR2 expression on their surface; Hp interaction induces calcium release that is reduced upon pretreatment with CCR2 antagonist; extracellular signal-regulated kinase (ERK)1/2, a signal transducer activated by CCR2, is phosphorylated following Hp treatment and this phosphorylation is reduced when cells are pretreated with a specific CCR2 inhibitor. Consistently, blocking the ERK1/2 pathway with U0126, the selective inhibitor of the ERK upstream mitogen-activated protein (MAP)-ERK kinase (MEK), results in a dramatic reduction (by almost 100%) of the capability of Hp to induce monocyte migration. Our data show that Hp is a novel monocyte chemoattractant and that its chemotactic potential is mediated, at least in part. by its interaction with CCR2.


Impact of Extended Primary Surgery on Suboptimally Operable Patients With Advanced Ovarian Cancer.
Extensive surgical efforts to achieve an optimal debulking (no residual tumor) in primary surgery of ovarian cancer are today's criterion standard in gyneco-oncologic surgery. However, it is controversial whether extensive surgery, including resections of metastases in the upper abdomen and bowel resections, is justifiable in patients with not completely operable lesions. All patients who had undergone surgery for ovarian cancer in the years 2002 to 2013 at our institution were viewed (n = 472). We retrospectively identified 278 operations for primary ovarian cancer. Ninety-six (35%) of the 278 patients showed postoperative tumor residuals and were included in this study. Fifty-five (57%) of 96 patients underwent bowel resection, showing significantly higher complication rates (64% vs 39% minor complications, P = 0.017; 31% vs 9.8% severe complications, P = 0.013) compared with patients without bowel resections as well as no improvement in progression-free or overall survival (median overall survival, 19.5 vs 32.9; P = 0.382). Multiple anastomoses (≥2) were associated with higher rates for anastomotic leakage (16.7% vs 2.6%, P = 0.02) and a higher mortality (16.7% vs 0%, P = 0.04) compared with patients with only 1 anastomosis. Extensive surgery of the upper abdomen was not associated with a significant increase in complication rates. Because of the increased morbidity of bowel resections without any evidence for improvement of survival, we suggest to restrain from further resection of intestines if an optimal debulking seems not feasible after removal of the major tumor bulk.


Burden of undiagnosed hypertension in sub-saharan Africa: a systematic review and meta-analysis.
The burden of hypertension in Sub-Saharan Africa has been increasing over the past few decades. However, a large proportion of the population with hypertension remains undiagnosed, untreated, or inadequately treated, contributing to the rising burden of cardiovascular disease in the region. We conducted a systematic review and meta-analysis to assess the recent burden of hypertension in Sub-Saharan Africa, based on studies published between 2000 and 2013. We pooled data from 33 surveys involving over 110 414 participants of mean age 40 years. Hypertension prevalence varied widely across the studies (range 15%-70%), partly because of differences in participant mean ages (31-76 years). The predicted prevalence of hypertension at mean participant ages of 30, 40, 50, and 60 years were 16%, 26%, 35%, and 44%, respectively, with a pooled prevalence of 30% (95% confidence interval, 27%-34%). Of those with hypertension, only between 7% and 56% (pooled prevalence: 27%; 95% confidence interval, 23%-31%) were aware of their hypertensive status before the surveys. Overall, 18% (95% confidence interval, 14%-22%) of individuals with hypertension were receiving treatment across the studies, and only 7% (95% confidence interval, 5%-8%) had controlled blood pressure. This review found a high prevalence of hypertension, as well as low percentage of hypertension awareness, treatment, and control in Sub-Saharan Africa, highlighting the need for implementation of timely and appropriate strategies for diagnosis, control, and prevention.


Comparative analysis of subtyping methods against a whole-genome-sequencing standard for Salmonella enterica serotype Enteritidis.
A retrospective investigation was performed to evaluate whole-genome sequencing as a benchmark for comparing molecular subtyping methods for Salmonella enterica serotype Enteritidis and survey the population structure of commonly encountered S. enterica serotype Enteritidis outbreak isolates in the United States. A total of 52 S. enterica serotype Enteritidis isolates representing 16 major outbreaks and three sporadic cases collected between 2001 and 2012 were sequenced and subjected to subtyping by four different methods: (i) whole-genome single-nucleotide-polymorphism typing (WGST), (ii) multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA), (iii) clustered regularly interspaced short palindromic repeats combined with multi-virulence-locus sequence typing (CRISPR-MVLST), and (iv) pulsed-field gel electrophoresis (PFGE). WGST resolved all outbreak clusters and provided useful robust phylogenetic inference results with high epidemiological correlation. While both MLVA and CRISPR-MVLST yielded higher discriminatory power than PFGE, MLVA outperformed the other methods in delineating outbreak clusters whereas CRISPR-MVLST showed the potential to trace major lineages and ecological origins of S. enterica serotype Enteritidis. Our results suggested that whole-genome sequencing makes a viable platform for the evaluation and benchmarking of molecular subtyping methods.


Crystal structure of Clostridium acetobutylicum Aspartate kinase (CaAK): An important allosteric enzyme for amino acids production.
Aspartate kinase (AK) is an enzyme which is tightly regulated through feedback control and responsible for the synthesis of 4-phospho-L-aspartate from L-aspartate. This intermediate step is at an important branch point where one path leads to the synthesis of lysine and the other to threonine, methionine and isoleucine. Concerted feedback inhibition of AK is mediated by threonine and lysine and varies between the species. The crystal structure of biotechnologically important Clostridium acetobutylicum aspartate kinase (CaAK; E.C. 184.108.40.206; Mw=48,030Da; 437aa; SwissProt: Q97MC0) has been determined to 3Å resolution. CaAK acquires a protein fold similar to the other known structures of AKs despite the low sequence identity (<30%). It is composed of two domains: an N-terminal catalytic domain (kinase) domain and a C-terminal regulatory domain further comprised of two small domains belonging to the ACT domain family. Pairwise comparison of 12 molecules in the asymmetric unit helped to identify the bending regions which are in the vicinity of ATP binding site involved in domain movements between the catalytic and regulatory domains. All 12 CaAK molecules adopt fully open T-state conformation leading to the formation of three tetramers unique among other similar AK structures. On the basis of comparative structural analysis, we discuss tetramer formation based on the large conformational changes in the catalytic domain associated with the lysine binding at the regulatory domains. The structure described herein is homologous to a target in wide-spread pathogenic (toxin producing) bacteria such as Clostridium tetani (64% sequence identity) suggesting the potential of the structure solved here to be applied for modeling drug interactions. CaAK structure may serve as a guide to better understand and engineer lysine biosynthesis for the biotechnology industry.


AMP kinase acts as a negative regulator of RANKL in the differentiation of osteoclasts.
AMP-activated protein kinase (AMPK) has been reported to stimulate differentiation and proliferation of osteoblasts, but the role of AMPK in the physiology of osteoclasts has not been investigated. Osteoclasts were differentiated from mouse BMMϕs. TRAP-positive multinucleated cells were considered to be osteoclasts using TRAP staining, and resorption area was determined by incubation of cells on dentine discs. Signaling pathways were investigated using Western blotting and RT-PCR. RANKL induced phosphorylation/activation of AMPK-α in BMMϕs and stimulated formation of TRAP-positive multinucleated cells. Pharmacological inhibition of AMPK with compound C and siRNA-mediated knockdown of AMPK-α1, the predominant α-subunit isoform in BMMϕs, increased RANKL-induced formation of TRAP-positive multinucleated cells and bone resorption via activation of the downstream signaling elements p38, JNK, NF-κB, Akt, CREB, c-Fos, and NFATc1. STO-609, an inhibitor of CaMKK, completely blocked the RANKL-induced activation of AMPK-α, but KN-93, an inhibitor of CaMK, did not. siRNA-mediated TAK1 knockdown also blocked RANKL-induced activation of AMPK-α. The AMPK activators metformin, (-)-epigallocatechin-3-gallate, berberine, resveratrol, and α-lipoic acid dose-dependently suppressed formation of TRAP-positive multinucleated cells and bone resorption. AMPK negatively regulates RANKL, possibly by acting through CaMKK and TAK1. Thus, the development of AMPK activators may be a useful strategy for inhibiting the resorption of bone that is stimulated under RANKL-activated conditions.


Neonatal exposure to sevoflurane expands the window of vulnerability to adverse effects of subsequent exposure to sevoflurane and alters hippocampal morphology via decitabine-sensitive mechanisms.
Deficiencies in neurocognitive function have been found in late childhood or adolescence in patients who had prolonged and/or repeated early-life general anesthesia. Animal studies suggest that anesthetic-induced impairment in the neuron-specific K+-2Cl- (Kcc2) Cl- exporter expression, which regulates developmental maturation of GABA type A receptor (GABAAR) signaling from excitatory to inhibitory, may play a mediating role. We tested whether the DNA methyltransferase (DNMT) inhibitor decitabine ameliorates the anesthetic's adverse effects. Sprague-Dawley male rats were injected with vehicle or decitabine 30 min before 2.1 % sevoflurane exposure for 5 h on postnatal day 5 (P5). On P19, P20, or P21, electroencephalography-detectable seizures were measured during 1 h of sevoflurane exposure, followed by collection of the trunk blood and brain tissue samples. Other rats were evaluated for changes in hippocampal CA1 dendrite morphology and gene expressions on ≥ P120. Rats in the vehicle plus sevoflurane group responded to sevoflurane exposure on P19, P20 or P21 with electroencephalography-detectable seizures and stress-like corticosterone secretion and had altered hippocampal dendrite morphology in adulthood. These rats had expressions of Kcc2 and Dnmt genes downregulated and upregulated, respectively, in the P19 - P21 cortex and hypothalamus and the ≥ P120 hippocampus. All measured parameters in the sevoflurane-exposed rats that were pretreated with decitabine were not different from those in the control group. Neonatal exposure to sevoflurane sensitizes rats to adverse effects of repeated exposure to the anesthetic. The anesthetic-caused changes in the decitabine-sensitive mechanisms may play a mediating role in the developmental effects of early-life anesthesia.


Horizontal transfer of diatomaceous earth and botanical insecticides in the common bed bug, Cimex lectularius L.; hemiptera: cimicidae.
Horizontal transfer of insecticide occurs when insects contact or ingest an insecticide, return to an aggregation or a nest, and transfer the insecticide to other conspecific insects through contact. This phenomenon has been reported in a number of insects including social insects, however it has not been reported in bed bugs. Since horizontal transfer can facilitate the spread of insecticide into hard to reach spaces, it could contribute greatly to the management of these public health pests. To demonstrate horizontal transfer of diatomaceous earth and botanical insecticides in C. lectularius, an exposed (donor) bed bug, following a 10-minute acquisition period, was placed with unexposed (recipient) bed bugs. Mortality data clearly demonstrates that diatomaceous earth (DE 51) was actively transferred from a single exposed bug to unexposed bugs in a concentration dependent manner. LC50 values varied from 24.4 mg at 48 h to 5.1 mg at 216 h when a single exposed bed bug was placed with 5 unexposed bed bugs. LT50 values also exhibited a concentration response. LT50 values varied from 1.8 days to 8.4 days when a 'donor' bug exposed to 20 and 5 mg of dust respectively was placed with 5 'recipient' bugs. Dust was also actively transferred from adult bed bugs to the nymphs. In addition we observed horizontal transfer of botanical insecticides including neem, ryania, and rotenone to varying degrees. Our data clearly demonstrate horizontal transfer of diatomaceous earth and botanical insecticides in the common bed bug, C. lectularius. Use of a fluorescent dust provided visual confirmation that contaminated bed bugs transfer dust to untreated bed bugs in harborage. This result is important because bedbugs live in hard-to-reach places and interaction between conspecifics can be exploited for delivery and dissemination of management products directed at this public health pest.


Bibliometric analysis of medicine-related publications on poverty (2005-2015).
Poverty is a global problem. The war against poverty requires not only financial support, but also poverty-related research to pinpoint areas of high need of intervention. In line with international efforts to fight poverty and negative consequences, we carried out this study to give a bibliometric overview of medicine-related literature on poverty. Such a s study is an indicator of the extent of interaction of various international key players on the war against poverty-related health problems. Scopus was used to achieve the objective of this study. The time span set for this study was 2005-2015. Poverty-related articles under the subject area "Medicine" were used to give bibliometric indicators such as annual growth of publications, international collaboration, highly cited articles, active countries, institutions, journals, and authors. The total number of retrieved articles was 1583. The Hirsh-index of retrieved articles was 56. A modest and fluctuating increase was seen over the study period. Visualization map of retrieved articles showed that "HIV", infectious diseases, mental health, India, and Africa were most commonly encountered terms. No significant dominance of any particular author or journal was observed in retrieved articles. The United States of America had the largest share in the number of published articles. The World Health Organization and Centers for Disease Prevention and Control were among top active institutions/organizations. International collaboration was observed in less than one third of publications. Top cited articles focused on three poverty-related health issues, mainly, infectious diseases, malnutrition, and child development/psychology. Most of top articles were published in high impact journals. Data indicated that articles on poverty were published in high influential medical journals indicative of the importance of poverty as a global health problem. However, the number publications and the extent of international collaborations was lower than expected given the huge burden of poverty-related health problems.


Low back pain.
Low back pain affects individuals of all ages and is a leading contributor to disease burden worldwide. Despite advancements in assessment and treatment methods, the management of low back pain remains a challenge for researchers and clinicians alike. One reason for the limited success in identifying effective treatments is the large variation in the manifestations, possible causes, precipitating and maintaining factors, course, prognosis and consequences in terms of activity interference and quality of life. However, despite these challenges, steady progress has been achieved in the understanding of back pain, and important steps in the understanding of the psychological and social risk factors, genetics and brain mechanisms of low back pain have been made. These new findings have given impetus to the development of new diagnostic procedures, evidence-based screening methods and more targeted interventions, which underscore the need for a multidisciplinary approach to the management of low back pain that integrates biological, psychological and social aspects.


Coxsackie B infection in a Scottish general practice.
The authors describe 38 cases of protracted illness, characterized by varied multisystem symptomatology. Significant antibody titres to Coxsackie B viruses, suggesting recent infection, were found in these 38 patients (47 per cent of 81 cases investigated). The syndrome has many of the features of myalgic encephalomyelitis (ME), a condition which has frequently been found in closely clustered epidemics but seldom in sporadic or endemic form. The reported cases emerged over a four-year period and fresh cases continue to appear.


Neurochemical evidence for differential effects of acute and repeated oxytocin administration.
A discrepancy in oxytocin's behavioral effects between acute and repeated administrations indicates distinct underlying neurobiological mechanisms. The current study employed a combination of human clinical trial and animal study to compare neurochemical changes induced by acute and repeated oxytocin administrations. Human study analyzed medial prefrontal metabolite levels by using 1H-magnetic resonance spectroscopy, a secondary outcome in our randomized, double-blind, placebo-controlled crossover trial of 6 weeks intranasal administrations of oxytocin (48 IU/day) and placebo within-subject design in 17 psychotropic-free high-functioning men with autism spectrum disorder. Medial prefrontal transcript expression levels were analyzed in adult male C57BL/6J mice after intraperitoneal injection of oxytocin or saline either once (200 ng/100 μL/mouse, n = 12) or for 14 consecutive days (200 ng/100 μL/mouse/day, n = 16). As the results, repeated administration of oxytocin significantly decreased the medial prefrontal N-acetylaspartate (NAA; p = 0.043) and glutamate-glutamine levels (Glx; p = 0.001), unlike the acute oxytocin. The decreases were inversely and specifically associated (r = 0.680, p = 0.004 for NAA; r = 0.491, p = 0.053 for Glx) with oxytocin-induced improvements of medial prefrontal functional MRI activity during a social judgment task not with changes during placebo administrations. In wild-type mice, we found that repeated oxytocin administration reduced medial frontal transcript expression of N-methyl-D-aspartate receptor type 2B (p = 0.018), unlike the acute oxytocin, which instead changed the transcript expression associated with oxytocin (p = 0.0004) and neural activity (p = 0.0002). The present findings suggest that the unique sensitivity of the glutamatergic system to repeated oxytocin administration may explain the differential behavioral effects of oxytocin between acute and repeated administration.


Insulin differentially influences brain glucose and lactate in traumatic brain injured patients.
Hypo- and hyperglycemia must be avoided to prevent additional brain damage following traumatic brain injury (TBI). However, the optimal blood glucose range requiring insulin remains unknown. Cerebral microdialysis is helpful in unmasking signs of metabolic impairment, thereby identifying deleterious blood glucose levels. A retrospective analysis of prospectively collected cerebral microdialysis samples obtained from 20 non-diabetic patients with severe TBI treated at the trauma surgical intensive care unit at the University Hospital Zürich, Switzerland. The impact of different arterial blood glucose values and concomitant insulin administration on cerebral interstitial glucose and lactate levels was investigated. In addition, energetic impairment was determined by calculating lactate-to-glucose ratios. Insulin administration was associated with significantly reduced cerebral glucose concentrations and significantly increased lactate-to-glucose ratios with arterial blood glucose levels <5 mM. At arterial blood glucose levels >7 mM, insulin administration was associated with significantly increased interstitial glucose values, significantly decreased lactate concentrations, and markedly diminished lactate-to-glucose ratios. Insulin exerts differential effects that depend strongly on the underlying arterial blood glucose concentrations. To avoid energetic impairment, insulin should not be administered at arterial blood glucose levels <5 mM. However, at arterial blood glucose levels >7-8 mM, insulin administration appears to be encouraged to increase extracellular glucose concentrations and decrease energetic impairment reflected by reduced interstitial brain lactate and decreased lactate-to-glucose ratios. Nevertheless, frequent analysis is required to minimize the risk of inducing impaired brain metabolism.


Myocardial transport of hexakis(2-methoxyisobutylisonitrile) and thallium before and after coronary reperfusion.
Effects of no-flow ischemia (NFI) and reperfusion (RPF) on myocardial extraction and retention of technetium-99m hexakis(2-methoxyisobutylisonitrile) (sestamibi) and thallium-201 were investigated in 12 isolated, blood-perfused rabbit hearts with isotope dilution studies at constant coronary perfusion. After a control injection of tracers, NFI was induced for 30-60 minutes. After coronary reflow, repeat tracer injections were given at early RPF (5-15 minutes of RPF) and late RPF (40-60 minutes of RPF). After NFI-RPF, maximal fractional extraction and capillary permeability-surface area product increased for sestamibi (from +39% to 69%) and decreased for thallium (from -14% to -68%). Net extraction was 33% lower for sestamibi than for thallium at control, 13% lower at early RPF, and 90% higher than thallium at late RPF. Interstitial-myocyte exchange estimates were always higher for sestamibi than for thallium and increased for both with NFI-RPF (sestamibi, from 57.4 to 122.4 ml/min/g; thallium, from 3.1 to 22.3 ml/min/g). Intramyocyte volumes of distribution were higher for sestamibi than for thallium (greater than 200% at control, 800-1,000% with RPF), and NFI-RPF had opposite effects on the two tracers (late RPF vs. control: +28% for sestamibi, -50% for thallium). Our results suggest that sestamibi and thallium have different transport or sequestering mechanisms and that NFI-RPF had opposite effects on myocardial capillary-tissue exchange and tissue retention of sestamibi and thallium. Therefore, myocardial perfusion might be overestimated with sestamibi and underestimated with thallium during early RPF.


Gravity perception in cerebellar patients.
Patients with midline cerebellar atrophy typically suffer from impaired postural balance. We asked whether this deficit is, in part, caused by a deficient perception of body position relative to gravity, and therefore measured the subjective visual vertical (SVV) in patients with degeneration predominantly of the midline cerebellar structures (n=11) and age-matched healthy human subjects (n=9). On a motorized turntable, subjects were placed in different roll positions [0, 75 degrees right ear down (RED), 75 degrees left ear down (LED)] and had to align a luminous arrow with the perceived earth-vertical. Both SVV deviations and intra-individual SD of SVV adjustments were not significantly different between patients and age-matched controls. Our findings suggest that the perception of verticality in patients with cerebellar ataxia may only deteriorate in a more advanced stage of the disease. To which degree the deterioration of perceived vertical is caused by a collateral vestibular impairment, awaits further clarification.


Non-surgical treatments of primary, non-melanoma eyelid malignancies: a review.
The diagnosis and management of periocular cutaneous malignancies are essential components of an ophthalmologist's practice. Skin cancers comprise nearly one-third of newly diagnosed malignancies. Furthermore, the incidence of skin cancer appears to be increasing. Multiple treatment modalities exist for periocular cutaneous malignancy. Surgical extirpation, often with the combined expertise of a Mohs micrographic surgeon, is typically the first line therapy and is often curative in the periocular region, depending on a variety of factors, including tumour histology, specific location, depth of invasion and surgical technique. However, there are instances where a less invasive, non-surgical treatment option is warranted, including patients who are poor surgical risks or those with diffuse disease. The purpose of this article is to review the literature and describe the non-surgical treatment options, indications and efficacies for non-melanoma primary eyelid malignancies.


Epigallocatechin Gallate Is the Most Effective Catechin Against Antioxidant Stress via Hydrogen Peroxide and Radical Scavenging Activity.
BACKGROUND Hydrogen peroxide-induced neuronal oxidative stress is a serious threat to the nervous system. Catechins and related compounds are effective radical scavengers that protect against nerve cell damage. MATERIAL AND METHODS Here, we investigated the antioxidant property of various catechins in protecting against hydrogen peroxide, as well as their radical-scavenging activity. RESULTS We found that catechins treatment effectively protected HT22 cells against H₂O₂-induced cell viability by decreasing and attenuating reactive oxidative species production in different proportions. In addition, all tested catechins performed radical scavenging activity, and partially removed the free radicals. Among all investigated catechins, epigallocatechin gallate was the most effective against ROS production and had the strongest radical-scavenging activity. These results suggest that beneficial effects were strongly related with structure of catechins, mainly because of the hydroxyl and galloyl groups. CONCLUSIONS In conclusion, epigallocatechin gallate is the most effective antioxidant polyphenol against hydrogen peroxide and radical-scavenging activity.


Beyond scattering and absorption: Perceptual unmixing of translucent liquids.
Is perception of translucence based on estimations of scattering and absorption of light or on statistical pseudocues associated with familiar materials? We compared perceptual performance with real and computer-generated stimuli. Real stimuli were glasses of milky tea. Milk predominantly scatters light and tea absorbs it, but since the tea absorbs less as the milk concentration increases, the effects of milkiness and strength on scattering and absorption are not independent. Conversely, computer-generated stimuli were glasses of "milky tea" in which absorption and scattering were independently manipulated. Observers judged tea concentrations regardless of milk concentrations, or vice versa. Maximum-likelihood conjoint measurement was used to estimate the contributions of each physical component-concentrations of milk and tea, or amounts of scattering and absorption-to perceived milkiness or tea strength. Separability of the two physical dimensions was better for real than for computer-generated teas, suggesting that interactions between scattering and absorption were correctly accounted for in perceptual unmixing, but unmixing was always imperfect. Since the real and rendered stimuli represent different physical processes and therefore differ in their image statistics, perceptual judgments with these stimuli allowed us to identify particular pseudocues (presumably learned with real stimuli) that explain judgments with both stimulus sets.


Active Site Sequence Representations of Human Kinases Outperform Full Sequence Representations for Affinity Prediction and Inhibitor Generation: 3D Effects in a 1D Model.
Recent advances in deep learning have enabled the development of large-scale multimodal models for virtual screening and de novo molecular design. The human kinome with its abundant sequence and inhibitor data presents an attractive opportunity to develop proteochemometric models that exploit the size and internal diversity of this family of targets. Here, we challenge a standard practice in sequence-based affinity prediction models: instead of leveraging the full primary structure of proteins, each target is represented by a sequence of 29 discontiguous residues defining the ATP binding site. In kinase-ligand binding affinity prediction, our results show that the reduced active site sequence representation is not only computationally more efficient but consistently yields significantly higher performance than the full primary structure. This trend persists across different models, data sets, and performance metrics and holds true when predicting pIC50 for both unseen ligands and kinases. Our interpretability analysis reveals a potential explanation for the superiority of the active site models: whereas only mild statistical effects about the extraction of three-dimensional (3D) interaction sites take place in the full sequence models, the active site models are equipped with an implicit but strong inductive bias about the 3D structure stemming from the discontiguity of the active sites. Moreover, in direct comparisons, our models perform similarly or better than previous state-of-the-art approaches in affinity prediction. We then investigate a de novo molecular design task and find that the active site provides benefits in the computational efficiency, but otherwise, both kinase representations yield similar optimized affinities (for both SMILES- and SELFIES-based molecular generators). Our work challenges the assumption that the full primary structure is indispensable for modeling human kinases.


Genome-Wide Transcription Factor Binding in Leaves from C3 and C4 Grasses.
The majority of plants use C3 photosynthesis, but over 60 independent lineages of angiosperms have evolved the C4 pathway. In most C4 species, photosynthesis gene expression is compartmented between mesophyll and bundle-sheath cells. We performed DNaseI sequencing to identify genome-wide profiles of transcription factor binding in leaves of the C4 grasses Zea mays, Sorghum bicolor, and Setaria italica as well as C3 Brachypodium distachyon In C4 species, while bundle-sheath strands and whole leaves shared similarity in the broad regions of DNA accessible to transcription factors, the short sequences bound varied. Transcription factor binding was prevalent in gene bodies as well as promoters, and many of these sites could represent duons that influence gene regulation in addition to amino acid sequence. Although globally there was little correlation between any individual DNaseI footprint and cell-specific gene expression, within individual species transcription factor binding to the same motifs in multiple genes provided evidence for shared mechanisms governing C4 photosynthesis gene expression. Furthermore, interspecific comparisons identified a small number of highly conserved transcription factor binding sites associated with leaves from species that diverged around 60 million years ago. These data therefore provide insight into the architecture associated with C4 photosynthesis gene expression in particular and characteristics of transcription factor binding in cereal crops in general.


Bevacizumab-induced isolated oculomotor nerve palsy in glioblastoma multiforme.
Bevacizumab, a monoclonal antibody against the vascular endothelial growth factor receptor, is the standard treatment of recurrent glioblastoma multiforme. In addition to common systemic side effects of bevacizumab, there are rare cases of cranial nerve palsy. We report a case of transient oculomotor nerve palsy after systemic administration of bevacizumab. Twenty-four hours after the systemic infusion of bevacizumab, transient oculomotor nerve palsy developed in a 49-year-old male patient. In the cranial MRI, there was no malignancy-related progression. Bevacizumab treatment was discontinued. Methylprednisolone was started considering that bevacizumab increased the inflammatory response. Oculomotor nerve palsy resolved in 14 days. There are many side effects of bevacizumab whose mechanisms of action have not been fully explained. Cranial nerve involvement is rarely reported. Our case is the first reported case of bevacizumab-induced oculomotor nerve palsy.


Are patients after severe injury who drop out of a longitudinal study at high risk of mental disorder?
In longitudinal studies of traumatic stress, it is particularly important to examine the data for any differences between those who drop out and those who continue to participate, because reluctance to participate might reflect symptoms of avoidance frequently seen in posttraumatic stress disorder (PTSD). However, whether those who drop out are at high risk of PTSD remains unclear. Over a 25-month period, 188 consecutive patients with motor vehicle accident (MVA)-related injuries admitted emergently were enrolled and followed for 4 to 6 weeks. Baseline characteristics were compared between subjects who did and did not participate in the follow-up study. At 4 to 6 weeks, 66 (35.1%) of the participants had dropped out. Bivariate analyses revealed that those who dropped out were likely to be men, alcohol drinkers, smokers, and unconscious just after MVA and to have fewer years of education, less severe injuries, less posttraumatic symptoms, and lower cooperativeness as assessed by the Temperament and Character Inventory. Logistic regression analysis revealed that male sex, unconsciousness during MVA, low cooperativeness, and less severe injuries were significant predictors of dropout. The literature says that male sex and unconsciousness just after MVA might be protective factors against MVA-related PTSD, whereas low cooperativeness is a risk factor for general mental problems. To summarize, it is expected that those who drop from the follow-up are unlikely to have MVA-related PTSD, but might have mental problems independent of injury.


Generalizability of an adult-validated model for predicting increased physical activity in after-school program children: Effects of treatment formats on fitness indicators.
Inappropriately high weight in children is a persistent problem in the United States, and physical activity is often reduced in schools due to academic demands. Effective after-school programs could positively impact both physical activity and overweight/obesity, however previous reviews and meta-analyses have indicated minimal effects. Both 4- and 3-day/week versions of a social cognitive theory-driven physical activity/nutrition after-school program were evaluated against unstructured care to assess effects in children (overall Mage = 10.00 years, SD = .80). For changes over 9 months in body mass index (BMI), effects sizes (Cohen's d) were .68, .40, and .07 in the 4-day (n = 70), 3-day (n = 70), and unstructured (n = 50) groups, respectively. Similar patterns of effects were found for changes in free-time physical activity and cardiovascular endurance. Incorporating a theory-based prediction model previously supported in teens through older adults, with and without medical disorders and health-risk factors, improvements in exercise-related self-regulation and self-efficacy, and mood, significantly predicted increased free-time physical activity (R2 = .48). Effects significantly strengthened to R2 = .62 when completion/non-completion of the recommended 300 min/week of physical activity was also accounted for. Change in BMI was inversely related to physical activity change, β=-.14, p < .05. The present evaluation process indicated promising effects, and indicated theory-based targets to foster future program improvements.


Evaluation of the Activity of a Combination of Three Bacteriophages Alone or in Association with Antibiotics on Staphylococcus aureus Embedded in Biofilm or Internalized in Osteoblasts.
Staphylococcus aureus is responsible for difficult-to-treat bone and joint infections (BJIs). This is related to its ability to form biofilm and to be internalized and persist inside osteoblasts. Recently, bacteriophage therapy has emerged as a promising option to improve treatment of such infections, but data on its activity against the specific bacterial lifestyles presented above remain scarce. We evaluated the activity of a combination of three bacteriophages, recently used for compassionate treatment in France, against S. aureus HG001 in a model of staphylococcal biofilm and a model of osteoblasts infection, alone or in association with vancomycin or rifampin. The activity of bacteriophages against biofilm-embedded S. aureus was dose dependent. In addition, synergistic effects were observed when bacteriophages were combined with antibiotics used at the lowest concentrations. Phage penetration into osteoblasts was observed only when the cells were infected, suggesting a S. aureus-dependent Trojan horse mechanism for internalization. The intracellular bacterial count of bacteria in infected osteoblasts treated with bacteriophages as well as with vancomycin was significantly higher than in cells treated with lysostaphin, used as a control condition, owing to the absence of intracellular activity and the rapid killing of bacteria released after the death of infected cells. These results suggest that bacteriophages are both inactive in the intracellular compartment after being internalized in infected osteoblasts and present a delayed killing effect on bacteria released after cell lysis into the extracellular compartment, which avoids preventing them from infecting other osteoblasts. The combination of bacteriophages tested was highly active against S. aureus embedded in biofilm but showed no activity against intracellular bacteria in the cell model used.


Association between teething and independent walking in healthy children.
Developing teeth provide a reliable indication of maturation and biological age. The objective of this study was to establish whether there is any association between the time of emergence of the first primary tooth and the time when independent walking occurs. A cross-sectional study was conducted with a sample of 206 healthy children (95 girls and 111 boys) aged 12 to 60 (mean: 29.79±0.66) months who were able to walk independently. The study was conducted using a questionnaire that was filled out by the parents. The first primary tooth emerged at 6.86±0.14 (min: 3-max: 13) months; the mean independent walking time was 12.58±2.15 (min: 8.50-max: 24.00) months. There was no correlation between the first teething and independent walking times (r=0.045, p=0.523). Factors such as breastfeeding status, intake of vitamins, walker usage and body mass index were found not to affect the time of either emergence of the first deciduous tooth or independent walking. To the best of our knowledge, this is the first paper in literature to have researched the relationship between the time of emergence of the first deciduous tooth and that of independent walking. It should be explained to parents that there is no relationship between the two in order to resolve anxiety when their child acquires a tooth but does not walk, or vice versa.


Medical and oncological management of malignant mesothelioma.
Mesothelioma is an aggressive cancer, for which no curative oncological treatment currently exists. This article outlines the options for managing malignant pleural effusions, describes the developments in chemotherapy over the past 10 years and summarizes the evidence for prophylactic and palliative radiotherapy.


Assessing drivers of full adoption of test and treat policy for malaria in Senegal.
Malaria treatment policy has changed from presumptive treatment to targeted "test and treat" (T&T) with malaria rapid diagnostic tests (RDTs) and artemisinin combination therapy (ACT). This transition involves changing behavior among health providers, meaning delays between introduction and full implementation are recorded in almost every instance. We investigated factors affecting successful transition, and suggest approaches for accelerating uptake of T&T. Records from 2000 to 2011 from health clinics in Senegal where malaria is mesoendemic were examined (96,166 cases). The study period encompassed the implementation of national T&T policy in 2006. Analysis showed that adherence to test results is the first indicator of T&T adoption and is dependent on accumulation of experience with positive RDTs (odds ratio [OR]: 0.55 [P ≤ 0.001], 95% confidence interval [CI]: 0.53-0.58). Reliance on tests for malaria diagnosis (rather than presumptive diagnosis) followed after test adherence is achieved, and was also associated with increased experience with positive RDTs (OR: 0.60 [P ≤ 0.001], 95% CI: 0.58-0.62). Logistic models suggest that full adoption of T&T clinical practices can occur within 2 years, that monitoring these behavioral responses rather than RDT or ACT consumption will improve evaluation of T&T uptake, and that accelerating T&T uptake by focusing training on adherence to test results will reduce overdiagnosis and associated health and economic costs in mesoendemic regions.


Life-long impairment of glucose homeostasis upon prenatal exposure to psychostimulants.
Maternal drug abuse during pregnancy is a rapidly escalating societal problem. Psychostimulants, including amphetamine, cocaine, and methamphetamine, are amongst the illicit drugs most commonly consumed by pregnant women. Neuropharmacology concepts posit that psychostimulants affect monoamine signaling in the nervous system by their affinities to neurotransmitter reuptake and vesicular transporters to heighten neurotransmitter availability extracellularly. Exacerbated dopamine signaling is particularly considered as a key determinant of psychostimulant action. Much less is known about possible adverse effects of these drugs on peripheral organs, and if in utero exposure induces lifelong pathologies. Here, we addressed this question by combining human RNA-seq data with cellular and mouse models of neuroendocrine development. We show that episodic maternal exposure to psychostimulants during pregnancy coincident with the intrauterine specification of pancreatic β cells permanently impairs their ability of insulin production, leading to glucose intolerance in adult female but not male offspring. We link psychostimulant action specifically to serotonin signaling and implicate the sex-specific epigenetic reprogramming of serotonin-related gene regulatory networks upstream from the transcription factor Pet1/Fev as determinants of reduced insulin production.


The incidence of oesophageal cancer in Eastern Africa: identification of a new geographic hot spot?
The incidence of oesophageal cancer (OC) varies geographically, with more than 80% of cases and deaths worldwide occurring in developing countries. The aim of this study is to characterize the disease burden of OC in four urban populations in Eastern Africa, which may represent a previously undescribed high-incidence area. Data on all cases of OC diagnosed between 2004 and 2008 were obtained from four population-based cancer registries in: Blantyre, Malawi; Harare, Zimbabwe; Kampala, Uganda; and Nairobi, Kenya. Age-standardized incidence rates (ASRs) were calculated for each population, and descriptive statistics for incident cases were determined. In Blantyre, 351 male (59%) and 239 (41%) female cases were reported, with ASRs of 47.2 and 30.3. In Harare, 213 male (61%) and 134 (39%) female cases were reported, with ASRs of 33.4 and 25.3, respectively. In Kampala, 196 male (59%) and 137 female (41%) cases were reported, with ASRs of 36.7 and 24.8. In Nairobi, 323 male (57%) and 239 female (43%) cases were reported, with ASRs of 22.6 and 21.6. Median age at diagnosis was significantly different among the four populations, ranging from 50 years in Blantyre to 65 years in Harare (p<0.0001). Except in Nairobi, incidence among males was significantly higher than among females (p<0.01). Squamous cell OC was the predominant histologic subtype at all sites. ASRs at all four sites were remarkably higher than the mean worldwide ASR. Investigation to evaluate potential etiologic effects of dietary, lifestyle, environmental, and other factors impacting the incidence in this region is needed.