Differential effect of long-term drug selection with doxorubicin and vorinostat on neuroblastoma cells with cancer stem cell characteristics.
Numerous studies have confirmed that cancer stem cells (CSCs) are more resistant to chemotherapy; however, there is a paucity of data exploring the effect of long-term drug treatment on the CSC sub-population. The purpose of this study was to investigate whether long-term doxorubicin treatment could expand the neuroblastoma cells with CSC characteristics and histone acetylation could affect stemness gene expression during the development of drug resistance. Using n-myc amplified SK-N-Be(2)C and non-n-myc amplified SK-N-SH human neuroblastoma cells, our laboratory generated doxorubicin-resistant cell lines in parallel over 1 year; one cell line intermittently treated with the histone deacetylase inhibitor (HDACi) vorinostat and the other without exposure to HDACi. Cells' sensitivity to chemotherapeutic drugs, the ability to form tumorspheres, and capacity for in vitro invasion were examined. Cell-surface markers and side populations (SPs) were analyzed using flow cytometry. Differentially expressed stemness genes were identified through whole genome analysis and confirmed with real-time PCR. Our results indicated that vorinostat increased the sensitivity of only SK-N-Be(2)C-resistant cells to chemotherapy, made cells lose the ability to form tumorspheres, and reduced in vitro invasion and the SP percentage. CD133 was not enriched in doxorubicin-resistant or vorinostat-treated doxorubicin-resistant cells. Nine stemness-linked genes (ABCB1, ABCC4, LMO2, SOX2, ERCC5, S100A10, IGFBP3, TCF3, and VIM) were downregulated in vorinostat-treated doxorubicin-resistant SK-N-Be(2)C cells relative to doxorubicin-resistant cells. A sub-population of cells with CSC characteristics is enriched during prolonged drug selection of n-myc amplified SK-N-Be(2)C neuroblastoma cells. Vorinostat treatment affects the reversal of drug resistance in SK-N-Be(2)C cells and may be associated with downregulation of stemness gene expression. This work may be valuable for clinicians to design treatment protocols specific for different neuroblastoma patients.


Varying the diameter of aligned electrospun fibers alters neurite outgrowth and Schwann cell migration.
Aligned, electrospun fibers have shown great promise in facilitating directed neurite outgrowth within cell and animal models. While electrospun fiber diameter does influence cellular behavior, it is not known how aligned, electrospun fiber scaffolds of differing diameter influence neurite outgrowth and Schwann cell (SC) migration. Thus, the goal of this study was to first create highly aligned, electrospun fiber scaffolds of varying diameter and then assess neurite and SC behavior from dorsal root ganglia (DRG) explants. Three groups of highly aligned, electrospun poly-l-lactic acid (PLLA) fibers were created (1325+383 nm, large diameter fibers; 759+179 nm, intermediate diameter fibers; and 293+65 nm, small diameter fibers). Embryonic stage nine (E9) chick DRG were cultured on fiber substrates for 5 days and then the explants were stained against neurofilament and S100. DAPI stain was used to assess SC migration. Neurite length and SC migration distance were determined. In general, the direction of neurite extension and SC migration were guided along the aligned fibers. On the small diameter fiber substrate, the neurite length was 42% and 36% shorter than those on the intermediate and large fiber substrates, respectively. Interestingly, SC migration did not correlate with that of neurite extension in all situations. SCs migrated equivalently with extending neurites in both the small and large diameter scaffolds, but lagged behind neurites on the intermediate diameter scaffolds. Thus, in some situations, topography alone is sufficient to guide neurites without the leading support of SCs. Scanning electron microscopy images show that neurites cover the fibers and do not reside exclusively between fibers. Further, at the interface between fibers and neurites, filopodial extensions grab and attach to nearby fibers as they extend down the fiber substrate. Overall, the results and observations suggest that fiber diameter is an important parameter to consider when constructing aligned, electrospun fibers for nerve regeneration applications.


I SURVIVE: inter-rater reliability of three physical functional outcome measures in intensive care unit survivors.
We prospectively assessed inter-rater reliability of three physical function measures in Canadian intensive care unit (ICU) survivors in the inpatient setting. We enrolled patients who had an ICU length of stay of ≥ three days, were mechanically ventilated for ≥ 24 hr, and were ambulating independently before hospital admission. Weekly from ICU discharge to hospital discharge, two trained frontline clinicians, blinded to each other's findings, independently performed the Physical Function ICU Test-scored (PFIT-s; score out of 10), 30-sec sit-to-stand (30STS; # of stands), and two-minute walk test (2MWT; distance in m), all within 24 hr. We calculated the intraclass correlation coefficient (ICC), standard error of measurement (SEM), and minimal detectable change (MDC90). We enrolled 42 patients. PFIT-s: in 36 patients with 66 paired scores, the ICC was 0.78 (95% confidence interval [CI], 0.66 to 0.86), the SEM was 1.04, and the MDC90 was 2.42. 30STS: in 35 patients with 67 paired scores, the ICC was 0.85 (95% CI, 0.76 to 0.90), the SEM was 1.91, and the MDC90 was 4.45. 2MWT: in 35 patients with 58 paired scores, the ICC was 0.78 (95% CI, 0.66 to 0.87), the SEM was 20.87, and the MDC90 was 48.69. These three measures show good inter-rater reliability when used by trained frontline clinicians to assess physical function in ICU survivors in the inpatient setting.


Overall body fat and regional fat distribution in young women: quantification with MR imaging.
Overall body fat and its distribution in different regions are important predispositions to known aberrations in lipid and glucose metabolism. The accuracy of MR imaging in estimating overall body fatness and regional fat distribution at individual landmarks was determined by comparing it with well-accepted measures by deuterium-oxide (D2O) dilution and bioimpedance analysis. Fourteen normal young women (athletes and control subjects) were studied. A total of 308 axial, T1-weighted, spin-echo MR images over a specific region in the trunk (21-24 scans per subject) were obtained. Morphometric computer image analysis was performed to determine the subcutaneous, internal, and total fat volumes in each image. The data were analyzed in two ways: data from all slices were summed to assess overall body fatness, and six anatomic landmarks were chosen for regional comparisons. MR-determined estimates of overall body fatness strongly correlated with total body fat measures by D2O dilution in both total fat (r = .91) and subcutaneous fat (r = .92) determinations. Athletes in both the low- and high-intensity training phases had significantly lower values of MR-determined total body fatness than did control subjects. Parallel to total body fatness, athletes had significantly lower MR-determined ratios of total fat/total volume in four of six individual landmarks compared with control subjects. Our experience suggests that MR is an accurate method to quantify overall body fatness, when compared with D2O dilution and bioimpedance analysis. MR could also discriminate regional components of subcutaneous and internal body fat at individual landmarks.


Renal nerve stimulation leads to the activation of the Na+/H+ exchanger isoform 3 via angiotensin II type I receptor.
Renal nerve stimulation at a low frequency (below 2 Hz) causes water and sodium reabsorption via α1-adrenoreceptor tubular activation, a process independent of changes in systemic blood pressure, renal blood flow, or glomerular filtration rate. However, the underlying mechanism of the reabsorption of sodium is not fully understood. Since the sympathetic nervous system and intrarenal ANG II appear to act synergistically to mediate the process of sodium reabsorption, we hypothesized that low-frequency acute electrical stimulation of the renal nerve (ESRN) activates NHE3-mediated sodium reabsorption via ANG II AT1 receptor activation in Wistar rats. We found that ESRN significantly increased urinary angiotensinogen excretion and renal cortical ANG II content, but not the circulating angiotensinogen levels, and also decreased urinary flow and pH and sodium excretion via mechanisms independent of alterations in creatinine clearance. Urinary cAMP excretion was reduced, as was renal cortical PKA activity. ESRN significantly increased NHE3 activity and abundance in the apical microvillar domain of the proximal tubule, decreased the ratio of phosphorylated NHE3 at serine 552/total NHE3, but did not alter total cortical NHE3 abundance. All responses mediated by ESRN were completely abolished by a losartan-mediated AT1 receptor blockade. Taken together, our results demonstrate that higher NHE3-mediated proximal tubular sodium reabsorption induced by ESRN occurs via intrarenal renin angiotensin system activation and triggering of the AT1 receptor/inhibitory G-protein signaling pathway, which leads to inhibition of cAMP formation and reduction of PKA activity.


[Multiple forms of monoaminoxidase in the rat brain during experimental catatonia].
Catalytic properties of multiple forms (separated by affinity chromatography) of rats brain mitochondrial monoamine oxidase (MAO, MAO-I, MAO-11 alpha, MAO-11 beta, MAO-111) have been studied in the animals selected for propensity to development of catatonic syndrome considered as an experimental model of the catatonic syndrome occurring in schizophrenia. It was shown that in experimental catatonia (as compared with rats of the corresponding control group) there was a dramatic increase in the brain stem of the rate of oxidative deamination of beta-phenylethylamine catalyzed by MAO-III; there was also a statistically significant (albeit less expressed than in the experiments with beta-phenylethylamine) increase in the rate of deamination of tyramine and a decrease in the rate of deamination of serotonin. In the systems with MAO-II beta we detected statistically significant increase in the rates of deamination of tyramine and beta-phenylethylamine in experimental catatonia as compared with corresponding control. Studies of multiple forms of brain MAO provide more informative data than estimation of "total" MAO activity (without separation of the multiple forms of this enzyme).


An exciton-coupled circular dichroism protocol for the determination of identity, chirality, and enantiomeric excess of chiral secondary alcohols.
Chiral mono-ols are among the most sought after targets in asymmetric synthesis, and therefore, their chemical characterization and associated enantiomeric excess (ee) values are commonly reported. A simple optical method for determining alcohol identity and ee could be widely used. Toward this end, an in situ-generated multicomponent assembly that creates diastereomeric tris(pyridine) metal complexes incorporating chiral secondary alcohols was explored using exciton-coupled circular dichroism (ECCD). Qualitative models were proposed to predict the preferential diastereomer and its twist, and computational studies provided a rationalization of the CD spectra. Different ECCD spectra found for diastereomers formed in the self-assembled tris(pyridine) complexes were used to determine the absolute configurations of chiral mono-ols. Linear discriminant analysis was successfully employed to classify the alcohol analytes, thereby allowing identification of the alcohols. Conformational effects imparted by heteroatoms were also explored, further expanding the substrate scope. Finally, ee calibration curves allowed the determination of the ee of unknown samples of three chiral secondary alcohols with an average error of 3%. The assay described here is unique because no preparation of structurally elaborated chiral hosts is needed.


Nondeterministic self-assembly with asymmetric interactions.
We investigate general properties of nondeterministic self-assembly with asymmetric interactions, using a computational model and DNA tile assembly experiments. By contrasting symmetric and asymmetric interactions we show that the latter can lead to self-limiting cluster growth. Furthermore, by adjusting the relative abundance of self-assembly particles in a two-particle mixture, we are able to tune the final sizes of these clusters. We show that this is a fundamental property of asymmetric interactions, which has potential applications in bioengineering, and provides insights into the study of diseases caused by protein aggregation.


Monitoring, treatment and control of blood glucose and lipids in Ontario First Nations people with diabetes.
Indigenous people worldwide are disproportionately affected by diabetes and its complications. We aimed to assess the monitoring, treatment and control of blood glucose and lipids in First Nations people in Ontario. We conducted a longitudinal population-based study using administrative data for all people in Ontario with diabetes, stratified by First Nations status. We assessed age- and sex-specific rates of completion of recommended monitoring for low-density lipoprotein (LDL) and glycated hemoglobin (A1c) from 2001/02 to 2014/15. We used data from 2014/15 to conduct a cross-sectional analysis of rates of achievement of A1c and LDL targets and use of glucose-lowering medications. The study included 22 240 First Nations people and 1 319 503 other people in Ontario with diabetes. Rates of monitoring according to guidelines were 20%-50% for A1c and 30%-70% for lipids and were lowest for younger First Nations men. The mean age- and sex-adjusted A1c level was higher among First Nations people than other people (7.59 [95% confidence interval (CI) 7.57 to 7.61] v. 7.03 [95% CI 7.02 to 7.03]). An A1c level of 8.5% or higher was observed in 24.7% (95% CI 23.6 to 25.0) of First Nations people, compared to 12.8% (95% CI 12.1 to 13.5) of other people in Ontario. An LDL level of 2.0 mmol/L or less was observed in 60.3% (95% CI 59.7 to 61.6) of First Nations people, compared to 52.0% (95% CI 51.1 to 52.9) of other people in Ontario. Among those aged 65 or older, a higher proportion of First Nations people than other Ontarians were using insulin (28.1% v. 15.1%), and fewer were taking no medications (28.3% v. 40.1%). As of 2014/15, monitoring and achievement of glycemic control in both First Nations people and other people in Ontario with diabetes remained suboptimal. Interventions to support First Nations patients to reach their treatment goals and reduce the risk of complications need further development and study.


[Morphogenesis of mitral valve calcification in rheumatism and the significance of the petrifications in surgical practice].
Eighty-five calcified mitral valves were examined histologically and histochemically and 28 valves were examined in the scanning electron microscope. Different forms of calcification were discovered in rheumatic sclerosis of the mitral valve: dust-like, laminar petrifact, large-tuberous petrifact with protein apoplexy. The cardiac valve calcification in rheumatic fever is preceeded by local dystrophic changes of collagen fibers which undergo swelling, homogenization, and become picrinophilic. Qualitative changes in collagen predispose to calcium salts adsorption by collagen fibers. Scanning electron microscopy revealed the features of three-dimensional structure of petrifactions at various stages of their development. Dust-like petrifactions are microplates of lime with smooth surface, laminar petrifacts consist of accumulations of these microplates. The surface of large-tuberous petrifacts is irregular, ulcerated, covered with thrombotic masses.


The association between Dietary Inflammatory Index scores and the prevalence of colorectal adenoma.
The Dietary Inflammatory Index (DII)TM, which was developed to characterize the inflammatory potential of a person's diet, has been shown to be associated with inflammatory conditions such as cancer. The present study aimed to investigate the association between DII scores and colorectal adenoma (CRA), a pre-cancerous condition. Responses to baseline dietary questionnaires were used calculate DII scores. In a cross-sectional study design, the association between DII scores and CRA prevalence was determined in men and women separately using logistic regression models. Ten cancer screening centres across the USA. Participants were those included in the screening arm of the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial. Among the 44 278 individuals included in these analyses, men with diets in the most inflammatory quartile of DII scores had higher odds of all types of CRA (advanced, non-advanced and multiple (>1)) compared with those with diets in the least inflammatory quartile of DII scores. In fully adjusted models, compared with those with DII scores in quartile 1 (least inflammatory), males with DII scores in quartile 3 (adjusted odds ratio (aOR)=1·28; 95 % CI 1·12, 1·47) and quartile 4 (aOR=1·41; 95 % CI 1·23, 1·62) were more likely to have prevalent distal CRA. Higher DII scores, representing a more inflammatory diet, also were weakly associated with a higher prevalence of CRA in women. Implementing an anti-inflammatory diet may be an effective means of primary prevention of CRA, especially in men.


Annotating images by mining image search results.
Although it has been studied for years by the computer vision and machine learning communities, image annotation is still far from practical. In this paper, we propose a novel attempt at model-free image annotation, which is a data-driven approach that annotates images by mining their search results. Some 2.4 million images with their surrounding text are collected from a few photo forums to support this approach. The entire process is formulated in a divide-and-conquer framework where a query keyword is provided along with the uncaptioned image to improve both the effectiveness and efficiency. This is helpful when the collected data set is not dense everywhere. In this sense, our approach contains three steps: 1) the search process to discover visually and semantically similar search results, 2) the mining process to identify salient terms from textual descriptions of the search results, and 3) the annotation rejection process to filter out noisy terms yielded by Step 2. To ensure real-time annotation, two key techniques are leveraged-one is to map the high-dimensional image visual features into hash codes, the other is to implement it as a distributed system, of which the search and mining processes are provided as Web services. As a typical result, the entire process finishes in less than 1 second. Since no training data set is required, our approach enables annotating with unlimited vocabulary and is highly scalable and robust to outliers. Experimental results on both real Web images and a benchmark image data set show the effectiveness and efficiency of the proposed algorithm. It is also worth noting that, although the entire approach is illustrated within the divide-and conquer framework, a query keyword is not crucial to our current implementation. We provide experimental results to prove this.


Assessment and characterisation of yeast-based products intended to mitigate ochratoxin exposure using in vitro and in vivo models.
The aim of this paper was to evaluate the capacity of several yeast-based products, derived from baker's and brewer's yeasts, to sequester the mycotoxin ochratoxin A (OTA) and to decrease its rate of absorption and DNA adduct formation in vivo. The experimental protocol included in vitro binding studies using isotherm models, in vivo chicken experiments, in which the serum and tissue concentrations of OTA were analysed in the absence and presence of the test compounds, and the profile of OTA-derived metabolites and their associated DNA adducts were determined. Additionally in vitro cell culture studies (HK2 cells) were applied to assess further the effects for yeast cell product enriched with glutathione (GSH) or selenium. Results of the in vitro binding assay in a buffer system indicated the ability of the yeast-based products, as sequester of OTA, albeit at a different level. In the in vitro experiments in chickens, decreased serum and tissue concentrations of treated animals confirmed that yeast-based products are able to prevent the absorption of OTA. A comparison of the binding affinity in a standard in vitro binding assay with the results obtained in an in vivo chicken experiment, however, showed a poor correlation and resulted in a different ranking of the products. More importantly, we could show that yeast-based products actively modulate the biotransformation of OTA in vivo as well as in vitro in a cell culture model. This effect seems to be attributable to residual enzymatic activities in the yeast-based products. An enrichment of yeast cell wall products with GSH or selenium further modulated the profile of the generated OTA metabolites and the associated pattern of OTA-induced DNA adducts by increasing the conversion of OTA into less toxic metabolites such as OTA, OTB and 4-OH-OTA. A reduced absorption and DNA adduct formation was particularly observed with GSH-enriched yeast, whereas selenium-enriched yeasts could counteract the OTA-induced decrease in cell viability, but at the same time increased the OTA-DNA adducts formation. These findings indicate the need for an in-depth characterisation of yeast-based products used as mycotoxin-mitigating feed additives, in in vivo models with target animal species taking into account not only their ability to sequester toxins in the gastrointestinal tract but also their potential effects on the biotransformation of mycotoxins.


Strong effect of weak charging in suspensions of anisotropic colloids.
Suspensions of hard colloidal particles frequently serve as model systems in studies on fundamental aspects of phase transitions. But often colloidal particles that are considered as "hard" are in fact weakly charged. If the colloids are spherical, weak charging has only a weak effect on the structural properties of the suspension, which can be easily corrected for. However, this does not hold for anisotropic particles. We introduce a model for the interaction potential between charged ellipsoids of revolution (spheroids) based on the Derjaguin approximation of Debye-Hückel theory and present a computer simulation study on aspects of the system's structural properties and phase behaviour. In line with previous experimental observations, we find that even a weak surface charge has a strong impact on the correlation functions. A likewise strong impact is seen on the phase behaviour, in particular, we find stable cubatic order in suspensions of oblate ellipsoids.


Cortical and thalamic contributions to response dynamics across layers of the primary somatosensory cortex during tactile discrimination.
Tactile information processing in the rodent primary somatosensory cortex (S1) is layer specific and involves modulations from both thalamocortical and cortico-cortical loops. However, the extent to which these loops influence the dynamics of the primary somatosensory cortex while animals execute tactile discrimination remains largely unknown. Here, we describe neural dynamics of S1 layers across the multiple epochs defining a tactile discrimination task. We observed that neuronal ensembles within different layers of the S1 cortex exhibited significantly distinct neurophysiological properties, which constantly changed across the behavioral states that defined a tactile discrimination. Neural dynamics present in supragranular and granular layers generally matched the patterns observed in the ventral posterior medial nucleus of the thalamus (VPM), whereas the neural dynamics recorded from infragranular layers generally matched the patterns from the posterior nucleus of the thalamus (POM). Selective inactivation of contralateral S1 specifically switched infragranular neural dynamics from POM-like to those resembling VPM neurons. Meanwhile, ipsilateral M1 inactivation profoundly modulated the firing suppression observed in infragranular layers. This latter effect was counterbalanced by contralateral S1 block. Tactile stimulus encoding was layer specific and selectively affected by M1 or contralateral S1 inactivation. Lastly, causal information transfer occurred between all neurons in all S1 layers but was maximal from infragranular to the granular layer. These results suggest that tactile information processing in the S1 of awake behaving rodents is layer specific and state dependent and that its dynamics depend on the asynchronous convergence of modulations originating from ipsilateral M1 and contralateral S1.


Instantaneous estimation of high-order nonlinear heartbeat dynamics by Lyapunov exponents.
This paper introduces a novel methodology able to provide time varying estimates of the Lyapunov Spectrum within a point process framework. The algorithm is applied to ECG-derived data to characterize heartbeat nonlinear dynamics by using a cubic autoregressive point process model. Estimation of the model parameters is ensured by the Laguerre expansion of the Wiener-Volterra kernels along with a maximum local log-likelihood procedure. In addition to the instantaneous Lyapunov exponents, as well as indices related to higher order dynamic polyspectra, our method is also able to provide all the instantaneous time domain and frequency domain measures of instantaneous heart rate (HR) and heart rate variability (HRV) previously considered. Experimental results show that our method is able to track complex cardiovascular control dynamics during fast transitional gravitational changes.


Haplotype-based genome-wide association study identifies loci and candidate genes for milk yield in Holsteins.
Since milk yield is a highly important economic trait in dairy cattle, the genome-wide association study (GWAS) is vital to explain the genetic architecture underlying milk yield and to perform marker-assisted selection (MAS). In this study, we adopted a haplotype-based empirical Bayesian GWAS to identify the loci and candidate genes for milk yield. A total of 1 092 Holstein cows were sequenced by using the genotyping by genome reducing and sequencing (GGRS) method. After filtering, 164 312 high-confidence SNPs and 13 476 haplotype blocks were identified to use for GWAS. The results indicated that 17 blocks were significantly associated with milk yield. We further identified the nearest gene of each haplotype block and annotated the genes with milk-associated quantitative trait locus (QTL) intervals and ingenuity pathway analysis (IPA) networks. Our analysis showed that four genes, DLGAP1, AP2B1, ITPR2 and THBS4, have relationships with milk yield, while another three, ARHGEF4, TDRD1 and KIF19, were inferred to have potential relationships. Additionally, a network derived from the IPA containing one inferred (ARHGEF4) and all four confirmed genes likely regulates milk yield. Our findings add to the understanding of identifying the causal genes underlying milk production traits and could guide follow up studies for further confirmation of the associated genes, pathways and biological networks.


A Kohonen-like decomposition method for the Euclidean traveling salesman problem-KNIES/spl I.bar/DECOMPOSE.
In addition to the classical heuristic algorithms of operations research, there have also been several approaches based on artificial neural networks for solving the traveling salesman problem. Their efficiency, however, decreases as the problem size (number of cities) increases. A technique to reduce the complexity of a large-scale traveling salesman problem (TSP) instance is to decompose or partition it into smaller subproblems. We introduce an all-neural decomposition heuristic that is based on a recent self-organizing map called KNIES, which has been successfully implemented for solving both the Euclidean traveling salesman problem and the Euclidean Hamiltonian path problem. Our solution for the Euclidean TSP proceeds by solving the Euclidean HPP for the subproblems, and then patching these solutions together. No such all-neural solution has ever been reported.


Exposure of juvenile green frogs (Lithobates clamitans) in littoral enclosures to a glyphosate-based herbicide.
The majority of studies on the toxicity of glyphosate-based herbicides to amphibians have focused on larval life stages exposed in aqueous media. However, adult and juvenile amphibians may also be exposed directly or indirectly to herbicides. The potential for such exposures is of particular interest in the littoral zone surrounding wetlands as this is preferred habitat for many amphibian species. Moreover, it may be argued that potential herbicide effects on juvenile or adult amphibians could have comparatively greater influence on overall recruitment, reproductive potential and thus stability of local populations than effects on larvae. In this experiment, juvenile green frogs (Lithobates clamitans) were exposed to two concentrations (2.16 and 4.27 kg a.e./ha) of a glyphosate-based herbicide formulation (VisionMax®), which were based on typical application scenarios in Canadian forestry. The experimental design employed frogs inhabiting in situ enclosures established at the edge of small naturalized wetlands that were split in half using an impermeable plastic barrier. When analyzed using nominal target application rates, exposure to the glyphosate-based herbicide had no significant effect on survival, body condition, liver somatic index or the observed rate of Batrachochytrium dendrobatidis infection. However, there were marginal trends in both ANOVA analysis and post-hoc regressions regarding B. dendrobatidis infection rates and liver somatic index in relation to measured exposure estimates. Results from this study highlight the importance of field research and the need to include multiple endpoints when examining potential effects of a contaminant on non-target organisms.


PD-L1 expression in the Merkel cell carcinoma microenvironment: association with inflammation, Merkel cell polyomavirus and overall survival.
Merkel cell carcinoma (MCC) is a lethal, virus-associated cancer that lacks effective therapies for advanced disease. Agents blocking the PD-1/PD-L1 pathway have demonstrated objective, durable tumor regressions in patients with advanced solid malignancies and efficacy has been linked to PD-L1 expression in the tumor microenvironment. To investigate whether MCC might be a target for PD-1/PD-L1 blockade, we examined MCC PD-L1 expression, its association with tumor-infiltrating lymphocytes (TILs), Merkel cell polyomavirus (MCPyV), and overall survival. Sixty-seven MCC specimens from 49 patients were assessed with immunohistochemistry for PD-L1 expression by tumor cells and TILs, and immune infiltrates were characterized phenotypically. Tumor cell and TIL PD-L1 expression were observed in 49% and 55% of patients, respectively. In specimens with PD-L1(+) tumor cells, 97% (28/29) demonstrated a geographic association with immune infiltrates. Among specimens with moderate-severe TIL intensities, 100% (29/29) demonstrated PD-L1 expression by tumor cells. Significant associations were also observed between the presence of MCPyV DNA, a brisk inflammatory response, and tumor cell PD-L1 expression: MCPyV(-) tumor cells were uniformly PD-L1(-). Taken together, these findings suggest that a local tumor-specific and potentially MCPyV-specific immune response drives tumor PD-L1 expression, similar to previous observations in melanoma and head and neck squamous cell carcinomas. In multivariate analyses, PD-L1(-) MCCs were independently associated with worse overall survival (hazard ratio 3.12; 95% CI, 1.28-7.61; p=0.012). These findings suggest that an endogenous immune response promotes PD-L1 expression in the MCC microenvironment when MCPyV is present, and provide a rationale for investigating therapies blocking PD-1/PD-L1 for patients with MCC.


Polymeric nanocomposites: compounding and performance.
Polymeric nanocomposites (PNC) are binary mixtures of strongly interacting, inorganic platelets dispersed in a polymeric matrix. For full exfoliation, the thermodynamic miscibility is required. There are three basic methods of organically-modified clay dispersion that might result in PNC: (1) in polymer solution (followed by solvent removal), (2) in a monomer (followed by polymerization), and (3) in molten polymer (compounding). Most commercial PNC are produced by the second method, but it is the third one that has the greatest promise for the plastics industry. Similarly as during the manufacture of polymer blends, the layered silicates must be compatibilized by intercalation with organic salts and/or addition of functionalized macromolecules. Compounding affects the kinetics of dispersion process, but rarely the miscibility. Melt compounding is carried out either in a single-screw (SSE) or a twin-screw extruder (TSE). Furthermore, an extensional flow mixer (EFM) might be attached to an extruder. Two versions of EFM were evaluated: (1) designed for polymer homogenization and blending, and (2) designed for dispersing nano-particles. In this review, the dispersion of organoclay in polystyrene (PS), polyamide-6 (PA-6) or in polypropylene (PP) is discussed. The PNC based on PS or PA-6 contained two components (polymer and organoclay), whereas those based on PP in addition had a compatibilizer mixture of two maleated polypropylenes. Better dispersion was found compounding PNC's in a SSE + EFM than in TSE with or without EFM. The mechanical performance (tensile, flexural and impact) was examined.


A Multicentre Retrospective Review of Clinical Characteristics of Uterine Sarcoma.
Professional societies have recently urged gynaecologists to counsel patients about the risks of encountering uterine sarcoma at fibroid surgery especially when morcellation is used. Our objective was to learn the preoperative and postoperative characteristics of patients with uterine sarcoma to better counsel patients undergoing surgery for presumably benign fibroids. This is a multicentre, retrospective cohort study. Three academic tertiary cancer centres in Southern Ontario over a 13-year period (2001-2014). Patients diagnosed with leiomyosarcoma or endometrial stromal sarcoma were included after identification using pathology databases. A retrospective chart review was conducted to determine clinical characteristics and survival data. The study included 302 patients with uterine sarcomas (221 leiomyosarcomas, 81 endometrial stromal sarcomas). Mean age at diagnosis was 55 years, and 59% were postmenopausal. Sarcoma diagnosis was made following endometrial sampling (25%), hysterectomy (69% laparotomy, 2.7% laparoscopic/vaginal), and myomectomy (3.3%). Of all the patients who underwent endometrial sampling, 65% were diagnosed with a uterine sarcoma in this manner. A general gynaecologist performed the primary surgical procedure in 166 of 302 patients (55%). Tumour disruption at the time of primary surgery occurred in 57 of 295 patients (19%): subtotal hysterectomy (21), myomectomy (10), dissection of adherent tumour (17), and morcellation (9). Morcellation, to facilitate a minimally invasive approach, was performed with scalpel (2 at laparotomy, 5 vaginally) and with a laparoscopic electro-mechanical morcellator (2). At a median follow-up of 2.9 years, there was no significant difference in survival for stage I and II patients with tumour disruption (n = 32) compared with those without tumour disruption (n = 143), regardless of sarcoma type (P = 0.6). The majority of patients with uterine sarcomas were postmenopausal. Many can be diagnosed preoperatively with endometrial sampling. Forty-one percent of patients with uterine sarcomas had a high preoperative index of suspicion, resulting in intervention by an oncologist. Morcellation with laparoscopic electro-mechanical morcellator was rare.


Crown cementation and pulpal health.
A direct result of the liquid continuum within the pulpo-dentine complex is the effect of restorative dentistry on the health of the dental pulp. Better understanding of the role of the complex in relation to restorative dentistry enables strategies to be devised in preserving pulp vitality. A review of the literature produced good laboratory evidence to support the prophylactic sealing of crown preparations with dentine bonding agents.


Promoting health literacy.
This report reviews some of the extensive literature in health literacy, much of it focused on the intersection of low literacy and the understanding of basic health care information. Several articles describe methods for assessing health literacy as well as methods for assessing the readability of texts, although generally these latter have not been developed with health materials in mind. Other studies have looked more closely at the mismatch between patients' literacy levels and the readability of materials intended for use by those patients. A number of studies have investigated the phenomenon of literacy from the perspective of patients' interactions in the health care setting, the disenfranchisement of some patients because of their low literacy skills, the difficulty some patients have in navigating the health care system, the quality of the communication between doctors and their patients including the cultural overlay of such exchanges, and ultimately the effect of low literacy on health outcomes. Finally, the impact of new information technologies has been studied by a number of investigators. There remain many opportunities for conducting further research to gain a better understanding of the complex interactions between general literacy, health literacy, information technologies, and the existing health care infrastructure.


Activation of the JNK pathway is essential for transformation by the Met oncogene.
The Met/Hepatocyte Growth Factor (HGF) receptor tyrosine kinase is oncogenically activated through a rearrangement that creates a hybrid gene Tpr-Met. The resultant chimeric p65(Tpr-Met) protein is constitutively phosphorylated on tyrosine residues in vivo and associates with a number of SH2-containing signaling molecules including the p85 subunit of PI-3 kinase and the Grb2 adaptor protein, which couples receptor tyrosine kinases to the Ras signaling pathway. Mutation of the binding site for Grb2 impairs the ability of Tpr-Met oncoprotein to transform fibroblasts, suggesting that the activation of the Ras/MAP kinase signaling pathway through Grb2 may be essential for cellular transformation. To test this hypothesis dominant-negative mutants of Grb2 with deletions of the SH3 domains were introduced into Tpr-Met transformed fibroblasts. Cells overexpressing the mutants were found to be morphologically reverted and exhibited reduced growth in soft agar. Surprisingly, the Grb2 mutants blocked activation of the JNK/SAPK but not MAP kinase activity induced by the Tpr-Met oncoprotein. Additionally, cells expressing dominant-negative Grb2 mutants had reduced PI-3-kinase activity and dominant-negative mutants of Rac1 blocked both Tpr-Met-induced transformation and activation of JNK. These experiments reveal a novel link between Met and the JNK pathway, which is essential for transformation by this oncogene.


Proteasome inhibition enhances antitumour effects of gemcitabine in experimental pancreatic cancer.
The clinical benefit of gemcitabine, the standard systemic therapy of pancreatic cancer (PaCa), remains modest as a result of high chemoresistance. The proteasome inhibitor bortezomib has antitumour activity against PaCa in vitro and in vivo. We examined the antitumour activity of combination PaCa therapy with bortezomib and gemcitabine. Cell proliferation assays were performed using WST-1 reagent. Protein expression was determined by Western blotting. Efficacy of bortezomib and/or gemcitabine was tested in vivo in a survival study. Bortezomib at 10 microM caused 29% and 72% inhibition in AsPC-1 PaCa cell proliferation at 48 and 96 h incubation, respectively. Bortezomib was even more active against PaCa cell lines Panc-1 and MiaPaCa, with 80% inhibition of proliferation at 48 h. The combination of bortezomib and gemcitabine inhibited AsPC-1 proliferation more effectively compared with each single agent alone. Poly(ADP-ribose) polymerase (PARP) cleavage, an apoptotic indicator, reached 6.6-, 2- and 8.5-fold over controls for bortezomib, gemcitabine and the combination. The median survival was 31 (controls and bortezomib), 40 (gemcitabine) and 48 days (combination), respectively (P < 0.002). Bortezomib and gemcitabine demonstrate additive antitumour activity in vitro and in an experimental PaCa model, indicating the potential for clinical PaCa benefits of additional multiagent therapies that will be based upon the bortezomib and gemcitabine combination.


Epidemiology, Clinical Features, and Disease Severity in Patients With Coronavirus Disease 2019 (COVID-19) in a Children's Hospital in New York City, New York.
Descriptions of the coronavirus disease 2019 (COVID-19) experience in pediatrics will help inform clinical practices and infection prevention and control for pediatric facilities. To describe the epidemiology, clinical, and laboratory features of patients with COVID-19 hospitalized at a children's hospital and to compare these parameters between patients hospitalized with and without severe disease. This retrospective review of electronic medical records from a tertiary care academically affiliated children's hospital in New York City, New York, included hospitalized children and adolescents (≤21 years) who were tested based on suspicion for COVID-19 between March 1 to April 15, 2020, and had positive results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Detection of SARS-CoV-2 from a nasopharyngeal specimen using a reverse transcription-polymerase chain reaction assay. Severe disease as defined by the requirement for mechanical ventilation. Among 50 patients, 27 (54%) were boys and 25 (50%) were Hispanic. The median days from onset of symptoms to admission was 2 days (interquartile range, 1-5 days). Most patients (40 [80%]) had fever or respiratory symptoms (32 [64%]), but 3 patients (6%) with only gastrointestinal tract presentations were identified. Obesity (11 [22%]) was the most prevalent comorbidity. Respiratory support was required for 16 patients (32%), including 9 patients (18%) who required mechanical ventilation. One patient (2%) died. None of 14 infants and 1 of 8 immunocompromised patients had severe disease. Obesity was significantly associated with mechanical ventilation in children 2 years or older (6 of 9 [67%] vs 5 of 25 [20%]; P = .03). Lymphopenia was commonly observed at admission (36 [72%]) but did not differ significantly between those with and without severe disease. Those with severe disease had significantly higher C-reactive protein (median, 8.978 mg/dL [to convert to milligrams per liter, multiply by 10] vs 0.64 mg/dL) and procalcitonin levels (median, 0.31 ng/mL vs 0.17 ng/mL) at admission (P < .001), as well as elevated peak interleukin 6, ferritin, and D-dimer levels during hospitalization. Hydroxychloroquine was administered to 15 patients (30%) but could not be completed for 3. Prolonged test positivity (maximum of 27 days) was observed in 4 patients (8%). In this case series study of children and adolescents hospitalized with COVID-19, the disease had diverse manifestations. Infants and immunocompromised patients were not at increased risk of severe disease. Obesity was significantly associated with disease severity. Elevated inflammatory markers were seen in those with severe disease.


Electron-beam induced deposition and autocatalytic decomposition of Co(CO)3NO.
The autocatalytic growth of arbitrarily shaped nanostructures fabricated by electron beam-induced deposition (EBID) and electron beam-induced surface activation (EBISA) is studied for two precursors: iron pentacarbonyl, Fe(CO)5, and cobalt tricarbonyl nitrosyl, Co(CO)3NO. Different deposits are prepared on silicon nitride membranes and silicon wafers under ultrahigh vacuum conditions, and are studied by scanning electron microscopy (SEM) and scanning transmission X-ray microscopy (STXM), including near edge X-ray absorption fine structure (NEXAFS) spectroscopy. It has previously been shown that Fe(CO)5 decomposes autocatalytically on Fe seed layers (EBID) and on certain electron beam-activated surfaces, yielding high purity, polycrystalline Fe nanostructures. In this contribution, we investigate the growth of structures from Co(CO)3NO and compare it to results obtained from Fe(CO)5. Co(CO)3NO exhibits autocatalytic growth on Co-containing seed layers prepared by EBID using the same precursor. The growth yields granular, oxygen-, carbon- and nitrogen-containing deposits. In contrast to Fe(CO)5 no decomposition on electron beam-activated surfaces is observed. In addition, we show that the autocatalytic growth of nanostructures from Co(CO)3NO can also be initiated by an Fe seed layer, which presents a novel approach to the fabrication of layered nanostructures.


Primary care patient and provider preferences for diabetes care managers.
The collaborative care model, using care managers, has been shown to be effective in achieving sustained treatment outcomes in chronic disease management. Little effort has been made to find out patient preferences for chronic disease care, hence, we conducted a study aimed at identifying these. A 20-item questionnaire, asking for patients' and providers' preferences and perceptions, was mailed out to 1000 randomly selected patients in Olmsted County, Minnesota, identified through a diabetes registry to have type 2 diabetes mellitus, a prototypical prevalent chronic disease. Surveys were also sent to 42 primary care providers. There were 254 (25.4%) patient responders and 28 (66%) provider responders. The majority of patients (>70%) and providers (89%) expressed willingness to have various aspects of diabetes care managed by a care manager. Although 75% of providers would be comfortable expanding the care manager role to other chronic diseases, only 39.5% of patient responders would be willing to see a care manager for other chronic problems. Longer length of time from initial diagnosis of diabetes was associated with decreased patient likelihood to work with a care manager. Despite study limitations, such as the lack of validated measures to assess perceptions related to care management, our results suggest that patients and providers are willing to collaborate with a care manager and that both groups have similar role expectations of a care manager.


Validation of the Behavioural Regulation in Exercise Questionnaire-2 for adults with chronic musculoskeletal pain.
The Behavioural Regulation in Exercise Questionnaire-2 (BREQ-2) has demonstrated reliability and validity in the general population; yet the psychometric properties have not been tested among rehabilitation populations, such as persons experiencing chronic musculoskeletal pain. The objective of the current study was to examine the factorial structure and psychometric properties of the BREQ-2 in a sample of adults with chronic musculoskeletal pain. Adults with chronic musculoskeletal pain (n=211) were recruited from clinical and community networks in the United States. Data were collected using a cross-sectional online survey and analysed using confirmatory factor analysis, multiple indicators multiple causes analysis, and correlational techniques. Participants completed the BREQ-2 and other measures relevant to self-determination theory, including outcome expectancies, self-efficacy and family and friend support for physical activity and exercise. Results indicated the intercorrelated model fit the data well. The five factors of amotivation, external regulation, introjected regulation, identified regulation and intrinsic regulation toward physical activity and exercise demonstrated good reliability and construct validity. This study provides evidence of factorial and construct validity for the BREQ-2 among people with chronic musculoskeletal pain. Implications for rehabilitation practitioners and researchers are discussed.


GRAMM-X public web server for protein-protein docking.
Protein docking software GRAMM-X and its web interface (http://vakser.bioinformatics.ku.edu/resources/gramm/grammx) extend the original GRAMM Fast Fourier Transformation methodology by employing smoothed potentials, refinement stage, and knowledge-based scoring. The web server frees users from complex installation of database-dependent parallel software and maintaining large hardware resources needed for protein docking simulations. Docking problems submitted to GRAMM-X server are processed by a 320 processor Linux cluster. The server was extensively tested by benchmarking, several months of public use, and participation in the CAPRI server track.


Behavioral Economic Research in Addiction as an Area of Growth for the Experimental Analysis of Behavior.
Behavioral economics, a synthesis of the experimental analysis of behavior (EAB) and economics, seeks to determine the relative value of reinforcers as a function of various environmental constraints. Early animal and human studies often focused on drug reinforcement, and this has continued to the present. In particular, behavioral economic analyses of human and animal behavior in relation to nicotine and cigarette smoking have contributed to a greater understanding of this behavior, and to a greater reliance on these methods in the field of smoking cessation treatment, tobacco regulatory science and tobacco control. In this commentary, we briefly describe the history of behavioral economics in the context of EAB methods, the particular contribution of these methods to understanding cigarette smoking and the advance of tobacco regulation, as well as opportunities for growth and remaining challenges in this area. As behavioral economics continues to stimulate research and inform policy, we propose that the underlying elements of a rigorous analytic approach to understanding behavior are key contributors to the fruitfulness of this approach.


Ovarian aging: melatonin regulation of the cytometric and endocrine evolutive pattern.
We have investigated the effect of melatonin administration on the cytometric and endocrine functions of the ovary during aging. Young cyclic (3 months old), middle-aged pre-acyclic (13 months old), and old acyclic (22 months old) female Wistar rats were used for two months, in both control and melatonin-treated groups. Cell cycle by flow cytometry: the percentage of ovarian cells in the G0-G1 phase was the highest in both control and melatonin-treated rats. However, melatonin treatment significantly reduced (P< 0.05) the percentage of cells in the G0-G1 phase compared to control rats. This reduction of cells in the G0-G1 phase is derived to the S phase in cyclic and acyclic rats. The possibility that a tumoral process leads to a proliferative effect observed in young and acyclic melatonin-treated rats was ruled out because no significant differences were found for p53 and Ki67 expression levels between control and melatonin groups. Density of oocytes: the oocyte number per ovary unit volume was not affected by melatonin treatment in the three age ranges studied. Melatonin treatment in middle-aged (pre-acyclic) rats resulted in significantly higher (P< 0.05) ovarian volume; higher oocyte volume, without significant differences, and oocytes in circular form were significantly (P< 0.05) higher than in control rats. Melatonin treatment during pre-acyclic age range could resynchronize the estrous cycle periodicity. Melatonin treatment was able to maintain the same levels of estradiol in the pre-acyclic age groups studied as those observed in the young cyclic rats. The present results indicate that melatonin administration to middle-aged female rats produces beneficial effects that extend the reproductive function of the ovary.


Selfish: discovery of differential chromatin interactions via a self-similarity measure.
High-throughput conformation capture experiments, such as Hi-C provide genome-wide maps of chromatin interactions, enabling life scientists to investigate the role of the three-dimensional structure of genomes in gene regulation and other essential cellular functions. A fundamental problem in the analysis of Hi-C data is how to compare two contact maps derived from Hi-C experiments. Detecting similarities and differences between contact maps are critical in evaluating the reproducibility of replicate experiments and for identifying differential genomic regions with biological significance. Due to the complexity of chromatin conformations and the presence of technology-driven and sequence-specific biases, the comparative analysis of Hi-C data is analytically and computationally challenging. We present a novel method called Selfish for the comparative analysis of Hi-C data that takes advantage of the structural self-similarity in contact maps. We define a novel self-similarity measure to design algorithms for (i) measuring reproducibility for Hi-C replicate experiments and (ii) finding differential chromatin interactions between two contact maps. Extensive experimental results on simulated and real data show that Selfish is more accurate and robust than state-of-the-art methods. https://github.com/ucrbioinfo/Selfish.


Study of chronic kidney disease-mineral bone disorders in newly detected advanced renal failure patients: A Hospital-based cross-sectional study.
We aim to evaluate the disturbances in mineral metabolism, abnormalities in bone mineral density (BMD), and extraskeletal calcification in newly detected, untreated predialysis stage 4 and 5 chronic kidney disease (CKD) patients at a tertiary care hospital in North India. This is cross-sectional observational study. A total of 95 (68 males, 27 females) newly detected patients underwent clinical evaluation, biochemical assessment [serum calcium, phosphorus, alkaline phosphatase (ALP), albumin, creatinine, intact parathyroid hormone (iPTH), 25- hydroxyvitamin D (25(OH)D)], BMD measurement (at spine, hip, and forearm) by dual-energy X-ray absorptiometry (DXA), lateral abdominal radiograph [for abdominal aortic calcification (AAC)], skeletal survey (to look for any abnormality including fractures), and echocardiography [for any cardiac valvular calcification (CVC)]. Symptoms related to CKD-mineral bone disorder were seen in 33.6% of the study patients. Prevalence of hypocalcemia, hyperphosphatemia, hyperparathyroidism, and hypovitaminosis D was 64.2%, 81.1%, 49.5%, and 89.5%, respectively. CVC was seen in 22.1% of patients on echocardiography, mostly involving the mitral valve. Patients with CVC were more likely to be males and smokers. There was no significant difference in iPTH levels between patients with or without CVC. AAC was seen in 10.5% of patients on lateral abdominal X-ray. Patients with AAC had higher levels of iPTH, phosphorus, and ALP and lower levels of calcium compared to patients without AAC. BMD by DXA showed a low bone mass in 41.05% of our patients and was more prevalent in CKD stage 5. Most of the study patients had hyperparathyroidism and low 25(OH)D levels. Our study shows that newly detected, naïve Indian CKD patients have a high prevalence of disturbances of mineral metabolism including hyperparathyroidism, Vitamin D deficiency, abnormal BMD, and valvular and vascular calcification, even before initiating dialysis.


Oral Wine Texture Perception and Its Correlation with Instrumental Texture Features of Wine-Saliva Mixtures.
Unlike solid food, texture descriptors in liquid food are scarce, and they are frequently reduced to the term viscosity. However, in wines, apart from viscosity, terms, such as astringency, body, unctuosity and density, help describe their texture, relating the complexity and balance among their chemical components. Yet there is uncertainty about which wine components (and their combinations) cause each texture sensation and if their instrumental assessment is possible. Therefore, the aim of the present work was to study the effect of wine texture on its main components, when interacting with saliva. This was completed by using instrumental measurements of density and viscosity, and by using two types of panels (trained and expert). For that, six different model-wine formulations were prepared by adding one or multiple wine components: ethanol, mannoproteins, glycerol, and tannins to a de-alcoholised wine. All formulations were mixed with fresh human saliva (1:1), and their density and rheological properties were measured. Although there were no statistical differences, body perception was higher for samples with glycerol and/or mannoproteins, this was also correlated with density instrumental measurements (R = 0.971, p = 0.029). The viscosity of samples with tannins was the highest due to the formation of complexes between the model-wine and salivary proteins. This also provided astringency, therefore correlating viscosity and astringency feelings (R = 0.855, p = 0.030). No correlation was found between viscosity and body perception because of the overlapping of the phenolic components. Overall, the present results reveal saliva as a key factor when studying the wine texture through instrumental measurements (density and viscosity).


Reverse remodelling and myocardial recovery in heart failure.
Advances in medical and device therapies have demonstrated the capacity of the heart to reverse the failing phenotype. The development of normative changes to ventricular size and function led to the concept of reverse remodelling. Among heart failure therapies, durable mechanical circulatory support is most consistently associated with the largest degree of reverse remodelling. Accordingly, research to analyse human tissue after a period of mechanical circulatory support continues to yield a wealth of information. In this Review, we summarize the latest findings on reverse remodelling and myocardial recovery. Accumulating evidence shows that the molecular changes associated with heart failure, in particular in the transcriptome, metabalome, and extracellular matrix, persist in the reverse-remodelled myocardium despite apparent normalization of macrolevel properties. Therefore, reverse remodelling should be distinguished from true myocardial recovery, in which a failing heart regains both normal function and molecular makeup. These findings have implications for future research to develop therapies to repair fully the failing myocardium. Meanwhile, recognition by society guidelines of this new clinical phenotype, which is coming to be known as a state of heart failure remission, underscores the need to accurately define and identify reverse modelled myocardium for the establishment of appropriate therapies.


Contrasting effect of 1-butanol and 1,4-butanediol on the triggered micellar self-assemblies of C16-type cationic surfactants.
The self-assembly in aqueous solutions of three quaternary salt-based C16-type cationic surfactants with different polar head groups and identical carbon alkyl chain viz., cetylpyridinium bromide (CPB), cetyltrimethylammonium tosylate (CTAT), and cetyltriphenylphosphonium bromide (CTPPB) in the presence of 1-butanol (BuOH) and 1,4-butanediol (BTD) was investigated using tensiometry, 2D-nuclear Overhauser enhancement spectroscopy (2D-NOESY) and small angle neutron scattering (SANS) techniques. The adsorption parameters and micellar characteristics evaluated at 303.15 K distinctly showed that BuOH promotes the mixed micelle formation while BTD interfered with the micellization phenomenon. The SANS data fitted using an ellipsoid (as derived by Hayter and Penfold using the Ornstein-Zernike equation and the mean spherical approximation) and wormlike micellar models offered an insight into the micelle size/shape and aggregation number (Nagg) in the examined systems. The evaluated descriptors presented a clear indication of the morphology transition in cationic micelles as induced by the addition of the two alcohols. We also offer an investigation into the acceptable molecular interactions governing the differences in micelle morphologies, using the non-invasive 2D-NOESY technique and molecular modeling. The experimental observations elucidated from computational simulation add novelty to this work. Giving an account to the structural complexity in the three cationic surfactants, the molecular dynamics (MD) simulation was performed for CPB micelles in an aqueous solution of alcohols that highlighted the micelle solvation and structural transition, which is further complemented in terms of critical packing parameter (PP) for the examined systems.


Novel regulatory roles of small G protein GDP dissociation stimulator (smgGDS) in insulin secretion from pancreatic β-cells.
Emerging evidence implicates novel roles for small G protein GDP dissociation stimulator (smgGDS) in G protein activation and subsequent targeting to relevant subcellular compartments for effector regulation. Given the well-established roles of small G proteins in insulin secretion, we undertook this investigation to determine the putative roles of smgGDS in insulin secretion. Immunoblotting studies revealed that both splice variants of smgGDS are expressed in human islets, rat islets and INS-1 832/13 cells. A significant inhibition (-52%) of glucose-stimulated insulin secretion (GSIS) was observed in INS-1 832/13 cells following siRNA-mediated depletion of smgGDS. In addition, insulin secretion elicited by a membrane depolarizing concentration of KCl (via increased calcium influx), forskolin (via increased cAMP generation) or IBMX (via inhibition of phosphodiesterase) was inhibited by -49%, -27%, and -28%, respectively. Subcellular distribution studies revealed no significant alterations in the abundance of smgGDS in the cytosolic and membrane fractions during the 45-min exposure of INS-1 832/13 cells to an insulinotropic concentration of glucose. Together, we present the first evidence of expression of smgGDS in human islets, rodent islets, and clonal β-cells. We also demonstrate novel regulatory roles of these proteins in insulin secretion derived from glucose metabolic events, including calcium- and cAMP-dependent signaling steps.


The regulation of RGLG2-VWA by Ca2+ ions.
RGLG2, an E3 ubiquitin ligase in Arabidopsis thaliana, affects hormone signaling and participates in drought regulation. Here, we determined two crystal structures of RGLG2 VWA domain, representing two conformations, open and closed, respectively. The two structures reveal that Ca2+ ions are allosteric regulators of RGLG2-VWA, which adopts open state when NCBS1(Novel Calcium ions Binding Site 1) binds Ca2+ ions and switches to closed state after Ca2+ ions are removed. This mechanism of allosteric regulation is identical to RGLG1-VWA, but distinct from integrin α and β VWA domains. Therefore, our data provide a backdrop for understanding the role of the Ca2+ ions in conformational change of VWA domain. In addition, we found that RGLG2closed, corresponding to low affinity, can bind pseudo-ligand, which has never been observed in other VWA domains.


[One of the methods of treatment of affective disorders in patients with alcoholism].
It has been demonstrated by a double blind placebo-controlled study that transcranial electric treatment (TET) by means of combination of direct current and pulse current and pulse current at a frequency of 70-80 Hz is an effective method of correcting affective disorders (anxiety, depressions) in patients suffering from alcoholism. The therapeutic effects of TET are coupled with changes in GABA and monoamine metabolism rather than in beta-endorphin as well as with a decrease of the latent period of the occurrence of alpha-rhythm after eyes closing.


Peptide Orientation Strongly Affected by the Nanoparticle Size as Revealed by Sum Frequency Scattering Spectroscopy.
The orientation of proteins at interfaces has a profound effect on the function of proteins. For nanoparticles (NPs) in a biological environment, protein orientation determines the toxicity, function, and identity of the NP. Thus, understanding how proteins orientate at NP surfaces is a critical parameter in controlling NP biochemistry. While planar surfaces are often used to model NP interfaces for protein orientation studies, it has been shown recently that proteins can orient very differently on NP surfaces. This study uses sum frequency scattering vibrational spectroscopy of the model helical leucine-lysine (LK) peptide on NPs of different sizes to determine the cause for the orientation effects. The data show that, for low dielectric constant materials, the orientation of the helical LK peptide is a function of the coulombic forces between peptides across different particle volumes. This finding strongly suggests that flat model systems are only of limited use for determining protein orientation at NP interfaces and that charge interactions should be considered when designing medical NPs or assessing NP biocompatibility.


Coupling a sensory hair-cell bundle to cyber clones enhances nonlinear amplification.
The vertebrate ear benefits from nonlinear mechanical amplification to operate over a vast range of sound intensities. The amplificatory process is thought to emerge from active force production by sensory hair cells. The mechano-sensory hair bundle that protrudes from the apical surface of each hair cell can oscillate spontaneously and function as a frequency-selective, nonlinear amplifier. Intrinsic fluctuations, however, jostle the response of a single hair bundle to weak stimuli and seriously limit amplification. Most hair bundles are mechanically coupled by overlying gelatinous structures. Here, we assayed the effects of mechanical coupling on the hair-bundle amplifier by combining dynamic force clamp of a hair bundle from the bullfrog's saccule with real-time stochastic simulations of hair-bundle mechanics. This setup couples the hair bundle to two virtual hair bundles, called cyber clones, and mimics a situation in which the hair bundle is elastically linked to two neighbors with similar characteristics. We found that coupling increased the coherence of spontaneous hair-bundle oscillations. By effectively reducing noise, the synergic interplay between the hair bundle and its cyber clones also enhanced amplification of sinusoidal stimuli. All observed effects of coupling were in quantitative agreement with simulations. We argue that the auditory amplifier relies on hair-bundle cooperation to overcome intrinsic noise limitations and achieve high sensitivity and sharp frequency selectivity.


A knowledge audit of the managers of primary care organizations: top priority is how to use routinely collected clinical data for quality improvement.
Technology has provided improved access to the rapidly expanding evidence base and to computerized clinical data recorded as part of routine care. A knowledge audit identifies from within this mass of information the knowledge requirements of a professional group or organization, enabling implementation of an appropriately tailored knowledge-management strategy. The objective of the study is to describe perceived knowledge gaps and recommend an appropriate knowledge-management strategy for primary care. The sample comprised 18 senior managers of Primary Care Trusts: the Chairman, Chief Executive Officer, or Research and Development Lead. A series of interviews were recorded verbatim, transcribed and analysed. Knowledge requirements were broad, suggesting that a broadly based knowledge-management strategy is needed in primary care. The biggest gap in current knowledge identified is how to perform needs assessment and quality improvement using aggregated routinely collected, general practice computer data.


Toxicity and Gene Expression Profiling of Copper- and Titanium-Based Nanoparticles Using Air-Liquid Interface Exposure.
To assess the toxicity of nanomaterials, most in vitro studies have been performed under submerged conditions, which do not reflect physiological conditions upon inhalation. An air-liquid interface (ALI) exposure may provide more reliable data on dosimetry and prevent interactions with cell culture media components. Therefore, an ALI exposure was combined with a high-throughput RT-qPCR approach to evaluate the toxicological potential of CuO and TiO2 nanoparticles (NP) in A549 cells. While TiO2 NP did not show any cytotoxicity or other effects compromising genomic stability up to 25.8 μg/cm2, CuO NP revealed a dose-dependent cytotoxicity, starting at 4.9 μg/cm2. Furthermore, CuO NP altered distinct gene expression patterns indicative for disturbed metal homeostasis, stress response, and DNA damage induction. Thus, induction of metal homeostasis associated genes (MT1X, MT2A) at 0.4 μg/cm2 and higher suggested uptake and intracellular dissolution of CuO NP, which was verified by a dose-dependent increase in intracellular copper concentration. Starting at 4.9 μg/cm2, oxidative stress markers (HMOX1, HSPA1A) were induced dose-dependently, supported by elevated ROS levels. Furthermore, a dose-dependent induction of genes associated with DNA damage response (DDIT3, GADD45A) was observed, in concordance with an increase in DNA strand breaks. Finally, transcriptional data suggested the induction of apoptosis at high doses, while flow cytometric analysis revealed increased numbers of either late apoptotic or necrotic cells and clearly necrotic cells at the highest concentrations. Thus, an ALI cell culture system was successfully combined with a comprehensive high-throughput RT-qPCR system, allowing the quantification of NP deposition and their impact on genomic stability. For CuO NP, in principle the data confirm observations made under submerged conditions with respect to intracellular copper ion release, as well as oxidative and genotoxic stress response. However, the results derived from ALI exposure allow the assessment of dose-response-relationships as well as the comparison of relative toxic potencies of different NP.


Calciphylaxis.
Calciphylaxis remains a poorly understood vascular calcification disorder with predilection for patients with end-stage renal disease (ESRD). Recent data from large patient registries and databases have begun to provide information regarding incidence, risk factors, and outcomes in patients with calciphylaxis. The most recent estimate places the incidence of calciphylaxis at 3.5 new cases/1000 patient-years among the patients with ESRD on chronic hemodialysis. It is possible that misdiagnosis or subclinical disease may attribute to lower than the true incidence. There is a suggestion that the incidence is higher in peritoneal dialysis patients compared with patients with hemodialysis. Recent studies have identified a number of risk factors and point to the effects of vitamin K deficiency mediated impairment in Matrix Gla Protein carboxylation as one of the likely pathogenic mechanisms. The outcomes in calciphylaxis patients remain poor with mortality approaching 30% at 6 months and 50% at 12 months. The present review describes recent literature in the field of calciphylaxis. Calciphylaxis registries and specimen biorepositories promise to provide insights into the pathogenesis of calciphylaxis and will pave the way for much needed clinical trials.


Clinical update on oncology treatments and trends.
This article provides a clinical update on recent trends and developments for treating non-small-cell lung, breast, and colorectal cancers and several types of non-Hodgkin's lymphoma. Included in the discussions are results from the latest clinical trials and a look ahead at further research in development. Advances in early detection and screening techniques have helped physicians to detect cancer early and increase survival rates. Monoclonal antibodies have shown promise in treating some forms of cancer and have been a successful part of treatment when coupled with chemotherapy. Such developments are helping to improve patient care and lower mortality rates.


pmartR: Quality Control and Statistics for Mass Spectrometry-Based Biological Data.
Prior to statistical analysis of mass spectrometry (MS) data, quality control (QC) of the identified biomolecule peak intensities is imperative for reducing process-based sources of variation and extreme biological outliers. Without this step, statistical results can be biased. Additionally, liquid chromatography-MS proteomics data present inherent challenges due to large amounts of missing data that require special consideration during statistical analysis. While a number of R packages exist to address these challenges individually, there is no single R package that addresses all of them. We present pmartR, an open-source R package, for QC (filtering and normalization), exploratory data analysis (EDA), visualization, and statistical analysis robust to missing data. Example analysis using proteomics data from a mouse study comparing smoke exposure to control demonstrates the core functionality of the package and highlights the capabilities for handling missing data. In particular, using a combined quantitative and qualitative statistical test, 19 proteins whose statistical significance would have been missed by a quantitative test alone were identified. The pmartR package provides a single software tool for QC, EDA, and statistical comparisons of MS data that is robust to missing data and includes numerous visualization capabilities.


Urethral diverticula in the adult female.
Urethral diverticulum in the adult female is an uncommon lesion which is practically ignored in the radiologic literature. The important symptom is incontinence. Because there is such poor clinical differentiation between stress incontinence, urgency incontinence, and post-void dribbling incontinence, we feel that any patient with incontinence that is not classical stress incontinence or of a neurologic cause should be investigated for urethral diverticulum. Six such patients plus one patient with a false positive panendoscopic examination are presented. Our method of examination with cinefluorography and conventional spot filming is explained. The urethrographic examination was found to be more reliable than the panendoscopic examination. When a discrepancy occurs, further studies are indicated.


Designing and implementing a research integrity promotion plan: Recommendations for research funders.
Various stakeholders in science have put research integrity high on their agenda. Among them, research funders are prominently placed to foster research integrity by requiring that the organizations and individual researchers they support make an explicit commitment to research integrity. Moreover, funders need to adopt appropriate research integrity practices themselves. To facilitate this, we recommend that funders develop and implement a Research Integrity Promotion Plan (RIPP). This Consensus View offers a range of examples of how funders are already promoting research integrity, distills 6 core topics that funders should cover in a RIPP, and provides guidelines on how to develop and implement a RIPP. We believe that the 6 core topics we put forward will guide funders towards strengthening research integrity policy in their organization and guide the researchers and research organizations they fund.


The addition of exercise to a weight loss diet on inflammatory markers: a systematic review and Meta-analysis of controlled clinical trials.
This systematic review and meta-analysis of controlled clinical trials was performed to evaluate the effects of weight-loss diets plus exercise compared with weight-loss diets alone on inflammatory biomarkers in adults. PubMed, Scopus, EMBASE, Web of Science (ISI), and Google Scholar were searched up to April 2021. Overall effects were derived using a random effects model. The overall quality of evidence was assessed using Grading of Recommendations Assessment, Development, and Evaluation (GRADE). Twenty-nine trials were included in this systematic review and meta-analysis. The meta-analysis showed no significant difference in serum high sensitivity C-reactive protein (hs-CRP) [weighted mean difference (WMD) = -0.004 mg/l, 95% CI: -0.140, 0.132, P = 0.954], interleukin-6 (IL-6) (WMD = -0.039 pg/ml, 95% CI: -0.233 to 0.156, P = 0.696), and tumor necrosis factor-α (TNF-α) (WMD = 0.405 ng/ml, 95% CI: -1.036 to 0.226, P = 0.209) after adding exercise to a weight-loss diet compared with a weight-loss diet alone. Subgroup analysis revealed that exercise plus a weight-loss diet significantly reduced TNF-α in studies greater than 16 weeks in duration (WMD = -0.742 ng/ml, 95% CI: -1.384 to -0.1, P = 0.024). This systematic review and meta-analysis indicate that exercise in addition to a weight-loss diet does not have a significant effect on further reducing inflammatory markers compared to a weight loss diet alone.


Edge focusing.
Edge detection in a gray-scale image at a fine resolution typically yields noise and unnecessary detail, whereas edge detection at a coarse resolution distorts edge contours. We show that ``edge focusing'', i.e., a coarse-to-fine tracking in a continuous manner, combines high positional accuracy with good noise-reduction. This is of vital interest in several applications. Junctions of different kinds are in this way restored with high precision, which is a basic requirement when performing (projective) geometric analysis of an image for the purpose of restoring the three-dimensional scene. Segmentation of a scene using geometric clues like parallelism, etc., is also facilitated by the algorithm, since unnecessary detail has been filtered away. There are indications that an extension of the focusing algorithm can classify edges, to some extent, into the categories diffuse and nondiffuse (for example diffuse illumination edges). The edge focusing algorithm contains two parameters, namely the coarseness of the resolution in the blurred image from where we start the focusing procedure, and a threshold on the gradient magnitude at this coarse level. The latter parameter seems less critical for the behavior of the algorithm and is not present in the focusing part, i.e., at finer resolutions. The step length of the scale parameter in the focusing scheme has been chosen so that edge elements do not move more than one pixel per focusing step.


Why do women physicians work fewer hours than men physicians?
Because of the large public investment in medical education, it is important to understand why women physicians work significantly less than men physicians. National survey data on office-based private practice physicians were used to estimate (using two-stage least squares technique) hours and weeks worked for men and women physicians separately. Contrary to conventional wisdom, shorter work weeks for women physicians are not the result of child care responsibilities. Nor would higher earnings encourage women physicians to work longer hours. Instead, we found significant work reductions among married women physicians (but not men), implying subordination of careers by women where combined family incomes are high.


A novel software tool for semi-automatic quantification of thoracic aorta dilatation on baseline and follow-up computed tomography angiography.
A dedicated software package that could semi-automatically assess differences in aortic maximal cross-sectional diameters from consecutive CT scans would most likely reduce the post-processing time and effort by the physicians. The aim of this study was to present and assess the quality of a new tool for the semi-automatic quantification of thoracic aorta dilation dimensions. Twenty-nine patients with two CTA scans of the thoracic aorta for which the official clinical report indicated an increase in aortic diameters were included in the study. Aortic maximal cross-sectional diameters of baseline and follow-up studies generated semi-automatically by the software were compared with corresponding manual measurements. The semi-automatic measurements were performed at seven landmarks defined on the baseline scan by two operators. Bias, Bland-Altman plots and intraclass correlation coefficients were calculated between the two methods and, for the semi-automatic software, also between two observers. The average time difference between the two scans of a single patient was 1188 ± 622 days. For the semi-automatic software, in 2 out of 29 patients, manual interaction was necessary; in the remaining 27 patients (93.1%), semi-automatic results were generated, demonstrating excellent intraclass correlation coefficients (all values ≥ 0.91) and small differences, especially for the proximal aortic arch (baseline: 0.19 ± 1.30 mm; follow-up: 0.44 ± 2.21 mm), the mid descending aorta (0.37 ± 1.64 mm; 0.37 ± 2.06 mm), and the diaphragm (0.30 ± 1.14 mm; 0.37 ± 1.80 mm). The inter-observer variability was low with all errors in diameters ≤ 1 mm, and intraclass correlation coefficients all ≥ 0.95. The semi-automatic tool decreased the processing time by 40% (13 vs. 22 min). In this work, a semi-automatic software package that allows the assessment of thoracic aorta diameters from baseline and follow-up CTs (and their differences), was presented, and demonstrated high accuracy and low inter-observer variability.


The pharmacokinetics of alcohol excretion in human perspiration.
The elimination of ethanol (ETOH) as a gas in human insensible perspiration (PAC) was investigated following the oral administration of 0.5 g/kg or 0.75 g/kg of alcohol. Simultaneous breath alcohol analysis (AAC) was also carried out for comparison purposes. To determine reproducibility, one subject experienced five trials at each dose over a period of six months, in addition to a single dose (0.5 g/kg) trial with five other subjects. All ETOH concentrations were determined by gas chromatography (GLC). A one-compartment open model with first order absorption and pseudo-zero-order elimination was used to compute the pharmacokinetic parameters. The average first order absorption rate constant (K alpha) and pseudo-zero-order elimination rate constant (beta) for ten trials in a single subject were: K alpha AAC = 1.60 h-1 and 1.39 h-1, beta AAC = 0.154 g/l/h and 0.192 g/l/h. K alpha PAC = 1.27 h-1 and 1.11 h-1, beta PAC = 0.109 g/l/h and 0.141 g/l/h for 0.5 g/kg and 0.75 g/kg ETOH, respectively. In all experiments, AAC was consistently higher than PAC during active absorption, with significantly higher (p less than 0.05) average peak concentration (Cmax) and shorter (p less than 0.05) average time to peak (tmax). However, during the later stages of elimination, AAC falls below PAC, with a significantly faster (p less than 0.05) elimination rate (beta). The data indicates that ETOH elimination via perspiration does not parallel the breath and that the pharmacokinetic parameters are significantly different.


Therapeutic Potential and Predictive Pharmaceutical Modeling of Stilbenes in Cannabis sativa.
Cannabis sativa is a plant used for recreational and therapeutic purposes; however, many of the secondary metabolites in the plant have not been thoroughly investigated. Stilbenes are a class of compounds with demonstrated anti-inflammatory and antioxidant properties and are present in cannabis. Many stilbenes present in cannabis have been investigated for their therapeutic effects. Fourteen stilbenes have been identified to be present in cannabis, all of which are structurally dihydrostilbenoids, with half possessing a prenylated moiety. The stilbenes summarized in this analysis show varying degrees of therapeutic benefits ranging from anti-inflammatory, antiviral, and anti-cancer to antioxidant effects. Many of the identified stilbenes have been researched to a limited extent for potential health benefits. In addition, predictive in silico modeling was performed on the fourteen identified cannabis-derived stilbenes. This modeling provides prospective activity, pharmacokinetic, metabolism, and permeability data, setting the groundwork for further investigation into these poorly characterized compounds.


Acidimangrovimonas sediminis gen. nov., sp. nov., isolated from mangrove sediment and reclassification of Defluviimonas indica as Acidimangrovimonas indica comb. nov. and Defluviimonas pyrenivorans as Acidimangrovimonas pyrenivorans comb. nov.
A Gram-stain-negative, aerobic, non-motile, short-rod bacterium, strain MS2-2T, was isolated from mangrove sediment sampled at Jiulong River Estuary, Fujian province, PR China. 16S rRNA gene sequence similarity analysis showed that strain MS2-2T was most closely related to Defluviimonas indica 20V17T (97.41 %) and Defluviimonas pyrenivorans PrR001T (96.18 %). Phylogenetic trees based on 16S rRNA genes and genome sequences both revealed that strain MS2-2T formed a distinct cluster with D. indica 20V17T and D. pyrenivorans PrR001T within family Rhodobacteracea, quite separate from other type species in the genus Defluviimonas. The average nucleotide identity value between strain MS2-2T and D. indica 20V17T was 78.35 %. Growth of strain MS2-2T was observed at 16-41 ° C (optimum, 34 ° C), pH 3.6-7.5 (pH 6.0) and 0.5-10.0 % (w/v) NaCl (4.0 %). The major cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C16 : 0 and C18 : 0. Ubiquinone 10 was the sole quinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 67.9 mol%. The combined genotypic and phenotypic data show that strain MS2-2T represents a novel species of a novel genus in the family Rhodobacteraceae, for which the name Acidimangrovimonassediminis gen. nov., sp. nov. is proposed, with the type strain MS2-2T (=MCCC 1K02682T=NBRC 112978T). We also propose the reclassification of Defluviimonas indica as Acidimangrovimonas indica comb. nov. and Defluviimonas pyrenivorans as Acidimangrovimonas pyrenivorans comb. nov.


Outcomes of Valvular Endocarditis in Patients With and Without Pericardial Effusion: A National Inpatient Sample Study.
Over the last decade, hospitalizations for infective endocarditis (IE) have been steadily increasing, leading to a significant healthcare burden. Pericardial effusion (PCE) has been identified as a serious complication of IE, yet no significant association with mortality has been established. Our study aims to further analyze and understand the significance of PCE in patients with IE. We performed a retrospective analysis using the national inpatient sample database to identify all the hospital admissions with IE using ICD 10 codes and stratified them into 2 groups based on the presence of PCE. The outcomes of interest were inhospital mortality, inhospital complications, need for cardiac surgery, and length of stay. From 2015 Q4-2019, a total of 76,260 hospitalizations were included (weighted: 381,300), of which 2.7% included a PCE diagnosis. Hospitalizations with a PCE diagnosis included patients that were younger (51 vs 61, P < 0.001), as well as slightly more males (58.0% vs 55.2%, P = 0.011), and black patients (16.9% vs 12.9%, P < 0.001). PCE was associated with higher in-hospital death (12.7% vs 9.0%, P < 0.001), longer lengths of stay (12 days vs 7 days, P < 0.001), higher rates of cardiac surgery (22.4% vs 7.3%, P < 0.001). The rates of heart failure, heart block, renal failure, cardiogenic shock, and embolic stroke were higher on PCE group. We found that presence of PCE is associated with higher inhospital mortality, longer length of stay, and greater utilization of cardiac surgery, as well as presence of heart failure, heart block, cardiogenic shock, and embolic stroke.


What Is Slowing Us Down? Six Challenges to Accelerating Advances in Health Behavior Change.
Accelerating advances in health behavior change requires releasing the brake, as well as applying the throttle. This paper discusses six challenges or "brakes" that have slowed progress. We engage with six issues that limit investigators' ability to delineate and test the strategy-target and target-behavior relations that underlie effective interventions according to the experimental medicine approach. We discuss the need for guidance on how to identify the relevant mechanism of action (target) in an intervention and whether a periodic table of health behavior constructs might aid investigators. Experimental and correlational analyses (prospective surveys and behavior change techniques) have been used to test the validity of targets, and we present evidence that there is little agreement among the findings from different research designs. Whereas target engagement is typically analyzed in terms of increasing scores on constructs that impel behavior change, we discuss the role of impeding targets and the benefits of adopting a broader construal of potential targets and approaches to engagement. There is presently a paucity of competitive tests regarding which strategies best engage targets and we discuss empirical criteria and conceptual developments that could enhance the evidence base. Finally, we highlight the need to take "context" or conditional intervention effects more seriously by leveraging the interplay between questions about why interventions work and questions about when and for whom they work. Candid appraisal of the challenges facing research on health behavior change can generate new opportunities for theoretical development and offer direction and cumulative impetus for empirical work.


Global Trends in Proteome Remodeling of the Outer Membrane Modulate Antimicrobial Permeability in Klebsiella pneumoniae.
In Gram-negative bacteria, the permeability of the outer membrane governs rates of antibiotic uptake and thus the efficacy of antimicrobial treatment. Hydrophilic drugs like β-lactam antibiotics depend on diffusion through pore-forming outer membrane proteins to reach their intracellular targets. In this study, we investigated the distribution of porin genes in more than 2,700 Klebsiella isolates and found a widespread loss of OmpK35 functionality, particularly in those strains isolated from clinical environments. Using a defined set of outer-membrane-remodeled mutants, the major porin OmpK35 was shown to be largely responsible for β-lactam permeation. Sequence similarity network analysis characterized the porin protein subfamilies and led to discovery of a new porin family member, OmpK38. Structure-based comparisons of OmpK35, OmpK36, OmpK37, OmpK38, and PhoE showed near-identical pore frameworks but defining differences in the sequence characteristics of the extracellular loops. Antibiotic sensitivity profiles of isogenic Klebsiella pneumoniae strains, each expressing a different porin as its dominant pore, revealed striking differences in the antibiotic permeability characteristics of each channel in a physiological context. Since K. pneumoniae is a nosocomial pathogen with high rates of antimicrobial resistance and concurrent mortality, these experiments elucidate the role of porins in conferring specific drug-resistant phenotypes in a global context, informing future research to combat antimicrobial resistance in K. pneumoniaeIMPORTANCEKlebsiella pneumoniae is a pathogen of humans with high rates of mortality and a recognized global rise in incidence of carbapenem-resistant K. pneumoniae (CRKP). The outer membrane of K. pneumoniae forms a permeability barrier that modulates the ability of antibiotics to reach their intracellular target. OmpK35, OmpK36, OmpK37, OmpK38, PhoE, and OmpK26 are porins in the outer membrane of K. pneumoniae, demonstrated here to have a causative relationship to drug resistance phenotypes in a physiological context. The data highlight that currently trialed combination treatments with a carbapenem and β-lactamase inhibitors could be effective on porin-deficient K. pneumoniae Together with structural data, the results reveal the role of outer membrane proteome remodeling in antimicrobial resistance of K. pneumoniae and point to the role of extracellular loops, not channel parameters, in drug permeation. This significant finding warrants care in the development of phage therapies for K. pneumoniae infections, given the way porin expression will be modulated to confer phage-resistant-and collateral drug-resistant-phenotypes in K. pneumoniae.


Multishot rosette trajectories for spectrally selective MR imaging.
In nuclear magnetic resonance, different spectral components often correspond to different chemical species and as such, spectral selectivity can be a valuable tool for diagnostic imaging. In the work presented here, a multishot image acquisition method based upon rosette K-space trajectories has been developed and implemented for spectrally selective magnetic resonance imaging (MRI). Parametric forms for the gradient waveforms and design constraints are derived, and an example multishot gradient design is presented. The spectral behavior for this imaging method is analyzed in a simulation model. For frequencies that are near to the resonant frequency, this method results in a lower intensity, but undistorted image, while for frequencies that are off-resonance by a large amount, the object is incoherently dephased into noise. A method by which acquisitions are delayed by small amounts is introduced to further reduce the residual intensity for off-resonant signals. An image reconstruction method based on convolution gridding, including a correction method for small amounts of magnetic field inhomogeneity, is implemented. Finally, the spectral selectivity is demonstrated in vivo in a study in which both water and lipid images are generated from a single imaging data set.


Tuberculous splenic abscess in the immunocompetent host: a report and review of literature.
Tubercular splenic abscess is rare, particularly in immunocompetent patients. Diagnostic difficulties usually arise in patients with tubercular splenic abscess because of its non-specific presentation. We report an elderly male who presented with cough and fever and had pulmonary infiltrates suspicious of tuberculosis. Bronchoalveolar lavage microbiology including XpertMTB/Rif assay was non-contributory. Contrast enhanced computed tomography scan of abdomen revealed multiple non-enhancing lesions in the spleen. Ultrasound guided splenic aspirate revealed pus that was positive for Mycobacterium tuberculosis in XpertMTB/Rif assay confirming the diagnosis of tuberculosis.


Gastrointestinal nematode infection interferes with experimental allergic airway inflammation but not atopic dermatitis.
Some helminth infections are negatively associated with the prevalence of allergic disorders, arguing for a modulation of allergic reactions by the parasites, depending on the worm species, intensity and phase of infection and the type of disease. The aim of this study was to analyse the influence of a chronic infection with the gastrointestinal nematode Heligmosomoides polygyrus, in a murine model of allergic airway disease and of atopic dermatitis (AD), respectively. Mice were infected with H. polygyrus and systemically sensitized with the model allergen ovalbumin. Subsequently, the animals were challenged with the allergen either via the airways for induction of airway disease, or via skin patches for induction of dermatitis. Mice concomitantly infected with H. polygyrus showed diminished eosinophil and lymphocyte recruitment into the lungs and decreased allergen-specific IgE levels when compared with sensitized and airway challenged controls. In addition, animals showed a trend towards reduced airway hyper-reactivity. In contrast, no significant differences in the severity of eczematous skin lesions were observed between infected and control animals in the AD model. Although H. polygyrus infection reduced CD8+ and CD4+ T-cell infiltration into the skin and production of allergen-specific IgE, mast cell recruitment was significantly increased in worm-infected mice in the dermatitis model. The worm infection was associated with significantly elevated numbers of Foxp3+ regulatory T cells (Treg) in peribronchial lymph nodes in H. polygyrus-infected sensitized and airway challenged mice. In contrast, Treg cells were basically absent in eczematous skin and their number was not increased in skin-draining lymph nodes of mice with experimental dermatitis. Infection with the gastrointestinal nematode used in our study leads to significant inhibition of mucosa-associated but not cutaneous allergic reactions, pointing to a site specificity of the immunomodulation exerted by helminths. This finding might be an important aspect for future considerations of helminths for treatment of allergic diseases.


On the diversity of secreted phospholipases A(2). Cloning, tissue distribution, and functional expression of two novel mouse group II enzymes.
Over the last decade, an expanding diversity of secreted phospholipases A(2) (sPLA(2)s) has been identified in mammals. Here, we report the cloning in mice of three additional sPLA(2)s called mouse group IIE (mGIIE), IIF (mGIIF), and X (mGX) sPLA(2)s, thus giving rise to eight distinct sPLA(2)s in this species. Both mGIIE and mGIIF sPLA(2)s contain the typical cysteines of group II sPLA(2)s, but have relatively low levels of identity (less than 51%) with other mouse sPLA(2)s, indicating that these enzymes are novel group II sPLA(2)s. However, a unique feature of mGIIF sPLA(2) is the presence of a C-terminal extension of 23 amino acids containing a single cysteine. mGX sPLA(2) has 72% identity with the previously cloned human group X (hGX) sPLA(2) and displays similar structural features, making it likely that mGX sPLA(2) is the ortholog of hGX sPLA(2). Genes for mGIIE and mGIIF sPLA(2)s are located on chromosome 4, and that of mGX sPLA(2) on chromosome 16. Northern and dot blot experiments with 22 tissues indicate that all eight mouse sPLA(2)s have different tissue distributions, suggesting specific functions for each. mGIIE sPLA(2) is highly expressed in uterus, and at lower levels in various other tissues. mGIIF sPLA(2) is strongly expressed during embryogenesis and in adult testis. mGX sPLA(2) is mostly expressed in adult testis and stomach. When the cDNAs for the eight mouse sPLA(2)s were transiently transfected in COS cells, sPLA(2) activity was found to accumulate in cell medium, indicating that each enzyme is secreted and catalytically active. Using COS cell medium as a source of enzymes, pH rate profile and phospholipid headgroup specificity of the novel sPLA(2)s were analyzed and compared with the other mouse sPLA(2)s.