An image classification approach to analyze the suppression of plant immunity by the human pathogen Salmonella Typhimurium.
The enteric pathogen Salmonella is the causative agent of the majority of food-borne bacterial poisonings. Resent research revealed that colonization of plants by Salmonella is an active infection process. Salmonella changes the metabolism and adjust the plant host by suppressing the defense mechanisms. In this report we developed an automatic algorithm to quantify the symptoms caused by Salmonella infection on Arabidopsis. The algorithm is designed to attribute image pixels into one of the two classes: healthy and unhealthy. The task is solved in three steps. First, we perform segmentation to divide the image into foreground and background. In the second step, a support vector machine (SVM) is applied to predict the class of each pixel belonging to the foreground. And finally, we do refinement by a neighborhood-check in order to omit all falsely classified pixels from the second step. The developed algorithm was tested on infection with the non-pathogenic E. coli and the plant pathogen Pseudomonas syringae and used to study the interaction between plants and Salmonella wild type and T3SS mutants. We proved that T3SS mutants of Salmonella are unable to suppress the plant defenses. Results obtained through the automatic analyses were further verified on biochemical and transcriptome levels. This report presents an automatic pixel-based classification method for detecting "unhealthy" regions in leaf images. The proposed method was compared to existing method and showed a higher accuracy. We used this algorithm to study the impact of the human pathogenic bacterium Salmonella Typhimurium on plants immune system. The comparison between wild type bacteria and T3SS mutants showed similarity in the infection process in animals and in plants. Plant epidemiology is only one possible application of the proposed algorithm, it can be easily extended to other detection tasks, which also rely on color information, or even extended to other features.


Low-affinity spermine block mediating outward currents through Kir2.1 and Kir2.2 inward rectifier potassium channels.
The outward component of the strong inward rectifier K(+) current (I(Kir)) plays a pivotal role in polarizing the membranes of excitable and non-excitable cells and is regulated by voltage-dependent channel block by internal cations. Using the Kir2.1 channel, we previously showed that a small fraction of the conductance susceptible only to a low-affinity mode of block likely carries a large portion of the outward current. To further examine the relevance of the low-affinity block to outward I(Kir) and to explore its molecular mechanism, we studied the block of the Kir2.1 and Kir2.2 channels by spermine, which is the principal Kir2 channel blocker. Current-voltage relations of outward Kir2.2 currents showed a peak, a plateau and two peaks in the presence of 10, 1 and 0.1 microm spermine, respectively, which was explained by the presence of two conductances that differ in their susceptibility to spermine block. When the current-voltage relations showed one peak, like those of native I(Kir), outward Kir2.2 currents were mediated mostly by the conductance susceptible to the low-affinity block. They also flowed in a narrower range than the corresponding Kir2.1 currents, because of 3- to 4-fold greater susceptibility to the low-affinity block than in Kir2.1. Reducing external [K(+)] shifted the voltage dependences of both the high- and low-affinity block of Kir2.1 in parallel with the shift in the reversal potential, confirming the importance of the low-affinity block in mediating outward I(Kir). When Kir2.1 mutants known to have reduced sensitivity to internal blockers were examined, the D172N mutation in the transmembrane pore region made almost all of the conductance susceptible only to low-affinity block, while the E224G mutation in the cytoplasmic pore region reduced the sensitivity to low-affinity block without markedly altering that to the high-affinity block or the high/low conductance ratio. The effects of these mutations support the hypothesis that Kir2 channels exist in two states having different susceptibilities to internal cationic blockers.


Long non-coding RNA FENDRR inhibits migration and invasion of cutaneous malignant melanoma cells.
The present study aimed to investigate the effects of lncRNA FENDRR on the migration and invasion of malignant melanoma (MM) cells. The expression levels of FENDRR in MM tissues and MM cell lines were detected using qRT-PCR, followed by construction of FENDRR-knocked down and overexpressed stable cells. Then the effects of FENDRR on cell proliferation, migration and invasion were detected using MTT assay and Transwell assay. The protein expression levels of matrix metallopeptidase 2 (MMP2), MMP9, and related factors in JNK/c-Jun pathway were detected using Western blot. FENDRR was down-regulated in MM tissues and cell lines. Besides, its expression levels in different MM cells were diverse. Knockdown of FENDRR facilitated MM cells proliferation, migration and invasion in A375 cells, while overexpressing FENDRR had reverse results. In addition, MMPs and JNK/c-Jun pathway involved in the FENDRR-mediated regulation of MM cell proliferation, migration and invasion. Our results demonstrated that FENDRR mediated the metastasis phenotype of MM cells by inhibiting the expressions of MMP2 and MMP9 and antagonizing the JNK/c-Jun pathway.


Effect on cell surface hydrophobicity and susceptibility of Helicobacter pylori to medicinal plant extracts.
Effects on aqueous extracts of medicinal plants on ten Helicobacter pylori strains were studied by the salt aggregation test to determine the possibility to modulate their cell surface hydrophobicity and by an agar diffusion assay for detection of antimicrobial activity. It was established that aqueous extracts of bearberry and cowberry leaves enhance cell aggregation of all H. pylori strains tested by the salt aggregation test, and the extract of bearberry possessed a remarkable bacteriostatic activity. Pure tannic acid showed a result similar to that of bearberry and cowberry extracts which contained a large amount of tannins. In contrast, extracts of wild camomile and pineapple-weed, which blocked aggregation of H. pylori, contained small amounts of tannins and did not reveal any antimicrobial activity. Tannic acid seems to be the component of bearberry and cowberry aqueous extracts with the highest activity to decrease cell surface hydrophobicity as well as in antibacterial activity against H. pylori.


Mobile genetic elements from the maternal microbiome shape infant gut microbial assembly and metabolism.
The perinatal period represents a critical window for cognitive and immune system development, promoted by maternal and infant gut microbiomes and their metabolites. Here, we tracked the co-development of microbiomes and metabolomes from late pregnancy to 1 year of age using longitudinal multi-omics data from a cohort of 70 mother-infant dyads. We discovered large-scale mother-to-infant interspecies transfer of mobile genetic elements, frequently involving genes associated with diet-related adaptations. Infant gut metabolomes were less diverse than maternal but featured hundreds of unique metabolites and microbe-metabolite associations not detected in mothers. Metabolomes and serum cytokine signatures of infants who received regular-but not extensively hydrolyzed-formula were distinct from those of exclusively breastfed infants. Taken together, our integrative analysis expands the concept of vertical transmission of the gut microbiome and provides original insights into the development of maternal and infant microbiomes and metabolomes during late pregnancy and early life.


Use of portfolios for assessment of resident teaching skills.
Portfolios are effective instruments for assessment of teaching skills among professional teachers and have recently been adapted in medical education. However, scoring rubrics are needed to effectively guide assessors. Portfolio assessors reviewed and made assessment comments about the resident-as-teacher sections of 11 internal medicine residents' electronic portfolios and discussed their assessments in an assessor group discussion. We performed qualitative analyses of written and oral comments. Major themes were identified, and member checking and triangulation with the literature was performed to evaluate the trustworthiness of the qualitative analysis. Three faculty educators reviewed and commented on 241 uploaded e-portfolio documents accompanying reflections. Three major themes were identified: Application of Teaching Skills, Presentation Skills, and Insights as a Teacher. Themes and subthemes matched closely to several components of the conceptual framework of effective presentations formulated in Glassick standards for scholarly work, as well as themes found in assessments of professional teachers' portfolios. Assessments of portfolios by experienced faculty educators appear to be useful for identifying many important facets of formal teaching presentations and may be useful for creation of a scoring rubric.


Reduced global plant respiration due to the acclimation of leaf dark respiration coupled with photosynthesis.
Leaf dark respiration (Rd ) acclimates to environmental changes. However, the magnitude, controls and time scales of acclimation remain unclear and are inconsistently treated in ecosystem models. We hypothesized that Rd and Rubisco carboxylation capacity (Vcmax ) at 25°C (Rd,25 , Vcmax,25 ) are coordinated so that Rd,25 variations support Vcmax,25 at a level allowing full light use, with Vcmax,25 reflecting daytime conditions (for photosynthesis), and Rd,25 /Vcmax,25 reflecting night-time conditions (for starch degradation and sucrose export). We tested this hypothesis temporally using a 5-yr warming experiment, and spatially using an extensive field-measurement data set. We compared the results to three published alternatives: Rd,25 declines linearly with daily average prior temperature; Rd at average prior night temperatures tends towards a constant value; and Rd,25 /Vcmax,25 is constant. Our hypothesis accounted for more variation in observed Rd,25 over time (R2 = 0.74) and space (R2 = 0.68) than the alternatives. Night-time temperature dominated the seasonal time-course of Rd , with an apparent response time scale of c. 2 wk. Vcmax dominated the spatial patterns. Our acclimation hypothesis results in a smaller increase in global Rd in response to rising CO2 and warming than is projected by the two of three alternative hypotheses, and by current models.


Behind the Indolent Facade: Uncovering the Molecular Features and Malignancy Potential in Lung Minimally Invasive Adenocarcinoma by Single-Cell Transcriptomics.
The increased use of low-dose computed tomography screening has led to more frequent detection of early stage lung tumors, including minimally invasive adenocarcinoma (MIA). To unravel the intricacies of tumor cells and the immune microenvironment in MIA, this study performs a comprehensive single-cell transcriptomic analysis and profiles the transcriptomes of 156,447 cells from fresh paired MIA and invasive adenocarcinoma (IA) tumor samples, peripheral blood mononuclear cells, and adjacent normal tissue samples from three patients with synchronous multiple primary lung adenocarcinoma. This study highlights a connection and heterogeneity between the tumor ecosystem of MIA and IA. MIA tumor cells exhibited high expression of aquaporin-1 and angiotensin II receptor type 2 and a basal-like molecular character. Furthermore, it identifies that cathepsin B+ tumor-associated macrophages may over-activate CD8+ T cells in MIA, leading to an enrichment of granzyme K+ senescent CD8+ T cells, indicating the possibility of malignant progression behind the indolent appearance of MIA. These findings are further validated in 34 MIA and 35 IA samples by multiplexed immunofluorescence. These findings provide valuable insights into the mechanisms that maintain the indolent nature and prompt tumor progression of MIA and can be used to develop more effective therapeutic targets and strategies for MIA patients.


Enterococcus-like infections in Macrobrachium rosenbergii are exacerbated by high pH and temperature but reduced by low salinity.
Macrobrachium rosenbergii (10 to 15 g and 8 to 12 g at intermolt) were challenged with an enterococcus-like bacterium (strain KM002) previously isolated and identified as the causal agent of mortality. Challenge doses and conditions of pH, salinity and temperature were varied to determine the influence of environmental factors on the development of disease and mortality. Survival was 100% for the unchallenged control groups in all trials. In pH tests, the onset of mortality was earlier at pH 8.8 to 9.5 than at pH 4.6 to 5.2 and 7.5 to 7.7. Also, as pH 8.8 to 9.5, all challenged prawns died within 6 d in high dose challenge tests. By contrast, 20% of the prawns challenged at pH 4.6 to 5.2 and 7.5 to 7.7 survived. At low dose challenge (5 x 10(4) cfu prawn(-1)), survival increased significantly except at pH 8.8 to 9.5. In salinity tests at 2 challenge doses (1 x 10(6) and 2 x 10(7) cfu prawn(-1)), onset of mortality was earliest at 15 ppt and cumulative mortality was 100% at 15 ppt and 0 ppt. By contrast, survival was 80% at 5 and 10 ppt at the low dose challenge and 40% and 60%, respectively, at the high dose challenge. When the challenge dose was reduced to 5 x 10(4) cfu prawn(-1), survival was not significantly different at different salinity levels. In temperature tests at pH 7.2 to 7.5 and at 2 challenge doses (2 x 10(7) and 4 x 10(7) cfu prawn(-1)), the onset of mortality was earliest at 33 to 34 degrees C and total mortality occurred at 27 to 28 degrees C and 33 to 34 degrees C. By contrast, there were 40% and 20% survivors, respectively, for low and high challenge doses at 30 to 31 degrees C. Reducing the challenge dose to 5 x 10(4) cfu prawn(-1) gave higher survival in all groups. However, survival at 33 to 34 degrees C was still lowest. In similar temperature tests but at pH 8.8 to 9.5, onset of mortality was somewhat accelerated and there was 100% death for all the high challenge groups. At low challenge doses, mortality was lower but still highest in the 33 to 34 degrees C group. Results indicated that mortality of M. rosenbergii caused by this Enterococcus-like bacterium was exacerbated by environmental parameters of temperature and pH different from those known to be optimal for prawn growth. By contrast, low salinity appeared to have a beneficial effect on survival. Further work is needed to determine the mechanisms underlying these effects.


Campylobacter jejuni bacteremia and Helicobacter pylori in a patient with X-linked agammaglobulinemia.
We describe a 15-year-old patient with X-linked agammaglobulinemia who developed malabsorption and bacteremia due to infection of Helicobacter pylori and Campylobacter jejuni. The Campylobacter bacteremia was only recognized after subculturing of blood culture bottles that failed to signal in the automated system. After 2 weeks of treatment with meropenem and erythromycin for 4 weeks, the patient developed a relapse of bacteremia 10 months later with a high level erythromycin resistant C. jejuni. Sequencing revealed an A2058C mutation in the 23 S rRNA gene associated with this resistance. Treatment with doxycycline for 4 weeks finally resulted in complete eradication. This case report illustrates the importance for physicians to use adapted culture methods and adequate prolonged therapy in patients with an immunodeficiency. A summary of published case reports and series of patients with hypogammaglobulinemia or agammaglobulinemia with Campylobacter or Helicobacter bacteremia is given.


Associations of body composition with type 2 diabetes mellitus.
The aims of this study were to establish the associations of stature, body mass index, waist to hip ratio, and waist circumference with Type 2 (non-insulin-dependent) diabetes mellitus in a random sample of 5887 men and 7018 women aged 20-59 years in a cross-sectional study set in The Netherlands. The crude prevalence of Type 2 diabetes (overall 1.58% in men, 0.94% in women) was significantly (p < 0.01) higher in shorter subjects and those with high body mass index, high waist to hip ratio, and larger waist circumference. Odds ratios and 95% confidence intervals (95% CI) were adjusted for age, cigarette smoking, alcohol consumption, physical activity, and education. Compared to the tallest tertile of height, odds ratios for Type 2 diabetes were 4.4 (95% CI: 1.3 to 11.5) in men and 1.6 (95% CI: 0.8 to 3.2) in women whose height was in the shortest tertile. Compared to the lowest tertile, odds ratios for Type 2 diabetes were 18.4 (95% CI: 4.3 to 78.5) in men and 5.3 (95% CI: 2.0 to 14.0) in women with waist to hip ratio in the highest tertile, 4.1 (95% CI: 2.0 to 8.4) in men and 2.1 (95% CI: 1.0 to 4.2) in women with body mass index in the highest tertile, and 4.9 (95% CI: 2.1 to 11.7) in men and 2.7 (95% CI: 1.2 to 5.9) in women with waist circumference in the highest tertile. In conclusion, although in longitudinal studies waist is a powerful predictor of diabetes incidence, Type 2 diabetes in a cross-sectional survey is associated with shortness in stature, as well as large waist circumference and high body mass index, and particularly strongly with high waist to hip ratio, suggesting that the development of Type 2 diabetes may modify hip circumference independently of body fat.


Role of Melatonin in the Regulation of Pain.
Melatonin is a pleiotropic hormone synthesized and secreted mainly by the pineal gland in vertebrates. Melatonin is an endogenous regulator of circadian and seasonal rhythms. Melatonin is involved in many physiological and pathophysiological processes demonstrating antioxidant, antineoplastic, anti-inflammatory, and immunomodulatory properties. Accumulating evidence has revealed that melatonin plays an important role in pain modulation through multiple mechanisms. In this review, we examine recent evidence for melatonin on pain regulation in various animal models and patients with pain syndromes, and the potential cellular mechanisms.


KERNEL-PENALIZED REGRESSION FOR ANALYSIS OF MICROBIOME DATA.
The analysis of human microbiome data is often based on dimension-reduced graphical displays and clusterings derived from vectors of microbial abundances in each sample. Common to these ordination methods is the use of biologically motivated definitions of similarity. Principal coordinate analysis, in particular, is often performed using ecologically defined distances, allowing analyses to incorporate context-dependent, non-Euclidean structure. In this paper, we go beyond dimension-reduced ordination methods and describe a framework of high-dimensional regression models that extends these distance-based methods. In particular, we use kernel-based methods to show how to incorporate a variety of extrinsic information, such as phylogeny, into penalized regression models that estimate taxonspecific associations with a phenotype or clinical outcome. Further, we show how this regression framework can be used to address the compositional nature of multivariate predictors comprised of relative abundances; that is, vectors whose entries sum to a constant. We illustrate this approach with several simulations using data from two recent studies on gut and vaginal microbiomes. We conclude with an application to our own data, where we also incorporate a significance test for the estimated coefficients that represent associations between microbial abundance and a percent fat.


Sequence biases in large scale gene expression profiling data.
We present the results of a simple, statistical assay that measures the G+C content sensitivity bias of gene expression experiments without the requirement of a duplicate experiment. We analyse five gene expression profiling methods: Affymetrix GeneChip, Long Serial Analysis of Gene Expression (LongSAGE), LongSAGELite, 'Classic' Massively Parallel Signature Sequencing (MPSS) and 'Signature' MPSS. We demonstrate the methods have systematic and random errors leading to a different G+C content sensitivity. The relationship between this experimental error and the G+C content of the probe set or tag that identifies each gene influences whether the gene is detected and, if detected, the level of gene expression measured. LongSAGE has the least bias, while Signature MPSS shows a strong bias to G+C rich tags and Affymetrix data show different bias depending on the data processing method (MAS 5.0, RMA or GC-RMA). The bias in the Affymetrix data primarily impacts genes expressed at lower levels. Despite the larger sampling of the MPSS library, SAGE identifies significantly more genes (60% more RefSeq genes in a single comparison).


Triggering protein folding within the GroEL-GroES complex.
The folding of many proteins depends on the assistance of chaperonins like GroEL and GroES and involves the enclosure of substrate proteins inside an internal cavity that is formed when GroES binds to GroEL in the presence of ATP. Precisely how assembly of the GroEL-GroES complex leads to substrate protein encapsulation and folding remains poorly understood. Here we use a chemically modified mutant of GroEL (EL43Py) to uncouple substrate protein encapsulation from release and folding. Although EL43Py correctly initiates a substrate protein encapsulation reaction, this mutant stalls in an intermediate allosteric state of the GroEL ring, which is essential for both GroES binding and the forced unfolding of the substrate protein. This intermediate conformation of the GroEL ring possesses simultaneously high affinity for both GroES and non-native substrate protein, thus preventing escape of the substrate protein while GroES binding and substrate protein compaction takes place. Strikingly, assembly of the folding-active GroEL-GroES complex appears to involve a strategic delay in ATP hydrolysis that is coupled to disassembly of the old, ADP-bound GroEL-GroES complex on the opposite ring.


A novel collagen-binding peptide promotes osteogenic differentiation via Ca2+/calmodulin-dependent protein kinase II/ERK/AP-1 signaling pathway in human bone marrow-derived mesenchymal stem cells.
The intracellular signaling events controlling human mesenchymal stem cell (hMSC) differentiation into osteoblasts are poorly understood. Collagen-binding domain is considered an essential component of bone mineralization. In the present study, we investigated the regulatory mechanism of osteoblastic differentiation of hMSC by the peptide with a novel collagen-binding motif derived from osteopontin. The peptide induced influx of extracellular Ca2+ via calcium channels and increased intracellular Ca2+ concentration ([Ca2+]i) independent of both pertussis toxin and phospholipase C, and activated ERK, which was inhibited by Ca2+/calmodulin-dependent protein kinase (CaMKII) antagonist, KN93. The peptide-induced increase of [Ca2+]i is correlated with ERK activation in a various cell types. The peptide stimulated the migration of hMSC but suppressed cell proliferation. Furthermore, the peptide increased the phosphorylation of cAMP-response element-binding protein, leading to a significant increase in the transactivation of cAMP-response element and serum response element. Ultimately, the peptide increased AP-1 transactivation, c-jun expression, and bone mineralization, which are suppressed by KN93. Taken together, these results indicate that the novel collagen-binding peptide promotes osteogenic differentiation via Ca2+/CaMKII/ERK/AP-1 signaling pathway in hMSC, suggesting the potential application in cell therapy for bone regeneration.


The combination of hypomethylating agents and histone deacetylase inhibitors produce marked synergy in preclinical models of T-cell lymphoma.
T-cell lymphomas (TCL) are aggressive lymphomas usually treated with CHOP (cyclophsophamide, doxorubicin, vincristine, prednisolone)-like regimens upfront. Recent data suggest that TCL are driven by epigenetic defects, potentially rendering them sensitive to epigenetic therapies. We explored the therapeutic merits of a combined epigenetic platform using histone deacetylase inhibitors (HDACIs) and DNA methyltransferase inhibitors (DNMT) in in vitro and in vivo models of TCL. The 50% inhibitory concentration (IC50 ) values revealed romidepsin was the most potent HDACI, with an IC50 in the low nanomolar range. The combination with a hypomethylating agent produced synergy across all cell lines, which was confirmed in cytotoxicity and apoptosis assays. An in vivo xenograft study demonstrated inhibition of tumour growth in the combination cohort compared to the single agent. Gene expression array and global methylation profiling revealed differentially expressed genes and modulated pathways for each of the single treatment conditions and the combination. Most of the effects induced by the single agent treatment were maintained in the combination group. In total, 944 unique genes were modulated by the combination treatment, supporting the hypothesis of molecular synergism. These data suggest combinations of hypomethylating agents and HDACIs are synergistic in models of TCL, which is supported at the molecular level.


Risk Culture and COVID-19 Protective Behaviors: A Cross-Sectional Survey of Residents in China.
The COVID-19 outbreak caused by the Severe Acute Respiratory Syndrome CoronaVirus type 2 (SARS-CoV-2) has spread across the world. However, our understanding of the public responses, in particular in adopting protective behaviors, has been limited. The current study aimed to determine the level of protective behaviors adopted by the residents in China and its association with their cultural attributes. A national cross-sectional online survey was conducted in mainland China from 4th to 13th August 2020. Protective behaviors were assessed as a summed score (ranging from 0 to 40) measured by ten items. The self-report tendency of study participants toward the four cultural attributes (individualism, egalitarianism, fatalism, hierarchy) was rated on a seven-point Likert scale. A total of 17651 respondents returned a valid questionnaire, representing 47.9% of those who accessed the online survey. Most (89.8%) respondents aged between 18 and 45 years in the age range of and 47.7% were male. High levels of protective behaviors (34.04 ± 5.78) were reported. The respondents had high scores in the cultural attributes of hierarchy (Median = 5) and egalitarianism (Median = 5), compared with low scores in individualism (Median = 1) and fatalism (Median = 1). High levels of protective behaviors were associated a higher tendency toward egalitarianism (AOR = 2.90, 95% CI 2.67-3.15) and hierarchy (AOR = 1.66, 95% CI 1.53-1.81) and a low tendency toward fatalism (AOR = 1.79, 95% CI 1.63-1.97) and individualism (AOR = 2.62, 95% CI 2.41-2.85). The cultural attributes explained 17.3% of the variations in the protective behavioral scores. In conclusion, the adoption of protective behaviors is associated a risk culture characterized by high levels of hierarchy and egalitarianism and low levels of individualism and fatalism. Government actions and communication strategies need to adapt to the cultural characteristics of their target audience.


Effect of regular resistance exercise, vitamin D, and calcium supplements on the bone mineral content and density in postmenopausal model of rats: An experimental study.
Postmenopausal osteoporosis progressively occurs due to alteration in the estrogen level during the menopause period, and subsequently elevates the risk of fractures. To evaluate the effect of regular resistance exercise, vitamin D, and calcium supplements on bone mineral content and density, postmenopausal rats used. In this experimental study, 72 female Sprague-Dawley rats (8-10 wk: 250 ± 15 gr) were ovariectomized and randomly divided into nine groups (n = 8/each): control, placebo, exercise (EX), exercise with vitamin D supplement (EX + D), exercise with calcium (EX + Ca), exercise with calcium and vitamin D (EX + Ca + D), vitamin D administration (D), calcium administration (Ca), and calcium and vitamin D (Ca + D) groups. Finally, the tail, hip, and lumbar bone mineral content, bone mineral density, bone thickness, and bone cells were evaluated in each group. The tail, hip, and lumbar bone mineral density was increased significantly in the EX + Vit D group compared to the control group (p = 0.004, p = 0.007, p = 0.003, respectively). However, there were no significant changes in the bone mineral content of the hips and lumbar among the groups. Besides, bone thickness in the Ex + Vit D group was more than the other groups (p = 0.02). The number of osteoclast cells were decreased in the Ca + Vit D, Ex + Ca, Ex + Vit D, and Ex + Vit D + Ca groups compared to the control group. Osteocyte numbers were increased only in the Ex + Vit D group. Resistance exercise in combination with vitamin D and calcium have a positive effect on the bone mineral density and bone mineral content and might be able to prevent or delay the osteoporosis among elderly women. However, additional researches are needed to assess the molecular pathways of this process.


Severity of statin-induced adverse effects on muscle and associated conditions: data from the DAMA study.
Statins are generally well tolerated, but muscular adverse effects appear to be the most common obstacle limiting their use. Our objective was to describe the severity of muscle injury (myalgia, myositis and rhabdomyolysis) and associated conditions related to statin therapy that may be clinically significant. A cross-sectional one-visit, non-interventional, national multicenter study including patients of both sexes over 18 years of age referred for past or present muscle symptoms associated with statin therapy was conducted. Clinical, biochemical and drug therapy characteristics were obtained at the initial evaluation. 3,845 patients were recruited from a one-day record from 2,001 physicians. Myalgia was present in 78.2% of patients, myositis in 19.3% and rhabdomyolysis in 2.5%. The prevalence of different comorbidities such as diabetes, hypertension, atrial fibrillation, and coronary heart disease increased as the severity of myopathy rose. High-intensity statin therapy was used in 33.4% of patients. Concomitant drugs metabolized by the CYP450 3A4 pathway were taken by 9.3% of patients, and statins with this metabolic route by 75%. Independent variables associated with myositis or rhabdomyolysis compared with myalgia alone in the multivariate model were excessive alcohol consumption and pravastatin therapy. Myalgia was the most common muscle adverse effect associated with statin therapy. Excessive alcohol consumption and pravastatin were independently associated with myositis or rhabdomyolysis.


Site specific immobilization of a potent antimicrobial peptide onto silicone catheters: evaluation against urinary tract infection pathogens.
Bacterial colonization of urinary catheters is a common problem leading to Catheter Associated Urinary Tract Infections (CAUTIs) in patients, which result in high treatment costs and associated complications. Due to the advantages of antimicrobial peptides (AMPs) compared to most other antimicrobial molecules, an increasing number of AMP-coated surfaces is being developed but their efficacy is hindered by suboptimal coating methods and loss of peptide activity upon surface tethering. This study aims to address this issue by employing a methodic approach that combines a simple selective chemical immobilization platform developed on a silicone catheter with the choice of a potent AMP, Lasioglossin-III (Lasio-III), to allow site specific immobilization of Lasio-III at an effective surface concentration. The Lasio-III peptide was chemically modified at the N-terminal with a cysteine residue to facilitate cysteine-directed immobilization of the peptide onto a commercial silicone catheter surface via a combination of an allyl glycidyl ether (AGE) brush and polyethylene glycol (PEG) based chemical coupling. The amount of immobilized peptide was determined to be 6.59 ± 0.89 μg cm-2 by Sulfo-SDTB assay. The AMP-coated catheter showed good antimicrobial activity against both Gram positive and negative bacteria. The antimicrobial properties of the AMP-coated catheter were sustained for at least 4 days post-incubation in a physiologically relevant environment and artificial urine and prevented the biofilm growth of E. coli and E. faecalis. Adenosine tri-phosphate leakage and propidium iodide fluorescence studies further confirmed the membranolytic mode of action of the immobilized peptide. To the best of our knowledge, this is the first proof-of-concept study that reports the efficacy of AMP immobilization by sulfhydryl coupling on a real catheter surface.


Associations of the Oral Microbiota with Obesity and Menarche in Inner City Girls.
Alterations of the oral microbiome have been associated with obesity, possibly based on inflammatory processes mediated by bacteria. Specific bacterial strains have been associated with obesity and periodontal disease. Little is known about the oral microbiome in children. Understanding the relationship between oral health and childhood growth could help identify preventable factors contributing to obesity and related conditions, including onset of menarche which is associated with obesity. In this pilot study, we investigated the saliva microbiome among 25 girls 7-15 years old (mean 11.1) and their mothers in an inner city dental clinic in New York City. The main outcome measures were body size, presence or absence of menarche and dental practices. We examined associations of microbiome richness, diversity, and relative abundance with pubertal and demographic factors and oral health. Girls had good dental health and a typical rich oral microbiome, based on the Shannon Index of all species detected. Older girls flossed more often and younger girls had more frequent dental check-ups. Microbiome richness among girls was similar to their mothers', but diversity was greater among mothers than girls. Richness was reduced among mothers with gum bleeding, flossing and increased teeth brushing. Overweight girls had greater diversity and less richness than normal weight girls. Certain bacterial species differed in abundance with respect to whether girls had reached menarche (Flavobacteria, Actinobacteria), overweight (Megasphaera, Lactorbacillales, Lactobacillus) and gingivitis in the girls (Scardovia, Bifidobacteriales, Gemellaceae). Differences found in specific bacteria in the oral microbiome were related to body size and menarche. With increasing interest on studying microbiome variability related to the multifactorial etiology of obesity in children, saliva is capable of providing clinically informative markers of this and related conditions.


Effects of aerobic exercise training on brain structure and psychological well-being in young adults.
There is convergent evidence that exercise increases psychological well-being; however, the mechanism of this psychological effect of exercise is not yet completely understood. The purpose of this study was to examine the effects of aerobic exercise training on brain structure and psychological well-being in young adults. University students who had not regularly exercised were divided into training group (N.=15) and control group (N.=15). The training group performed a total 30 periods of aerobic exercise training, while the control group never performed. Whole-brain magnetic resonance imaging scans and mental health questionnaire examinations were performed before and after the exercise training period for all of the participants. A voxel-based morphometry (VBM) analysis was used to compare the changes in gray-matter volumes in the two groups. VBM is an objective whole-brain technique for characterization of regional cerebral volume and tissue concentration differences in structural magnetic resonance images. The results of VBM analysis revealed no change in gray-matter volume in the training group, although the gray-matter volume of the left insula was significantly decreased in the control group after the exercise training period. The training group exhibited significant improvement in some scores on the mental health questionnaire after the exercise training period, compared with the control group. These findings suggest that aerobic exercise training may inhibit gray-matter volume loss in the insula, and that a relationship may exist between preservation of insula gray-matter and improvement of psychological well-being by aerobic exercise training.


Multiple mechanisms are responsible for transactivation of the epidermal growth factor receptor in mammary epithelial cells.
The number of distinct signaling pathways that can transactivate the epidermal growth factor receptor (EGFR) in a single cell type is unclear. Using a single strain of human mammary epithelial cells, we found that a wide variety of agonists, such as lysophosphatidic acid (LPA), uridine triphosphate, growth hormone, vascular endothelial growth factor, insulin-like growth factor-1 (IGF-1), and tumor necrosis factor-alpha, require EGFR activity to induce ERK phosphorylation. In contrast, hepatocyte growth factor can stimulate ERK phosphorylation independent of the EGFR. EGFR transactivation also correlated with an increase in cell proliferation and could be inhibited with metalloprotease inhibitors. However, there were significant differences with respect to transactivation kinetics and sensitivity to different inhibitors. In particular, IGF-1 displayed relatively slow transactivation kinetics and was resistant to inhibition by the selective ADAM-17 inhibitor WAY-022 compared with LPA-induced transactivation. Studies using anti-ligand antibodies showed that IGF-1 transactivation required amphiregulin production, whereas LPA was dependent on multiple ligands. Direct measurement of ligand shedding confirmed that LPA treatment stimulated shedding of multiple EGFR ligands, but paradoxically, IGF-1 had little effect on the shedding rate of any ligand, including amphiregulin. Instead, IGF-1 appeared to work by enhancing EGFR activation of Ras in response to constitutively produced amphiregulin. This enhancement of EGFR signaling was independent of both receptor phosphorylation and PI-3-kinase activity, suggestive of a novel mechanism. Our studies demonstrate that within a single cell type, the EGFR autocrine system can couple multiple signaling pathways to ERK activation and that this modulation of EGFR autocrine signaling can be accomplished at multiple regulatory steps.


Hierarchy of Azimuthal Anisotropy Harmonics in Collisions of Small Systems from the Color Glass Condensate.
We demonstrate that the striking systematics of two-particle azimuthal Fourier harmonics v_{2} and v_{3} in ultrarelativistic collisions of protons, deuterons, and helium-3 ions off gold nuclei measured by the PHENIX Collaboration at the Relativistic Heavy Ion Collider (RHIC) is reproduced in the color glass condensate effective field theory. This contradicts the claim in C. Aidala et al. (PHENIX Collaboration), arXiv:1805.02973, that their data rule out initial state-based explanations. The underlying systematics of the effect, as discussed previously in K. Dusling, M. Mace, R. Venugopalan, Phys. Rev. D 97, 016014 (2018)PRVDAQ2470-001010.1103/PhysRevD.97.016014; Phys. Rev. Lett. 120, 042002 (2018)PRLTAO0031-900710.1103/PhysRevLett.120.042002; Proc. Sci., QCDEV2017 (2018) 039, arise from the differing structure of strong color correlations between gluon domains of size 1/Q_{S} at fine (p_{⊥}⪆Q_{S}) or coarser (p_{⊥}⪅Q_{S}) transverse momentum resolution. Further tests of the limits of validity of this framework can be carried out in light-heavy ion collisions at both RHIC and the Large Hadron Collider. Such measurements also offer novel opportunities for further exploration of the role of the surprisingly large short-range nuclear correlations measured at Jefferson Lab.


Analytical methods for quantifying environmental connectivity for the control and surveillance of infectious disease spread.
The sustained transmission and spread of environmentally mediated infectious diseases is governed in part by the dispersal of parasites, disease vectors and intermediate hosts between sites of transmission. Functional geospatial models can be used to quantify and predict the degree to which environmental features facilitate or limit connectivity between target populations, yet typical models are limited in their geographical and analytical approach, providing simplistic, global measures of connectivity and lacking methods to assess the epidemiological implications of fine-scale heterogeneous landscapes. Here, functional spatial models are applied to problems of surveillance and control of the parasitic blood fluke Schistosoma japonicum and its intermediate snail host Oncomelania haupensis in western China. We advance functional connectivity methods by providing an analytical framework to (i) identify nodes of transmission where the degree of connectedness to other villages, and thus the potential for disease spread, is higher than is estimated using Euclidean distance alone and (ii) (re)organize transmission sites into disease surveillance units based on second-order relationships among nodes using non-Euclidean distance measures, termed effective geographical distance (EGD). Functional environmental models are parametrized using ecological information on the target organisms, and pair-wise distributions of inter-node EGD are estimated. A Monte Carlo rank product analysis is presented to identify nearby nodes under alternative distance models. Nodes are then iteratively embedded into EGD space and clustered using a k-means algorithm to group villages into ecologically meaningful surveillance groups. A consensus clustering approach is taken to derive the most stable cluster structure. The results indicate that novel relationships between nodes are revealed when non-Euclidean, ecologically determined distance measures are used to quantify connectivity in heterogeneous landscapes. These connections are not evident when analysing nodes in Euclidean space, and thus surveillance and control activities planned using Euclidean distance measures may be suboptimal. The methods developed here provide a quantitative framework for assessing the effectiveness of ecologically grounded surveillance systems and of control and prevention strategies for environmentally mediated diseases.


Matching of muscle properties and motoneurone firing patterns during early stages of development.
In adults, muscle fibres match the functional requirements of the motoneurone that supplies them. During early stages of postnatal development of the rat neither muscle fibre properties, nor activity patterns of motoneurones supplying fast and slow muscles have completed their differentiation. Nevertheless, even at this early stage of development the muscles have characteristic properties that are well matched to the activity patterns of immature motoneurones. With further development differentiation of motoneurone activity and muscle fibre properties goes hand in hand. If during this period of linked differentiation, connections between the motoneurones and muscle fibres are disrupted, the development of fast muscles is permanently impaired.


In vitro correction of iduronate-2-sulfatase deficiency by adenovirus-mediated gene transfer.
Hunter syndrome is a lethal lysosomal storage disorder caused by the deficiency of iduronate-2-sulfatase and characterized by severe skeletal and neurological symptoms. Only symptomatic treatments are available and, although bone marrow transplantation has been suggested, no encouraging results have been obtained so far. Therefore, gene therapy might be a route to be pursued for treatment of the disease. In this respect, one major goal to achieve is the generation of an overexpressing vector able to correct, in particular, central nervous system (CNS) cells. Adenoviruses have been shown to infect CNS cells efficiently with minor or even absent immunological response. We describe the generation of a replication-defective adenoviral vector, AdRSVIDS, which is able to express in vitro high levels of iduronate-2-sulfatase. After infection, accumulation of mucopolysaccharides in treated Hunter cells was normalized. Furthermore, endocytosis of the transduced IDS did occur via the mannose-6-phosphate (M6P) receptor. Since no animal model for the disease is available, we developed a system based on the generation of derma-equivalents which enabled us to verify the expression of high levels of sulfatase up to 30 days after infection.


Efficient packaging of readthrough RNA in ALV: implications for oncogene transduction.
Readthrough viral transcripts are present at relatively high levels in cells infected with avian leukosis virus. It has been proposed that they can function as intermediates in the transduction of proto-oncogenes by retroviruses. It is shown here, by the analysis of viruses containing a mutation in the AAUAAA polyadenylation signal, that readthrough RNAs have the requisite properties to function as transduction intermediates: readthrough RNAs were polyadenylated and packaged as efficiently as normal viral RNA, RNAs nearly 11.2 kilobases (3.5 kilobases larger than wild-type avian leukosis virus genomes) were present in virions of the mutant virus, and virus particles containing both readthrough and normal genomes were most likely infectious.


Naloxone enhances myocardial responses to isoproterenol in dog isolated heart-lung.
Intracoronary injection of naloxone produces rapid local increases in myocardial performance. The role of beta-adrenergic activation in this response was investigated. Naloxone was injected into the left anterior descending artery (LAD) of anesthetized dogs with the adjacent circumflex territory serving as control. Naloxone increased the contractile state locally in the LAD region. Pretreatment with propranolol eliminated the regional inotropic response. An isolated heart-lung model was set up, and isoproterenol dose responses were determined. When repeated after naloxone, contractile (dP/dt) responses to isoproterenol were both augmented and prolonged. In a third study imipramine was used to determine whether naloxone might act by reducing neuronal uptake of the isoproterenol. Imipramine extended the effect of isoproterenol temporally but did not alter the peak response. Adding naloxone increased the peak response to isoproterenol and maintained it above the extended imipramine response. The data support the concept that the blockade of opiate receptors facilitates adrenergic activity mediated by myocardial beta 1-receptors 1) directly through postsynaptic mechanisms, 2) indirectly through beta 2-mediated release of norepinephrine, or 3) via interference with nonneuronal disposal mechanisms.


Proper and dark heroes as DADS and CADS : Alternative mating strategies in British Romantic literature.
Empirical tests described in this article support hypotheses derived from evolutionary theory on the perceptions of literary characters. The proper and dark heroes in British Romantic literature of the late eighteenth and early nineteenth centuries respectively represent long-term and short-term mating strategies. Recent studies indicate that for long-term relationships, women seek partners with the ability and willingness to sustain paternal investment in extended relationships. For short-term relationships, women choose partners whose features indicate high genetic quality. In hypothetical scenarios, females preferred proper heroes for long-term relationships. The shorter the relationship under consideration, the more likely women were to choose dark heroes as partners.


Underuse of yoga as a referral resource by health professions students.
Nearly 38% of U.S. adults use complementary and alternative medicine approaches to manage physical conditions (e.g., chronic pain, arthritis, cancer, heart disease, and high blood pressure) and psychological or emotional health concerns (e.g., post-traumatic stress disorder, anxiety, and depression). Research evidence has accumulated for yoga as an effective treatment approach for these conditions. Further, yoga has increased in popularity among healthcare providers and the general population. Given these trends, this study explored perceptions about yoga as a viable complementary treatment to which health professions students would refer patients. More than 1500 students enrolled in health professions programs at a Pacific Northwest school were enrolled; data were obtained from 478 respondents. The study assessed willingness to refer patients to yoga as a complementary and alternative medicine for 27 symptoms (identified in the literature as having evidence for yoga's utility), which were subsequently grouped into skeletal, physical, and psychological on the basis of factor analysis. Responses were assessed using a mixed-model analysis of variance with health profession and yoga practitioner as between-subjects variables and symptoms as a within-subjects factor. In descending order of likelihood to refer patients to yoga were students in occupational therapy, physician assistant program, psychology, physical therapy, pharmacy, dental hygiene, speech and audiology, and optometry. All groups perceived yoga's greatest utility for skeletal symptoms, followed by psychological and physical symptoms. Findings also revealed a significant positive relationship between level of personal yoga practice and willingness to refer patients to yoga. Although students expressed some openness to referring patients to yoga, ratings of appropriateness were not accurately aligned with extant evidence base. Personal experience seemed to be a salient factor for accepting yoga as a referral target. These findings suggest the importance of developing strategies to make health professionals more aware of the merits of yoga, regardless of whether they themselves are yoga practitioners.


Low-concentration tracer tests to measure air-water interfacial area in porous media.
The aqueous-based interfacial tracer method employing miscible-displacement tests is one method available for measuring air-water interfacial areas. One potential limitation to the method is the impact of tracer-induced drainage on the system. The objective of this study was to investigate the efficacy of a low-concentration tracer test method for measuring air-water interfacial area. Tracer concentrations and analytical methods were selected that allowed the use of tracer input concentrations that were below the threshold of tracer-induced drainage. Multiple tracer tests were conducted at different water saturations. Interfacial areas increased from 34.8 to 101 cm-1 with the decrease in saturation from 0.86 to 0.62. The method produced relatively robust measurements of air-water interfacial area, with coefficients of variation ranging from 6 to 26%. A variably saturated flow and transport model that accounts for the effects of tracer on interfacial tension, and the retention of tracer at the air-water and solid-water interfaces, was used to test for potential tracer-induced drainage. The simulations showed that the use of low tracer-input concentrations eliminated this phenomenon. This is consistent with the measured data for effluent-sample masses, which exhibited minimal change during the tests, and with the observation that the interfacial areas obtained with the low-concentration-tracer method were consistent with values measured with two methods that are not influenced by tracer-induced drainage. These results demonstrate that the low-concentration miscible-displacement tracer test method is an effective approach for measuring air-water interfacial areas in porous media.


Screening and Identification of Key Biomarkers in Lower Grade Glioma via Bioinformatical Analysis.
Lower-grade glioma (LGG) is a common type of central nervous system tumor. Due to its complicated pathogenesis, the choice and timing of adjuvant therapy after tumor treatment are controversial. This study explored and identified potential therapeutic targets for lower-grade. The bioinformatics method was employed to identify potential biomarkers and LGG molecular mechanisms. Firstly, we selected and downloaded GSE15824, GSE50161, and GSE86574 from the GEO database, which included 40 LGG tissue and 28 normal brain tissue samples. GEO and VENN software identified of 206 codifference expressed genes (DEGs). Secondly, we applied the DAVID online software to investigate the DEG biological function and KEGG pathway enrichment, as well as to build the protein interaction visualization network through Cytoscape and STRING website. Then, the MCODE plug is used in the analysis of 22 core genes. Thirdly, the 22 core genes were analyzed with UNCLA software, of which 18 genes were associated with a worse prognosis. Fourthly, GEPIA was used to analyze the 18 selected genes, and 14 genes were found to be a significantly different expression between LGGs and normal brain tumor samples. Fifthly, hierarchical gene clustering was used to examine the 14 important gene expression differences in different histologies, as well as analysis of the KEGG pathway. Five of these genes were shown to be abundant in the natural killer cell-mediated cytokines (NKCC) and phagosome pathways. The five key genes that may be affected by the immune microenvironment play a crucial role in LGG development.


Windows of Susceptibility and Consequences of Early Life Exposures to 17β-estradiol on Medaka (Oryzias latipes) Reproductive Success.
Estrogens and estrogen mimics are commonly found in surface waters and are associated with deleterious effects in fish populations. Impaired fertility and fecundity in fish following chronic exposures to estrogens and estrogen mimics during critical windows in development are well documented. However, information regarding differential reproductive effects of exposure within defined developmental stages remains sparse. In this study, reproductive capacity was assessed in Japanese medaka (Oryzias latipes) after exposure to two concentrations of 17β-estradiol (E2β; 2 ng/L and 50 ng/L) during four distinct stages of development: gonad development, gonad differentiation, development of secondary sex characteristics (SSC) and gametogenesis. Exposure to E2β did not adversely impact survival, hatch success, growth, or genotypic ratios. In contrast, exposure to 50 ng/L E2β during SSC development altered phenotypic ratios and SSC. Exposure to both E2β treatments reduced reproductive capacity (fertility, fecundity) by 7.3-57.4% in adult medaka breeding pairs, with hindrance of SSC development resulting in the largest disruption in breeding capacity (51.6-57.4% decrease) in the high concentration. This study documents differential effects among four critical stages of development and provides insight into factors (window of exposure, exposure concentration and duration of exposure period) contributing to reproductive disruption in fish.


[What are the main factors assisting general practitioners in the development of educational strategies?]
General practitioners (GP) are in a special position to conduct educational strategies for their patients with chronic disease. Despite the GP's desire to be involved in patient education, this approach remains underdeveloped.Objective: To determine the factors required support the development of educational strategies in general practice from the GPs' perspective.Methods: Qualitative study based on a "modified nominal group technique" carried out among 21 GPs (not specifically trained in patient education) participating as investigators in an interventional study testing a structured educational strategy. The interventional study based on processes and tools specifically developed to suit general practice was held during their usual consultations. Thirty one patients with COPD were followed for 9 months.Results: GPs identified 23 specific factors restructured after the clarification-reformulation step in 6 main factors required for the development of educational strategies in general practice. The consensus revealed that reinforcement of initial and continuing medical training was the most frequently cited factor and was considered to be a priority. The development of inter- and intra-professional cooperation was the second leading factor to be taken into account.Conclusion: The improvement of GP training and the promotion of areas of cooperation could help GPs to develop educational strategies during their consultations. These results may provide insight to regional health agencies concerning the deployment of patient education in primary care.


Application of the recent American practice resources for risk stratification system for patients presenting to a Japanese emergency department because of syncope.
The American College of Physicians (ACP) and the American College of Emergency Physicians (ACEP) recently published practice guidelines and recommendations for evaluation of patients with syncope based on historical, physical, and ECG findings. The objective of the present study was to determine if risk stratification using these practice resources is valid in a series of Japanese patients. A total of 912 consecutive patients brought to our emergency department between 1988 and 1997 because of syncope were identified. Follow-up information about mortality was obtained for 707 patients by means of mailed questionnaires and from medical records, and the mortality data were analyzed by the actuarial life-table method. A total of 187 patients who fulfilled the admission criteria according to the ACP guidelines were found to have higher overall and cardiac mortality than the other 520 patients (P < 0.0001), and 153 patients who fulfilled the admission criteria according to the ACEP recommendations also had higher overall and cardiac mortality than the other 554 patients (P < 0.0001). The recent American practice recommendations can be used for risk stratification of syncope patients in Japan. Historical, physical, and ECG findings available on presentation can be used to stratify the risk of mortality in patients brought to Japanese emergency departments because of syncope.


gamma-Aminobutyric acid (GABA)- and barbiturate-mediated 36Cl- uptake in rat brain synaptoneurosomes: evidence for rapid desensitization of the GABA receptor-coupled chloride ion channel.
"Desensitization" of the gamma-aminobutyric acid (GABA) receptor-coupled chloride ion channel was studied using an in vitro method for measuring chloride (Cl-) permeability in brain vesicles (synaptoneurosomes). Muscimol, a GABA agonist, stimulated 36Cl- uptake in rat cerebral cortical synaptoneurosomes in a concentration-dependent manner (EC50 7.3 +/- 0.5 microM), whereas pentobarbital stimulated 36Cl- uptake in a biphasic manner, indicated by a bell-shaped concentration-response relationship, with a maximal response at 500 microM (EC50 271 +/- 17 microM). Higher concentrations of pentobarbital led to progressively smaller stimulation of 36Cl- uptake and blocked muscimol-stimulated 36Cl- uptake. Lower concentrations of pentobarbital (100-200 microM), when added with muscimol, produced an additive effect in stimulating 36Cl- uptake, whereas even lower (subthreshold) concentrations of pentobarbital (50 microM) potentiated muscimol-stimulated 36Cl- uptake. Following continuous exposure of synaptoneurosomes (up to 20 min) to muscimol (50 microM) or pentobarbital (500 microM), the 36Cl- uptake response diminished to a new steady state level with a t1/2 of approximately 6 sec and 30 sec, respectively. The decrement in response to these agonists was dependent on both concentration and length of exposure. No decrement was observed in the ability of subthreshold concentrations of pentobarbital to enhance muscimol-stimulated 36Cl- uptake following prolonged (20 min) incubation. "Heterologous desensitization" between muscimol and pentobarbital was observed in experiments where either muscimol or pentobarbital was added to the vesicles following pretreatment with the other. These findings suggest that "desensitization" of the GABA receptor/Cl- ion channel may involve both the GABA and barbiturate recognition sites or a common effector component such as the ionophore itself.


Phasing diffuse scattering. Application of the SIR2002 algorithm to the non-crystallographic phase problem.
A new phasing algorithm has been used to determine the phases of diffuse elastic X-ray scattering from a non-periodic array of gold balls of 50 nm diameter. Two-dimensional real-space images, showing the charge-density distribution of the balls, have been reconstructed at 50 nm resolution from transmission diffraction patterns recorded at 550 eV energy. The reconstructed image fits well with a scanning-electron-microscope (SEM) image of the same sample. The algorithm, which uses only the density modification portion of the SIR2002 program, is compared with the results obtained via the Gerchberg-Saxton-Fienup HiO algorithm. The new algorithm requires no knowledge of the object's boundary and proceeds from low to high resolution. In this way, the relationship between density modification in crystallography and the HiO algorithm used in signal and image processing is elucidated.


Structure-function relationship of avian eggshell matrix proteins: a comparative study of two major eggshell matrix proteins, ansocalcin and OC-17.
The role of individual matrix proteins in avian eggshell calcification is poorly understood despite numerous attempts to characterize and localize their presence in the eggshell matrix. Ansocalcin, the major matrix protein from goose eggshell, was found to induce the formation of calcite crystal aggregates under in vitro. Owing to its high similarity with the chicken eggshell matrix protein ovocleidin 17 (OC-17), a comparative investigation has been carried out to understand the structure-function relationship. RP-HPLC shows that ansocalcin is the major component in extracts of goose eggshells before and after bleach treatment. However, OC-17 was observed in minute quantities in the extract of bleach-treated chicken eggshells. In vitro crystal growth experiments showed that OC-17 and ansocalcin interact differently with the calcite crystals formed. Circular dichroism, intrinsic tryptophan fluorescence, and dynamic light scattering studies showed that, under the conditions used in our experiments, OC-17 does not aggregate in solution or induce the nucleation of calcite aggregates in the concentration range used. These observations indicate that OC-17 and ansocalcin play different roles in the eggshell calcification. To our knowledge, this is the first report on the comparison of properties of homologous eggshell proteins that belong to the same phylogeny.


D-glucose-sensitive neurosecretory cells of the crab Cancer borealis and negative feedback regulation of blood glucose level.
We studied the effects of glucose on cultured X-organ neurons of the crab Cancer borealis using single-electrode current- and voltage-clamp techniques. A subpopulation of the cells responded to D-glucose with a hyperpolarization. These cells, but not glucose-insensitive cells, showed immunoreactivity to crustacean hyperglycemic hormone (CHH), the hormone responsible for the elevation of blood glucose levels in crustaceans. Glucose-sensitive cells were also inhibited by serotonin and gamma-aminobutyric acid but were not affected by dopamine and Leu-enkephalin. The response was specific for D-glucose, with an EC50 of 0.25 mmoll-1. No response was seen to L-glucose, sucrose, galactose, mannose or fructose. The glucose response persisted in the absence of extracellular Na+ and in low-Ca2+/Mn2+ saline. In voltage-clamp experiments, D-glucose evoked a small current with a reversal potential close to that of voltage-dependent K+ currents. We conclude that D-glucose activates a K+ current in CHH-immunoreactive cells that, in normal saline, induces a hyperpolarization. We propose that this enables glucose to regulate directly the release of CHH into the hemolymph, thus constituting a negative feedback mechanism regulating hemolymph glucose concentration.


Dependence of exponents on text length versus finite-size scaling for word-frequency distributions.
Some authors have recently argued that a finite-size scaling law for the text-length dependence of word-frequency distributions cannot be conceptually valid. Here we give solid quantitative evidence for the validity of this scaling law, using both careful statistical tests and analytical arguments based on the generalized central-limit theorem applied to the moments of the distribution (and obtaining a novel derivation of Heaps' law as a by-product). We also find that the picture of word-frequency distributions with power-law exponents that decrease with text length [X. Yan and P. Minnhagen, Physica A 444, 828 (2016)PHYADX0378-437110.1016/j.physa.2015.10.082] does not stand with rigorous statistical analysis. Instead, we show that the distributions are perfectly described by power-law tails with stable exponents, whose values are close to 2, in agreement with the classical Zipf's law. Some misconceptions about scaling are also clarified.


A training session in a clinical simulation laboratory for the acquisition of clinical skills by newly recruited medical interns.
In organized orientation programs for newly recruited medical interns of the Nippon Medical School Hospital, the working committee of the clinical simulation laboratory introduced a laboratory training session that was designed to improve the clinical skills of the medical interns. The session consisted of 6 training courses, comprising internal examination, tracheal intubation, auscultation of heart sounds, bandaging and the collection of samples of venous and arterial blood. Medical interns rotated to a new course every 30 minutes and did practical trainings in each of the 6 skills. A total of 36 newly recruited medical interns participated in the training session. The majority of medical interns took part in the practical training actively and positively. The session was efficiently carried out from the standpoints of human resources and the teaching hours involved. A post training questionnaire survey, completed by the medical interns, revealed that many of them valued the sessions for comprehensibility of the instructions, the descriptions in the manual and the content of the training; however, only 21% thought that they had successfully acquired the clinical skills. Medical interns must continually engage in self-training to steadily acquire basic clinical skills. The convenience of a clinical simulation laboratory, together with the reinforcement of the education of clinical skills during internship, is necessary to strengthen the educational benefits of the training session.


Influence of angle between the nozzle and skin surface on the heat flux and overall heat extraction during cryogen spray cooling.
High speed video imaging and an inverse heat conduction problem algorithm were used to observe and measure the effect of the angle between the nozzle and surface of a skin phantom on: (a) surface temperature; (b) heat flux q; and (c) overall heat extraction Q during cryogen spray cooling (CSC). A skin phantom containing a fast-response temperature sensor was sprayed with 50 ms cryogen spurts from a commercial nozzle placed 30 mm from the surface. The nozzle was systematically positioned at angles ranging from 5 to 90 degrees (perpendicular) with respect to the phantom surface. It is shown that angles as low as 15 have an insignificant impact on the surface temperature, q and Q. Only exaggerated angles of 5 show up to 10% lower q and 30% lower Q with respect to the maximal values measured when nozzles are aimed perpendicularly. This study proves that the slight angle that many commercial nozzles have does not affect significantly the CSC efficiency.


Experiences Using Pre-Exposure Prophylaxis for Safer Conception Among HIV Serodiscordant Heterosexual Couples in the United States.
Antiretroviral pre-exposure prophylaxis (PrEP) is a promising HIV prevention strategy for HIV serodiscordant couples (HIV-infected male, uninfected female) seeking safer conception. However, most research on PrEP for safer conception has focused on couples in sub-Saharan Africa; little is known about the perspectives or experiences of heterosexual couples in the United States. We conducted qualitative interviews with six couples (six women and five of their male partners) receiving PrEP for conception services at an urban safety net hospital in the US Northeast. In-depth interview guides explored couple relationships and contextual factors and attitudes, perceptions, and decision-making processes surrounding PrEP for safer conception. Thematic analyses focused on identifying the following emergent themes. We found that couple relationships were situated within broader social and cultural contexts of immigration, family, and community that shaped their experiences with HIV and serodiscordant relationship status. Despite strong partner support within relationships, HIV stigma and disapproval of serodiscordant relationships contributed to couples' feelings of social isolation and subsequent aspirations to have "normal" families. By enabling "natural" conception through condomless sex, PrEP for safer conception provided a sense of enhanced relationship intimacy. Couples called for increasing public awareness of PrEP through positive messaging as a way to combat HIV stigma. Findings suggest that relationship dynamics and broader social contexts appear to shape HIV serodiscordant couples' fertility desires and motivations to use PrEP. However, increased public awareness of PrEP for safer conception may be needed to combat HIV stigma at the community level.


Predictors of Developmental Patterns of Obesity in Young Children.
Introduction: The current study characterizes longitudinal patterns in obesity in young children and their prediction from developmental programming and social determinant hypotheses. Materials and Methods: The data are based on the Family Life Project, a prospective longitudinal study of 1,292 families recruited from low-income, racially diverse, rural communities in Pennsylvania, and North Carolina. Pre-natal, peri-natal, and post-natal risks for childhood obesity were collected from 2 months of age; in-person assessments of child growth were used to identity obesity on multiple occasions from 24 to 90 months of age. Results: Two major novel findings emerged. First, longitudinal analyses identified four distinct obesity development profiles: stable obesity, later-onset obesity, moderate/declining obesity, and non-obese; these groups had distinct risk profiles. Second, prediction analyses favored developmental programming explanations for obesity, including evidence even in early childhood that both low- and high birth weight was associated with stable obesity. There was no indication that pre- and peri-natal and post-natal factors predicted obesity differently in non-minority and minority children. Discussion: Factors derived from the developmental programming model of obesity overlapped with, but predicted early onset obesity independently from, risks associated with social determinant models of obesity.


Neutral pi-associated porphyrin [2]catenanes.
A series of neutral porphyrin-containing catenanes has been synthesised, consisting of a zinc porphyrin strapped by a polyethylene glycol chain containing four or six ethylenoxy-units and incorporating a central naphthoquinol unit, interlinked with a naphthalene diimide macrocycle. The napthalene diimide precursor units exhibit only weak binding with the strapped porphyrins (Ka between 8 and 0.02 M(-1)), but good yields of the catenanes were obtained by Glaser coupling of the alkynyl napthalene diimide precursors in the presence of the porphyrins. Structures and solution conformations were determined by mass spectral and detailed 1H NMR studies. For the longer strapped porphyrins, the diimide macrocycle rotates around the central naphthoquinol unit at 420-450 times per second, while rotation is virtually prevented in the tighter strapped derivatives. A second dynamic process occurring in both sets of catenanes and described as 'yawing' leads to inequivalence in the naphthalene moieties. UV-Visible spectra indicate charge transfer interactions and electronic communication between the two components of the catenane.


Epothilone B induces glioblastoma cell death via survivin down-regulation.
The clinical resistance of glioblastomas to chemotherapeutic agents can be attributed to drug efflux pumps, such as P-glycoprotein, which contributes to reduce drug efficacy. The present study examined the utility of epothilone B, which is not a substrate for P-glycoprotein, on glioblastoma cells. In vitro methods with glioblastoma cells varying in p53 status were used to assess the efficacy of epothilone B to induce anti-neoplastic responses. Immunofluorescence and ELISA procedures were used to examine levels of tubulin and survivin in epothilone B treated glioblastoma cells, while acridine orange labeling was used to detect the mode of epothilone B induced cell death. A clinically achievable concentration of epothilone B induced a cytotoxic response in p53 mutant glioblastoma cells, as a consequence of survivin down-regulation and tubulin redistribution, while a cytostatic response was observed in p53 null glioblastoma cells with a modest increase in survivin expression post-epothilone B treatment. However, p53 wild-type glioblastoma cells did not sustain a positive anti-tumorigenic response to epothilone B. Epothilone B, induced positive differential responses in glioblastoma cells with abnormal p53 status, but not in p53 wild-type cells. This suggests that epothilone B is a potential alternative to classic microtubule inhibiting agents (ie vincristine, paclitaxel) used to treat clinical glioblastomas with p53 mutations.


Stress response in tardigrades: differential gene expression of molecular chaperones.
Semi-terrestrial tardigrades exhibit a remarkable tolerance to desiccation by entering a state called anhydrobiosis. In this state, they show a strong resistance against several kinds of physical extremes. Because of the probable importance of stress proteins during the phases of dehydration and rehydration, the relative abundance of transcripts coding for two alpha-crystallin heat-shock proteins (Mt-sHsp17.2 and Mt-sHsp19.5), as well for the heat-shock proteins Mt-sHsp10, Mt-Hsp60, Mt-Hsp70 and Mt-Hsp90, were analysed in active and anhydrobiotic tardigrades of the species Milnesium tardigradum. They were also analysed in the transitional stage (I) of dehydration, the transitional stage (II) of rehydration and in heat-shocked specimens. A variable pattern of expression was detected, with most candidates being downregulated. Gene transcripts of one Mt-hsp70 isoform in the transitional stage I and Mt-hsp90 in the anhydrobiotic stage were significantly upregulated. A high gene expression (778.6-fold) was found for the small alpha-crystallin heat-shock protein gene Mt-sHsp17.2 after heat shock. We discuss the limited role of the stress-gene expression in the transitional stages between the active and anhydrobiotic tardigrades and other mechanisms which allow tardigrades to survive desiccation.


Insights into the dependent-scattering contributions to the extinction coefficient in highly scattering suspensions.
We study theoretically the extinction of collimated light in random systems of highly scattering particles embedded in nonabsorbing media. We aim to provide rough guidelines on the behavior of the extinction coefficient in the so-called dependent-scattering regime. We base our analysis on Keller's second order perturbative approximation to the effective propagation constant. To gain physical insight, we also analyze a simple model based on the physical notion that particles in a dense system scatter light in an effective medium.


Cell Therapies for Spinal Cord Injury: Trends and Challenges of Current Clinical Trials.
Cell therapies have the potential to revolutionize the treatment of spinal cord injury. Basic research has progressed significantly in recent years, with a plethora of cell types now reaching early-phase human clinical trials, offering new strategies to repair the spinal cord. However, despite initial enthusiasm for preclinical and early-phase clinical trials, there has been a notable hiatus in the translation of cell therapies to routine clinical practice. Here, we review cell therapies that have reached clinical trials for spinal cord injury, providing a snapshot of all registered human trials and a summary of all published studies. Of registered trials, the majority have used autologous cells and approximately a third have been government funded, a third industry sponsored, and a third funded by university or healthcare systems. A total of 37 cell therapy trials have been published, primarily using stem cells, although a smaller number have used Schwann cells or olfactory ensheathing cells. Significant challenges remain for cell therapy trials in this area, including achieving stringent regulatory standards, ensuring appropriately powered efficacy trials, and establishing sustainable long-term funding. However, cell therapies hold great promise for human spinal cord repair and future trials must continue to capitalize on the exciting developments emerging from preclinical studies.


Decoding working memory of stimulus contrast in early visual cortex.
Most studies of the early stages of visual analysis (V1-V3) have focused on the properties of neurons that support processing of elemental features of a visual stimulus or scene, such as local contrast, orientation, or direction of motion. Recent evidence from electrophysiology and neuroimaging studies, however, suggests that early visual cortex may also play a role in retaining stimulus representations in memory for short periods. For example, fMRI responses obtained during the delay period between two presentations of an oriented visual stimulus can be used to decode the remembered stimulus orientation with multivariate pattern analysis. Here, we investigated whether orientation is a special case or if this phenomenon generalizes to working memory traces of other visual features. We found that multivariate classification of fMRI signals from human visual cortex could be used to decode the contrast of a perceived stimulus even when the mean response changes were accounted for, suggesting some consistent spatial signal for contrast in these areas. Strikingly, we found that fMRI responses also supported decoding of contrast when the stimulus had to be remembered. Furthermore, classification generalized from perceived to remembered stimuli and vice versa, implying that the corresponding pattern of responses in early visual cortex were highly consistent. In additional analyses, we show that stimulus decoding here is driven by biases depending on stimulus eccentricity. This places important constraints on the interpretation for decoding stimulus properties for which cortical processing is known to vary with eccentricity, such as contrast, color, spatial frequency, and temporal frequency.


Automatic Relocalization and Loop Closing for Real-Time Monocular SLAM.
Monocular SLAM has the potential to turn inexpensive cameras into powerful pose sensors for applications such as robotics and augmented reality. We present a relocalization module for such systems which solves some of the problems encountered by previous monocular SLAM systems--tracking failure, map merging, and loop closure detection. This module extends recent advances in keypoint recognition to determine the camera pose relative to the landmarks within a single frame time of 33 ms. We first show how this module can be used to improve the robustness of these systems. Blur, sudden motion, and occlusion can all cause tracking to fail, leading to a corrupted map. Using the relocalization module, the system can automatically detect and recover from tracking failure while preserving map integrity. Extensive tests show that the system can then reliably generate maps for long sequences even in the presence of frequent tracking failure. We then show that the relocalization module can be used to recognize overlap in maps, i.e., when the camera has returned to a previously mapped area. Having established an overlap, we determine the relative pose of the maps using trajectory alignment so that independent maps can be merged and loop closure events can be recognized. The system combining all of these abilities is able to map larger environments and for significantly longer periods than previous systems.


Biomarker-driven trial in metastatic pancreas cancer: feasibility in a multicenter study of saracatinib, an oral Src inhibitor, in previously treated pancreatic cancer.
Src tyrosine kinases are overexpressed in pancreatic cancers, and the oral Src inhibitor saracatinib has shown antitumor activity in preclinical models of pancreas cancer. We performed a CTEP-sponsored Phase II clinical trial of saracatinib in previously treated pancreas cancer patients, with a primary endpoint of 6-month survival. A Simon MinMax two-stage phase II design was used. Saracatinib (175 mg/day) was administered orally continuously in 28-day cycles. In the unselected portion of the study, 18 patients were evaluable. Only two (11%) patients survived for at least 6 months, and three 6-month survivors were required to move to second stage of study as originally designed. The study was amended as a biomarker-driven trial (leucine rich repeat containing protein 19 [LRRC19] > insulin-like growth factor-binding protein 2 [IGFBP2] "top scoring pairs" polymerase chain reaction [PCR] assay, and PIK3CA mutant) based on preclinical data in a human pancreas tumor explant model. In the biomarker study, archival tumor tissue or fresh tumor biopsies were tested. Biomarker-positive patients were eligible for the study. Only one patient was PIK3CA mutant in a 3' untranslated region (UTR) portion of the gene. This patient was enrolled in the study and failed to meet the 6-month survival endpoint. As the frequency of biomarker-positive patients was very low (<3%), the study was closed. Although we were unable to conclude whether enriching for a subset of second/third line pancreatic cancer patients treated with a Src inhibitor based on a biomarker would improve 6-month survival, we demonstrate that testing pancreatic tumor samples for a biomarker-driven, multicenter study in metastatic pancreas cancer is feasible.


Role and regulation of IL-12 in the in vivo response to staphylococcal enterotoxin B.
Injection of a staphylococcal superantigen (SAg) such as staphylococcal enterotoxin B (SEB) in adult mice results in cytokine production and cell proliferation which can lead to septic shock. The aim of the present work was to identify the cytokines and co-stimulatory molecules regulating the in vivo systemic release of IFN-gamma, a cytokine known to play an important role in the pathophysiology associated with bacterial infections. We demonstrate in this study that (i) in contrast to lipopolysaccharide (LPS), SEB administration induces high levels of the p70, bioactive form, of IL-12; (ii) IL-12 production in response to SEB requires both CD40-dependent signals and IFN-gamma secretion; (iii) the early systemic release of IFN-gamma (3 h post-treatment) in response to SEB is IL-12 independent, while the sustained, late response (6-9 h post-treatment) requires endogenous IL-12 production; (iv) IL-12 produced during the primary SEB response (day 0) is responsible for priming cells in vivo to high IFN-gamma production upon secondary challenge (day 2); (v) the priming effect of IL-12 is TCR unrelated, as SEB-primed animals secrete high levels of IFN-gamma in response to both staphylococcal enterotoxin A and LPS administered 48 h later. The ability of bacterial SAg to induce septic shock and to modulate the immune response to unrelated antigens may therefore be related to their unique capacity to induce systemic IL-12 production in vivo. These observations also help to explain why SEB-primed animals, known to express an anergic phenotype 48 h post-treatment (as judged by defective IL-2 production and proliferation), nevertheless display an increased capacity to secrete the inflammatory cytokine IFN-gamma.


Presenteeism Among Nurses in Switzerland and Portugal and Its Impact on Patient Safety and Quality of Care: Protocol for a Qualitative Study.
Nurses dispense direct care in a wide variety of settings and are considered the backbone of the health care system. They often work long hours, face emotional stress, and are at a high risk of psychosocial and somatic illnesses. Nurses sometimes fall sick but work regardless, leading to presenteeism and subsequent risks to quality of care and patient safety due to the increased likelihood of patients falling, medication errors, and staff-to-patient disease transmission. This study aims to understand presenteeism among frontline nurses and nurse managers in acute, primary, and long-term health care settings and to contribute to the development of future interventional studies and recommendations. A qualitative study based on online focus group discussions will explore the perceptions of, attitudes to, and experiences with presenteeism among frontline nurses and nurse managers. Using a pilot-tested interview guide, 8 focus group discussions will involve nurses working in acute care hospitals, primary care settings, and long-term residential care facilities in Switzerland's French-speaking region and Portugal's Center region. The data collected will be examined using a content analysis approach via NVivo 12 QSR International software. The University of Applied Sciences and Arts Western Switzerland's School of Health Sciences and the Polytechnic of Leiria's School of Health Sciences in Portugal have both approved funding for the study. The research protocol has been approved by ethics committees in both countries. Study recruitment commenced in February 2021. The results of the data analysis are expected by September 2021. This present study aims to gain more insight into the dilemmas facing nurses as a result of all causes of presenteeism among frontline nurses and nurse managers in different health care settings. The researchers will prepare manuscripts on the study's findings, publish them in relevant peer-reviewed journals, exhibit them in poster presentations, and give oral presentations at appropriate academic and nonscientific conferences. Regarding further knowledge transfer, researchers will engage with stakeholders to craft messages focused on the needs of nurses and nurse managers and on disseminating our research findings to deal with the issue of nursing presenteeism. PRR1-10.2196/27963.


[Epidemic strains of different plasmidovars of bacteria from Shigella genus].
Plasmid analysis of 46 strains of Shigella species confirmed the view about heterogeneity and variability of plasmids composition in these microorganisms. On the other hand,the presence of low-molecular, multiplicated extrachromosomal replicons, which were stable, was shown. Using them, it was possible to divide the Shigella isolates into two major plasmidovars, characterize them according to repertoire of the most stable plasmids and determine epidemically significant variants during sporadic and outbreak incidence of shigellosis in towns Nizhny Novgorod and Kstovo (Nizhny Novgorod region).


Incident sexually transmitted infection as a biomarker for high-risk sexual behavior after diagnosis of acute HIV.
Sexually transmitted infection (STI) diagnosis after diagnosis of acute HIV infection (AHI) indicates ongoing high-risk sexual behavior and possible risk of HIV transmission. We assessed predictors of STI acquisition and the effect of time since care entry on STI incidence in patients with AHI in care and receiving consistent risk-reduction messaging. Data on incident gonorrhea, chlamydia, trichomoniasis, primary/secondary syphilis, demographic, and clinical risk factors were abstracted from medical charts for patients diagnosed as having AHI and engaged in care. Poisson regression models using generalized estimating equations were fit to estimate incidence rates (IRs), IR ratios, and robust 95% confidence intervals. Among 185 patients with AHI, 26 (14%) were diagnosed as having at least 1 incident STI over 709.4 person-years; 46 STIs were diagnosed during follow-up (IR, 6.8/100 person-years). The median time from HIV care entry to first STI diagnosis was 609 days (range, 168-1681 days). Men who have sex with men (P = 0.03), a shorter time between presentation to medical care and AHI diagnosis (P = 0.06), and STI diagnosis before AHI diagnosis (P = 0.0003) were predictors of incident STI. Sexually transmitted infection IR greater than 1 year after entering care was double that of patients in care 1 year or less (IR ratio, 2.0; 95% confidence interval, 0.8-4.9). HIV viral load was above the limits of detection within 1 month of 11 STI diagnoses in 6 patients (23.1%) (median, 15,898 copies/mL; range, 244-152,000 copies/mL). Despite regular HIV care, STI incidence was high among this primarily young, men who have sex with men AHI cohort. Early antiretroviral initiation may decrease HIV transmission given ongoing risk behaviors despite risk-reduction messaging.


Epidural local anaesthetics versus opioid-based analgesic regimens on postoperative gastrointestinal paralysis, PONV and pain after abdominal surgery.
Gastrointestinal paralysis, nausea and vomiting, and pain, are major clinical problems following abdominal surgery. Anaesthetic and analgesic techniques that reduce pain and postoperative nausea and vomiting (PONV), and prevent or reduce postoperative ileus, may reduce postoperative morbidity, duration of hospitalisation and hospital costs. To compare effects of postoperative epidural local anaesthetic with regimens based on systemic or epidural opioids, on postoperative gastrointestinal function, postoperative pain, PONV and surgical/anaesthetic complications. Trials were identified by computerised searches of the Cochrane Controlled Trials Register, MEDLINE, EMBASE and by checking the reference lists of trials and review articles. Randomised controlled trials comparing the effects of postoperative epidural local anaesthetic with systemic or epidural opioids. Collected data included treatment in active (local anaesthetic) and control (opioid based) groups, time to first postoperative stool, time to first postoperative flatus, gastric emptying measured by the paracetamol absorption test, duration of the passage of barium sulphate, pain assessments, use of supplementary analgesics, nausea, vomiting and surgical/anaesthetic complications. Most studies in this review involved a small number of patients. Furthermore half of the studies indicated a poor level of methodology in particular regarding blinding and report of withdrawals. Heterogeneity of included studies was substantial. Results consistently showed reduced time to return of gastrointestinal function in the epidural local anaesthetic group compared with groups receiving systemic or epidural opioid (37 hours and 24 hours, respectively). Postoperative pain was comparable. Two studies compared the effect of epidural local anaesthetic with a combination of epidural local anaesthetic and opioid on gastrointestinal function. One study favoured epidural local anaesthetic and one study was indifferent. A meta analysis of five of eight studies comparing the effect of epidural local anaesthetic with a combination of epidural local anaesthetic and opioid on postoperative pain, yielded a reduction in VAS pain scores (0-100 mm) on the first postoperative day of 15 mm, in favour of the combination. No significant differences in PONV were observed between epidural local anaesthetic and opioid based regimens. Administration of epidural local anaesthetics to patients undergoing laparotomy reduce gastrointestinal paralysis compared with systemic or epidural opioids, with comparable postoperative pain relief. Addition of opioid to epidural local anaesthetic may provide superior postoperative analgesia compared with epidural local anaesthetics alone. The effect of additional epidural opioid on gastrointestinal function is so far unsettled. Randomized, controlled trials comparing the effect of combinations of epidural local anaesthetic and opioid with epidural local anaesthetic alone on postoperative gastrointestinal function and pain are warranted.


Regulation and identity of florigen: FLOWERING LOCUS T moves center stage.
The transition from vegetative to reproductive growth is controlled by day length in many plant species. Day length is perceived in leaves and induces a systemic signal, called florigen, that moves through the phloem to the shoot apex. At the shoot apical meristem (SAM), florigen causes changes in gene expression that reprogram the SAM to form flowers instead of leaves. Analysis of flowering of Arabidopsis thaliana placed the CONSTANS/FLOWERING LOCUS T (CO/FT) module at the core of a pathway that promotes flowering in response to changes in day length. We describe progress in defining the molecular mechanisms that activate this module in response to changing day length and the increasing evidence that FT protein is a major component of florigen. Finally, we discuss conservation of FT function in other species and how variation in its regulation could generate different flowering behaviors.


Everolimus for Treatment of Pseudomyogenic Hemangioendothelioma.
Pseudomyogenic hemangioendothelioma (PMH) is a recently described vascular neoplasm that occurs most commonly in the soft tissue of the distal extremities of young adults. Metastatic PMH can be fatal and there are no effective medications. We describe a case of a 15-year-old boy with metastatic PMH, who responded to treatment with everolimus, a mammalian target of rapamycin inhibitor. Immunohistochemistry showed that mammalian target of rapamycin was expressed in PMH biopsy specimens, which may explain the reduction in PMH tumor size following treatment.


Anatomically Contoured Anterior Plating for Isolated Tibiotalar Arthrodesis: A Systematic Review.
We performed a systematic review of the published literature to characterize patient demographic, surgical techniques, and functional outcomes to elucidate the complication and revision rates following isolated tibiotalar arthrodesis with anatomically contoured anterior plating. A comprehensive literature search was performed. Inclusion criteria were peer-reviewed studies in English, after 1990, at least 10 patients, and reporting clinical outcomes following contoured anterior plating and with follow-up of at least 80% and 1 year. Primary outcomes were fusion rate, time to fusion, return to activities, satisfaction, and functional outcome scores. Complication rates, reoperation, and revision were also extracted. Eight primary studies with 164 patients met the inclusion criteria. The average sample size was 21 ± 10.0 patients and average age was 49.2 years with 61.6% male. Posttraumatic arthritis (49.4%) was the most common operative indication, followed by primary osteoarthrosis (18.9%). The average follow-up was 21.1 months. At this time, 97.6% of patients went on to uneventful union at a weighted average time of 18.7 weeks postoperatively. AOFAS scores improved significantly ( P < .05). 25% complication rate was reported with wound complication (7.9%) and hardware irritation (6.7%) most common. Overall, 21.3% of patients underwent reoperation; 4 for revision arthrodesis following nonunion. Isolated tibiotalar arthrodesis utilizing anatomically contoured anterior plating demonstrates excellent clinical and functional outcomes at short-term follow-up. Overall, 97.6% of patients went on to fusion and functional outcomes consistently improved following surgery. Furthermore, while one-quarter of patients experienced complications, wound complications were relatively uncommon and less than one-quarter of these required surgical intervention. Level IV: Systematic Review.


Initial studies of holmium:YAG laser creation of spinal defects in fetal rabbits: model for urologic effects of myelomeningocele.
Myelomeningocele (MMC) is characterized by paraplegia and incontinence, often necessitating surgery. Current models of MMC in sheep and primates create a spinal defect long after anomalous neural tube closure ordinarily occurs. An ideal model of MMC would allow creation of the defect at the earliest age in a low-cost species with a short gestation. We present a method utilizing the holmium laser to create spinal defects in rabbits in utero for the study of the pathophysiology and repair of MMC. Pregnant rabbits of 22 to 23 days' gestational age were prepared and draped in sterile fashion for laparotomy under general anesthesia. The abdomen was opened, and both uterine horns were inspected. Double opposing pursestring sutures were placed to secure the chorioamniotic membranes over the fetal lumbar spine. Amniotic fluid was removed with a needle and saved. Electrocautery was used to open the uterus within the pursestring suture, exposing the fetal dorsum. The spine was exposed by laser dissection of the fetal dorsal musculature. Posterior laminectomy was accomplished with laser incisions of each side of the spinous process, leaving the underlying dura and cord exposed. The pursestring was then cinched, amniotic fluid was returned, and the uterus and trocar sites were closed. Cesarian section was performed at 30 to 31 gestational days, and the pups were examined and then humanely sacrificed for histologic evaluation of the lesion. The rabbit is an inexpensive species with a short gestation (33-35 days), and four or more fetuses may be operated on per litter, with the remainder serving as controls. Utilization of minimally invasive techniques including holmium:YAG laser dissection facilitates creation of spinal defects at an early age in this small-animal model.