Identification of differentially expressed genes in seeds of two Brassica napus mutant lines with different oleic acid content

The regulation of seed oleic acid synthesis in rapeseed is largely unknown. In this study, gene expression pattern during seed development between two Brassica napus mutants was compared. Using immature seeds 27 days after pollination, differentially expressed cDNA clones were identified bysubtractive suppression hybridization (SSH). A total of 480 cDNA clones corresponding to 88 genes were found up-regulated and 18 genes down-regulated in seeds with high oleic acid content. Most ofthe differentially expressed genes are related to metabolism and regulation. The possible role of these genes in seeds was discussed. Further analysis of the function of these genes may provide novel targets for manipulation of fatty acid composition in rapeseed

Antioxidant treatment enhances human mesenchymal stem cell anti-stress ability and therapeutic efficacy in an acute liver failure model

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EFEITOS DA INTOXICAÇÃO AGUDA COM ETANOL SOBRE A EXSUDAÇÃO DE NEUTRÓFILOS PARA A CAVIDADE PERITOENAL DE EM CAMUNDONGOS INOCULADOS COM Staphilococcus aureus.

Introdução. Há demonstrações que uma intoxicação aguda pelo etanol tem efeitos antiinflamatórios, com redução da exsudação de neutrófilos e com aumento da susceptibilidade a bactérias, especialmente Streptococcus pneumoniae. No entanto não se conhece o tempo durante o qual persistem os efeitos inibidores da exsudação celular e da redução da atividade microbicida de leucócitos, após a intoxicação etílica aguda. Objetivos. Avaliar o tempo após uma intoxicação etílica aguda no qual persistem os efeitos inibidores da exsudação de neutrófilos e a redução da capacidade microbicida do exsudato inflamatório em um modelo de peritonite induzida por Staphylococcus aureus. Métodos. Camundongos C57BL/6 receberam, por gavagem, uma dose de 7mg de etanol/g peso corporal em solução a 40%. Uma, 12,24, 48 e 72 horas após, receberam uma inoculação intraperitoneal de Staphylococcus aureus (0,5 ml contendo 6 a 9x108 UFC/mL; cepa ATCC 25923). Seis horas depois, os animais eram eutanasiados e a cavidade peritoneal lavada com PBS/EDTA 0,01M; uma alíquota era utilizada para contagem do número de UFC e a outra para contagem global e específica das células do exsudato utilizando câmara de Neubauer e citocentrífuga para confecção de esfregaços, corados por corante hematológico rápido (Dipquick). A contagem de UFC foi feita pelo método de diluição seriada com semeadura em placas de ágar Müller-Hinton. Resultados. Todos os animais que receberam etanol apresentaram sinais de embriaguês, que chegou a um estado de letargia profunda do qual todos os animais se recuperavam em, no máximo, 45 minutos. A dose de etanol utilizada induziu involução do timo, evidente 24 horas após a alcoolização, mas com recuperação após 120 horas. Nos animais alcoolizados que receberam o inóculo do estafilococo houve redução significativa da exsudação celular, devido a redução da exsudação de neutrófilos, até 24 horas após a ingestão do etanol. A análise dos esfregaços mostrava maior quantidade de bactérias fora das células no grupo etanol e o número de UFC foi maior no período avaliado, mas a diferença não foi estatisticamente significativa. Conclusão. Confirma-se os efeitos antiinflamatórios da intoxicação etílica aguda, com redução significativa no exsudato de leucócitos, até 24 horas após a exposição ao etanol; essa redução decorre especialmente da redução da exsudação de neutrófilos, já que o número de mononucleares exsudados no período avaliado foi semelhante nos dois grupos experimentais. O poder microbicida da cavidade peritoneal frente aos estafilococos foi menor no grupo etanol embora sem significância estatística, possivelmente porque os macrófagos residentes foram menos afetados pelos efeitos do etanol. Palavras-chaves: Alcoolismo agudo, neutrófilos, inflamação

N-wasp Is Required For Structural Integrity Of The Blood-testis Barrier.

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Perfluorooctanesulfonate (PFOS)-induced Sertoli cell injury through a disruption of F-actin and microtubule organization is mediated by Akt1/2

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Microbial fuel cells: a green and alternative source for bioenergy production

Microbial fuel cell (MFC) represents one of the green technologies for the production of bioenergy. MFCs using microalgae produce bioenergy by converting solar energy into electrical energy as a function of metabolic and anabolic pathways of the cells. In the MFCs with bacteria, bioenergy is generated as a result of the organic substrate oxidation. MFCs have received high attention from researchers in the last years due to the simplicity of the process, the absence in toxic by-products, and low requirements for the algae growth. Many studies have been conducted on MFC and investigated the factors affecting the MFC performance. In the current chapter, the performance of MFC in producing bioenergy as well as the factors which influence the efficacy of MFCs is discussed. It appears that the main factors affecting MFC’s performance include bacterial and algae species, pH, temperature, salinity, substrate, mechanism of electron transfer in an anodic chamber, electrodes materials, surface area, and electron acceptor in a cathodic chamber. These factors are becoming more influential and might lead to overproduction of bioenergy when they are optimized using response surface methodology (RSM)

Independent and Combined Effects of Dietary Weight Loss and Exercise on Leukocyte Telomere Length in Postmenopausal Women

Objective: Investigate the effects of 12 months of dietary weight loss and/or aerobic exercise on leukocyte telomere length in postmenopausal women. Design and Methods 439 overweight or obese women (50–75 y) were randomized to: i) dietary weight loss (N=118); ii) aerobic exercise (N=117), iii) diet + exercise (N=117), or iv) control (N=87). The diet intervention was a group-based program with a 10% weight loss goal. The exercise intervention was 45 mins/day, 5 days/week of moderate-to-vigorous aerobic activity. Fasting blood samples were taken at baseline and 12 months. DNA was extracted from isolated leukocytes and telomere length was measured by quantitative-polymerase chain reaction (qPCR). Mean changes were compared between groups (intent-to-treat) using generalized estimating equations. Results: Baseline telomere length was inversely associated with age (r=−0.12 p<0.01) and positively associated with maximal oxygen uptake (r=0.11, p=0.03), but not with BMI or %body fat. Change in telomere length was inversely correlated with baseline telomere length (r=−0.47, p<0.0001). No significant difference in leukocyte telomere length was detected in any intervention group compared to controls, nor was the magnitude of weight loss associated with telomere length at 12 months. Conclusions: Twelve-months of dietary weight loss and exercise did not change telomere length in postmenopausal women

High throughput mutagenesis for identification of residues regulating human prostacyclin (hIP) receptor

The human prostacyclin receptor (hIP receptor) is a seven-transmembrane G protein-coupled receptor (GPCR) that plays a critical role in vascular smooth muscle relaxation and platelet aggregation. hIP receptor dysfunction has been implicated in numerous cardiovascular abnormalities, including myocardial infarction, hypertension, thrombosis and atherosclerosis. Genomic sequencing has discovered several genetic variations in the PTGIR gene coding for hIP receptor, however, its structure-function relationship has not been sufficiently explored. Here we set out to investigate the applicability of high throughput random mutagenesis to study the structure-function relationship of hIP receptor. While chemical mutagenesis was not suitable to generate a mutagenesis library with sufficient coverage, our data demonstrate error-prone PCR (epPCR) mediated mutagenesis as a valuable method for the unbiased screening of residues regulating hIP receptor function and expression. Here we describe the generation and functional characterization of an epPCR derived mutagenesis library compromising >4000 mutants of the hIP receptor. We introduce next generation sequencing as a useful tool to validate the quality of mutagenesis libraries by providing information about the coverage, mutation rate and mutational bias. We identified 18 mutants of the hIP receptor that were expressed at the cell surface, but demonstrated impaired receptor function. A total of 38 non-synonymous mutations were identified within the coding region of the hIP receptor, mapping to 36 distinct residues, including several mutations previously reported to affect the signaling of the hIP receptor. Thus, our data demonstrates epPCR mediated random mutagenesis as a valuable and practical method to study the structurefunction relationship of GPCRs. © 2014 Bill et al

Support vector machine model for diagnosis of lymph node metastasis in gastric cancer with multidetector computed tomography: a preliminary study

<p>Abstract</p> <p>Background</p> <p>Lymph node metastasis (LNM) of gastric cancer is an important prognostic factor regarding long-term survival. But several imaging techniques which are commonly used in stomach cannot satisfactorily assess the gastric cancer lymph node status. They can not achieve both high sensitivity and specificity. As a kind of machine-learning methods, Support Vector Machine has the potential to solve this complex issue.</p> <p>Methods</p> <p>The institutional review board approved this retrospective study. 175 consecutive patients with gastric cancer who underwent MDCT before surgery were included. We evaluated the tumor and lymph node indicators on CT images including serosal invasion, tumor classification, tumor maximum diameter, number of lymph nodes, maximum lymph node size and lymph nodes station, which reflected the biological behavior of gastric cancer. Univariate analysis was used to analyze the relationship between the six image indicators with LNM. A SVM model was built with these indicators above as input index. The output index was that lymph node metastasis of the patient was positive or negative. It was confirmed by the surgery and histopathology. A standard machine-learning technique called k-fold cross-validation (5-fold in our study) was used to train and test SVM models. We evaluated the diagnostic capability of the SVM models in lymph node metastasis with the receiver operating characteristic (ROC) curves. And the radiologist classified the lymph node metastasis of patients by using maximum lymph node size on CT images as criterion. We compared the areas under ROC curves (AUC) of the radiologist and SVM models.</p> <p>Results</p> <p>In 175 cases, the cases of lymph node metastasis were 134 and 41 cases were not. The six image indicators all had statistically significant differences between the LNM negative and positive groups. The means of the sensitivity, specificity and AUC of SVM models with 5-fold cross-validation were 88.5%, 78.5% and 0.876, respectively. While the diagnostic power of the radiologist classifying lymph node metastasis by maximum lymph node size were only 63.4%, 75.6% and 0.757. Each SVM model of the 5-fold cross-validation performed significantly better than the radiologist.</p> <p>Conclusions</p> <p>Based on biological behavior information of gastric cancer on MDCT images, SVM model can help diagnose the lymph node metastasis preoperatively.</p