Cloning of two Atlantic salmon growth hormone receptor isoforms and <Emphasis Type="Italic">in vitro</Emphasis> ligand-binding response

Two isoforms of the full-length cDNA of the growth hormone receptor (GHR) of the Atlantic salmon (Salmo salar; ss) were cloned by a PCR approach using RACE. Respectively, the cDNA sequences of ssGHR isoforms 1 and 2 are 2654 and 2608 nucleotides long, with 1782 and 1773 nucleotide ORFs. The resulting coded proteins are 594 and 590 aa long, with 19 and 20 aa signal peptides. The two isoforms share 86% protein and 87% cDNA sequence similarity. Isoform 1 is most similar to other salmonid GHR isoforms 1 while isoform 2 is most similar to salmonid GHR isoforms 2 (93–95%). Similarity with other teleost species was lower (37–44%). The bioactivity of the cloned ssGHR was tested by transfecting the ssGHR isoform 1 cDNA into CHO-K1 hamster cells, incubating with recombinant salmon GH (sGH) or native ovine prolactin (oPRL), and measuring cell proliferation by the MTT assay. The ssGHR-transfected cells significantly increased proliferation when stimulated by sGH at all concentrations. oPRL stimulated ssGHR-transfected cells at higher concentrations due to receptor cross reaction. ssGHR isoforms 1 and 2 contain a single transmembrane domain and the typical conserved motifs found in other teleost GHRs, including four paired cysteine residues and five potential N-glycosylation sites in the extracellular domain, Box I and Box II, as well as seven potential tyrosine phosphorylation sites in the intracellular domain. However, in salmonids, these motifs differ from those of other teleosts, and could be responsible for differentiated hormone binding, signal transduction and response.     

盖子红七彩神仙形态指标分析及其雌雄鉴别模型建立

测定和分析了138尾盖子红七彩神仙的形态学性状,并建立了雌雄个体的判别模型。对22项直观测量性状的主成分分析和21项标准化比例性状的R聚类分析,两者均显示盖子红七彩神仙的个体差异主要集中在体型大小、头部宽度、鱼鳍长度和宽度方面。对21项标准化比例性状进行T-检验,结果表明在雌雄间这些性状均差异不显著。以21项标准化比例性状为预选参数,通过逐步判别,建立了判别方程,对群体识别的正确率为80.2%。用32尾盖子红七彩神仙对这一判别方程进行识别验证,识别的正确率为84.4%。     

Chemical structure and thermal properties of lignin modified with polyethylene glycol during steam explosion

Thermoplastic processing of lignin is restricted by its high glass transition temperature (T _g). In this study, lignin was modified with polyethylene glycol (PEG) during steam explosion to improve its thermoplastic properties, and the effects of steam explosion and PEG on the chemical structure and thermal properties of lignin were investigated. Structure characterization using Fourier transform infrared spectroscopy showed that hydroxyl and ether functional groups increased and the activity of lignin was improved by steam explosion. In addition, steam explosion treatment was more effective than heat treatment for promoting the reaction of PEG with lignin. Solid-state ¹³C NMR revealed that PEG was grafted onto lignin. The T _g of raw lignin was 164.1 °C; after steam explosion, lignin exhibited more than one T _gs. The T _g of lignin was reduced when the steam explosion temperature increased and decreased further, to around 60 °C, when PEG was used to modify lignin. Therefore, this work provides an effective approach to reducing the high T _g of lignin.     

Low-Molecular-Weight Fucoidan Induces Endothelial Cell Migration via the PI3K/AKT Pathway and Modulates the Transcription of Genes Involved in Angiogenesis

Low-molecular-weight fucoidan (LMWF) is a sulfated polysaccharide extracted from brown seaweed that presents antithrombotic and pro-angiogenic properties. However, its mechanism of action is not well-characterized. Here, we studied the effects of LMWF on cell signaling and whole genome expression in human umbilical vein endothelial cells and endothelial colony forming cells. We observed that LMWF and vascular endothelial growth factor had synergistic effects on cell signaling, and more interestingly that LMWF by itself, in the absence of other growth factors, was able to trigger the activation of the PI3K/AKT pathway, which plays a crucial role in angiogenesis and vasculogenesis. We also observed that the effects of LMWF on cell migration were PI3K/AKT-dependent and that LMWF modulated the expression of genes involved at different levels of the neovessel formation process, such as cell migration and cytoskeleton organization, cell mobilization and homing. This provides a better understanding of LMWF’s mechanism of action and confirms that it could be an interesting therapeutic approach for vascular repair.     

大豆抗菌核病的全基因组关联研究

找到大豆与抗菌核病强关联的候选位点或候选基因,为抗病基因克隆和抗病分子标记开发提供借鉴,服务大豆抗菌核病育种。对126个加拿大大豆品种的基因组DNA用ApekⅠ酶消化后Illumina Hiseq2000平台测序进行基因分型,供试的126个材料用棉垫接种核盘菌菌丝体进行表型鉴定。采用Structure 2.3.4、SPSS 20.0、TASSEL 5.0和PLINKv 1.07软件分别模拟群体遗传结构、二元主成分分析、邻接法聚类,进行SNP-phenotype和Haplotype-phenotype的关联分析(只考虑加性效应)。最小等位基因频率0.01过滤,得到30 125个SNPs。主成分及群体结构聚类结果中度一致,将126个供试材料划分为2个组群,Kappa聚类一致度检验K=0.44。邻接法(The neighbor-joining algorithm,NJ)聚为3个组群。α≤0.05时,在单个SNP-phenotype的关联研究中,最强关联在3号染色体物理位置34387780、34387823和34387841处(P值都为8.669E-7),可分别解释表型变异的17.80%,其次在20,1,4,17号染色体上。Haplotype-phenotype的关联分析中,最强关联在17号染色体物理位置5575883/5647814/5648648/5734897处(P值为1.038E-6),可解释表型变异的17.56%。200 kb范围内,3号染色体上的候选基因有Glma.03g129100、Glma.03g129200、Glma.03g129300、Glma.03g129500、Glma.03g129800、Glma.03g129900。17号染色体上为Glma.17g071300、Glma.17g072200、Glma.17g073300。     

Chronic otitis externa with heat shock protein 70-positive intranuclear inclusion bodies in the ceruminous gland epithelium of a Chihuahua dog

A Chihuahua dog showed persistent itching in the right ear canal. Anti-inflammatory medicines and prednisolone were ineffective and total ear canal ablation was performed. Histological diagnosis was chronic otitis externa. Eosinophilic intranuclear inclusion bodies (Cowdry type A and full-type) were occasionally observed in the ceruminous gland epithelium. The inclusion bodies were negative for nucleic acid and ultrastructurally composed of fibrous structures (approximately 10 nm in width). Viral infection was initially suspected, but polymelase chain reaction tests did not detect the expected viral genes. Immunohistochemistry revealed that the inclusion bodies were positive for heat shock protein 70 (HSP70), suggesting that these bodies could be protein aggregates including HSP70. The etiology of this lesion has not been elucidated, but chronic inflammation may influence the cytoplasm-to-nuclear transportation of HSP70. To the best of our knowledge, this is the first report of canine chronic otitis externa with HSP70-positive intranuclear inclusion bodies.     

The effects of vitamin E on non-specific immune response of the juvenile soft-shelled turtle <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>

ABSTRACT:   This study was conducted to determine the effect of dietary vitamin E supplements on non-specific immune responses in juvenile soft-shelled turtles. Turtles were fed diets supplemented with vitamin E at 0 (control), 50, 250, 500, 1000 and 5000 mg/kg, respectively, for 4 weeks. Results showed that blood cell phagocytosis and serum bactericidal activity were significantly improved in turtles from two diets (250 and 500 mg/kg vitamin E supplementation), while had no significant improvement in turtles from the three remaining diets (50, 1000 and 5000 mg/kg vitamin E supplementation) when compared to turtles from the control group. Serum bacteriolytic activity in turtles from diets with 50, 250, 500 and 1000 mg/kg vitamin E supplementation were higher than activity levels quantified for the control group, but no significant difference was observed between the 5000 mg/kg vitamin E supplementation diet and the control diet. The results suggest that vitamin E has an upper and lower threshold for improving non-specific immune function in soft-shelled turtles, and the optimal supplementation may be between 250 and 500 mg/kg.     

The effects of less-expensive plant protein sources replaced with soybean meal in the juvenile diet of grass carp (Ctenopharyngodon idella): growth, nutrient utilization and body composition

In this study, the effects of less-expensive protein sources replaced with soybean meal on growth performance, nutrient utilization and body composition of juvenile grass carp were investigated. For this purpose, a control diet containing 42% soybean meal was prepared. Cotton seed meal (CSM), sunflower meal (SFM) and corn meal (CM) were added to the experimental diets by replacing 100% of the soybean meal protein used in the control diet. All diets were iso-nitrogenous (40% crude protein) and iso-caloric (19.5?MJ?kg^?1 gross energy). Each treatment had three replicates, and 20 juvenile grass carps (3.5?±?0.1?g, initial weight) were located in per replicate. The fish were fed to satiety three times in a day during 3?months at 26°C of water temperature in glass aquarium (45 l). The results of the study showed that the weight gain (308.6?C448.6%), specific growth rate (1.56?C1.89%/day), feed conversion rate (1.2?C1.9), protein efficiency ratio (1.32?C2.08), and apparent digestibility coefficients of protein (80.5?C89%) or sum of amino acids (80.5?C88.9%) were not significantly affected by the experimental diets, except diet CM. All of these values obtained for the diet CM were significantly lower than those of fish fed on the control, CSM and SFM diets. However, there were no significant differences between the lipid digestibility of the diets, and crude protein, lipid, and gross energy contents in the whole body of fish in experiments. In conclusion, CSM and SFM diets were highly utilized by juvenile grass carp, but not CM diet. These results combined with the remarkable acceptability of diets containing high levels of plant protein ingredients with identical growth performances of juvenile grass carp show clearly that dietary soybean meal level can be considerably reduced without any adverse consequence in terms of somatic growth and nitrogen utilization.     

The relationship between water intake rates,paddy ponding depth,and farmers’ water management techniques

Japanese farmers manage their irrigation water based on their past experiences and preferences, considering such factors as weather and available water (hereafter defined as empirical water management). They elaborately control the intake and drainage rates of their own paddy fields to maintain optimal ponding depths. But these well-managed systems will drastically change because of the decreasing number of farmers. Therefore, it is necessary to clarify if the optimal ponding depth will be maintained within the limits of traditionally-allowed water intake rate from the main river. The first objective of this study was the quantification of actual water use in the paddy fields, resulting from the farmers water management on the basis of their experience. The significance of the present water intake rate under empirical water management was studied for a paddy field command area of about 230 ha. Water intake rates and the water requirements of the whole area were investigated by measuring the flow rate at 17 points of irrigation and drainage canals. Characteristics of the farmers empirical water management were investigated by measuring the hourly changes in inflow and outflow rates for a sub-area using an automatic measurement system, and an inferential method of determining water management patterns for the paddy fields was proposed. The newly-proposed inferential method was introduced in the tank model, which expresses the characteristics of water management in the command area. The Shuffled Complex Evolution Algorithm (SCE-UA) method was used for optimizing the model parameters. It was proven that the model accuracy improved when the farmers empirical water management was taken into account. The optimal amount of water to be applied to the command area was quantified by the simulation. The second objective was to predict the effect of the decreasing number of farmers on future water use conditions. The simulated result indicates the difficulty of maintaining optimal ponding depth for the whole command area when the farmers empirical water management is not maintained. In other words, results indicated that efficient water use requires an automatic water management system or a new pipeline system to replace the farmers present empirical water management.