Characterization of an intravenous lipopolysaccharide inflammation model in calves with respect to the acute-phase response

Our objective was to develop a lipopolysaccharide (LPS) inflammation model in calves to evaluate the acute-phase response with respect to the release of pro-inflammatory cytokines and acute-phase proteins, fever development and sickness behaviour. Fourteen 4-week-old male Holstein Friesian calves were included and randomly assigned to a negative control group (n = 3) and an LPS-challenged group (n = 11). The latter received an intravenous bolus injection of 0.5 μg of LPS/kg body weight. Blood collection and clinical scoring were performed at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 12, 18, 24, 28, 32, 48, 54 and 72 h post LPS administration (p.a.). In the LPS group, the following clinical signs were observed successively: tachypnoea (on average 18 min p.a.), decubitus (29 min p.a.), general depression (1.75 h p.a.), fever (5 h p.a.) and tachycardia (5 h p.a.). Subsequent to the recovery from respiratory distress, general depression was prominent, which deteriorated when fever increased. One animal did not survive LPS administration, whereas the other animals recovered on average within 6.1 h p.a. Moreover, the challenge significantly increased plasma concentrations of tumour necrosis factor-α, interleukin 6, serum amyloid A and haptoglobin, with peaking levels at 1, 3.5, 24 and 18 h p.a., respectively. The present LPS model was practical and reproducible, caused obvious clinical signs related to endotoxemia and a marked change in the studied inflammatory mediators, making it a suitable model to study the immunomodulatory properties of drugs in future research.     

Differential response of zucchini varieties to low storage temperature

Several parameters related to cold damage in zucchini have been measured in fruits from different varieties cultivated in the province of Almería (south-eastern Spain). The purpose of this work is to correlate physiological parameters with commercial quality and chilling damage, in order to establish reliable criteria for selecting varieties having better genetic adaptation to low-temperature storage. The results show that, for varieties harvested during February, the variety Natura was the best adapted to storage at 4 °C, as its fruits suffered less from chilling injuries, had a lower weight loss, and the levels of metabolites malonyldialdehyde and H₂O₂ were not as high as in the other varieties. Genetic variation was detected for some of the parameters analysed. The positive correlation among the levels of MDA, H₂O_2, and chilling injury, as well as the negative correlation among catalase and chilling injury, makes these parameters good indicators of chilling damage in our system.     

浅谈室内山墙窗口圈梁构造柱防结露措施

本文论述了室内山墙、窗口、圈梁、构造拄、结露的产生原因及预防措施。     

不同处理对西藏牦牛超数排卵的影响研究

本研究选用了埋植CIDR栓法、埋植PRID栓法、PG法以及PMSG+PG法四套方案对西藏41头母牦牛进行了超数排卵试验。试验结果:埋植CIDR法得到的平均卵母细胞数为7.4枚/头,埋植PRID法得到的平均卵母细胞数为6.5枚/头,PG法得到的平均卵母细胞数为4枚/头,PMSG+PG法得到的平均卵母细胞数为4.8枚/头,各组之间差异极显著(P<0.01);而经过卵母细胞成熟培养之后,埋植CIDR法得到的平均可用卵母细胞数为6.2枚/头,埋植PRID法得到的平均可用卵母细胞数也为6.2枚/头,PG法得到的平均可用卵母细胞数为4枚/头,PMSG+PG法得到的平均可用卵母细胞数为4.6枚/头,CIDR+FSH法与PRID+FSH法两组、两次PG法与PMSG+PG法两组差异不显著(P>0.05),但是CIDR+FSH法和PRID+FSH法组与两次PG法和PMSG+PG法组之间差异都极显著(P<0.01);用CIDR法和PRID法进行西藏牦牛超排效果较好,并可获得较多的可用卵母细胞。     

水稻纹枯病菌PG的分离纯化及其理化性质研究

水稻纹枯病菌产生的多聚半乳糖醛酸酶(Polygalacturonase,PG)是其重要的致病因子之一。用丙酮法提取PG粗蛋白,分别经DEAE-Sepharose Fast Flow离子交换柱、Phenyl-Sepharose 6 Fast Flow疏水柱、Sephadex G-75凝胶柱和DE52离子交换柱层析纯化得到一种具有较高活性的PG纯蛋白。该蛋白分子量为39.81 kD;等电点为4.58;含有糖基,含糖量为1.48%;含有α氨基酸,但不含芳香族氨基酸;不含脂基。这种蛋白在pH4~12范围内均具有活性,pH5时活性最大;对热不稳定,100℃下水浴20 min,活性完全丧失;对胰蛋白酶和蛋白酶K敏感,酶处理后其活性只有对照的35.0%和35.2%;对紫外线和氯仿亦敏感,处理后活性仅为对照的40.0%和51.7%。     

桃PG基因启动子克隆及序列分析

为获得桃多聚半乳糖醛酸酶( PG)基因启动子并分析调控元件,以桃品种大久保基因组DNA为模板,采用染色体步移技术克隆了PG基因的部分序列及其5′侧翼区,并分析其调控元件组成。结果表明,克隆产物长986 bp,3′侧翼序列与 GenBank中的PG基因序列的5′端部分序列同源性为96%,登录号为FJ940722。在789~839 bp位置存在基础启动子序列,转录起始位点位于828 bp的碱基C,其可能性为0.98。除含有CAAT-Box、TATA-Box等基本启动子元件外,还存在众多参与光响应元件、干旱诱导MYB结合位点、水杨酸响应元件等应答激素和胁迫信号元件,表明PG基因的转录和表达可能受到激素、干旱、光照等外界环境的胁迫以及衰老的调控。     

油菜cDNA表达文库构建及核盘菌致病因子互作蛋白的筛选和鉴定

核盘菌 [Sclerotinia sclerotiorum (Lib.) de Bary] 是引起油菜菌核病的病原真菌。核盘菌侵染植物细胞早期,可分泌一些细胞壁降解酶, 如不同类型的多聚半乳糖醛酸酶(polygalacturonase, PG)。这些酶对病菌在植物组织上的定植起关键作用, 同时酶活性的高低也决定病原菌的毒性或致病性。PG可以激活细胞的防卫反应而与其酶活性无关。为了解PG的信号转导途径, 利用RT-PCR方法克隆了核盘菌的PG基因, 将PG的编码区与酵母GAL4的DNA结合功能区融合, 构建到酵母诱饵蛋白表达载体PGBKT7中; 然后在酵母中构建了油菜cDNA表达文库。通过酵母双杂交方法在油菜cDNA表达文库中对与PG互作的蛋白进行了筛选, 分离到一个与PG互作的蛋白, 测序及BLAST分析表明该蛋白含有一个与钙离子结合的结构域, 称为C2结构域(C2-domain), 推测该蛋白是一种钙依赖的膜结合蛋白。该蛋白与拟南芥中一个含C2结构域功能未知蛋白的氨基酸同源性高达80.24%。利用半定量PCR分析了核盘菌诱导后该基因在油菜花、茎、叶中的表达, 结果表明该基因在叶片中表达量最高。克隆到的C2蛋白与其他物种中的C2蛋白比较发现, 与钙离子结合的天冬氨酸残基很保守。     

湘东黑山羊同期发情技术的研究

应用PG+PMSG处理对54只湘东黑山羊在2～5天内集中发情并配种.结果:(1)在处理后的2～3天.有49头发情(其中同期自然发情2头),同期处理的发情率87.0%[47/54],同期发情率87.5%[49/56].(2)发情母羊的初配率为83.9%[47/56],待第一个情期(21天)后采用发情鉴定法进行初步妊娠诊断,有返情表现的羊只为7只,发情配种成功率85.1%[40/47],初步确定初配受胎率为71.4%[40/56].     

PG,ACC,乙烯对番茄果实成熟的影响

番茄果实在成熟过程中硬度下降迅速，ＰＧ活性急剧上升，表现出显著的负相关；用ＡＣＣ和乙烯利处理绿熟期果实，结果乙烯促进了正常番茄和ａｌｃ番茄果实成熟进程，其ＰＧ活性也升高，而外源乙烯和ＡＣＣ不影响ｎｏｒ基因突变体果实的成熟进程。ＡＣＣ和ＰＧ无直接关系。     

怀孕母山羊传染性胸膜肺炎PG3垂直感染试验

取健康易感怀孕母山羊3只，气管接种丝状支原体山羊亚种PG3，第28天，其中1头产出羊羔1只，第32天，剖杀3头母羊及4日龄羊羔，均见肺部有明显的山羊传染性胸膜肺炎病变，无菌抽取羊水，并剖取胎羊的肺，肝和4日龄羔羊的胸水，肺和肺门淋巴结，均培养分离出多形性支原体，用该支原体培养物作生长抑制试验均为PG3阳性，Y98阳性，Y98阴性，与接种株PG3－致，说明出羊传染性胸膜肺炎PG3可通过胎盘垂直感染胎羊。