Variations in parameters of liver function and plasma progesterone related to underfeeding and ketosis in a dairy herd

Twenty-eight Norwegian Red Cattle dairy cows were fed silage ad libitum and restricted amounts of concentrates. Blood samples were collected before morning feeding, once or twice weekly, from 2 weeks before to 12 weeks after calving. Parameters of liver function, carbohydrate status and fertility were recorded in order to assess their interrelationships. Eight cows were treated for clinical ketosis. Four of these had to be treated 2 or 3 times. Aspartate aminotransferase and bilirubin showed the highest within-animal coefficients of correlation with acetoacetate. Analysis of variance revealed a significant effect of carbohydrate status (indicated by plasma acetoacetate levels) on the levels of aspartate aminotransferase, glutamate dehydrogenase and sorbitol dehydrogenase, though only a small part of the total variation was explained by this factor. The estimated volume density of liver fat in the 4th week of lactation averaged 6.0 +/- 6.4% (+/- SD) ranging from 0.1-25.1%. Liver fat content at this stage of lactation was not significantly correlated with other indicators of liver function or carbohydrate status. Cows treated for clinical ketosis had significantly lower plasma progesterone values at the time of first ketosis treatment than untreated multiparous cows. The frequency of high progesterone values (greater than 3 ng/ml) being significantly lower in treated than in untreated cows during the period from 3-5 weeks post partum, though not at later stages. In conclusion, the results revealed a significant relationship between carbohydrate status and liver function, and also between clinical ketosis and luteal function.     

D-Dimer Improves the Prognostic Value of Combined Clinical and Laboratory Data in Equine Gastrointestinal Colic

The discriminating ability of 15 parameters alone or in combinations, including results from analysis of plasma endotoxin, the Nycomed plasma D-Dimer test and phospholipase A₂, were analyzed to predict morbidity and mortality in equine gastrointestinal colic. Endotoxaemia was a characteristic feature of the colic horses. The problem of adequately predicting non-survivors among colic horses required several parameters to be included in the logistic model: if the “classical parameters”, (heart rate, respiratory rate, PCV, anion gap) were included in the model, addition of plasma D-dimer, phospholipase A₂, and Cl^- significantly improved the predictive value of the logistic model. Increasing heart rate and D-dimer together with decreasing chloride was a risk factor for nonsurvival. The sensitivity of this three-parameter logistic model to predict nonsurvival was 78% and specificity 77%. The Nycomed D-Dimer test is recommended as a horse-site test to predict disseminated intravascular coagulation and nonsurvival in equine colic.     

The potential use of plasma glutamate dehydrogenase activity for the evaluation of hepatic disease in the cockatiel (Nymphicus hollandicus)

A reference interval for plasma glutamate dehydrogenase (GD) (E.C. 1.4.1.3) activity, of 0-8 IU/L, was established for the cockatiel (Nymphicus hollandicus). An automated modification of a commercial manual assay was used. This enzyme is considered liver specific in humans and numerous domestic animals, due to its organ distribution. A similar distribution was found in cockatiels in this study. Maximal enzyme activity was recovered from liver and kidney homogenate supernatants. Minimal activity was detected in skeletal muscle preparations. These results suggest a potential use for plasma GD activity in the evaluation of hepatocelluar injury/necrosis in cockatiels.     

组氨醇脱氢酶抑制剂的研究动态

组氨醇脱氢酶(HDH)是组氨酸生物合成过程中最后两步反应的催化酶。对组氨醇脱氢酶的结构、催化机理、组氨醇脱氢酶抑制剂等方面的研究动态进行了较为详细的综述。     

Generation and analysis of expressed sequence tag sequences from a soft‐seeded pomegranate cDNA library

The cultivation of soft‐seeded pomegranate is an important direction in pomegranate breeding. To comprehensively understand the molecular mechanisms involved in the formation of soft‐seeded pomegranate (Punica granatum. var. Hongmanaozi), we established an expressed sequence tag (EST) resource. Two thousand valid sequences were generated, from which 907 unigenes were obtained after initial assembly using the clustalx program. Among these unigenes, 51 showed no similarity to any protein in the public databases, 433 matched with proteins of unknown function, and 423 matched with proteins of known or putative functions. The 423 unigenes were further classified into 13 categories. Among these categories, protein synthesis, cell structure, protein destination and storage, secondary metabolism, signal transduction and transporters accounted for 8%, 8%, 4%, 7%, 6% and 17%, respectively. We also successfully developed 10 highly polymorphic expressed sequence tag‐simple sequence repeat (EST‐SSR) markers for pomegranate. The results provide a new tool for future activities in pomegranate breeding.     

棉花根系和叶片质外体汁液分离方法的改进

质外体汁液(apoplastic washing fluid, AWF)在植物生长发育和抵抗生物及非生物逆境方面发挥着重要作用。目前普遍采用真空渗透离心法提取质外体汁液,但具体提取流程和条件却因植物培养条件、种类和器官等不同而不同。本试验以营养液培养的棉花幼苗为材料,改进了棉花根系和叶片AWF分离的取样方法、渗透条件和离心参数等。结果表明,相对于通常的非整体取样,改良后整体取样简单易操作且显著降低了质外体与共质体的苹果酸脱氢酶(malate dehydrogenase, MDH)活性比值,更适宜开展质外体汁液组分研究。根据真空渗透鲜重增加和AWF稀释因子,进一步证明根系不用真空渗透;而叶片需真空渗透,适宜真空强度/时间为-60 kPa/1min,真空后恢复到正常压强约110s。证明叶片颜色变深面积可作为判定真空渗透强度或时间是否适宜的一种简易可行方法。最后,AWF体积、质外体与共质体的可溶性蛋白含量比值和MDH活性比值综合分析表明,棉花根系适宜离心力大小/时间长度为800×g/10～20 min,叶片为400×g/5 min。本改进方法为棉花AWF组分,如蛋白质组学和代谢组学等研究结果的准确性和可靠性奠定了基础,为其他作物AWF分离方法的优化提供了参考。     

艾纳香脱氢酶基因家族的生物信息学分析

采用RT-PCR和RACE（Rapid amplification of cDNA ends）技术,从艾纳香（Blumea balsamifera (L.) DC.）叶片中克隆到4条编码艾纳香脱氢酶（BbADH1、BbADH2、BbADH3、BbADH4）基因的cDNA序列,并对4条核苷酸及其编码的氨基酸序列进行生物信息学分析。结果显示:4条艾纳香脱氢酶序列开放阅读框均在900 bp左右,蛋白质等电点（pI）值在5.0~9.0之间,含量最多的氨基酸为亮氨酸（Leu）,最少的为色氨酸（Try）,具有明显的疏水区和亲水区,N端未发现信号肽,且无跨膜区;同源性比对结果显示,艾纳香BbADH蛋白与其他植物中ADH蛋白具有高度的相似性,且具有脱氢酶的特征功能域;系统发育分析表明,艾纳香（B. balsamifera (L.) DC.）BbADH1和BbADH3同处于一个分支,且与胡杨（Populus euphratica Oliv.）PeADH物种亲缘关系较近,而艾纳香BbADH4和BbADH2处于不同分支,且分别与葡萄（Vitis vinifera）VvADH和烟草（Nicotiana tabacum）NtADH物种亲缘关系较近。     

Uneven ripening in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) berries in India is linked to seed viability

Uneven ripening (UR) is a physiological disorder of unknown origin in ‘Bangalore blue’ grape (Vitis vinifera × Vitis labrusca) leading to wine of inferior quality. A preliminary study found wide variations in total dehydrogenase activity (TDH) of seeds from unevenly ripe berries. In our experiments, gibberellin (GA₃) applied to young grapes increased seed TDH activity and reduced the incidence of uneven ripening to 2% compared with 35% in the control. In contrast, paclobutrazol (PBZ) decreased TDH activity and increased the incidence of the disorder to 58%. GA₃-treated berries had higher concentrations of sucrose and TDH activity in seed representing mature seeds with high viability. In contrast, PBZ-treated and control berries had higher concentrations of glucose and lower TDH activity, indicating immature seeds with low viability. These results suggested that competition among developing berries can lead to differences in seed gibberellin content, seed viability and the rate of berry growth resulting in green, purple, and black berries at harvest. The study established the role of seed viability in uneven ripening and demonstrated that the incidence of the disorder is reduced by the application of GA₃ to immature berries.     

黄瓜低霜霉威残留性相关基因SDH克隆及表达

通过Solexa技术和比较基因组学方法鉴定霜霉威胁迫下黄瓜果实差异表达基因,筛选黄瓜低霜霉威残留性相关基因SDH。研究利用PCR克隆获得SDH基因全长,命名为Cs SDH。构建亚细胞定位表达载体,利用农杆菌浸润法注射烟草叶片后激光共聚焦显微镜下观察;分别用荧光定量RT-PCR方法和酶联免疫法研究高、低农残品系D9320和D0351中Cs SDH基因在农药霜霉威处理后,不同时间点、不同组织部位时空表达特征和琥珀酸脱氢酶活性。结果表明,Cs SDH基因在烟草叶片细胞中被定位在细胞膜上。黄瓜低农残品种D0351中,Cs SDH基因在果实受霜霉威胁迫后反应迅速,主要在处理前期响应表达强烈,琥珀酸脱氢酶活性显著增加,且该基因表达量显著高于其在高农残品种D9320中表达量。Cs SDH果、叶中表达量较茎中高,且各组织部位表达量高低顺序稳定,为叶果茎。黄瓜高农残品种D9320中,Cs SDH基因表达量在0.5、1、3、12 h呈现上调表达,6、9 h基因表达量差异不显著,48 h出现上调表达高峰。说明克隆得到的Cs SDH基因属农药处理前期响应基因,且该基因可能在降低黄瓜农药残留过程中发挥重要作用。试验通过研究黄瓜琥珀酸脱氢酶基因(SDH)在霜霉威胁迫下表达情况,为挖掘黄瓜低农药残留性功能基因以及探明其作用分子机制奠定基础。     

Molecular cloning,purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein

The Brucella mdh gene was successfully cloned and expressed in E. coli. The purified recombinant malate dehydrogenase protein (rMDH) was reactive to Brucella-positive bovine serum in the early stage, but not reactive in the middle or late stage, and was reactive to Brucella-positive mouse serum in the late stage, but not in the early or middle stage of infection. In addition, rMDH did not react with Brucella-negative bovine or mouse sera. These results suggest that rMDH has the potential for use as a specific antigen in serological diagnosis for early detection of bovine brucellosis.