Glucose-6-Phosphate Dehydrogenase Activity and Glutathione Stability in Fetal and Adult Bovine Erythrocytes

Previously, we found a substantially higher glucoses-phosphate dehydrogenase (G6PD) activity and a slightly higher 6-phosphogluconate dehydrogenase (6PGD) activity in bovine fetal erythrocytes than in bovine adult erythrocytes (Steensgaard 1968). Now, we have investigated whether these differences in dehydrogenase activities were followed by characteristic differences in glutathione (GSH) stability and glutathione concentration. The results are shown in Table 1, which also gives the results of the same investigations on normal and G6PD deficient human erythrocytes.     

On The Normal Activities of Glucose-6-Phosphate Dehydrogenase and 6-Phospho-Gluconate Dehydrogenase in Bovine Erythrocytes

The activities of glucose-6-phosphate dehydrogenase and of 6-phos-pho-gluconate dehydrogenase have been determined in cows, heifers, bulls and calves. The material consisted of 80 RDM, 46 SDM, 31 Jersey animals older than 3 months, 16 newborn RDM calves and 29 fetuses of various breeds.

1. In RDM averages of 322 ± 56 units G6PD and 52.8 ± 12.2 units 6PGD per 100 ml PGV were found. In SDM 330 ± 43 units G6PD and 47.1 ± 9.2 units 6PGD were found, and in Jersey 315 ± 39 units G6PD and 49.2 ± 8.0 units 6PGD per 100 ml PGV. The activities of both dehydrogenases were shown to be normally distributed in all three breeds.
2. The 6PGD activities in SDM were significantly lower than in RDM, but no other breed differences between the mean activities were detected.
3. There is no relation between the age (apart from fetuses and newborn calves), and milk yield of the animals and the activities of G6PD and 6PGD in the erythrocytes.
4. Statistical treatment of the results demonstrates that there is no relation between the G6PD and 6PGD activities in the blood samples, and that the enzyme activities in units per 100 ml PGV is not correlated with MCHG.
5. Further, the statistical treatment shows that when measuring G6PD and 6PGD activities in erythrocytes, PGV is to be preferred to hemoglobin concentration as parameter.
6. In bovine fetuses G6DP activities more than twice as high as in adult erythrocytes were found, while the 6PGD activities were only slightly higher than in adult erythrocytes. After birth the high G6PD activities decreased over a period of 40 days to the normal level for mature cattle. The 6PGD activity also attains the normal level for older animals in approximately 40 days after birth.

     

副干酪乳杆菌6-磷酸山梨醇脱氢酶(gutF)基因的克隆和超表达

为研究6-磷酸山梨醇脱氢酶基因在乳酸菌中超表达的影响,本试验根据副干酪乳杆菌的基因序列设计引物,用PCR的方法扩增6-磷酸山梨醇脱氢酶(gutF)基因,将其连接到乳酸菌表达载体pMG36e,并将重组质粒转化到副干酪乳杆菌及乳酸乳球菌中获得重组菌株;对重组菌株表达产物进行蛋白质水平的电泳检测,检测出表达了大约28kD的蛋白;使用山梨醇替代培养基中的葡萄糖,以未转化基因菌株做阴性对照,进行生长曲线的测定,结果表明,转化菌株能够利用在以山梨醇作为碳源的条件下,重组菌株的生长速度和最终菌浓度均优越于对照菌株。本研究通过超表达6-磷酸山梨醇脱氢酶蛋白,对于进一步研究乳酸菌的耐受性具有十分重要的理论基础。     

海南藏羊红细胞钾型的研究

采用火焰光度法对海南州９０只藏羊的红细胞钾型进行了研究。结果：高钾型羊６３只（７０％）,低钾型羊２７只（３０％）,Ｋｈ和ＫＬ等位基因频率分别为０．８３６７和０．１６３３     

Glucose-6-phosphate dehydrogenase and leptin are related to marbling differences among Limousin and Angus or Japanese Black x Angus steers

This work investigated the metabolic basis for the variability of carcass and i.m. adiposity in cattle. Our hypothesis was that the comparison of extreme breeds for adiposity might allow for the identification of some metabolic pathways determinant for carcass and i.m. adiposity. Thus, 23- to 28-mo-old steers of 3 breeds, 2 with high [Angus or Japanese Black x Angus (J. Black cross)] and 1 with low (Limousin) i.m. and carcass adiposity, were used to measure activities or mRNA levels, or both, of enzymes involved in de novo lipogenesis [acetyl-coA carboxylase, fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme], circulating triacylglycerol (TAG) uptake (lipoprotein lipase), and fatty acid esterification (glycerol-3-phosphate dehydrogenase), as well as the mRNA level of leptin, an adiposity-related factor. In a first study, enzyme activities were assayed in the s.c. adipose tissue (AT), the oxidative rectus abdominis, and the glycolytic semitendinosus muscles from steers finished for 6 mo. Compared with Angus or J. Black cross, Limousin steers had a 27% less (P = 0.003) rib fat thickness, and 23 and 29% less (P < or = 0.02) FAS and G6PDH activities in s.c. AT. In rectus abdominis and semitendinosus, the 75% less (P < 0.001) TAG content was concomitant with 50% less (P < 0.001) G6PDH activity. In a second study, enzyme activities plus mRNA levels were assayed in an oxido-glycolytic muscle, the longissimus thoracis (LT), in the i.m. AT dissected from LT, and in s.c. AT from the same Limousin steers and from Angus steers finished for 10 mo. Compared with Angus, the 50% less (P < 0.001) rib fat thickness in Limousin contrasted with the 1.1- to 5.8-fold greater (P < or = 0.02) mRNA levels or activities, or both, of acetyl-coA carboxylase, G6PDH, lipoprotein lipase, and glycerol-3-phosphate dehydrogenase in s.c. AT. Conversely, the 90% less (P < 0.001) TAG content in Limousin LT was concomitant to the 79 and 83% less (P < or = 0.002) G6PDH activity and leptin mRNA level. Such differences could arise from a greater number of adipocytes in LT from Angus steers because no difference was found between Limousin and Angus for G6PDH activity and leptin mRNA in i.m. AT. We conclude that FAS and G6PDH in s.c. AT could be involved in differences in carcass adiposity, but this relationship disappeared when the fatness increased strongly. Leptin and G6PDH are related to the expression of marbling whatever the body condition and thus could be relevant indicators of marbling in beef cattle.     

青海细毛羊血红蛋白和红细胞蛋白质多态性的研究

采用聚丙烯酰胺凝胶电泳法对４７９只青海细毛羊的血红蛋白和红细胞蛋白质的多态性进行研究。结果发现：①青海细毛羊的血红蛋白基因座上有ＨＢＡＡ，ＨＢＡＢ，ＨＢＡＣ和ＨＢＢＢ四种基因型。ＨＢ＾Ａ，ＨＢ＾Ｃ等位基因频率分别为０．２６４１，０．７３４９和０．００１０；②血红蛋白基因座的基因型频率和基因频率没有性别间差异；③青海细毛羊ＥＰ－１和ＥＰ－２是单态性蛋白质。     

Effects of Selenium Administration on Erythrocyte and Blood Plasma Glutathione Peroxidase Activity in Goats

The activity of glutathione peroxidase (GSH-Px, E.G. 1.11.1.9.) was determined in heparinized whole blood, blood plasma and washed erythrocytes from goats before and up to 4 weeks after the administration of selenium (0.4 mg/10 kg BW) and vitamin E (20 mg/10 kg BW) or only vit. E (20 mg/10 kg BW). It was found that Se administration caused a significant increase in enzyme activity in whole blood and washed erythrocytes first detected 2 weeks after the intramuscular injection of Se. No changes were observed in plasma from the treated animals. Minor and insignificant changes were seen in the vit. E treated control animals. It is concluded that GSH-Px activity in blood plasma or serum is of no value as a short-term indicator of the selenium status of goats but whole blood is a good indicator of the long-term status.     

乌蒙半细毛羊“锌缺乏症”的研究

试验旨在研究乌蒙半细毛羊"锌缺乏症"的病因。在赫章县兴发养殖场选取20只锌（Zn）缺乏的乌蒙半细毛羊作为试验组,在威宁县凉水沟种羊场选取20只健康的乌蒙半细毛羊作为对照组。采用电感耦合等离子体原子发射光谱法分析土壤、牧草和动物血液及肝脏中微量元素的含量,应用全自动血细胞分析仪测定血液参数,应用全自动生化分析仪测定血清生理生化指标。结果发现,试验牧场土壤及牧草中锌含量均极显著低于对照牧场（P<0.01）;试验组乌蒙半细毛羊血液和肝脏中Zn含量极显著低于对照组（P<0.01）,两组间其他元素含量均无显著差异（P>0.05）;试验组乌蒙半细毛羊血液中红细胞数（RBC）、淋巴细胞（LY）和中性粒细胞（NE）均极显著低于对照组（P<0.01）;血清中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）、乳酸脱氢酶（LDH）、碱性磷酸酶（AKP）活性均极显著低于对照组（P<0.01）,丙二醛（MDA）含量极显著高于对照组（P<0.01）,而两组间血红蛋白（Hb）、红细胞压积（PCV）、白细胞数（WBC）、血清尿素氮（BUN）含量和谷草转氨酶（AST）、谷丙转氨酶（ALT）、谷氨酰转肽酶（γ-GT）活性均无显著差异（P>0.05）。综上所述,乌蒙半细毛羊"锌缺乏症"主要由土壤和牧草锌含量过低引起,且在日粮中添加适量的硫酸锌可以防治乌蒙半细毛羊"锌缺乏症"的发生。     

链球菌Gapc基因检测及其序列分析

根据已发表的猪链球菌2型3-磷酸甘油醛脱氢酶(Gapc)基因保守区域设计并合成1对特异性引物,对10株不同来源链球菌Gapc基因进行PCR扩增,并选择PCR产物进行克隆并测序分析,结果在10株不同来源链球菌菌株中有8株可扩增出与预期大小(1011 bp)相一致的DNA片段;选取4株细菌PCR产物进行克隆测序,发现其片段大小为1009或1012 bp,与GenBank参考菌株Gapc基因序列的核苷酸同源性为85.3%~98.3%,氨基酸同源性为87.2%~99.4%;生物学软件分析结果显示Gapc基因在链球菌属细菌中相对保守,且具有较多的潜在抗原表位。本研究为链球菌检测技术和核酸疫苗的研究奠定了基础。     

顶复门原虫3-磷酸甘油醛脱氢酶功能及其应用研究进展

糖酵解途径广泛存在于各类生物中,是顶复门原虫的主要供能方式。3-磷酸甘油醛脱氢酶是糖酵解途径的重要酶,与顶复门原虫的生存密切相关,可以作为抗寄生虫药物研发的重要靶标。文章主要从顶复门原虫糖酵解途径、3-磷酸甘油醛脱氢酶的基因分析、作用机理及应用等方面进行综述。     

Congenital Hemolytic Anemia in the Basenji Dog Due to Erythrocyte Pyruvate Kinase Deficiency

Congenital hemolytic anemia in the Basenji dog resembles pyruvate kinase (PK) deficiency in man as it is characterized by an abbreviated erythrocyte life span, an intense reticulocytosis, type II autohemolysis and splenomegaly. Glucose utilization and lactate production were inadequate with respect to the immature cell population. Analysis of enzymes involved in erythrocyte glycolysis revealed a deficiency of pyruvate kinase.     

戈壁草原蒙古羊春季羔羊铜缺乏症的研究

[目的] 确定内蒙古苏尼特左旗北部地区春季发生的戈壁草原蒙古羊羔羊发病、死亡现象的主要原因。[方法] 选取3户羔羊发病情况最为严重的牧户采样,每户各采集5只发病羔羊的血液样本、放牧草场的饮用水样本,以及羔羊所处环境中3种主要牧草（红砂、珍珠猪毛菜和盐爪爪）样本和土壤样本,同时采集当地种羊场15只相同品种健康羔羊的样本作为对照,测定上述样本中的10种矿物质元素（钙、磷、铁、锰、铜、锌、镁、钼、硒、钴）的含量。[结果] 发病羔羊血液中铜和镁的含量显著（P<0.05）低于同品种健康羔羊,钼的含量显著（P<0.05）高于同品种健康羔羊,其他矿物质元素的含量差异不显著（P>0.05）。发病羔羊所处环境中饮用水的镁和钙含量较高。发病羔羊所处环境中红砂的钼、铁、锰和硒含量均高于文献报道值,其他矿物质元素含量均处于正常范围;盐爪爪的钼、铁、锰、硒、镁和钴的含量高于文献报道值,其他矿物质元素含量均处于正常范围;珍珠猪毛菜、红砂和盐爪爪的铜钼比分别为15.96∶1、0.23∶1和2.34∶1。发病羔羊所处环境中土壤样本的铜、铁、锰、硒、钴含量均低于文献报道值,镁、钙和磷含量均高于文献报道值,钼和锌含量在正常范围内。[结论] 该地区戈壁草原蒙古羊羔羊发病、死亡的原因主要是放牧羊摄入铜不足、钼超标、铁和锰等矿物质元素摄入量过多,导致羔羊对铜的吸收发生障碍,最终引起继发性铜缺乏症。     

Glutathione Peroxidase Activity and Erythrocyte Lipid Peroxidation as Indices of Selenium and Vitamin E Status in Young Pigs

A randomized, blocked 2³ factorial experiment was conducted with 48 pigs from sows fed a diet low in selenium and vitamin E. From 3 to 12 weeks of age the piglets were kept in single pens and fed a basic diet consisting mostly of barley, dried skim milk, soybean meal and dried yeast, and containing 55 µg selenium and 3 mg vitamin E per kg. The treatment factors — i.e. feed supplements — were 2 levels of Se (nil, 60 µg/kg), 2 levels of vitamin E (nil, 50 mg/kg), and 2 levels of the feed antioxidant ethoxyquin (nil, 150 mg/kg). Blood samples, collected at termination of the experiment, were examined for glutathione peroxidase activity (GSH-Px) and resistance against erythrocyte lipid peroxidation (ELP) to evaluate Se and vitamin E status, respectively. Analysis of variance showed the GSH-Px activity to be litter-dependent (P < 0.001) and influenced by selenium supplementation (P < 0.001) but not by the other supplements or by interactions between supplements. Resistance against ELP was influenced only by vitamin E supplementation (P < 0.001). GSH-Px and ELP thus seem to be valuable and simple methods for evaluating, respectively, Se status and vitamin E status in growing pigs.     

半胱胺对藏绵羊断奶羔羊瘤胃pH值、总脱氢酶及TVFA的影响

试验研究了半胱胺(Cs)对藏绵羊断奶羔羊瘤胃pH值、总脱氢酶及TVFA的影响。将10只(公母各半)80日龄左右、体重平均为(16.00±0.31)kg、装有永久性瘤胃瘘管的藏绵羊断奶羔羊随机分为2组(试验组和对照组),在日粮精料中添加300 mg/(kg·BW)的Cs,隔日添加1次,试验期42 d。正式试验当日及试验第7、14、21、28、35天的7:00、9:00、11:00、13:00、15:00、17:00、19:00、21:00、23:00、1:00、3:00、5:00于颈静脉采血样10 mL,同时采集瘤胃液10 mL,测定瘤胃pH值、总脱氢酶的活性和TVFA。检测结果表明,试验组精料中添加Cs使藏绵羊断奶羔羊瘤胃液pH值升高;血清尿素氮浓度降低,与对照组相比差异极显著(P0.01);瘤胃液总脱氢酶活力提高极其明显(P0.01);整个试验期内,半胱胺对瘤胃TVFA的影响极显著(P0.01)。     

三河马6-磷酸葡萄糖脱氢酶及葡萄糖磷酸异构酶的多态性

采用淀粉凝胶电脉及琼脂覆盖技术对三河马血液红细胞6－磷酸葡萄糖脱氢酶和葡萄糖磷酸异构入酶的电泳变异进行了检测。在6－磷酸葡萄糖脱氢酶位点发现3个表现型，即FF、FS和SS，在葡萄糖磷酸异构酶位点共发现2个表现型，即Ⅱ,FI。等位基因频率直接通过表型计算得出：PGD^F为0．840，PGD^为0．160，而PI^F为0．140，GPI^I为0．860。父权否定概率在6－PGD和GPI位点分别为0．1163和0．1059。