New features of site-specific horseradish peroxidase (HRP) glycosylation uncovered by nano-LC-MS with repeated ion-isolation/fragmentation cycles

Horseradish peroxidase (HRP) is widely used in biomedical research as a reporter enzyme in diagnostic assays. In addition, it is of considerable interest as a model glycoprotein with core-xylosylated and -(alpha1-3)-fucosylated N-glycans that form antigenic elements of plant allergens and parasitic helminths. Using a combination of techniques comprising (1) nano-liquid chromatography (LC)-mass spectrometry (MS)/MS with multiple selection/fragmentation cycles of HRP tryptic (glyco-)peptides, (2) nano-electrospray MS of intact HRP, and (3) carbohydrate linkage analysis, it was revealed that most of the HRP N-glycosylation sites can be occupied with an alternative Fuc(1-3)GlcNAc-disaccharide. Two main variants of HRP occur: The major population (approximately 60%) has eight glycosylation sites carrying core(1-3)fucosylated, xylosylated, trimannosyl N-glycans, with the ninth potential N-glycosylation site Asn316 not occupied. Another group of HRP carries seven of the above-mentioned N-glycans, with an eighth N-glycosylation site carrying the alternative Fuc(1-3)GlcNAc-unit (approximately 35%). In addition, minor subsets of HRP were found to contain a xylosylated, trimannosyl N-glycan lacking core-fucosylation as a ninth N-glycan attached to Asn316, which has hitherto been assumed to be unoccupied. The finding of these new features of glycosylation of an already exceptionally well-studied glycoprotein underscores the potential of the nano-LC-MS(n) based analytical approach followed.     

Structure based approach to the design of bicyclic-1H-isoindole-1,3(2H)-dione based androgen receptor antagonists

A novel series of isoindoledione based compounds were identified as potent antagonists of the androgen receptor (AR). Co-crystallization of members of this family of inhibitors was successfully accomplished with the T877A AR LBD. A working model of how this class of compounds functions to antagonize the AR was created. Based on this model, it was proposed that expanding the bicyclic portion of the molecule should result in analogs which function as effective antagonists against a variety of AR isoforms. In contrast to what was predicted by the model, SAR around this new series was dictated by the aniline portion rather than the bicyclic portion of the molecule.     

西藏冷杉原始林林隙对草本植物和灌木树种多样性的影响

研究了西藏东南部亚高山原始冷杉林林隙对草本植物和灌木树种多样性的影响．结果表明,西藏东南部亚高山原始冷杉林林隙与非林隙林分内草本植物和灌木树种的种类组成、数量、密度和物种多样性均存在较大的差异．由于林隙内光照条件的改善,草本植物和灌木树种更加丰富;林隙内有喜光的草本植物和灌木,也有耐荫性较强的草本植物和灌木,而非林隙林分内仅有耐荫和较耐荫的草本植物和灌木;大部分林隙内的草本植物和灌木树种的物种多样性和均匀度大于非林隙林分;林隙内与非林隙林分的群落相似系数较小,表明林隙与非林隙林分草本植物和灌木群落组成存在差异．林隙大小在174．2～562．2m^2之间,群落相似系数与林隙大小无明显关联．林隙促进了草本植物和灌木树种的繁衍,在维持藏东南亚高山原始冷杉林的物种多样性方面起着重要作用．     

环境因子对豆科共生固氮影响的研究进展

环境因子的限制一直是豆科植物一根瘤菌共生固氮体系没有在农业生产中充分发挥作用的重要原因之一。目前,研究涉及的环境因子主要行水分、矿质营养元素、温度、重金属、钠盐、CO2、土壤类型以及pH等。水分胁迫会导致豆科植物根瘤减少和固氮效率低下;矿质元素方面,除氮磷钾外,微量死素对固氮影响也很明显;不适的温度会对豆科植物的结瘤固氮产生一定的限制;重金属能从不同方面直接和间接地影响共生同氮,寻找适合作尾矿先锋植物的豆科植物是当前的一个研究热点。本文除详细阐述了这方面开展的研究以外,还浅析了这方而研究目前国内外存在的一些主要问题和发展趋势。     

水分胁迫和氮肥对花生根系形态发育及叶片生理活性的影响

以"花育22号"花生为试验材料,在中度干旱胁迫和充足灌水两个水分条件下,分别设置不施氮肥(N0)、中氮(N1,90 kg·hm-2)、高氮(N2,180 kg·hm-2)3个施氮水平,研究不同土壤水分和氮肥条件对花生叶片生理活性及根系形态发育特征的影响.结果表明:与不施氮肥处理相比,两个水分条件下中氮处理均显著增加花生产量,但对收获指数无显著影响.干旱胁迫条件下,中氮处理对总根系生物量和总根长无显著影响,但显著增加花生总根系表面积;中氮和高氮处理均显著增加20~40 cm土层内根长和根系表面积,且高氮处理显著增加40 cm以下土层内根系生物量和根系表面积;施用氮肥显著提高叶片过氧化氢酶(CAT)和过氧化物酶(POD)活性,而丙二醛(MDA)含量随施氮量的增加而降低.正常供水条件下,施用氮肥显著降低了花生根系表面积和40 cm以下土层内根系生物量、根长和根系表面积,中氮处理可提高叶片保护酶活性.相关性分析表明,20~40 cm土层内根长和叶片超氧化物歧化酶(SOD)、CAT、POD活性与产量呈显著相关.     

少数民族医学生医学微生物学实验教学的改革与实践——以酸奶制备为例

医学微生物学实验教学中基本操作是双基训练的重要部分,学生对实验技术的掌握程度决定了实验教学的成败。通过酸奶制备的一系列实验过程,把实验内容由验证性改变为设计性,激发了学生的学习兴趣,动手能力提高,实验教学效果良好。     

The influence of season on oxidant-antioxidant status in trained and sedentary subjects

The association between oxidative stress and cardiovascular diseases is a widely accepted fact today. Generally, men have a higher risk of cardiovascular incidents and mortality from acute myocardial infarction and strokes. We have examined sport-associated circannual rhythms of oxidant and antioxidant processes by measuring plasma LPO, erythrocyte SOD, CAT, Gpx activity and plasma hormonal status in both sedentary and long-term trained men and women. We have shown seasonal variations in both oxidant and antioxidant status in all examined groups. The largest difference was observed in the oxidant status between sedentary men and women during autumn and winter, which is considered a period of high coronary risk for men. Sport decreased LPO in trained men in autumn, while the same effect in trained women was shifted towards summer. These data state that regular, long-term physical exercise training induces adaptive responses that confer protection against oxidative stress, as well as the beneficial effect of exercise with regard to season, particularly in men during a period of high coronary risk (autumn and winter, respectively) and in women during summer.     

IPSE/alpha-1, a major secretory glycoprotein antigen from schistosome eggs, expresses the Lewis X motif on core-difucosylated N-glycans

Schistosomes are parasitic flatworms that infect millions of people in (sub)tropical areas around the world. Glycoconjugates of schistosomes play a critical role in the interaction of the different developmental stages of the parasite with the host. In particular, glycosylated components of the eggs produced by the adult worm pairs living in the bloodstream are strongly immunogenic. We have investigated the glycosylation of interleukin-4-inducing factor from schistosome eggs (IPSE/alpha-1), a major secretory egg antigen from Schistosoma mansoni that triggers interleukin-4 production in human basophils, by MS analysis of tryptic glycopeptides. Nanoscale LC-MS(/MS) and MALDI-TOF(/TOF)-MS studies combined with enzymatic degradations showed that monomeric IPSE/alpha-1 contains two N-glycosylation sites, which are each occupied for a large proportion with core-difucosylated diantennary glycans that carry one or more Lewis X motifs. Lewis X has been reported as a major immunogenic glycan element of schistosomes. This is the first report both on the expression of Lewis X on a specific schistosome egg protein and on a protein-specific glycosylation analysis of schistosome eggs.     

Met-enkephalin modulation of age-related changes in red cell antioxidant status

Opioid peptides have been recognized as modulators of reactive oxygen species (ROS) in mouse macrophages and human neutrophils. Since the effect cannot be ascribed to its direct scavenger properties, in this study, we tested the hypothesis that methionine-enkephalin (MENK) modulates ROS by alteration of antioxidant enzyme activity (AOE). For this purpose superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) are measured in red blood cells of 1, 4, 10, and 18-month-old CBA mice of both sexes injected with 10 mg/kg MENK. The results indicate that MENK-affected antioxidant enzyme activity of red blood cells is age- but not sex-related. The most abundant effects were observed at the reproductive stage. Increased sensitivity to oxidative stress by opioid peptides was in both sexes mainly due to increased SOD activity followed by GPX decrease. Thus, the damage ascribed to opioid peptides might be, at least partly, ascribed to deleterious effects of accumulated hydrogen peroxide (H2O2).