The mechanism for the effect of selenium supplementation on immunity

Lipid peroxide (LPO) in lymphocytes from mice was evaluated by measuring substances reactive to thiobarbituric acid (TBA). The product resulting from the reaction of TBA with lymphocytes was extracted with n-butyl and fluorescence intensity was determined. The degree of lipid peroxidation, expressed as fluorescence intensity f547, was assessed for stimulation of lymphocytes with concanavalin A (Con A), and was related to lymphocyte proliferation in response to Con A if Se was administered. The lymphocyte proliferation was determined by [³H]thymidine incorporation, expressed as cpm. The effect of superoxide dismutase (SOD), added to cell culture on lymphocyte proliferation was also evaluated. It was found that LPO in lymphocytes before Con A stimulation was significantly less than that after stimulation (p<0.001), and that SOD promoted lymphocyte proliferation dose dependently. The addition of Na₂SeO₃ to lymphocyte culture or supplementation in drinking water to mice decreased the produced LPO in lymphocyte in response to Con A. In the presence of Se, there is an inverse correlation between the levels of LPO in lymphocyte and the stimulated proliferation (r=−0.8902,r=−0.9439). In conclusion, active oxygen species scavenging was proposed as one of the mechanisms for Se to promote immunity.     

原生质体电融合酵母多倍体生理特性的研究

对原生质体电融合技术获得的11株酵母多倍体融合株,进行了一系列生理特性的分析比较．结果发现,在电融合过程中,融合株的细胞体积及DNA含量并不随着核倍性呈线性递增．而是有其特殊的变化规律。当糖化酵母sta1、sta2和sta3在细胞核中各自纯合时,明显表现出表达的剂量效应。其中sta1、sta2纯合的融合株．在YEPS培养中,GA分泌可被淀粉大量诱导,表现出一定的二次生长特性。从分析结果推测,在亲株5301-14D及HU-TY—1A中可能有对GA分泌不利的因素。     

Oxidative damage to 5-methylcytosine in DNA.

Exposure of pyrimidines of DNA to ionizing radiation under aerobic conditions or oxidizing agents results in attack on the 5,6 double bond of the pyrimidine ring or on the exocyclic 5-methyl group. The primary product of oxidation of the 5,6 double bond of thymine is thymine glycol, while oxidation of the 5-methyl group yields 5-hydroxymethyluracil. Oxidation of the 5,6 double bond of cytosine yields cytosine glycol, which decomposes to 5-hydroxycytosine, 5-hydroxyuracil and uracil glycol, all of which are repaired in DNA by Escherichia coli endonuclease III. We now describe the products of oxidation of 5-methylcytosine in DNA. Poly(dG-[3H]dmC) was gamma-irradiated or oxidized with hydrogen peroxide in the presence of Fe3+ and ascorbic acid. The oxidized co-polymer was incubated with endonuclease III or 5-hydroxymethyluracil-DNA glycosylase, to determine whether repairable products were formed, or digested to 2'-deoxyribonucleosides, to determine the total complement of oxidative products. Oxidative attack on 5-methylcytosine resulted primarily in formation of thymine glycol. The radiogenic yield of thymine glycol in poly(dG-dmC) was the same as that in poly(dA-dT), demonstrating that 5-methylcytosine residues in DNA were equally susceptible to radiation-induced oxidation as were thymine residues.     

植物蛋白质S-亚硝基化修饰的检测与分析

S-亚硝基化是一种重要的蛋白质翻译后修饰方式, 是指一氧化氮(NO)基团共价连接至靶蛋白特定半胱氨酸残基的自由巯基, 从而形成S-亚硝基硫醇(SNO)的过程。S-亚硝基化修饰广泛存在于各有机体中, 通过改变蛋白质生化活性、稳定性、亚细胞定位以及蛋白质-蛋白质相互作用等机制而调控不同的生物学过程或信号通路。在蛋白质S-亚硝基化检测分析方法中, 最为广泛使用的是生物素转化法(biotin switch assay), 其基本原理是首先封闭未被修饰的自由巯基, 进而将被修饰的SNO基团特异地还原为自由巯基并使用生物素将其特异标记。被生物素标记的半胱氨酸残基(即被修饰位点)可进一步通过蛋白质免疫印迹和/或质谱等方法进行检测分析。该文详细描述了植物蛋白质样品的体内和体外生物素转化法的实验流程, 并对实验过程中的注意事项进行了讨论。     

用亲和层析技术分离纯化天冬氨酸酶

<正> 亲和层析技术做为现代生化实验室常用的提纯方法,具有专一性强,活力损失小,分离步骤少等优点。天冬氨酸酶是重要的酶制剂,以往见报的分离纯化工艺步骤繁琐,活力损失大。本文报道的新工艺是用聚乙二醇(PEG)/磷酸钾双水相抽提;DEAE-sophadex A-50柱层析除PEG和部分杂蛋白;环氧氯丙烷活化载体,底物作配基的亲和层析技术获得了电泳纯的天冬氨酸酶。     

草原生态学研究方法学术讨论会在成都召开

中国植物学会于1981年11月21日至27日在四川省成都市召开了有61名代表参加的草原生态学研究方法学术讨论会。大会收到包括植物群落结构调查研究、第一性生产力测定、第二性生产力测定、光合作用测定、物质与水分循环、热值测定、数学生态等有关内容的研究方法论文和报告34篇。其中有18篇论文在大会上作了报告。在小组讨论中,代表们就第一性生产力测定、水分与物质循环、光合作用测定     

Incorporation of the transmembrane hydrophobic domain of glycophorin into small unilamellar phospholipid vesicles Ion Flux studies

Human erythrocyte glycophorin is one of the best characterized integral membrane proteins. Reconstitution of the membrane-spanning hydrophobic segment of glycophorin (the tryptic insoluble peptide released when glycophorin is treated with trypsin) with liposomes results in the production of freeze-fracture intrabilayer particles of 80 Å diameter (Segrest, J.P., Gulik-Krzywicki, T. and Sardet, C. (1974) Proc. Natl. Acad. Sci. U.S.A. 71, 3294–3298), with particles appearing at or above a tryptic insoluble peptide concentration of 4 mmol per mol phosphatidylcholine. In the present study, increasing concentrations of tryptic insoluble peptide were added to sonicated small unilamellar egg phosphatidylcholine vesicles and the rate of efflux of ²²Na⁺ was examined by rapid (30 s) gel filtration on Sephadex G-50. Below a concentation of 3–5 mmol tryptic insoluble peptide/mol phosphatidylcholine, ²²Na⁺ efflux occurs at a constant slow rate at given tryptic insoluble peptide concentrations. Above a concentration of 3–5 mM, the rate of efflux is biphasic at given tryptic insoluble peptide concentrations, exhibiting both an initial fast and a subsequent slow component. On the basis of graphic and computer curve-fitting analysis, with increasing tryptic insoluble peptide concentration, the rate of the slow component reaches a plateau at a tryptic insoluble peptide concentration of 3–5 mM and remains essentially constant until much higher concentrations are reached; the fast component increases linearly with increasing tryptic insoluble peptide concentration well beyond 5 mM. The most consistent interpretation of this data is as follows. The slow ²²Na⁺ efflux component is due to perturbations of small unilamellar vesicle integrity by tryptic insoluble peptide monomers. At a tryptic insoluble peptide concentration of 3–5 mmol/mol, a critical concentration is reached following which there is intrabilayer tryptic insoluble peptide self-association. The fast ²²Na⁺ efflux component is due to the increasing presence of tryptic insoluble peptide self-associated multimers the 80-Å particles seen by freeze-fracture electron microscopy) which results in a significantly larger bilayer defect than do tryptic insoluble peptide monomers. The failure of complete saturation of efflux by the fast component is ascribed to the presence of two populations of small unilamellar vesicles, some of which contain tryptic insoluble peptide multimers and some of which do not.Addition of cholesterol to the tryptic insoluble peptide/phosphatidylcholine vesicles decreases the rate of ²²Na⁺ efflux by inhibiting primarily the fast component. Freeze-fracture electron microscopy indicates that the presence of cholesterol has no effect on the size, number or distribution of 80-Å intra-bilayer particles in the tryptic insoluble peptide/phosphatidylcholine vesicles. These results are consistent with a mechanism to explain the fast Na⁺ efflux component involving protein-lipid boundary perturbations.Efflux of ⁴⁵Ca²⁺ from phosphatidylcholine vesicles is also enhanced by incorporation of tryptic insoluble peptide, but only if divalent cations (Ca²⁺ or Mg²⁺) are present in the external bathing media as well as inside the sonicated vesicles. If monovalent Na⁺ only is present in the bathing media no ⁴⁵Ca²⁺ efflux is seen. Under conditions where ⁴⁵Ca²⁺ efflux is seen, both a fast and a slow component are present, although both appear lower than corresponding rate constants for ²²Na⁺ efflux. These results suggest a coordinated mechanism for ion efflux induced by tryptic insoluble peptide and, together with the ²²Na⁺ efflux studies, may have mechanistic implications for the transbilayer phospholipid exchange (flip-flop) suggesed to be induced at glycophorin/phospholipid interfaces (de Kruiff, B., van Zoelen, E.J.J. and van Deenen, L.L.M. (1978) Biochim. Biophys. Acta 509, 537–542).