GABAB Receptor-Mediated Effects in Synaptosomes of Lethargic (lh/lh) Mice

Abstract: Previously, we have shown a significant increase in number of GABA_B receptor binding sites in neocortex and thalamus of lethargic ( lh/lh ) mice, a mutant strain exhibiting absence seizures. This study was performed to test our hypothesis that presynaptic GABA_B receptors would inhibit [³H]GABA release to a greater degree in lh/lh mice compared with their nonepileptic littermates (designated +/+). Synaptosomes isolated from neocortex and thalamus of age-matched male lh/lh and +/+ mice were similar in uptake of [³H]GABA. In the neocortical preparation, baclofen dose-dependently inhibited [³H]GABA release evoked by 12 m M KCl, an effect mediated by GABA_B receptors. The maximal inhibition ( I _max) value was significantly greater (80%) in lh/lh than +/+ mice, whereas the IC₅₀ (3 µ M ) was unchanged. In the thalamic preparation, the effect of baclofen (50 µ M ) was 58% less robust in lh/lh mice. Other effects mediated by GABA_B receptors (inhibitions in Ca²⁺ uptake and cyclic AMP formation) were also significantly reduced in thalamic synaptosomes from lh/lh mice. These data suggest a greater presynaptic GABA_B receptor-mediated effect in neocortex and a reduced effect in thalamic nuclei of lh/lh mice. It is possible that selective effects of presynaptic GABA_B receptors on GABA release in neocortex and thalamic nuclei of lh/lh mice may contribute to mechanisms underlying absence seizures.     

Brassinosteroid-induced rice lamina joint inclination and its relation to indole-3-acetic acid and ethylene

Brassinosteroid (BR)-induced rice (Oriza sativa L.) lamina joint (RLJ) inclination and its relationship to indole-3-acetic acid (IAA) and ethylene were investigated using BR isolated from beeswax. The effect of BR on RLJ inclination was time- and concentration-dependent. Etiolated lamina were more sensitive to BR than green lamina. The BR-induced inclination was accompanied by increased lamina fresh weight, total water content, free-water content, proton extrusion and ethylene production, and decreased bound-water content. Lamina dry weight was not changed. The inclination was due to greater expansion of the adaxial cells relative to the dorsal cells in the lamina joint. This response was caused by BR and/or BR-induced signal(s) that were transported from the leaf sheath to the leaf blade. Both BR-induced RLJ inclination and ethylene production were inhibited by cobalt chloride (CoCl₂), an inhibitor of ACC oxidase. BR-induced inclination was much higher than that of IAA, and was inhibited by high concentration of 2,3,5-triiodobenzoic acid (TIBA), an inhibitor of IAA transport. A synergistic effect was observed between BR and IAA. These results suggest that the effects of BR on RLJ inclination and pulvinus cell expansion may be resulted from BR-increased water potential and proton extrusion in the lamina. The BR-induced RLJ inclination may involve the action of ethylene but may be independent of IAA.Abbreviations BR brassinolide or brassinosteroid(s) - IAA indole-3-acetic acid - TIBA 2,3,5-triiodobenzoic acid - RLJ rice lamina joint     

地衣芽孢杆菌β－甘露聚糖酶的纯化及酶学性质

地衣芽孢杆菌（Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ）ＮＫ－２７菌株发酵产生的β－甘露聚糖酶（β－ｍａｎｎａｎａｓｅ）经硫酸铵盐析沉淀,两次ＤＥＡＥ纤维素和ＳｅｐｈａｄｅｘＧ－１００离子交换柱层析以及制备ＰＡＧＥ筹步骤,获得了凝胶电泳均一的样品。用ＳＤＳ－凝胶电泳测得纯化后的β－甘露聚糖酶分子量为２６ｋＤ,用凝胶聚焦电泳测得等电点ＰＩ为５．０。酶反应的最适ｐＨ为９．０,最后温度为６０℃,稳定ｐＨ为６．０—９．０,稳定温度为４０℃。金属离子中Ｍｇ￣（２＋）、Ｃａ￣（２＋）、Ｆｅ￣（２＋）、Ｎｉ￣（２＋）对该酶有一定的激活作用;而Ｓｎ￣（２＋）、Ｚｎ￣（２＋）、Ａｌ￣（３＋）、Ａｇ￣＋和Ｈｇ￣（２＋）对该酶有强烈的抑制作用。ＮＫ－２７菌株的β－甘露聚糖酶对魔芋葡萄甘露聚糖和角豆胶半乳甘露聚糖的Ｋｍ值分别为７．１４和５．５６ｍｇ·ｍｌ￣（－１）;Ｖ＿（ｍａｘ）分别为２００．５３和１５７．４５μｍｏｌ·ｍｇ￣（－１）·ｍｉｎ￣（－１）。     

Predictions of protein backbone bond distances and angles from first principles

A procedure is described, based on a spline-function representation of ab initio peptide conformational geometry maps, that allows one to predict backbone bond distances and angles of proteins as functions of the peptide ?(N-C^α)/Ψ(C^α-C′) torsions with an accuracy comparable to that of high-resolution protein crystallography. For example, for the more than 40 residues of crambin, the rms deviation between predicted and crystallographic values of N-C^α-C′ is 1.9° for the 1.5 Å resolution structure and 1.9° for the 0.83 Å resolution structure, compared with angle variations of < 10°. Accurate information on protein backbone geometries is important for establishing dictionaries of flexible geometry functions for use in empirical peptide and protein modeling. © 1995 John Wiley & Sons, Inc.     

丁酸钠对CHO-EPO工程细胞株rhEPO表达量的影响

以稳定整合有ｐＥＤＥＰＯ的ＣＨＯＥＰＯ工程细胞株为研究对象,在无血清条件下,系统观察了０５、１０、２５和５０ｍｍｏｌ／Ｌ４个浓度的丁酸钠作用于该细胞株的情况,结果表明：丁酸钠对ＣＨＯＥＰＯ工程细胞的生长有明显的抑制作用;影响ＣＨＯＥＰＯ工程细胞ＥＰＯ表达,浓度１０ｍｍｏｌ／Ｌ可提高ＥＰＯ表达量２５倍左右,并可持续较长的一段时间;延缓ＣＨＯＥＰＯ工程细胞在无血清培养时的细胞脱落;提高ＣＨＯＥＰＯ工程细胞ＥＰＯｍＲＮＡ水平     

Partial purification of two forms of Choline kinase and separation of Choline kinase from sphingosine kinase of rat brain

Choline kinase of rat brain was purified approximately 200,000 fold using acid precipitation, ammonium sulphate fractionation, Q-Sepharose, Octyl-Sepharose and AH-Sepharose chromatography. The ability of this enzyme to catalyze the phosphorylation of choline, ethanolamine (Etn), monomethylethanolamine (MeEtn), dimethylethanolamine (Me₂Etn) and sphingosine was investigated. Choline kinase was separated from sphingosine kinase. The fraction with highly purified choline kinase had four major polypeptides with different molecular masses and possessed activities towards choline, Etn, MeEtn and Me₂Etn. Two forms of choline kinase were obtained when the enzymatically active fractions eluted from the Q-Sepharose column were subjected to a horizontal isoelectrofocusing electrophoresis. One form focused around pH 4.7 and is able to phosphorylate choline, Etn, MeEtn and Me₂Etn. The other form focused around pH 10 and possessed only choline kinase activity. The latter form of choline kinase did not display classical Michaelis-Menten's mechanism but revealed a positive co-operative pattern for two choline binding sites. This form was purified to apparent homogeneity with a approximate molecular mass of 14.4 kDa.Abbreviations Etn ethanolamine - MeEtn N-monomethylethanolamine - Me₂Etn N, N-dimethylethanolamine     

应用CTB基因启动子及信号肽序列构建分泌性表达系统

利用霍乱毒素B亚基基因的启动子、信号肽序列及ctx操纵子的转录终止信号构建了分泌性表达的质粒载体pMCOSS。Β－半乳糖苷酶基因克隆至霍乱毒素B亚基基因的信号肽序列下游后能得到高效分泌性表达。不同的宿主菌和培养基成分中对β-半乳糖苷酶的表达产量有较大的影响,以MM2为宿主菌、在玉米浆培养基中β－半乳糖苷酶的表达产量达4 lOOu／ml,产物的大部分分泌至细胞的周质,活力测定的结果与SDS—PAGE电泳测定结果基本一致,说明表达的β－半乳糖苷酶绝大部分都具有酶活性。构建的蛋白质分泌性表达的载体－宿主系统及合适的培养条件为易形成包含体的蛋白质的高效表达提供了一条新的途径。     

氮磷对污水净化中藻类叶绿素含量的影响

在室内模拟生物净化槽中比较研究了氮磷对污水中藻类叶绿素含量和污水净化的影响。污水中磷含量均为７ｍｇ／Ｌ左右,氮含量分别为７７．４、４４．４、２４．８和１３．５ｍｇ／Ｌ,结果发现ＴＮ／ＴＰ＝７７．４／７．０１ｍｇ／Ｌ组,污水经７ｄ净化,藻类叶绿素含量最高,污水净化效果较好。四个实验组比较,叶绿素含量随ＴＮ／ＴＰ比例的上升而上升,呈显著正相关。     

Improved resolution with one-dimensional polyacrylamide gel electrophoresis: myofibrillar proteins from typed single fibers of human muscle

Standard procedures for one-dimensional discontinuous sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and silver staining were modified to give more effective separation and an improved resolution of human skeletal muscle proteins. In this system, an electrophoresis buffer composed of 100 mM L-isoleucine, 25 mM Tris base, and 0.1% SDS was used. The separating gel consisted of 16% acrylamide with N,N'-methylenebisacrylamide as a crosslinker (1:23), 0.4% SDS, 1.5 M Tris-HCl, pH 8.8. By the present procedure, the slow and the fast forms of myosin light chains (LCs, LCf) and other contractile proteins from human muscle could be better separated. The silver stain is based on a combination of methods previously described. The modified method requires a small fragment of a single fiber to observe as few as 10 ng of myofibrillar muscle proteins. The described simplifications made it possible to assay and compare up to 40 single fibers in the same electrophoretic run. Improved separation of other proteins migrating at basic pH could be achieved by a similar approach.     

机械损伤对9种水生植物叶片的影响

水生植物叶片的功能性状特征与陆生植物有所不同,同时叶脉类型也显著影响叶片的功能性状。本研究选取9种具有不同叶脉类型的水生植物,通过对叶脉进行直接损伤,分析叶片性状（形态、色素含量和叶绿素荧光指标）在叶脉受损后的变化程度与叶脉类型的关系。结果显示：具有平行脉的3种水生植物对叶脉损伤具有较强的耐受性;具羽状脉的4种植物主脉受损后显著抑制叶片生长,而侧脉受损的影响在不同物种间有所不同,具有物种特异性。本研究可为大型湖泊水生植物修复的水生物种筛选提供参考。