Monoclonal antibodies specific for type 3 protein kinase C recognize distinct domains of protein kinase C and inhibit in vitro functional activity

Monoclonal antibodies (mAbs) which distinguish Type 3 protein kinase C (PKC) from Types 1 and 2 have been obtained from mice immunized with purified Type 3 PKC from rabbit brain cytosol. Most of these mAbs (seven out of eight) selectively recognize Type 3 versus Types 1 and 2 PKC in both enzyme-linked immunosorbent and immunoblot assays. Trypsin treatment of Type 3 PKC reduced the immunoreactivity with 82-kDa PKC and generated immunoreactive fragments of 45 and 35 kDa. The mAbs can be divided into two classes based on their ability to recognize the 45-kDa catalytic fragment (5/8) or the 35 kDa regulatory domain fragment (3/8). Each of the mAbs inhibits phosphorylation of histone or lipocortin by PKC, although the extent of the inhibition varied. Only those mAbs that recognize the 35-kDa regulatory domain inhibited phorbol ester binding. The inhibition of both kinase and binding activities by this group of mAbs was sensitive to the concentration of phospholipid used in the assay. This functional inhibition suggests that these mAbs may be useful for defining the phospholipid binding domain(s) of Type 3 PKC. The mAbs recognized 82-kDa PKC in a variety of cell types; the presence of smaller molecular weight fragments was not consistently found. Distinct immunofluorescence staining patterns were observed with mAbs directed toward different epitopes, suggesting that there may be heterogeneity in the subcellular localization of PKC. The type specificity of these mAbs will make them valuable tools for studying activation and regulation of Type 3 PKC in cell culture model systems.     

Studies on the phospholipid composition of pathogenic cell membranes of Mycoplasma hyopneumoniae

The membrane phospholipid and fatty acid compositions of Mycoplasma hyopneumoniae, a pathogen of porcine enzootic pneumoniae isolated in China, was studied by thin-layer chromatography and gas chromatography. The results showed that membrane phospholipids consisted predominantly of diphosphatidylglycerol. The percentage of C16 - C18 fatty acids comprised 79% of the total fatty acids, of which oleic acid as well as palmitic acid are the major fatty acids. Some differences were shown in fatty acid composition as compared with membranes of other species of Mycoplasma.     

血友病患者血清中淋巴腺病病毒/人T细胞Ⅲ型病毒抗体检测

对浙江省1982～1984年注射了美国产浓缩Ⅶ因子制剂的18例血友病患者,用酶联免疫吸附法(ELTSA)检测了血清中淋巴腺病病毒/人T细胞Ⅲ型病毒(LAV/HTLV-Ⅲ)抗体,发现4例阳性,并经免疫荧光试验和Western印迹法证实。2例应用了国产浓缩Ⅷ因子者抗体阴性。一例从美国来华旅游死于艾滋病者,LAV/HTLV-Ⅲ抗体阳性。本研究证明,LAV/HTLVⅢ病毒巳通过美国生产的Ⅷ因子制剂传入中国。     

HBsAg合成肽段的免疫源性和抗肽血清与天然HBsAg的结合测定

我们将三种乙型肝炎表面抗原的合成肽段在联结载体或不接载体的情况下,分别制备成4种免疫原对兔子进行了免疫。大部分兔子都产生了抗肽抗体。其中抗P_(122-48)(auw)与抗P_(122-148)(adw)都能与天然HBsAg反应,而抗P_(122-148)(adw)的抗HBs活力比文献报道的都高,通过结构分析表明:含有天然蛋白上免疫显性区域的免疫原是合成疫苗的理想选者。     

体外分化的人体巨噬细胞抑制天然杀伤(NK)细胞的研究

本文继先前工作后,进一步应用正常健康人外周血单个核细胞(PBMNC)经塑料培皿粘附技术把单核细胞分离出来,经培养进一步纯化,随后动态观察培养0,2,4,6和8天的单核-巨噬细胞的形态变化和对新鲜分离同种异基因个体PBMNC中NK活性的影响。实验表明,体外分化6天和8天的巨噬细胞质/核比例和胞浆内空泡显著增加,细胞直径约为0天时的2倍。这些细胞和PBMNC之比为0.5:1时,引起了NK细胞活性的50%以上抑制(4小时~(51)Cr标记K 562肿瘤的同位素释放试验)。这种抑制效应不为过氧化氢酶(Catalase 4000单位/毫升)和前列腺素合成酶的抑制剂(Indom 1×10~(-5)M)所阻断。实验证明,同种异基因个体的NK细胞不能识别巨噬细胞表面抗原,从而排除了巨噬细胞和K562肿瘤抗原竞争的可能性。实验还表明,巨噬细胞对NK活性的抑制是不受HLA约束的。应用高频超声振荡破碎巨噬细胞膜方法和免疫调变技术进一步提示,人体巨噬细胞对NK活性的抑制与巨噬细胞体积无关,而与体外分化所赋有的固有特性和它们分泌的免疫调节分子有关。     

大萼香茶菜丁素的化学结构

从大萼香茶菜叶中又分得一个具有细胞毒活性的新的二萜类化合物,命名为大萼香茶菜丁素(macrocalyxin D)。根据光谱和化学数据鉴定其化学结构为[3]。     

百合花粉母细胞间细胞融合期间腺苷三磷酸酶活性的细胞化学定位及其与染色质胞间转移的关系

用标准的磷酸铅沉淀的细胞化学方法,对百合花粉母细胞间染色质穿壁运动期间及其前后三个时期中的腺苷三磷酸酶(ATP 酶)活性进行了超微结构的定位。结果表明:(1)在穿壁前,ATP 酶活性主要定位于质膜、胞间连丝及细胞间隙;在内质网、高尔基体、质体和某些局部的基质(groundplasm)中,也表现有 ATP 酶活性反应的产物;但在染色质和核仁中,一般都没有这种反应。(2)在穿壁时,染色质从一个细胞穿壁转移到另一个相邻细胞,同时看到染色质和核仁内出现密集的 ATP 酶活性反应产物;在内质网和高尔基体的腔内以及质体的片层上也产生明显的 ATP 酶活性反应;而在质膜、胞间连丝及细胞间隙内 ATP 酶活性明显降低,甚至看不到明显的活性反应。(3)在穿壁后,质膜及细胞间隙中又产生明显的 ATP 酶活性反应产物,但核内染色质上的 ATP 酶活性则显著降低,而核仁内则仍有较高的活性。同前二个时期一样,内质网、高尔基体和质体上的 ATP 酶仍表现明显的活性反应。最后讨论了三个不同发育时期 ATP 酶活性及其分布部位的改变与染色质胞间转移的关系。     

四川自贡大山铺蜀龙动物群——简报Ⅴ 两栖类

本文对扁头中国短头鲵(新属新种) (Sinobrachyops placenticephalus gen. et sp. nov.) 的形态特征和分类位置进行简述.标本得自著名的恐龙化石产地——自贡大山铺,产出时代为中侏罗世.中国短头鲵是目前迷齿类中在地史上最年轻的一个属.它的发现使迷齿类在地球上生存时代的上限推移到中侏罗世.     

青海贵德、共和两盆地晚新生代哺乳动物

本文记述了青海省贵德及共和两盆地的几个含晚新生代哺乳动物化石地点的地层剖面及采自18个地点的26种(代表20个属)哺乳动物化石.对其中的 Myospalax arvicolinus, Microtus, Anancus, Leptobos crassus gonghenensis subsp. nov., L. vallisarni, Bison (Superbison) crassicornis, Boopsis breviceps作了较详细的描述和讨论.通过对动物群的分析初步确定了盆地主要堆积物的时代——贵德盆地:上新世—早更新世;共和盆地:早—中更新世,并简略探讨了两盆地的发育历史.     

新疆石河子地区印度麻花叶病植株病毒分离物的鉴定

1984年,从新疆石河子农学院实验站印度麻花叶病植株上,分离到一株病毒分离物Sc-1,经汁液摩擦接种试验表明,它可以侵染10种豆科植物和2种藜科植物。在印度麻、蚕豆、豌豆、箭舌豌豆、扁豆、山藜豆、田菁和红三叶草上引起系统花叶,在豇豆上产生局部枯斑和系统花叶,在苋色藜、昆诺藜上表现为系统黄斑。失毒温度为55～60℃,稀释限点10~(-3)～10~(-4),体外保毒期3～4天。可经汁液和蚜虫传播,不通过种子传毒。病毒粒体为线条状,大小为13～15×750nm。光学显微镜检查可见,病叶表皮细胞内形成不定形的内含体。电镜下可见风轮状、环状内含体。分离物Sc-1与菜豆黄色花叶病毒(BYMV)抗血清呈阳性反应。我们将Sc-1归为菜豆黄色花叶病毒(BYMV),且为豇豆株系。