A Mitosis-specific Phosphorylation of the Gap Junction Protein Connexin43 in Human Vascular Cells: Biochemical Characterization and Localization

Western blotting studies revealed that connexin43 (Cx43), one of the major gap junction proteins in human vascular endothelial cells, is posttranslationally modified during mitosis. This mitosis-specific modification results in a Cx43 species that migrates as a single protein band and was designated Cx43_m. Cx43_m was shown to be the result of additional Ser/Thr phosphorylation as indicated by: (a) the increased gel mobility induced by both alkaline phosphatase and the Ser/ Thr-specific protein phosphatase-2A (PP2A) and (b) the removal of virtually all ³²P_i from Cx43_m by PP2A. Immunofluorescent confocal microscopy of mitotic cells revealed that Cx43 is intracellularly located, while in nonmitotic cells Cx43 is located at regions of cell–cell contact. Dye coupling studies revealed that mitotic endothelial cells were uncoupled from each other and from nonmitotic cells. After cytokinesis, sister cells resumed cell coupling independent of de novo protein synthesis. The mitosis-specific phosphorylation of Cx43 correlates with the transient loss of gap junction intercellular communication and redistribution of Cx43, suggesting that a protein kinase that regulates gap junctions is active in M-phase.     

Esophageal PCO2 as a monitor of perfusion failure during hemorrhagic shock

Sato, Yoji, Max Harry Weil, Wanchun Tang, Shijie Sun,Jianlin Xie, Joe Bisera, and Hidehiro Hosaka. EsophagealPCO₂ as a monitor of perfusionfailure during hemorrhagic shock. J. Appl.Physiol. 82(2): 558-562, 1997.Measurement ofgastric wall PCO₂(Pg_CO2) bytonometric method has emerged as an attractive option for estimatingvisceral perfusion during circulatory shock. However, gastric acidsecretion obfuscates the tonometric measurement. We, therefore,investigated the option of measuringPCO₂ in the esophagus to minimizethese restraints. Hemorrhagic shock was induced in five Sprague-Dawleyrats, and five rats served as sham controls.Pg_CO2 wasmeasured with an ion-sensitive field effect transistor that wassurgically implanted into the gastric wall. Esophageal luminalPCO₂(Pe_CO2) wasmeasured by a second ion-sensitive field effect transistor sensor.During hemorrhagic shock, mean aortic pressure declined from 150 to 50 mmHg. Gastric blood flow decreased from 58 to 12 ml ^· min^{1 ·} 100 g¹ (21% of preshock) andesophageal blood flow from 44 to 7 ml ^· min^{1 ·} 100 g¹ (16% of preshock).Pg_CO2simultaneously increased from 47 to 116 Torr andPe_CO2 from 47 to 127 Torr. The increases inPg_CO2 werehighly correlated with increases inPe_CO2(r = 0.90). Esophageal tonometry may,therefore, serve as a practical alternative to gastric tonometry.

     

Yersinia pseudotuberculosis in China

Thirty strains of Yersinia pseudotuberculosis were isolated from rabbits (17 strains), wild rats (9 strains) and house rats (4 strains) in China between 1990 and 1993. The biochemical properties of these isolates were identical with those of Y. pseudotuberculosis and no special characteristics were found in these strains. Serologically, serogroups 4b and 5b were identical to isolates found in Japan, and a new serogroup 1c and unclassified strains have also been detected. The existence of virulence-associated properties were different among strains. The pYV plasmid was detected from 6 strains of 30 isolates. This report documents the presence of Y. pseudotuberculosis in China, providing important epidemiological information.     

囊萼紫草属与滇紫草属花粉形态比较研究

本文借助光学显微镜和扫描电镜研究了囊萼紫草属3种和滇紫草属12种植物的花粉形态。囊萼紫草属的花粉为哑铃形或茧形,中等大小,P／E比为1．6一1．67,三孔沟,内孔横长;具小刺状纹饰。滇紫草属的花粉为近长球形或近卵球形,P／E为l—1．23;三孔沟或三合沟孔,内孔一般纵长,具皱波状纹饰,在皱波上具密集的小瘤或微颗粒。从花粉形态的角度,本文支持把囊萼紫草属从滇紫草属(广义)中分离出来的观点。值得注意的是,在滇紫草属的花粉中首次观察到了一种比较少见且特化的花粉即单极三合沟孔的花粉。     

phbB,phbC基因克隆,序列分析及植物表达载体的构建

利用聚合酶链式反应技术,从真养产碱杆菌Ａｌｃａｌｉｇｅｎｅｓ ｅｕｔｒｏｐｈｕｓ Ｈ１６染色体ＤＮＡ中的主增并克隆了调控聚－β－羧基丁酸生物合成的两个关键酶基因;依赖ＮＡＤＰＨ的乙酰乙酰ＣｏＡ还原酶基因和ＰＨＢ合成酶基因。限制性内切酶图谱和核苷酸序列分析证实了克隆结果,并表明所克隆的基因与国外报道的有很高的同源性。     

薏苡胚乳传递细胞的超微结构

传粉后１０ ｄ,薏苡（Ｃｏｉｘ ｌａｃｒｙｍ ａ－ｊｏｂｉＬ．）颖果基部胚乳最外层传递细胞具长而多的壁内突,第二层细胞的壁内突较第一层的短而少,均具瓣裂的细胞核、丰富的线粒体、粗糙内质网、核糖体、产生小泡的高尔基体及与壁内突质膜相连的、含深色物质的囊泡。线粒体分布于壁内突附近或其间。授粉后２５ ｄ,第一、二层细胞壁内突发达,几乎充满了细胞,但细胞器可见。第四层传递细胞具树枝状及网状的壁内突,大量线粒体、具质体膜的淀粉粒、脂体存在壁内突附近或壁内突的间隙内。高尔基体常见,仅见很少的片段内质网。第五层传递细胞具短的壁内突、较大的淀粉粒及许多小蛋白质体。两个时期的第一、二层细胞内均未观察到胞间连丝。授粉后２５ ｄ,第四层及以上的传递细胞的细胞壁和呈网状的壁内突均含有胞间连丝。还讨论了各种细胞器的作用及各层传递细胞的功能     

水稻线粒体DNA雄性不育有关特异片段的克隆及序列分析

应用任意单引物聚合酶链反应技术,从水稻ＷＡ 型雄性不育系的线粒体ＤＮＡ 中得到一个特异的扩增片段Ｒ２－６３０ ＷＡ。以该片段为探针进行Ｓｏｕｔｈｅｒｎ 杂交分析检测到在雄性不育胞质与正常可育胞质间存在的线粒体ＤＮＡ 多态性。不育系珍汕９７Ａ 和其Ｆ１ 杂种的杂交图谱相同。而保持系珍汕９７Ｂ和恢复系明恢６３ 的杂交图谱一样。序列测定该片段全长６２９ ｂｐ,其碱基组成Ａ＋ Ｔ＝ ５４．１％ ,同源性比较结果显示, 该片段与１２３６ 个已报道的植物基因（包括１６ 个水稻线粒体基因）序列的同源性均小于５０％ 。序列内含有一个长度为１０ ｂｐ 的反向重复序列５－ＡＣＣＡＴＡＴＧＧＴ－３,位于２６２—２７２ 区段。另外,其３７９—４３９ 区段可编码一个含２０个氨基酸残基的短肽。上述结果表明,Ｒ２－６３０ ＷＡ 片段确与水稻野败型雄性不育密切相关。推测反向重复序列５－ＡＣＣＡＴＡＴＧＧＴ－３在细胞质雄性不育性状形成中,可能起着重要作用     

脱落酸对线粒体Na^＋—K^＋ATPase活性的影响

在提取大豆（Ｇｌｙｃｉｎｅ ｍ ａｘ）黄化子叶的线粒体时,于清洗和悬浮介质中加入ＡＢＡ,发现４０ μｍ ｏｌ／Ｌ（±） ＡＢＡ 对线粒体膜结合Ｎａ＋ －Ｋ＋ ＡＴＰａｓｅ 活性及线粒体吸氧速率有明显的促进作用。ＡＢＡ 可使１６℃与２７℃培养的大豆子叶线粒体膜结合Ｎａ＋ －Ｋ＋ ＡＴＰａｓｅ Ａｒｒｈｅｎｉｕｓ图的折点温度分别下降６．３℃和５．９℃,并将该酶的底物动力学曲线由正协同曲线转变为非协同曲线。表明ＡＢＡ 可降低线粒体的膜相变温度     

基因转移与肿瘤基因工程疫苗

综述了近年来有关利用基因转移技术修饰肿瘤细胞制备肿瘤基因工程疫苗的最新研究进展,着重阐述了逆转录病毒载体介导的基因转移及其安全性;归纳了目前可用于肿瘤基因工程疫苗的各种目的基因的特点及作用并对这类肿瘤疫苗制备过程中所存在的问题进行了分析.     

一种用聚乙二醇制备微粒体的方法

介绍一种用聚乙二醇(PEG)制备微粒体的方法.大鼠肝匀浆经聚乙二醇-6000凝聚,及两次高速离心即可得到微粒体组分,与超速离心方法比较,可省去超速离心步骤,又缩短了分离制备的时间,是一种比较简单易行的方法．