Isolation and Characterization of the Plasma Membrane by Two-Phase Partitioning from the Alkalophilic Cyanobacterium Spirulina platensis

Partition in aqueous dextran-polyethylene glycol two-phase systemwas used to isolate the plasma membranes from the alkalophiliccyanobacterium Spirulina platensis. The upper phase containeda colorless membranes obtained in relatively short time, 3–4h. This fraction had a different protein profile than that ofthe thylakoid fraction obtained in the lower phase. It did notcontain cytochrome c-oxidase activity, but retained characteristicMg²⁺-ATPase activity that is sensitive to vanadate, stimulatedby K⁺, and has a pH optimum near 8.5. These data support ourassumption that the upper phase of the gradient consist of theplasma membrane of S. platensis. (Received November 25, 1993; Accepted April 12, 1994)     

Transfection of the mouse ICAM-1 gene into murine neuroblastoma enhances susceptibility to lysis,reduces in vivo tumorigenicity and decreases ICAM-2-dependent killing

We determined the expression of intercellular adhesion molecules (ICAM) on neuro-2a cells in order to evaluate whether they were involved in cytolysis of murine neuroblastoma. Fluorescence-activated cell sorting analysis revealed that the control neomycin-resistance-genetransduced line (neuro-2a/LN) had poor expression of ICAM-1 (mean channel fluorescence, MCF=3.7). An ICAM-1-positive transfectant of neuro-2a (neuro-2a/ICAM-1⁺) (CMF=64.3) was generated to evaluate directly the role of this adhesion molecule in cytolysis. Neuro-2a/ICAM-1⁺ was more sensitive to LAK killing (69.7% at an effector-to-target ratio of 1001) compared to neuro-2a/LN (48.6%) (P<0.001). Blocking of neuro-2a/LN and neuro-2a/ICAM-1⁺ lysis with anti-ICAM-1 monoclonal antibodies (mAbs) did not account for all the LFA-1-dependent killing. These data indicate that even in neuro-2a/ICAM-1⁺ cells, other LFA-1 ligands participated in the effector-target interaction. Therefore, we examined these cell lines for ICAM-2 expression. Both neuro-2a/LN and neuro-2a/ICAM-1⁺ lines expressed ICAM-2 (MCF=16.4 and 16.5). ICAM-2 accounted for the majority of the LFA-1-dependent killing in the ICAM-1-negative target, neuro-2a/LN, while ICAM-1 played a primary role in the cytolysis of the ICAM-1⁺ transfectant. Inhibition of lysis in the presence of anti-ICAM-1 and ICAM-2 mAbs was comparable to that seen with the addition of anti-LFA-1 mAb, indicating that other LFA-1 ligands were not involved in this system. ICAM-1 expression was associated with decreased in vivo tumorigenicity; mice inoculated with neuro-2a/ICAM-1⁺ cells had a significantly longer survival compared to those receiving neuro-2a/LN cells (median survival time 35.5 versus 24.5 days) (P<0.001). It is important to note that ICAM-1 transfection of murine neuroblastoma did not alter its metastatic potential. We conclude that transfection of mouse neuroblastome with ICAM-1 increases its sensitivity to in vitro lysis and reduces its in vivo tumorgenicity. In ICAM-1-negative murine neuroblastoma cells, ICAM-2 plays a primary role in cell-mediated lysis.This work was supported in part by the Children's Cancer Research Fund, the Minnesota Medical Foundation, the Viking Children's Fund and NIH grants PO1-CA-21737, NO1-AI-85002. E. K. is a recipient of the Irvine McQuarrie Research Scholar Award and B. R. B. a recipient of the Edward Mallinkrodt Foundation Scholar Award     

滇蜀豹子花核型及其变异研究

本文详细报道了滇蜀豹子花的核型,发现居群中存在两种细胞型,即A型和B型。A型参考核型为2n = 24＝2m(2SAT)+2sm+8st(4SAT)+12t(2SAT),其第3号两条同源染色体长臂均无居间随体：B型参考核型为2n=24＝2m(2SAT)+2sm+8st(2SAT)+12t(3SAT)+0—1b,其第3号一条同源染色体长臂紧靠着丝点处有一大而明显的居间随体,而另一条同源染色体则无,构成明显的3号染色体的结构杂合性。统计表明,居群中二者的比例近似为1A;2B。研究还发现了大量的体细胞染色体结构变异核型,表明滇蜀豹子花核型尚未趋于稳定,还处于强烈分化之中,高频率的体细胞染色体结构变异是其种内分化不可忽视的一种进化要素。     

油松鳞叶中细胞核穿壁现象的初步研究

利用薄切片和超薄切片技术对油松衰老鳞叶中细胞核穿壁运动进行了观察研究。首次,细胞核内的染色质逐渐收缩集中,形成染色深的球状体,以后,细胞核逐渐与细胞壁接近。接着紧贴细胞壁的细胞核通过细胞壁上的通道或胞间连丝转移到相邻的细胞内,此外,还发现有多种其它的转移方式存在。研究结果表明细胞核的穿壁运动现象同时存在于连子植物和裸植物中,电镜观察进一步证明细胞壁道在细胞核穿壁之前已形成。     

蜜蜂脑中5—HT，GABA，Ach免疫反应阳性神经元分布的比较研究

通过免疫细胞化学方法－ＰＡＰ法,研究了中华峰（Ａｐｉｓｓｉｎｅｎｓｉｓ）和意大利峰（ＡｐｉｓｍｅｌｌｉｆｅｒａＬ）脑和视叶中乙酰胆碱（Ａｃｈ）、５－羟色胺（５－ＨＴ）、γ－氨基丁酸（ＧＡＢＡ）免疫反应阳性神经元胞体和纤维的形态、染色强度、分布和各种神经丛之间的联系．一方面将这些阳性分布系统和蜜蜂脑中相应的神经元类型进行比较,另一方面也分析了在一些特殊解剖学通路中存在的可能的神经递质及其作用．     

Complex duplications in maize lines

Rp1 is a disease resistance complex and is the terminal morphological marker on the short arm of maize chromosome 10. Several restriction fragment length polymorphisms (RFLPs), which map within 5 map units of Rp1, were examined to determine if they are also complex in structure. Two RFLP loci, which mapped distally to Rp1, BNL3.04 and PIO200075, existed in a single copy in all maize lines examined. These two loci cosegregated perfectly in 130 test cross progeny. Two RFLP loci that map proximally to Rp1 had unusual structures, which have not yet been reported for maize RFLPs; the loci were complex, with variable numbers of copies in different maize lines. One of the loci, NPI285, occasionally recombined in meiosis to yield changes in the number of copies of sequences homologous to the probe. The other proximal locus, detected by the probes NPI422, KSU3, and KSU4, was relatively stable in meiosis and no changes in the number of restriction fragments were observed. The similarity in map position between Rp1 and the complex RFLP loci indicate there may be genomic areas where variable numbers of repeated sequences are common. The structure of these complex loci may provide insight into the structure and evolution of Rp1.     

Logistic曲线参数的一个最佳估计方法

本文提出用O．628优选法和SAS/STAT软件NLIN模块中的DUD法,对Logistic曲线中的参数k,a和b可得到最佳估计。     

PCR检测HPV－18DNA

采用聚合酶链式反应（ＰＣＲ）技术,对ＨＰＶ－１８序列中引物ＨＰ＿１、ＨＰ＿２之间的片段（Ｆ）进行扩增,通过两组阴、阳性对照实验证明扩增片段的特异性。用不同Ｍｇ浓度的缓冲系统进行ＰＣＲ反应发现,缓冲系统中Ｍｇ浓度高低是影响ＨＰＶ－１８／ＨＰ＿１、ＨＰ＿２特异扩增的重要因素,高浓度Ｍｇ导致扩增特异性降低。对１７例宫颈癌组织ＤＮＡ进行ＰＣＲ检测,有９例检出Ｆ片段,其检出率是５３％,为ＨＰＶ－１８与宫颈癌的相关性提供证据。     

零下低温对杂交杨树皮层膜脂组成的影响

以不耐寒的美洲黑杨（Ｐｏｐｕｌｕｓｄｅｌｔｏｉｄｅｓｃｖ．“Ｌｕｘ”Ｉ－６９／５５,父本）和耐寒性较强的欧美杨（Ｐ．ｅｕｒａｍｅｒｉｃａｎａｃｌｃｖ．Ｉ－４５／５１,母本）的４个杂交Ｆ＿１代无性系（９５杨、５５９杨、６００杨和１３８１杨）为材料,分析了零下低温寒潮前后枝条皮层的脂质组成。结果表明,寒潮影响下,皮层中磷脂含量增加而组成基本不变,膜脂脂肪酸组成的变化规律是：寒潮前脂肪酸不饱和指数（ＩＵＦＡ）值大的无性系,寒潮前后的ＩＵＦＡ值变化量小;寒潮前ＩＵＦＡ值较小的无性系,寒潮前后ＩＵＦＡ值变化量较大。本文借用力学概念,提出相对抗性概念,给出杨树无性系的相对抗性序列。序列表明Ｆ＿１代无性系的耐寒性已较不耐寒的父本提高,这与田间观察基本一致。