Micro-scission of YAC tumor cells during antibody-dependent cellular cytotoxicity mediated by human neutrophils

The cellular events accompanying neutrophil-mediated antibody-dependent cellular cytotoxicity (ADCC) directed against YAC erythroleukemic target cells have been studied by time-lapse fluorescence-intensified microscopy. The YAC plasma membrane and cytosol were labeled with the fluorescent probes diC18Icc and eosin Y, respectively. Fluorescently labeled and IgG-opsonized YAC cells were incubated at 37 degrees C while observed by optical microscopy. During temporal studies of neutrophil-YAC conjugates, the cytosol of YAC cells accumulated in tubular and spherical compartments of the neutrophils' vacuolar apparatuses. To distinguish between several possible mechanisms of target cytosol uptake, diC18Icc-labeled YAC cells were observed during identical conditions. The membrane label diC18Icc was found to accumulate within neutrophils in an identical fashion. At roughly 30 min, 25 and 38% of neutrophils in apparent conjugates had internalized tumor cell cytosol or plasma membrane, respectively, within a vesicular compartment. The IgG-dependent uptake of eosin Y and diC18Icc by neutrophils was diminished by exposure to 2.5 mM sodium azide. When cells were exposed to 5.5 mM sodium azide, 1 mM iodoacetamide, or 4 degrees C, conjugate formation and uptake of eosin Y or diC18Icc were abolished. An artifactual accumulation of eosin Y or diC18Icc in neutrophils was further ruled out by control studies. Non-specific exchanges of eosin Y and diC18Icc labels of YAC cells with tannic acid-treated red blood cells (RBCs) and normal neutrophils were studied. Since hemoglobin binds tightly to eosin Y, RBCs can easily detect eosin Y leakage. No exchange of eosin Y or diC18Icc from YAC cells into bound tannic acid-treated erythrocytes was found.(ABSTRACT TRUNCATED AT 250 WORDS)     

Yeast regulatory protein LEU3: a structure-function analysis.

Eleven mutations resulting in partially deleted or truncated LEU3 protein were generated by linker insertion or other modifications at restriction sites, deletion of restriction fragments, or oligonucleotide-directed mutagenesis. Functional studies of these mutants showed the following: (i) A specific DNA binding region is contained within the 173 N-terminal residues, but other regions of the protein are required for optimal binding. (ii) Activation of LEU2 expression depends on the C-terminal 113 residues of the LEU3 protein. (iii) Deletion of part or all of a central section of LEU3 eliminates the ability of the LEU3 protein to respond to the co-activator alpha-isopropylmalate, i.e. creates an unmodulated activator. (iv) Overproduction of unmodulated activator slows down cell growth. (v) Specific deletion of two short acidic regions, including one with net charge - 19, has only minor effects on activation and modulation.     

人体小卫星DNA探针的制备

根据人体小卫星DNA核心顺序,化学合成长23碱基寡核苷酸探针,筛选人体基因组文库,旨在获得能用作遗传分析探针的小卫星顺序。结果得到15个含小卫星的阳性重组子。随机取其一(C_(35.9))作探针,试做群体分析。所有个体均可检出多条杂交带。其中某些带具有多态性。在一定检测条件下,检出的DNA图谱在有限的个体内具有个体特异性。结果表明筛选文库得到的小卫星顺序可用于小卫星多态性的检测。其它小卫星探针的筛选和应用性研究正在进行。     

烟草抗黑胫病突变体的细胞筛选

经实验我们成功地建立了在细胞水平上筛选烟草抗黑胫病突变体的筛选体系。该体系的主要内容为:γ-射线500—2000拉德诱变高度感病品种的花药后用50—80％的黑胫病菌粗毒素为选择压力,筛选出抗毒素花粉植株,用离体叶片法测定选出抗病植株,再从后代鉴定中选出抗病性能够稳定遗传的突变系。γ-射线及高浓度毒素处理均能得到抗病植株。选自感病品种的花粉植株中约有9—50％是真正抗病的。这些抗病植株中有一部分的抗病性能够稳定遗传。用该法已从感病优质品种小黄金1025及乔庄黑苗中选出6个突变系。并自N.C.628(抗)×小黄金1025(感)及N.C.628(抗)×庆胜2号(感)的F_1花粉植株中选出4个抗病系。所有的抗病系经3—4代后均表现出稳定抗性。其中一个突变体(R400)的抗性似由不完全显性多基因控制。     

Noninvasive determination of ulnar stiffness from mechanical response--in vivo comparison of stiffness and bone mineral content in humans

An approach referred to as Mechanical Response Tissue Analysis (MRTA) has been developed for the noninvasive determination of mechanical properties of the constituents of the intact limb. Of specific interest in the present study is the bending stiffness of the ulna. The point mechanical impedance properties in the low frequency regime, between 60 and 1,600 Hz are used. The procedure requires a proper design of the probe for good contact of the skin at midshaft and proper support of the proximal and distal ends of the forearm to obtain an approximation to "simple support" of the ulna. A seven-parameter model for the mechanical response is then valid, which includes the first mode of anterior-posterior beam bending of the ulna, the damping and spring effect of the soft tissue between probe and bone, and the damping of musculature. A dynamic analyzer (HP3562A) provides in seconds the impedance curve and the pole-zero curve fit. The physical parameters are obtained from a closed-form solution in terms of the curve-fit parameters. The procedure is automated and is robust and analytically reliable at about the five percent level. Some 80 human subjects have been evaluated by this mechanical response system and by the Norland single photon absorptiometer, providing for the first time in vivo, a comparison of elastic bending stiffness (ulna) and bone mineral content (radius). Three functional parameters of potential clinical value are the cross-sectional bending stiffness EI, the axial load capability Pcr (Euler buckling load) and the bone "sufficiency" S, defined as the ratio of Pcr to body weight. The correlation between EI and bone mineral (r = 0.81) is only slightly less than previous in vitro results with both measurements on the same bone (r = 0.89). When sufficiency is taken into consideration, the correlation of Pcr and bone mineral content is improved (r = 0.89). An implication is that "quality" of bone is a factor which is not indicated by bone mineral content but which is indicated by stiffness. Bone mineral is necessary for proper stiffness but not sufficient. Therefore mechanical measurement should provide a new dimension to be used toward a better understanding of the factors related to bone health and disease.     

Expression of an ouabain-resistant Na,K-ATPase in CV-1 cells after transfection with a cDNA encoding the rat Na,K-ATPase alpha 1 subunit

We have used a gene transfer system to investigate the relationship between expression of the rat Na,K-ATPase alpha 1 subunit gene and ouabain-resistant Na,K-ATPase activity. A cDNA clone encoding the entire rat Na,K-ATPase alpha 1 subunit was inserted into the expression vector pSV2neo. This construct (pSV2 alpha 1) conferred resistance to 100 microM ouabain to ouabain-sensitive CV-1 cells. Hybridization analysis of transfected clones revealed the presence of both rat-specific and endogenous Na,K-ATPase alpha 1 subunit DNA and mRNA sequences. A single form of highly ouabain-sensitive 86Rb+ uptake was detected in CV-1 cells, whereas two distinct classes of ouabain-inhibitable uptake were observed in transfectants. One class exhibited the high ouabain sensitivity of the endogenous monkey Na,K-ATPase, while the second class showed the reduced ouabain sensitivity characteristic of the rodent renal Na,K-ATPase. Examination of the ouabain-sensitive, sodium-dependent ATPase activity of the transfectants also revealed a low affinity component of Na,K-ATPase activity characteristic of the rodent kidney enzyme. These results suggest that expression of the rat alpha 1 subunit gene is directly responsible for ouabain-resistant Na,K-ATPase activity in transfected CV-1 cells.     

限制性内切酶诱发的姊妹染色单体互换

用限制性内切酶PstⅠ,SalⅠ,PvuⅡ和BamHⅠ处理CHO细胞后,发现其SCE率升高,与对照相比,前三种酶具有显著性差异。但这些酶诱导SCE的效应与其致染色体畸变效应相比则较弱,提示引起DNA双链断裂的限制性内切酶不是SCE的强刺激物。实验结果表明,BrdU取代胸苷不能消除限制酶对底物DNA的识别及裂解。     

油菜潜叶蝇春季为害的预测

油菜潜叶蝇(Phytomyza atricornis Meigen)是油菜食叶的重要害虫。由于体型小,生活隐蔽,钻蛀破坏为害普遍而严重,对油菜生产影响极大。在湖南安仁县,是当前油菜生产上急 作者于1967年开始在安仁县农作物病虫测报站设点,进行观祭。1969年到1970年注意该虫的发生规律,并进行药剂防治试验。1971年以后,继续进行此项试验研究,探索有效的防洽措施,为油菜增产提供科学依据。现将1967年到1979年的初步研究结果报道如下。     

山西忻州中新世鼠兔科化石

本文记述了山西忻州发现的跑兔属—新种.该种以个体较小, P~2 较简单,无前沟, M~2 后叶无后突, P_3 小,无前内沟与前外沟等特征与属型种 B. forsythmajori 相区别,命名为杨胡跑兔 B. yanghuensis sp. nov..新种在进化水平上较属型种稍原始,但两者显示出较近的系统关系.根据 B. yanghuensis sp. nov. 的进化水平,推断其地质时代可能为中中新世早期,约相当于欧洲 Orleanian 期.