Polysomes and protein synthesis in cells infected with a DNA virus

In HEp-2 cells infected with herpes simplex virus the rate of protein synthesis at first decline, is stimulated between 4 and 8 hours after infection, and progressively and irreversibly declines from 9 to 16 hours later. The increase and decrease in rates coincide with corresponding changes in the amounts of cytoplasmic polysomes and amounts of labeled amino acids in nascent peptides bound to polysomes. The data indicate that (i) early and late inhibition and intervening stimulation of protein synthesis are due to the corresponding breakdown and formation of polysomes, and (ii) the bulk of viral proteins is probably made on cytoplasmic polysomes.     

G1/S transition in normal human T-lymphocytes requires the nuclear protein encoded by c-myb

Exposure of peripheral blood mononuclear cells (PBMC) to an 18-base c-myb antisense oligomer before mitogen or antigen stimulation resulted in almost complete inhibition of c-myb messenger RNA and protein synthesis and blockade of T lymphocyte proliferation. Expression of early and late activation markers, interleukin-2 receptor and transferrin receptor, respectively, by PBMC was unaffected by antisense oligomer exposure as was the expression of c-myc messenger RNA. In contrast, histone H3 messenger RNA levels and DNA content were selectively decreased. These results suggest that c-myb protein deprivation does not perturb T lymphocyte activation or early molecular events that may prepare the cell for subsequent proliferation. Rather, it appears to specifically block cells in late G1 or early S phase of the cell cycle.     

鸡TIGAR基因真核表达质粒的构建及其抗凋亡功能评价

【背景】 TP53诱导的糖酵解和凋亡调节因子（TP53-induced glycolysis and apoptosis regulator,TIGAR）是p53下游的靶基因,具有调节糖酵解水平和去除活性氧（Reactive oxygen species,ROS）并降低由活性氧诱发的细胞凋亡水平。【目的】 构建鸡源TIGAR基因的真核表达质粒,并检测TIGAR基因在DF1细胞中的抗凋亡作用,为建立稳定表达鸡TIGAR基因的细胞系做准备。【方法】根据GenBank（登录号：XM_417232.6）中预测基因设计引物,利用RT-PCR的方法从SPF鸡脾脏中扩增鸡TIGAR基因,将扩增产物克隆至载体（Flag-CMV14）后送公司测序验证;随后构建进化树对鸡TIGAR基因与其他哺乳动物及水生动物的TIGAR基因进行同源性分析。将重组质粒（Flag-TIGAR）转染入DF1细胞24 h后,使用新城疫病毒诱导细胞凋亡,利用Western Blot检测重组质粒表达情况以及Poly ADP-Ribose Polymerase（PARP）裂解情况。此外还将重组质粒（Flag-TIGAR）转染入DF1细胞,并于收样前2 h使用Staurosporine刺激细胞发生凋亡,分别在转染后24、48 h收集样品,并用流式细胞仪检测细胞凋亡情况。【结果】 RT-PCR扩增TIGAR基因,在843 bp处出现目的条带与预测相符,构建的TIGAR真核表达质粒（Flag-TIGAR）经测序,结果显示其序列与GenBank上预测的基本一致。Western Blot结果显示在30 h、36 h收集的样品中PARP均被裂解且转染重组质粒（Flag-TIGAR）的实验组与未转染质粒（MOCK）组或转染空载体（Flag-CMV14）组的样品相比,裂解的PARP表达量明显降低,且差异极显著（P<0.01）。流式结果显示：24 h 检测转染Flag-CMV14后细胞总凋亡率为11%（早期凋亡7.8%,晚期凋亡3.2%）,而转染Flag-TIGAR后细胞总凋亡率仅为4%（早期凋亡3.7%,晚期凋亡0.3%）,转染Flag-CMV14的早期凋亡率和晚期凋亡率均高于转染Flag-TIGAR组,且差异显著（P<0.05）。48 h转染Flag-CMV14后细胞总凋亡率为20.3%（早期凋亡14.3%,晚期凋亡6.0%）,而转染Flag-TIGAR后细胞总凋亡率为6.4%（早期凋亡4.8%,晚期凋亡1.6%）,转染Flag-CMV14组的细胞早期凋亡和晚期凋亡率均高于转染Flag-TIGAR的组,且差异极显著（P<0.01）。【结论】成功扩增出鸡TIGAR基因,并构建了其真核表达质粒,通过Western Blot和流式实验均证实过表达TIGAR后可降低细胞的凋亡程度并有利于细胞存活。     

Inhibition of endothelial regeneration by type-beta transforming growth factor from platelets

Damage to the vessel wall is a signal for endothelial migration and replication and for platelet release at the site of injury. Addition of transforming growth factor-beta (TGF-beta) purified from platelets to growing aortic endothelial cells inhibited [3H]thymidine incorporation in a concentration-dependent manner. A transient inhibition of DNA synthesis was also observed in response to wounding; cell migration and replication are inhibited during the first 24 hours after wounding. By 48 hours after wounding both TGF-beta-treated and -untreated cultures showed similar responses. Flow microfluorimetric analysis of cell cycle distribution indicated that after 24 hours of exposure to TGF-beta the cells were blocked from entering S phase, and the fraction of cells in G1 was increased. The inhibition of the initiation of regeneration by TGF-beta could allow time for recruitment of smooth muscle cells into the site of injury by other platelet components.     

Erythropoietin retards DNA breakdown and prevents programmed death in erythroid progenitor cells

The mechanism by which erythropoietin controls mammalian erythrocyte production is unknown. Labeling experiments in vitro with [3H]thymidine demonstrated DNA cleavage in erythroid progenitor cells that was accompanied by DNA repair and synthesis. Erythropoietin reduced DNA cleavage by a factor of 2.6. In the absence of erythropoietin, erythroid progenitor cells accumulated DNA cleavage fragments characteristic of those found in programmed cell death (apoptosis) by 2 to 4 hours and began dying by 16 hours. In the presence of erythropoietin, the progenitor cells survived and differentiated into reticulocytes. Thus, apoptosis is a major component of normal erythropoiesis, and erythropoietin controls erythrocyte production by retarding DNA breakdown and preventing apoptosis in erythroid progenitor cells.     

Caffeine-induced uncoupling of mitosis from the completion of DNA replication in mammalian cells

Caffeine was shown to induce mitotic events in mammalian cells before DNA replication (S phase) was completed. Synchronized BHK cells that were arrested in early S phase underwent premature chromosome condensation, nuclear envelope breakdown, morphological "rounding up," and mitosis-specific phosphoprotein synthesis when they were exposed to caffeine. These mitotic responses occurred only after the cells had entered S phase and only while DNA synthesis was inhibited by more than 70 percent. Inhibitors of protein synthesis blocked these caffeine-induced events, while inhibitors of RNA synthesis had little effect. These results suggest that caffeine induces the translation or stabilizes the protein product (or products) of mitosis-related RNA that accumulates in S-phase cells when DNA replication is suppressed. The ability to chemically manipulate the onset of mitosis should be useful for studying the regulation of this event in mammalian cells.     

Mycoplasma inhibition of phytohemagglutinin stimulation of lymphocytes

Goat lymphocytes were cultured in vitro with phytohemagglutinin and nonviable mycoplasmas. Addition of the mycoplasmas, even as late as 45 hours after adding phytohemagglutinin, completely inhibited the increase in synthesis of DNA and RNA normally induced in lymphocytes by the mitogen. The suppression of synthesis did not result from killing of the cells by the mycoplasmas, combination of the organisms with phytohemagglutinin, or competition for combining sites on the cell surface, which indicates that some other mechanism of inhibition was operative. A similar depression of response to phytohemagglutinin in lymphocytes in culture has been observed in human diseases associated with an immune defect. The present demonstration that at least certain mycoplasmas can profoundly affect lymphocyte function in vitro suggests that thay may alter the immune response in vivo.     

cdc2 gene expression at the G1 to S transition in human T lymphocytes

The product of the cdc2 gene, designated p34cdc2, is a serine-threonine protein kinase that controls entry of eukaryotic cells into mitosis. Freshly isolated human T lymphocytes (G0 phase) were found to have very low amounts of p34cdc2 and cdc2 messenger RNA. Expression of cdc2 increased 18 to 24 hours after exposure of T cells to phytohemagglutinin, coincident with the G1 to S transition. Antisense oligodeoxynucleotides could reduce the increase in cdc2 expression and inhibited DNA synthesis, but had no effect on several early and mid-G1 events, including blastogenesis and expression of interleukin-2 receptors, transferrin receptors, c-myb, and c-myc. Induction of cdc2 required prior induction of c-myb and c-myc. These results suggest that cdc2 induction is part of an orderly sequence of events that occurs at the G1 to S transition in T cells.     

Chromosomal and nucleolar RNA synthesis in root tips during mitosis

Comparative rates of RNA synthesis in chromatin and nucleolar fractions during mitosis in root-tip cells of Allium and Nigella were studied by pulse-labeling of cells with tritiated cytidine. Although the rate of RNA synthesis decreases in the condensing chromosomes during prophase, it remains normal in the nucleolar fraction as long as nucleoli are maintained. RNA synthesis stops in mitotic cells lacking distinct nucleoli. In the late telophase or very early interphase cells, RNA synthesis resumes at a faster rate in the pronucleolar bodies than in the chromatin.     

Checkpoints: controls that ensure the order of cell cycle events

The events of the cell cycle of most organisms are ordered into dependent pathways in which the initiation of late events is dependent on the completion of early events. In eukaryotes, for example, mitosis is dependent on the completion of DNA synthesis. Some dependencies can be relieved by mutation (mitosis may then occur before completion of DNA synthesis), suggesting that the dependency is due to a control mechanism and not an intrinsic feature of the events themselves. Control mechanisms enforcing dependency in the cell cycle are here called checkpoints. Elimination of checkpoints may result in cell death, infidelity in the distribution of chromosomes or other organelles, or increased susceptibility to environmental perturbations such as DNA damaging agents. It appears that some checkpoints are eliminated during the early embryonic development of some organisms; this fact may pose special problems for the fidelity of embryonic cell division.     

Hydroxyurea: suppression of two-stage carcinogensis in mouse skin

Hydroxyurea, a selective cytotoxic agent for cells in DNA synthesis, injected intraperitoneally at 24 and 48 hours after the first painting with 1 percent croton oil, significantly reduced the tumor yield in the two-stage chemical carcinlogenesis in mouse skin. A comparable group of mice receiving hydroxyurea only once at 24 hours had a tumor induction similar to that in controls.     

Hydroxyurea: differential lethal effects on cultured mammalian cells during the cell cycle

Hydroxyurea has a differential lethal effect on cultured Chinesehamster cells that are at different stages in their cell cycle. Cells synthesizing DNA at the time of exposure to the drug are lethally damaged. Cells in the phase of growth preceding DNA synthesis (G(1)) survive but are prevented from beginning DNA synthesis. Cells in the phase after DNA synthesis (G(2)) survive and appear to progress until just before the beginning of the next period of DNA synthesis. This differential lethal and inhibitory effect of hydroxyurea may be useful for synchronizing asynchronous cell populations and explaining effects of the drug in human therapy.     

DNA synthesis in differentiating skeletal muscle cells: initiation by ultraviolet light

As skeletal muscle cells differentiate, they fail to initiate DNA synthesis. This rigid regulation, which persists even after cells are fully developed, does not extend to "repair" DNA synthesis, in that ultraviolet light initiates DNA synthesis in 99 percent of the muscle nuclei exposed. The rate of "repair" DNA synthesis in these nuclei, however, drops over 50 percent at the time of cell differentiation.     

黄姑鱼正常二倍体和雌核发育体胚胎发育及早期生长的比较研究

对黄姑鱼正常二倍体（N）、雌核发育二倍体（G）和单倍体（H）的胚胎发育进行观察,并对其早期生长情况进行比较。结果显示：（1）受精率N>H>G,孵化率N>G>H,畸形率H>G>N,72 h成活率N>G>H;（2）正常二倍体胚胎经21 h 10 min孵化出膜,雌核发育二倍体和单倍体孵化出膜分别用时23 h 10 min和23 h 30 min;雌核发育二倍体发育滞后主要出现于原肠早期和孵出期,单倍体滞后出现于原肠晚期。从形态上看,单倍体胚胎呈现典型的单倍体综合症,雌核发育二倍体和正常二倍体胚胎发育形态正常且两者无明显差异。各组死亡高峰均出现在原肠晚期,单倍体组开口前全部死亡;（3）60日龄之前雌核发育二倍体生长速度明显慢于正常二倍体且个体间差异较大。     

平贝母体细胞胚胎发生早期核酸和蛋白质合成动态的研究

在长期平贝母(Frifiiavia ussuviensis Maxim)组织培养中,建立比较稳定的体细胞胚胎发生体系,应用放射性同位素~3H 的液体闪烁技术测得,体细胞胚 RNA、蛋白质和 DNA 合成分别在诱导培养第5,7,8b 达到高峰。组织细胞学观察表明,体细胞胚胎发生早期的核酸与蛋白质合成的变化与愈伤组织中细胞的胚性化和胚性细胞的增殖相关。     

Cytochalasin B inhibits lymphotoxin production by antigen-stimulated lymphocytes

Lymph node cells of rats sensitized with hen ovalbumin produced lymphotoxin after 6 to 12 hours of exposure to specific antigen. Lymphotoxin was assayed by its cytotoxicity for fibroblasts from syngeneic embryos during a 72-hour incubation. Cytochalasin B inhibited lymphotoxin production, as well as later DNA synthesis, at concentrations (0.1 to 5.0 micrograms per milliliter) comparable to those which affect microfilament function and cell motility in other systems, and this inhibition was reversible. Binding of antigen was not affected.     

稻秆潜蝇的生物学与生态学特性研究

稻秆潜蝇在重庆市巳由次要害虫上升为主要害虫,该虫在重庆地区1年发生3代,以第3代幼虫越冬。越冬代．幼虫在3月下旬化蛹,4月中旬至下旬为第1代成虫期,5月上旬为产卵期。5月上旬至6月上旬为第1代幼虫期,6月上旬至下旬为蛹期。第2代成虫期为6月下旬至7月中旬,7月上、中旬为产卵期,7月中旬至8月上旬为幼虫期,8月上旬至下旬为蛹期。第3代成虫期为9月上旬至下旬,并迁至越冬寄主上产卵,10月初至11月底孵化为幼虫进入越冬期。成虫羽化后1—2h即可交配,但多数于羽化后1—3d才交配。更尾后越冬代于4—9d,第2代于3—7d后产卵,一般1叶仅产1卵,偶有数粒。单雌产卵量2—81粒不等。成虫寿命为20d左右。     

Autoradiographic investigations of incorporation of H3-thymidine into cells of the rat and mouse

The application of H(3)-thymidine results in labeling of those nuclei of cells in which deoxyribonucleic acid (DNA) is synthesized during the interval between application and the sacrifice of tne animal (1-3). This paper reports autoradiographic investigation with H(3)-thymidine of rats and mice. This method permits a more exact statement of the number of dividing cells than does the microscopic estimate of mitosis. The latter method is practically impossible in tissues with small fusiform cells. Moreover, it is possible to obtain information about the relative time of DNA synthesis in different cells.     

Interleukin-1 mitogenic activity for fibroblasts and smooth muscle cells is due to PDGF-AA

Both interleukin-1 (IL-1) and platelet-derived growth factor (PDGF) induce proliferation of cultured fibroblasts and smooth muscle cells. These polypeptide mediators are released by activated macrophages and other cell types in response to injury and are thought to have a role in tissue remodeling and a number of pathologic processes. Analysis of the kinetics of [3H]thymidine incorporation by cultured fibroblasts demonstrated that the response to IL-1 is delayed approximately 8 hours relative to their response to PDGF. IL-1 transiently stimulated expression of the PDGF A-chain gene, with maximum induction after approximately 2 hours. Subsequent synthesis and release of PDGF activity into the medium was detected as early as 4 hours after IL-1 stimulation, and downregulation of the binding site for the PDGF-AA isoform of PDGF followed PDGF-AA secretion. Antibodies to PDGF completely block the mitogenic response to IL-1. Therefore, the mitogenic activity of IL-1 for fibroblasts and smooth muscle cells appears to be indirect and mediated by induction of the PDGF A-chain gene.