Cell-free protein synthesis: effects of age and state of ribosomal aggregation

In cell-free extracts derived from Streptococcus faecalis, protein synthesis directed by endogenous messenger RNA increases as the culture ages. The increased activity is accompanied by an increase in the percentage of membranebound ribosomes and by a decrease in ribosomal monomers and subunits. These changes progress against a background of structural and compositional modifications in the membrane. Membrane modifications possibly related to endogenously directed protein synthesis in cell-free extracts include: (i) decreased specific activity of a membrane-associated polynucleotide phosphorylase capable of polysome degradation, and (ii) increased concentrations of certain phospholipids.     

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。     

Collagen synthesis by cells synchronously replicating DNA

Replication and the performance of a differentiated function have been considered antagonistic processes. When cells in culture are partially tially synchronized with 5-fluoro-2'-deoxyuridine (FUdR), the synthesis of the specialized protein (collagen) is not reduced during chromosomal replication (S period). Collagen synthesis varies with general protein synthesis through the S period.     

Fish culture in the United States

The culture of channel catfish, trout, and crayfish is a well-established, profitable enterprise, but aquaculture provides only a fraction of the protein consumed by Americans, who prefer red meat. This situation could change, since pond-raised fish require less energy for protein synthesis than land animals, the supply of ocean food fish no longer appears inexhaustible, and fish culture can utilize resources unsuitable for other agriculture.     

Regenerating fish optic nerves and a regeneration-like response in injured optic nerves of adult rabbits

Regeneration of fish optic nerve (representing regenerative central nervous system) was accompanied by increased activity of regeneration-triggering factors produced by nonneuronal cells. A graft of regenerating fish optic nerve, or a "wrap-around" implant containing medium conditioned by it, induced a response associated with regeneration in injured optic nerves of adult rabbits (representing a nonregenerative central nervous system). This response was manifested by an increase of general protein synthesis and of selective polypeptides in the retinas and by the ability of the retina to sprout in culture.     

Chloramphenicol: effects on mouse myeloma cells in tissue culture

Within 36 hours of being administered, chloramphenicol (50 micrograms per milliliter) inhibits by 50 percent the rate of protein synthesis in mouse myeloma cells grown in suspension culture. Although there is a decrease in the amount of globulin synthesized, the rate of synthesis per cell is unchanged; the observed decrease is traced to the inhibition of cell proliferation caused by chloramphenicol.     

High-frequency C-type virus induction by inhibitors of protein synthesis

When inhibitors of protein synthesis are added to BALB/c mouse cells in culture, induction of naturally integrated C-type RNA virus occurs in a high percentage of cells. The action of protein synthesis inhibitors differs from that of halogenated pyrimidines, another class of virus inducers, in their effects on biologically distinguishable viruses. The use of such inhibitors to study integrated virus expression provides a means for studying gene regulation in mammalian cells.     

Euglena gracilis: formation of giant mitochondria

The addition of antimycin A (0.5 microgram per milliliter) to cultures of a bleached strain of Euglena gracilis in the logarithmic phase of growth on succinate as a carbon source results in (i) an interruption of growth for 24 hours and (ii) an increase in whole-cell respiration and the emergence of a novel succinoxidase activity within 2 to 4 hours. After 3 to 5 hours, the mitochondria enlarge, fuse, and form a sheathlike structure situated close to the periphery of the cell.     

Vitamin D3: induction of calcium-binding protein in embryonic chick intestine in vitro

Induction of the synthesis of calcium-binding protein in chick embryonic intestine maintained in vitro was accomplished by simply adding vitamin D(3) to the culture medium. Accompanying the induction of this protein, there was enhanced radiocalcium uptake by the intestine. These observations represent the first demonstration of an in vitro physiological effect of vitamin D(3) on the calcium absorptive mechanism of the intestine.     

重金属胁迫培养对微生物蛋白质含量的影响

[目的]研究重金属胁迫培养对微生物蛋白质含量的影响。[方法]选择4种典型微生物（大肠杆菌、枯草芽孢杆菌、啤酒酵母、链霉菌）为试验材料,采用传统微生物培养方法,在不同浓度的Hg2＋、Cd2＋、Cr6＋和Pb2＋重金属离子下进行胁迫培养,通过测定4种典型微生物的蛋白质含量,探讨重金属胁迫对4种微生物蛋白质合成的影响。[结果]低浓度的重金属对4种微生物蛋白质的合成有一定的促进作用,随着重金属浓度的增加,微生物蛋白质的合成均受到不同程度的抑制。枯草芽孢杆菌对这4种重金属的耐受性比其他3种微生物要强,4种重金属离子在比较大的质量浓度范围内（5～50 mg/L）对枯草芽孢杆菌蛋白质的合成有促进作用。低浓度Cr6＋对大肠杆菌蛋白质的合成有较强的促进作用;Pb2＋对大肠杆菌蛋白质合成有明显的抑制作用,对其他3种微生物蛋白质合成在一定浓度范围内有促进作用;低浓度的Cd2＋对4种微生物蛋白质的合成都有促进作用。[结论]该研究为探讨微生物对重金属胁迫的生理响应提供了理论依据。     

Embryonic chick intestine in organ culture: response to vitamin D 3 and its metabolites

Embryonic chick intestine maintained in organ culture responded to vitamin D(3) and its metabolites 25-hydroxycholecalciferol and 1,25-dihydroxycholecalciferol by synthesis of calcium-binding protein and enchanced calcium-45 uptake. The dihydroxy metabolite was by far the most potent inducer of the protein and also acted more rapidly than vitamin D(3) to stimulate isotope uptake. Despite its lower potency, vitamin D(3) itself was effective.     

稀土元素对冬凌草细胞合成冬凌草甲素的影响

[目的]探讨稀土元素对冬凌草细胞合成冬凌草甲素的影响。[方法]以冬凌草悬浮培养细胞为材料,研究不同浓度的2种稀土离子镧和亚铈对冬凌草细胞分泌和释放冬凌草甲素的影响。[结果]添加不同种类、不同浓度的稀土离子均可促进冬凌草甲素的合成。添加镧离子的细胞合成的冬凌草甲素明显高于添加铈离子的细胞。当细胞处于延滞期时,添加稀土元素并未产生明显的影响;悬浮培养指数期和稳定期加入稀土元素,均可促进冬凌草细胞对冬凌草甲素的合成,且在稳定期添加稀土元素的效果更加显著。在稳定期加入500μl溶液的培养液,获得的冬凌草甲素含量最高,可达28．89mg／g。[结论]该研究为冬凌草细胞的工业化生产奠定了基础。     

Dormin (Abscisin II), inhibitor of plant DNA synthesis?

Dormin (abscisin II), inhibits growth of Lemna minor cultures. At 1 part per million (3.8x10(-6)M), the culture appears nearly completely dormant but can be revived readily by transferring it to fresh medium free of dormin. The cytokinin benzyladenine, but not auxin or gibberellin, will counteract the dormin effect. Quantitative restoration of normal growth by cytokinin, however, can be achieved only if the dormin concentration does not exceed a critical level. Separation, after phenol-detergent extraction, of nucleic acids on methylated albumin kieselguhr columns showed suppression of nucleic acid synthesis by dormin in all fractions. Inhibition of the synthesis of the DNA fraction seems to precede that of RNA. Cytokinins reverse the process. They promote synthesis of all nucleic acid fractions, but again DNA seems to lead. Further work on the interaction of dormin with growth-promoting hormones might be facilitated by adopting the Monod model of allosteric transition, with, for example, DNA polymerase as the protein, dormin as the inhibitor, and cytokinin or other growth promoters as activators.     

Temperature effect on protein synthesis in a heat-synchronized protozoan treated with actinomycin D

Protein synthesis was studied in the ciliated protozoan Tetrahymena pyriformis GL after actinomycin D was added to the culture medium. When the temperature rose above that optimum for growth, there were significant reductions in protein synthesis. Lipid biosynthesis under the same conditions was slightly stimulated, an indication that the eflect was not due to an underlying reduction in energy sources. The phenomenon appears to be unique to the protein synthesizing system. Correlation with previous data suggests that it is due to nontranslational destruction of template RNA.     

匍枝筋骨草悬浮培养中β-蜕皮甾酮的积累与调控

植物源蜕皮激素——β-蜕皮甾酮(β-EC)是一种细胞生长调节激素,在医疗和生物农药等方面有极高的应用价值。利用匍枝筋骨草细胞中含有较高β-EC这一特性,为从改善细胞悬浮培养、监测及加入添加物等途径找出一个可以高效、快速生产β-EC的生物学方法,通过紫外分光光度法和高效液相色谱等研究方法在前期已建立的匍枝筋骨草细胞悬浮培养体系的基础上,进一步研究了细胞培养过程中的营养消耗、次生代谢产物生成的动力学曲线;并探索了向细胞悬浮培养体系中添加L-苯丙氨酸(L-PAL)、甲羟戊酸(MVA)、α-蒎烯、松油醇、NO(硝普钠SNP)等来提高匍枝筋骨草细胞中β-EC含量。结果表明:匍枝筋骨草细胞生长符合逻辑斯蒂方程曲线“S”型,生长周期共19 d并在第11天进入细胞生长稳定期;电导率与细胞干质量、蜕皮甾酮的积累呈负相关,因而,可利用电导率的变化确定最佳细胞收获时间;多次继代会弱化细胞生产蜕皮甾酮的能力;通过将α-蒎烯、MVA、SNP联合添加于弱化的细胞中,可以显著提高细胞悬浮培养体系中β-EC的含量,联合添加的适宜组合和最优浓度为:α-蒎烯50 μL/L、MVA 10 mg/L、SNP 80 μmol/L。      

Fate of alpha-bungarotoxin bound to acetylcholine receptors of normal and denervated muscle

In organ culture, alpha-[(125)I]bungarotoxin bound to extrajunctional receptors of denervated muscle is lost from the tissue at a more rapid rate than the toxin bound to the junctional receptors of normal muscle. The rapid loss of toxin from denervated muscle can be blocked by inhibitors of energy production and protein synthesis, and may reflect turnover of the toxin-receptor complex in the membrane.     

Autocrine induction of collagenase by serum amyloid A-like and beta 2-microglobulin-like proteins

Two autocrine proteins of 14 and 12 kilodaltons that induce the synthesis of rabbit fibroblast collagenase were identified. The proteins were purified from serum-free culture medium taken from rabbit synovial fibroblasts stimulated with phorbol myristate acetate. The amino-terminal sequences of the 14- and 12-kilodalton species were approximately 60 to 80 percent homologous with serum amyloid A and beta 2 microglobulin, respectively. The polyacrylamide gel-eluted proteins retained the ability to induce collagenase synthesis in rabbit and human fibroblasts. These autocrine proteins may provide a means to modulate collagenase synthesis in normal remodeling as well as in inflammation and disease states.     

Regulation of the erythropoietin gene: evidence that the oxygen sensor is a heme protein

Erythropoietin (Epo), the hormone that stimulates red blood cell production, is synthesized in the kidney and liver in response to hypoxia. The human hepatoma cell line Hep3B regulates its production of Epo in a physiologic manner. Either hypoxia or cobalt chloride markedly increases expression of Epo mRNA as well as production of biologically active and immunologically distinct Epo protein. New protein synthesis is required before the induction of increased levels of hypoxia- or cobalt-induced Epo mRNA. Hypoxia, cobalt chloride, and nickel chloride appear to stimulate Epo production through a common pathway. The inhibition of Epo production at low partial pressures of oxygen by carbon monoxide provides evidence that a heme protein is integrally involved in the oxygen-sensing mechanism. This hypothesis is further supported by the finding that when heme synthesis is blocked, hypoxia-, cobalt-, and nickel-induced Epo production are all markedly inhibited. A model is proposed in which a ligand-dependent conformational change in a heme protein accounts for the mechanism by which hypoxia as well as cobalt and nickel stimulate the production of Epo.     

Electroshock effects on brain protein synthesis: relation to brain seizures and retrograde amnesia

The effects of electroshock on brain seizure activity and brain protein synthesis were studied in male mice. A significant but short-lasting inhibition of brain protein synthesis and an increase in the amount of free leucine were produced by electroshock at intensities above the brain seizure threshold. Electroshock at intensities below the brain seizure threshold did not affect brain protein synthesis.     

Neurotrophic protein regulates muscle acetylcholinesterase in culture

Skeletal muscles lose acetylcholinesterase in culture as a result of denervation. A protein fraction isolated from peripheral nerves maintained the level of acetylcholinesterase in cultures of aneural embryonic muscle or denervated adult chicken muscle. These results indicate that trophic regulation of muscle acetylcholinesterase might be mediated by a protein produced by nerves.