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[Effect of actinomycin D and cycloheximide on lipoproteins in blood serum and liver cytosol of rats under oxidative stress].

The influence of protein synthesis inhibitors on rat lipoprotein content and composition under oxidative stress caused by cobalt chloride injection has been investigated in the present work. It has been concluded that apoprotein synthesis is very important process influencing on adaptive reactions under free-radical oxidation activation conditions. Co-administration of cobalt chloride and actinomycin D or cycloheximide (the inhibitors of the protein synthesis) has realy prevented hyperlipoproteinemia in many cases but hasn't influenced on lipoprotein oxidation. Pre-beta- and beta-lipoproteins were discussed to have mRNA pool in hepatocytes.

Animals↗

[Study of the ultrastructural and functional expression of the hepatocyte genome under conditions of prolonged depression of protein biosynthesis by cycloheximide. III].

Dynamics of changes in mtRNA synthesis and mitochondria ultrastructure is strictly dependent on the level of inhibition of biosynthesis of cytoplasm proteins and "soluble" proteins of mitochondria by cycloheximide in hepatocytes: 1-6 hrs later a progressive weakening of protein synthesis is accompanied by a drop in mtRNA synthesis and essential destruction of mitochondria; from 12 to 24 hrs a partial restoration of protein biosynthesis induces the processes of the above-mentioned indexes normalization.

Animals↗

[Cycloheximide induced decrease in frost resistance of winter wheat is mediated by inhibition of adaptive rearrangement of the cell and chloroplast structure].

Cycloheximide in the concentration inhibiting protein synthesis and frost resistance of plants proved to prevent formation of cell and chloroplast structures specific for the plants adapted to frost. Important role of structural rearrangement of the cells during hardening preventing intracellular ice nucleation is proposed.

Adaptation, Physiological↗

[Ultrastructural study of the pseudopregnant rat luteal cell following the effect of luteinizing hormone and of 2 inhibitors of synthesis (amino-glutethimide and cycloheximide)].

Incubation of ovaries from pseudopregnant rats have been performed with the gonadotropin LH and with two inhibitors, aminoglutéthimide and cycloheximide for periods comprised between 1 and 6 hours. Then the luteal cells have been studied with electron microscope. Measurements in have been used to express the variations in volume of two organels, mitochondria and lipid droplets. The principal modifications induced by these compounds bear on the mitochondria. Treatment with LH is followed by an increase in the volume of individual mitochondria resulting probably of fusion of organels. Treatments with the inhibitors are followed by reduction in the volume of individual mitochondria and in the total volume of mitochondrial apparatus. Mixed treatments using LH and an inhibitor allow to obtain organels of intermediate size. Volume of lipids is increased by incubation with LH and probably decreased by the inhibitors. Smooth endoplasmic reticulum shows rearrangements with the various treatments. It seems not convenient to extend incubations over 4 hours in order to avoid cytolysis.

Aminoglutethimide↗

Effect of cycloheximide on viral precursor protein B in Sindbis virus-infected BHK cells.

The possible role of B protein, a postulated precursor of viral envelope proteins was studied in Sindbis virus-infected BHK cells. Although tryptic digestion of the B protein produces tryptic peptides of the viral envelope proteins it seems unlikely that B molecules accumulated in the cytoplasm of infected cells are subsequently cleaved into structural proteins. We suggest that if B is really the precursor of E1 and PE2 viral envelope proteins, the cleavage takes place on the ribosomes during the synthesis of the second part of the B protein. Low concentrations of cycloheximide completely blocked the synthesis of B protein but did not inhibit the synthesis of viral structural proteins.

Animals↗

The effect of cycloheximide on butyrylcholinesterase activity in vivo.

The paper describes the catalytic activity of butyrylcholinesterase (BuChE) measured in plasma, liver, white adipose tissue, heart, and brain of rats intraperitoneally administered a single non-lethal dose of cycloheximide (2.0 mg/kg body weight; CHM). The BuChE assay was performed on rats of both sexes either administered CHM or saline (controls), and killed 2, 3, 4, 5, 10 days later. A significant decrease of BuChE catalytic activity was observed in all tested tissues except plasma. In animals of both sexes, the lowest BuChE catalytic activity was found in the liver (2-6%), while it was higher in white adipose tissue, heart, and brain. However, the respective values remained significantly different from controls (33-67%, 49-62%, and 14-71% in males, and 24-82%, 72-86%, and 33-67% in females). Since there was no effect of CHM on BuChE catalytic activity in plasma, the data suggest that CHM inhibits the synthesis of BuChE rather than its active site.

Animals↗

Effects of cycloheximide treatment on in-vitro development of porcine parthenotes and somatic cell nuclear transfer embryos.

This study aimed to verify the beneficial effect of cycloheximide (CHX) treatment on the development of porcine somatic cell nuclear transfer (NT) embryos, which were constructed with enucleated oocytes and cumulus cells by using a single direct current (DC) pulse. In the first experiment, a single DC pulse applied to the induction of fusion and activation of NT embryos gave a high fusion rate. However, cleavage and subsequent development of fused couplets (NT embryos) to the blastocyst stage were poor. Experiment 2 was conducted to determine whether CHX treatment could enhance metaphase II (M II) oocyte activation and improve the subsequent parthenogenetic development. After giving the DC pulse and incubation with or without CHX, M II oocytes incubated with CHX showed higher cleavage and development to blastocysts compared with those incubated without CHX (P < 0. 05). Experiment 3 was carried out to verify the beneficial effect of CHX on the development of NT embryos. The NT embryos treated with the DC pulse and CHX treatment showed higher cleavage and subsequent development compared with those treated with the DC pulse alone (P < 0.05) . The present study demonstrates that CHX treatment enhances the electrical stimulus-induced activation of oocytes and NT embryos, and improves the subsequent development of parthenotes and NT embryos. The results indicate that protein synthesis inhibition treatment required for the induction of oocyte activation promotes the development of NT embryos.

Animals↗

[Wallerian degeneration of the rat sciatic nerve. Lack of effect of cycloheximide and of actinomycin D on the increase of radiophosphate incorporation in acid soluble phosphates observed after section of the nerve].

Plasma inorganic phosphate incorporation into inorganic and organic acid-soluble phosphate fractions of the rat sciatic nerve increases markedly during Wallerian degeneration. Administration of cycloheximide and actinomycin D remains without effect on this specific increase which appears thus independent of cytoplasmic protein neosynthesis.

Animals↗

[Study on a synchronized population of the antimitotic effect of cycloheximide at various phases of the cell cycle in Allium sativum L].

The effect on the entry in mitosis, induced in synchronized meristematic cells of Allium sativum L. by a treatment with cycloheximide (10 mug/ml-30 min) at different phases of the cell cycle has been investigated. In all cases cells enter mitosis after a long delay but it appears that the delay induced on G1 cells is longer. Incorporation of 3H leucine is immediately inhibited at any stage of the cell cycle.

Cell Division↗

A moderate dose of cycloheximide does not prevent the febrile response to endotoxin but interfere with induction of pyrogenic tolerance in rabbit.

The purpose of the present study was to examine the effect of cycloheximide (Cx)--inhibitor of protein synthesis, on the development of pyrogenic tolerance to LPS. It has been observed that Cx at a dose of 1 mg/kg given intravenously 1 h prior to LPS did not prevent fever response, however it modified the induction of pyrogenic tolerance. It was manifested in existence of the second phase of fever after the following administrations of LPS into rabbits pretreated with Cx. In control group of rabbits the induction of pyrogenic tolerance was accompanied with decaying of the second peak of fever visible as early as the second dose of LPS.

Animals↗

[Structural changes of chromatin during proto-oncogene activation by cycloheximide: dose-effect].

The level of expression of cellular proto-oncogens c-myc and c-fos in rat liver has been studied as a function of protein synthesis rate (cycloheximide dose). Activation of proto-oncogens has been established to be initiated by 50% inhibition of nuclear protein synthesis. This promotes a certain level in chromatin structural rearrangements which is manifested, in particular, in decreasing activity of chromatin cleavage by Ca2+, Mg(2+)-DNAase and increasing degree of chromatin condensation. A role of topoisomerase II in chromatin structural rearrangements during proto-oncogen activation is postulated.

Animals↗

[Induction of apoptosis in FAF28 cells by the protein synthesis inhibitor cycloheximide].

We have studied the influence of translation of inhibitor cycloheximide (CHI) on the Chinese hamster fibroblast line FAF28. This treatment inhibited the cell cycle and induced apoptosis. A short-term treatment and the following removal of CHI reduced the number of apoptotic cells up to the normal level. Under changed conditions (change in CO2 proportion and pH of cultural medium), the ability of CHI to induce apoptosis increases. The study of cell ultrastructure demonstrates three groups of apoptotic cells differing from each other in the state of cytoplasmic organelles: 1) cells with many well-identified organelles; 2) cells with a scanty number of cytoplasmic organelles; 3) cells lacking any recognizable organelles in the cytoplasm. Presumable causes of this morphological variety of apoptotic cells are discussed.

Animals↗

Rapid induction of apoptosis in rat liver by cycloheximide.

A single administration of the inhibitor of protein synthesis cycloheximide results in the occurrence of apoptosis in rat liver. The presence of intracellular apoptotic bodies was detected as early as 2 hours after treatment. No evidence of cell necrosis could be observed by histologic and biochemical analysis. Apoptosis was followed by an increased expression of testosterone-repressed prostate message-2 RNA, a gene whose activity has been associated to apoptotic cell death in involuting rat prostate. The finding of in vivo induction of apoptosis in nonproliferating cells by an inhibitor of protein synthesis, together with the rapidity and synchrony in the occurrence of cell death make this model potentially useful for the analysis of the kinetics of the apoptotic cycle and in exploring some of the mechanisms of regulation of genes possibly involved in this type of cell death.

Animals↗

Influence of cycloheximide on apoptosis in CHO cells, induced by ethane 1,2-dimethanesulphonate (EDS).

Ethane 1,2-dimethanesulphonate (EDS) causes apoptotic death of Leydig cells. Additionally, EDS causes damage of Chinese Hamster Ovary (CHO) cells but the occurrence of apoptosis is not such plentiful. The present study tested whether the inhibition of protein synthesis by cycloheximide (CHX) would influence apoptosis of CHO cells, induced by EDS. The study compounds induced morphological changes in CHO cell typical for apoptosis. An active form of caspase-9 and an alternation of mitochondrial transmembrane potential were also observed. In our study, a more cumulative effect of the CHX and EDS on apoptosis was observed, when both compounds were simultaneously employed. The obtained results indicated that synthesis of antiapoptotic proteins plays a very important role in the inhibition of apoptosis.

Animals↗

Effect of cycloheximide on the adaptive response induced by low dose radiation.

Human lymphocytes pre-exposed to 10 mGy or 50 mGy of X-rays become less sensitive to subsequent large dose irradiation, exhibited lower rate of chromosome aberration than expected. This adaptive response could be inhibited by cycloheximide, a protein synthesis inhibitor for successive 2 h period ranging from 0.5h before to 4h after the low dose exposure, indicating that the adaptive response was directly related with the protein synthesis.

Adaptation, Physiological↗

Induction by cycloheximide of the glycoprotein hormone alpha-subunit gene in human tumor cell lines and identification of a possible negative regulatory factor.

The steady-state level of mRNA encoding the glycoprotein hormone alpha-subunit is increased about 4-fold in HeLa cells by cycloheximide (CHX) or puromycin at concentrations that inhibit protein synthesis. This effect is observed in a number of cell lines that ectopically produce alpha-subunit, including ChaGo (brochogenic carcinoma), FL (amnion), and HeLa (cervical carcinoma). No increase in alpha-subunit mRNA is evident in two choriocarcinoma cell lines (JAr, JEG-3) that produce alpha-subunit as an eutopic product. The half-life of alpha-subunit mRNA is unchanged in the presence of CHX, but nuclear run-on assays demonstrate a 2.6-fold greater loading of RNA polymerase on the alpha-subunit gene in nuclei from CHX-treated cells. These results suggest that inhibition of protein synthesis results in higher transcription rates and not in decreased mRNA turnover. A nuclear protein (Mr 50,000) that binds to a DNA fragment located 5' proximal to the alpha-subunit gene but not to more distal 5'-flanking sequence or to the alpha-subunit cDNA has been identified in HeLa but not in JEG-3 cell lines. The p50 DNA binding activity in HeLa cells decreases in the presence of CHX at a rate similar to that at which alpha-subunit mRNA increases. Moreover, in a series of HeLa cell clones, the levels of p50 are directly proportional to the magnitude of induction produced by CHX. These data are consistent with a model for alpha-subunit gene regulation involving a labile repressor and constitute yet another level of differential regulation of the alpha-subunit gene in cells that produce the hormone subunit in an ectopic versus eutopic manner.

Cell Line↗

[Effects of cycloheximide on protein, RNA and DNA synthesis in cultures of CHO cells and human diploid fibroblasts].

In CHO cell line and primary human diploid fibroblasts culture an incorporation of protein, RNA and DNA biosyntheses precursors was investigated under different conditions of inhibition of translation by cycloheximide (CHM). Both CHO and human fibroblasts transitory treatment by CHM in the serumfree medium resulted in inhibition of protein and DNA syntheses during S-period while RNA synthesis increased up to 130% (CHM concentration from 0.003 to 2 Mg/ml), as well as in Go--an incorporation of 3H-U increased to 200% (CHM concentration-100 Mg/ml). Long-term treatment (48 hours) in the serum-free medium resulted in decreased uptake of 3H-T and 3H-L during first 6 hours of experiment, while incorporation of 3H-U increased to 160%. By 16-th hour of treatment characters of protein, RNA and DNA syntheses came back to control levels.

Animals↗

[Comparison of lipid metabolism in rat liver cells and nuclei during cycloheximide-induced superinduction of nuclear oncogenes].

Using a model of cycloheximide (CHI)-induced expression of nuclear oncogens, a comparative study of metabolism of the major lipid classes in rat liver nuclei and cells was carried out. A short-term activation of sphingomyelinase which preceded on a time scale the maximal accumulation of c-fos and c-myc transcripts was observed both in the cells and in the nuclei. In contrast with the whole cell, the level of phospholipase C activity in the nuclei did not change under conditions of oncogene activation. It was found that the maximal expression of nuclear oncogens coincided in time with cyclic changes in the content of practically all phospholipids and neutral lipids with simultaneous activation of their synthesis both in the cells and in the nuclei. However, in the nuclei the sphingomyelin metabolism activation was predominant. It is concluded that in the nucleus sphingomyelin and its metabolites may influence oncogene expression via nuclear protein kinase C.

Animals↗