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Biomedical subjects

M Orunesu

Publications and source records attributed to M Orunesu.

At least 37 records · Page 2Linked to original sources
PubMed1986

Effect of glutathione and N-acetylcysteine on hepatocellular modifications induced by 2-acetylaminofluorene.

The exposure of rats to a dietary regimen containing 2-acetylaminofluorene induces a sequence of hepatocellular alterations leading to the development of preneoplastic nodules. Groups of 2-acetylaminofluorene-treated rats were given glutathione or N-acetylcysteine to evaluate the effects of these different thiols on the sequence of events that originate transformed cells. It is well known that intracellular thiols protect biological macromolecules from scavenging free radicals and electrophilic compounds produced by the metabolism of chemical agents. Male Wistar rats were maintained on a feeding regimen containing 0.05% 2-acetylaminofluorene. The diet of 2 groups of 2-acetylaminofluorene-treated animals was supplemented with either 0.1% glutathione or N-acetylcysteine. The effects in the liver of the exogenously supplied thiols during 2-acetylaminofluorene treatment were assessed evaluating DNA damage, glutathione levels, activity of marker enzymes glucose-6-phosphatase, gamma-glutamyltranspeptidase, and glutathione-S-transferase, survival rates, and development of salivary gland tumors. Our results demonstrate that the mortality due to 2-acetylaminofluorene exposure was reduced or completely abolished by thiols and that the development of salivary gland tumors was inhibited. Exogenously supplied thiols significantly reduced DNA damage as assessed by alkaline elution. At the doses employed, glutathione and N-acetylcysteine induce early stimulation of glutathione-S-transferase, had little effect on the loss of glucose-6-phosphatase activity and scanty influence on the net increase in gamma-glutamyltranspeptidase activity.

2-Acetylaminofluorene↗
PubMed1985

Triiodothyronine-stimulated RNA synthesis in primary cultures of adult rat hepatocytes.

We have studied the effect of triiodothyronine (T3) on RNA synthesis by primary cultures of rat hepatocytes in order to ascertain whether hepatocyte transcriptional activity is directly stimulated by this hormone. The results demonstrate that T3 stimulates RNA synthesis as measured by [3H]orotic acid incorporation into RNA and by RNA polymerase activity. The responsiveness of cultured hepatocytes to T3 becomes evident only after a fairly long latency period required for the recovery of T3 nuclear binding sites. The response of RNA synthesis to T3 was absent unless the hepatocytes were simultaneously exposed to insulin and dexamethasone, indicating a permissive role of these factors in the action of T3 on RNA synthesis.

Animals↗
PubMed1985

Membrane phosphatase activities in isolated and cultured adult rat hepatocytes.

Membrane bound phosphohydrolysing enzymes, such as Na-K-ATPase, Mg-ATPase, ALPase and G-6-Pase were assayed in intact liver, in freshly isolated cells and in cultured hepatocytes to evaluate the effects of the isolation procedure and culture on these enzyme activities. Na-K-ATPase and Mg-ATPase are significantly reduced following cell dispersion while ALPase and G-6-Pase are nearly unaffected. During culture, Na-K-ATPase is restored to the "in vivo" level within the first two days, but rapidly declines in the following days. The Na dependent, energy requiring AIB uptake shows a similar pattern; Mg-ATPase is practically unmodified. A significant increase in ALPase activity and a net decrease of G-6-Pase activity, as a function of the culture time has been observed.

Adenosine Triphosphatases↗
PubMed1984

Biochemical and morphological comparison of microsomal preparations from rat, quail, trout, mussel, and water flea.

Differential centrifugation methods already in use were applied to purify rat, quail, and trout liver microsomes and modified as necessary to purify microsomes from mussel digestive gland and whole water flea. All these microsomal preparations were comparatively characterized with respect to protein and RNA content, levels of markers of subcellular contaminants, ultrastructural morphology, differential spectra of cytochromes P-450 and b5, monoxygenase activity, and in vitro metabolism of p-dichlorobenzene. Yields of microsomal proteins of the tested organisms differed widely, with mussel showing the lowest yield. Very low levels of nuclear and mitochondrial contaminants were detected in all microsomal preparations, but cell membrane contaminants were clearly present in most preparations. Daphnia microsomes were significantly contaminated by plasma membranes, and hepatopancreas microsomes contained significant amounts of partially disrupted secretory granules and plasma membrane. From a qualitative standpoint differential spectra of cytochrome b5 were very similar for all the preparations, whereas cytochrome P-450 spectra were largely dependent on the microsomal preparation as well as on the assay method used. The content of cytochrome P-450 was highest for rat liver microsomes and very low or absent in Daphnia and mussel preparations; the range of cytochrome b5 contents was much narrower. Significant differences were observed among monoxygenase activities of the different preparations. In Daphnia and mussel microsomes, aniline hydroxylase was absent and benzo[a]pyrene hydroxylase activity was much lower than in rat and quail microsomes. Benzo[a]pyrene hydroxylase activity of trout liver microsomes was similar to that of rat and quail microsomes, whereas hydroxylation of substrates which in rat liver are preferentially metabolized by phenobarbital-inducible forms of cytochrome P-450 was much lower in trout microsomes.

Animals↗
PubMed1984

Collagenase perfusion of rat liver induces DNA damage and DNA repair in hepatocytes.

Evidence is presented that the collagenase perfusion of adult rat liver results in significant damage to nuclear DNA as evaluated by the alkaline elution technique. The extent of the damage is related to the perfusion time as well as to the clostridial enzyme preparation used. The DNA structure of isolated cells is almost completely repaired within 12 h of their culture in chemically defined medium.

Animals↗
PubMed1981

Increased levels of rat liver RNA polymerase I(A) and I(B) following the administration of triiodothyronine.

The levels of the transcribing RNA polymerase I(B) in the nucleus and of the non-transcribing RNA polymerase I(A) in the cytoplasm are both approximately doubled 24 h after a single i.p. injection of triiodothyronine into thyroidectomized rats. This suggests that the triiodothyronine-induced stimulation of ribosomal RNA synthesis is associated with an increase in the total RNA polymerase I content of rat liver cells.

Animals↗
PubMed1980

Inhibition of isolated rat liver RNApolymerases I and II by aminoacridines.

9-Aminoacridine and 2 derivatives which contain hydroxyalkylic or aminoalkylic side chains in the 9-position totally inhibit the transcription of calf thymus DNA by rat liver RNA polymerases I and II. This inhibitory action does not always appear to be completely related to the ability of aminoacridines to intercalate into the DNA template.

Aminacrine↗
PubMed1979

[Influence of temperature on the preferential extraction of RNA polymerase I from hepatic nuclei of the rat].

RNA polymerase I has been extracted from rat liver nuclei by three consecutive washings at 0 degrees C with a medium of relatively low ionic strength (0.15 M KCl) containing Mg++ rather than by incubating the organelles at 37 degrees C in the same medium, as originally proposed by Chesterton and Butterworth. The modified technique, which has the advantage of preventing a temperature-mediated conversion of form IB to IA, gives similar yields of RNA polymerase I and retains the capacity of preferentially extracting the enzyme with respect to the other forms of nuclear RNA polymerase.

Animals↗
PubMed1979

[Reduced catalytic effectiveness of RNA polymerase I in hepatocytes of rats treated with cycloheximide].

Rat liver RNA polymerase I solubilized from isolated nuclei and present in a soluble form in the cytoplasmic fraction has been analyzed by phosphocellulose chromatography 3 hours after the administration of cycloheximide. The antibiotic did not induce any change in the chromatographic properties of both nuclear and cytoplasmic RNA polymerase I. They appeared to remain in the IB and IA forms, characteristic of the transcribing (IB) and non-transscribing (IA) enzyme. While the level of the nuclear enzyme was not modified, the level of the cytoplasmic one appeared significantly increased. These results support previous ones indicating that the cycloheximide-induced inhibition of ribosomal RNA synthesis cannot be merely explained by a decrease in the nuclear or cellular level of RNA polymerase I. The cellular level of RNA polymerase I, taking into account the relative proportion of the enzyme found in nuclei and cytoplasm, appeared to be slightly increased. Cycloheximide administration did not seem to result in the appearance, in intact nuclei, of enzyme molecules in a free form or as blocked transcription complexes. It is concluded that the antibiotic affects the catalytic efficiency rather than the number of RNA polymerase I molecules actually engaged in the transcription of ribosomal cistrone.

Animals↗
PubMed1977

Increased activity of rat liver nucleolar protein kinase following triiodothyronine administration.

Triiodothyronine (T3) administration to thyroidectomized rats induces a significant increase in the nucleolus-associated protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) activity. The general properties of the protein kinase solubilized from liver nucleoli have been investigated. Mg2+ (20 mM) is essential for the reaction and an appropriate concentration of NaCl (100 mM) is required to achieve maximal phosphorylation rates. The optimal pH for casein phosphorylation is 7.6. The kinase phosphorylates casein more efficiently than phosvitin and displays an almost undetectable activity towards histones and protamine. No significant stimulation of the kinase activity by cyclic AMP has been detected. The apparent Km values for casein and ATP are 1.5 mg/ml and 1.5-10(-5) M, respectively, and are not affected by the hormone administration.

Animals↗
PubMed1977

Increased RNA polymerase activity in isolated liver nucleoli from thyroidectomized rats treated with triiodothyronine.

In isolated liver nucleoli from thyroidectomized rats the activity of the two RNA polymerase I populations, one of which is active and the other inactive towards the endogenous chromatin template, is greatly enhanced 10 and 24h after a single ip injection triiodothyronine (T3). When the nucleolar enzyme is solubilized and assayed with exogenous DNA as template, it retains, after T3 treatment, the same increase in activity as observed in intact nucleoli. On the contrary, the template availability, as judged by the binding capacity of isolated nucleoli for [3H]actinomycin D, does not appear to be modified by the hormone. These observations support the conclusion that the enhanced nucleolar RNA synthesis following T3 administration is due to an increased activity of the RNA polymerase I itself rather than to a greater availability of ribosomal RNA cistrons. The hormonal stimulation of both nucleolar RNA polymerase activities depends on continuous protein synthesis since it is almost completely abolished by the administration of cycloheximide.

Animals↗
PubMed1975

Sequential stimulation of nuclear RNA polymerase activities in livers from thyroidectomized rats treated with triiodothyronine.

A single ip injection of triiodothyronine (T3; 30 mug/100 g BW) to thyroidectomized rats markedly stimulates RNA synthesis in isolated liver nuclei. The increased level of RNA synthesized in vitro by isolated nuclei does not depend on a reduced degradation of the nascent RNA molecules, since ribonuclease activities are not affected by the administration of T3. In addition, our results have confirmed previous findings of Tata et al. that the increase in nucleolar alpha-amanitin-resistant RNA polymerase I activity at low ionic strength always preceded the rise of the nucleoplasmic alpha-amanitin-sensitive RNA polymerase II activity at high ionic strength. Moreover, it has been found that a significant increase in an alpha-amanitin-resistant activity at high ionic strength occurs as early as 10 h after hormone injection. This enzyme, which forms RNA with a U to G ratio significantly higher than that of RNA synthesized by the nucleolar alpha-amanitin-resistant enzyme, is probably nucleoplasmic RNA polymerase III which is though to synthesize 5S and transfer RNAs. The possible role and the mechanism(s) of the early and concomitant increase in nucleolar and nucleoplasmic alpha-aminitin-resistant activities, and of the subsequent rise of RNA polymerase II activity following T3 administration are discussed.

Amanitins↗