[Investigation of properties of the thermostable protein complex obtained from the myocardium left ventricles of adult white rats].

A thermostable protein complex (TSPC) obtained from the myocardium ventricules of adult rats inhibits mitotic and transcriptional activity of cardiomyocytes. At the same time this complex is more active on early stages of the postnatal ontogenesis in rats, aged 1 and 5 days. Following its action on RNA synthesis, the TSPC reveals tissue specificity only in cells with terminal differentiation, and is determined by its nuclear membranes. We continue studies for identifying the molecular weight and chemical nature of the TSPC, and the role of its different fractions in regulation of proliferation processes. Besides, it is planned to produce antibodies against TSPC fractions with the purpose to block its inhibitory effect on myocyte regeneration in the damaged myocardium.

Changes in nuclear macromolecules and nucleolar volume associated with selective ribosomal cistron activation by ethylene.

Pieces of leaf base tissue from the equatorial region of the third turgid onion (Allium cepa L.; yellow, sweet Spanish, quiescent tissue) leaf base with two visible major nucleoli (volume = 0.51 microns3) and two inactive minor nucleolar organizer regions (NORs) in the outer epidermal cells were placed in shallow water (time = T0 control). After 3 h (T3 control), the two visible nucleoli enlarged (178% of T0) and changed shape (from 88% round and 12% oval at T0 to 5% round, 68% oval, 20% elongated-oval, and 7% dumbbell). The minor NORs remained inactive. Nuclear RNA and non-histone protein (nHP) increased (RNA = 128% and nHP = 134%, compared with T0 data, set at 100%). The content of DNA and histone protein (H) in nuclei remained unchanged. Major nucleoli in pieces of onion placed in 10(-5) M ethylene enlarged and minor nucleoli became visible in 33% of the nuclei; i.e., ethylene acted as a selective ribosomal cistron regulator. The nucleolar volume (212% of T0) was statistically greater than those for T0 and T3. DNA and H remained unchanged (not statistically different) but RNA (159% of T0) and nHP (169% of T0) were greater (statistically significant) than those for T0 and T3 controls. Cobalt chloride (10(-5) M Co) alone or in 10(-5) M ethylene (Co + E) inhibited the enlargement of some of the major nucleoli and completely inhibited the appearance of minor nucleoli. The nucleolar volumes for the cobalt and for cobalt-ethylene treatments were significantly less than those for T3 controls and ethylene treatment at T3. DNA and histone did not change.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence for the preventive effect of the polyunsaturated phytol side chain in tocotrienols on 17beta-estradiol epoxidation.

BACKGROUND: We found that 17beta-estradiol (E2) could be activated by epoxidation to bind DNA and to inhibit nuclear RNA synthesis. Vitamin E compounds are powerful antioxidants and chain-breaking free radical scavengers. The chromanol ring in Vitamin E is believed to be involved in these reactions. METHODS: Here, we examined the preventive effect of alpha-tocopherol, alpha-, gamma- and delta-tocotrienols on E2 activation. RESULTS: We found that when any one of these Vitamin E compounds was mixed with E2 for epoxidation by the epoxide-forming oxidant dimethyldioxirane (DMDO), alpha-tocopherol was the least effective as compared with the tocotrienols against the formation of E2 epoxide as reflected by the loss of the ability of E2 to inhibit nuclear RNA synthesis. This conclusion was further confirmed by the binding studies of [3H] labeled E2 to DNA using either DMDO or liver microsomes activation system. CONCLUSIONS: Since the chromanol ring is shared by both tocopherols and tocotrienols and the only difference between these two subgroups of Vitamin E is the phytol side chain, we conclude that the polyunsaturated phytol group in tocotrienols plays a key preventive role in E2 epoxidation. This is the first report showing that the polyunsaturated phytol side chain in tocotrienols is involved in an antioxidative activity and it may also have a preventive effect against the E2 epoxide induced breast cancer carcinogenesis at the initiation.

Phospholipase C digestion induces the removal of nuclear RNA: a cytochemical quantitative study.

It has been reported that the incubation of isolated rat liver nuclear matrices with phospholipase C causes the digestion of the matrix-bound phospholipids and the release of most matrix-linked RNAs (Cocco et al., 1980). In this paper, the presence of phospholipids in nuclear substructures and the effects of their removal by phospholipase C digestion have been investigated by means of enzyme-colloidal gold cytochemistry. The nuclear phospholipids appear to be localized in the interchromatin areas and in the nucleolus and are virtually absent in the heterochromatin, when labelled with phospholipase C-colloidal gold. The double labelling test with ribonuclease A and phospholipase C conjugated with gold particles of different diameters shows that the nuclear phospholipids are co-localized with RNA-containing structures. The enzymatic digestion of phospholipids on thin sections of either isolated nuclei or pancreas embedded in LR White resin results in the decrease of the RNase-A colloidal gold labelling of nuclear RNA-containing structures, but not of the rough endoplasmic reticulum. The data confirm the presence of phospholipids in the nucleus in the absence of possible translocation due to isolation procedures and strengthen the hypothesis that they are involved in interactions between nucleic acids and proteins of the nuclear matrix.

Transcriptional induction of hepatic NADPH: cytochrome P450 oxidoreductase by thyroid hormone.

Studies were carried out to elucidate the mechanism whereby thyroid hormone (T3) induces NADPH:cytochrome P450 oxidoreductase (P450R) mRNA in rat liver in vivo. Northern blot analysis revealed that T3 treatment increases unspliced liver nuclear P450R RNA 4-fold within 8 h and that this induction precedes the induction of mature, cytoplasmic P450R RNA. Unspliced nuclear P450R RNA was suppressed below basal levels 24 h after T3 treatment, despite the continued presence of elevated circulating T3 levels. To determine whether the T3-stimulated increase in nuclear P450R RNA reflects an increase in P450R transcription initiation, nuclear run-on transcription assays were carried out. T3 induced a 6- to 8-fold increase in P450R transcription rate within 12 h, sufficient to account for the observed increase in nuclear P450R precursor RNA, followed by a decrease back to basal transcription levels at 24 h, consistent with the nuclear RNA profile. Similar transcriptional increases were observed in nuclear run-on transcription studies using hybridization probes corresponding to nine different fragments of the P450R gene, spanning exon 2 to exon 16. Thus, P450R transcription initiation, not transcription elongation, is the T3-regulated event. Similar results were obtained during short (5 min) compared with long (45 min) nuclear run-on transcription assays, suggesting that changes in nuclear RNA processing or regulated degradation do not contribute to the overall RNA induction. This finding was confirmed by the ability of the RNA polymerase inhibitor actinomycin D, administered in vivo, to block T3 induction of P450R transcriptional activity. We conclude that P450R transcription, rather than nuclear RNA processing or mRNA stabilization, is the primary mechanism whereby T3 induces hepatic P450R mRNA.

Aberrance and modification of alpha-1- and alpha-2-globin gene expression in human and mouse erythroleukemia cells.

In contrast to normal human erythroid tissues where the alpha 2:alpha 1-globin mRNA ratio is about 72:28, in human erythroleukemia K562 cells this ratio was found to be quite low, i.e. about 8:92. The ratio was moderately increased by hemin induction and approached almost normal levels after chromosomal transfer from K562 to the mouse erythroleukemia (MEL) cells. We suggest that operationally positive regulatory factors may exist in erythroleukemia cells, modifying the relative alpha 1- and alpha 2-globin gene expression by events following induction and by the adult MEL environment. These factors may act by altering the relative rate of alpha 1- and alpha 2-globin mRNA synthesis.