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V Mares

Publications and source records attributed to V Mares.

At least 19 recordsLinked to original sources
PubMed1992

Spontaneous hemorrhage in the cerebral cortex of immature rats.

We report the presence of spontaneous hemorrhages in the brain of normal neonatal rats. The quantitative evaluation showed that the hemorrhaging was most frequent in 4- to 5-day-old rats. Nearly all the lesions were found in the cerebral cortex of the lateral part of the forebrain hemisphere, particularly in the middle cortical layers. Their average incidence amounted to 6.71 +/- 2.95 per six 5-microns sections from 5-day-old rats. The lesions were nearly absent in the newborn and 7-day-old animals. We suggest that vascular wall immaturity and some extravascular factors, like developmental increase of blood pressure, may account for the perinatal hemorrhaging.

Animals↗
PubMed1992

Accumulation of boron-10 (10B) in cell cultures exposed to mercaptododecaborate (Na2H(11)10B12SH) used for the neutron capture therapy of brain tumors.

Toxicity of mercaptoundecahydro-closo-dodecaborate (MHB, Na2H(11)10B12SH) and accumulation of MHB-derived 10B were studied in E7 neuroblastoma, C6 glioma, HeLa cells and embryonic lung LEP 19 fibroblasts in culture in exponential and stationary phases of growth (2- and 7-day-old cultures, respectively). The pilot study of acute toxicity, performed on C6 glioma cells, showed good tolerance of the drug up to 1000 micrograms/ml (4.8 x 10(-3) M), when cell growth slowed and a small part of the population was lethally damaged (8.3%, 20-h incubation interval). The changes became more extensive and appeared sooner (toward 5 h) at 2000 micrograms MHB/ml (9.5 x 10(-3) M). None of the four cell lines used was found to be affected in gross morphology or growth by 200 micrograms MHB/ml within a 5-day culture interval. When exposed to this dose for 4 h, the amount of 10B accumulated in cell lines at the exponential growth phase ranged from 0.51 to 4.4 ng/micrograms protein; in the stationary cultures of the corresponding cell phenotype, the 10B values were 3 to 10 times lower (0.12-1.2 ng/micrograms protein). Irrespective of the growth phase, the values achieved in C6 glioma cells were several times higher than in the other cell lines. Furthermore, in the glioma cells, particularly in the exponential phase of growth, accumulation of 10B proceeded against the marked concentration gradient. The data provide a new indication for the use of MHB for boron neutron capture therapy of brain tumors.

Animals↗
PubMed1991

Changes in cell cycle and chromatin distribution in C6 glioma cells treated by dibutyryl cyclic AMP.

C6 glioma cells in culture were treated with 1 mM dibutyryl cyclic AMP (Db-cAMP) for 5, 8, 24 and 72 h. The cells were labelled with [3H]-thymidine before either the end, or the beginning, of the Db-cAMP treatment. The cell cycle passage was monitored by the simultaneous determination of DNA content and DNA synthesis in propidium iodide stained autoradiograms. The data revealed an early (t less than or equal to 3-8 h) and moderate inhibitory effect of Db-cAMP on all phases of the cell cycle except mitosis; some cells (2%) were completely blocked in the S phase. Later (8 less than t less than 24-72 h), the cycling of a substantial part of the population became inhibited in G1 phase. Microdensitometric texture analysis of Feulgen-stained nuclei, performed 24 h after administration of Db-cAMP, showed a higher inhomogeneity of the DNA distribution in cell nuclei, caused by the condensation of a part of the chromatin. This may reflect either changes in genome expression taking part in the process of cAMP induced differentiation or transit of some cells into quiescent G0 or S0 phases.

Animals↗
PubMed1991

Thy 1 expression in the brain of nude mice.

The expression and cell distribution of Thy 1 antigen was studied in the brain of both normal and athymic (nude) young adult mice of the BALB/c strain by immunochemistry. In nude animals Thy 1 fluorescence was less intense and less regularly distributed in the molecular layer of the cerebellum and hippocampus. Thy 1 content determined by ELISA was lower by 10-16% in the cerebellum and 20-25% in the olfactory bulbs of nude mice. The total wet weight of the brain was lower by 16% than in control animals; the deficit in body weight ranged from 34-45%. It is supposed that the changes in Thy 1 expression in nude animals are caused mainly by the underdevelopment of late developing brain regions due to thermoregulatory problems and other postnatal strains occurring in the mutants.

Animals↗
PubMed1990

Premitotic DNA synthesis in the brain of the adult frog (Rana esculenta L.): an autoradiographic 3H-thymidine study.

Replicative synthesis of DNA in the brain of the adult frog was studied by light microscope autoradiography. Animals collected during the active period (May-June) and in hibernation (January) were used. In active frogs, 3H-thymidine labelling occurred mainly in the ependymal cells which line the ventricles. The mean labelling index (LI%) was higher in the ependyma of the lateral and fourth ventricles than in the ependyma of the lateral diencephalon and tectal parts of the mesencephalon. In the recessus infundibularis and preopticus the number of labelled cells (LCs) was several times greater than in the lateral parts of the third ventricle. LCs were seen subependymally only occasionally. The incidence of LCs in the parenchyma of the brain was much lower in most regions than in the ventricular ependyma; LCs were mainly small and, from their nuclear morphology, they were glial cells. The LI% reached the highest value in the septum hippocampi and in the nucleus entopeduncularis. In these locations, LCs were larger and closer in size to the nerve cells of these regions. From comparison with data obtained earlier in the brain of mammals, it is evident that the distribution of proliferating cells in the olfactory and limbic system is phylogenetically conservative. The occurrence of pyknotic cells in the same areas which contain LCs, suggests that cell division reflects in part the process of cell renewal observed in mammals. However, proliferating cells could also be linked to the continuous growth observed in non-mammalian vertebrates. In hibernating frogs, LCs and pyknoses were not seen or were found occasionally, which further indicates the functional significance of both processes.

Animals↗
PubMed1989

Na+, K+-ATPase activity in cultured C6 glioma cells.

Rat C6 glioma cells were cultured for 4 days in MEM medium supplemented with 10% bovine serum and Na+, K+-ATPase activity was determined in homogenates of harvested cells. Approximately 50% of enzyme activity was attained at 1.5 mM K+ and the maximum (2.76 +/- 0.13 mumol Pi/h/mg protein) at 5 mM K+. The specific activity of Na+, K+-ATPase was not influenced by freezing the homogenates or cell suspensions before the enzyme assay. Ten minutes' exposure of glioma cells to 10(-4) or 10(-5) M noradrenaline (NA) remained without any effect on NA+, K+-ATPase activity. Neither did the presence of NA in the incubation medium, during the enzyme assay, influence the enzyme activity. The nonresponsiveness of Na+, K+-ATPase of C6 glioma cells to NA is consistent with the assumption that alpha (+) form of the enzyme may be preferentially sensitive to noradrenaline. Na+, K+-ATPase was inhibited in a dose-dependent manner by vanadate and 50% inhibition was achieved at 2 x 10(-7) M concentration. In spite of the fact that Na+, K+-ATPase of glioma cells was not responsive to NA, the latter could at least partially reverse vanadate-induced inhibition of the enzyme. Although the present results concern transformed glial cells, they suggest the possibility that inhibition of glial Na+, K+-ATPase may contribute to the previously reported inhibition by vanadate of Na+, K+-ATPase of the whole brain tissue.

Animals↗
PubMed1988

Intranuclear differences in the response of Purkinje cell DNA of the rat cerebellum to bleomycin. A microphotometric and autoradiographic study.

A single dose of the DNA-binding cytostatic agent bleomycin (100 micrograms/g body weight, subcutaneously) was given to 10-day-old rats to study unscheduled repair DNA synthesis in nucleolar and in bulk nuclear chromatin of postmitotic Purkinje neurons. The Feulgen reaction and Hoechst 33342 staining were used for quantitative evaluation of nuclear DNA content and chromatin structure. The repair synthesis of DNA was detected by 3H-thymidine autoradiography. The data showed a lesser staining of Purkinje as well as granule cell DNA by Hoechst 33342 in bleomycin-treated animals than in controls, but there was no difference in staining with the Feulgen reaction. The mechanisms of DNA staining by both cytochemical methods suggest that bleomycin reacted preferentially with AT-rich and single stranded DNA in cerebellar cells in vivo. Weak 3H-thymidine labelling was found in Purkinje cells of both control and treated rats, but in the latter group the labelling was more pronounced near or over the nucleolus. The enhanced unscheduled DNA synthesis in the nucleolar region of Purkinje cells of treated animals may be due to greater damage of DNA in this region or may indicate a greater ability of the nucleolar chromatin to repair its DNA.

Animals↗
PubMed1987

Purkinje cell maturation in the light of RNA synthesis.

Wistar rats 1- to 90-day-old received an injection of 3H-uridine and were killed 20 min to 44 h later. Autoradiographic examination revealed the highest grain count densities in Purkinje cell nuclei around postnatal day (PD) 6 while the incidence of labelled nuclei stayed at the peak values till PD 15. Silver staining of Purkinje cell nuclei showed that the expression of nucleolar r-RNA coding genes is maximal at PD 15; in some cells it even slightly exceeds adult values. After PD 15, the percentage of labelled Purkinje cell nuclei declined; this was more pronounced in the nucleolar region than outside the nucleolus. The percentage of cells with cytoplasmic labelling culminated on PD 15. The highest grain counts were found in Purkinje cell cytoplasm on PD 6 at 44 h p.i. interval. Reversal in nuclear grain counts at 2 and 6 h p.i. intervals observed between PD 15 and PD 25 suggests faster degradation, or processing and export, of a newly synthesized nuclear RNA in these age groups. Frequency distribution analysis of grain count densities revealed a small group of Purkinje cells with higher incorporation of 3H-uridine both in the nucleolar region and the whole nucleus at PD 15. In situ hybridization of 3H-r-RNA revealed a slight binding excess to DNA of some Purkinje cell nuclei but not in granule cells of 1-month-old rats. These data, together with those published recently by Brodsky et al. (1985), indicate an uneven structural organization and partial overexpression of the genom coding r-RNA synthesis in the population of Purkinje cells.

Aging↗
PubMed1985

DNA content in nerve-cell nucleus. A biochemical and cytophotometric study of the rat cerebrum.

The cerebral nuclei of 30-day-old rats were separated by a two-step gradient centrifugation into the fractions of large and small nuclei. The DNA content per nucleus was determined biochemically and cytophotometrically in these fractions as well as in the non-separated cerebellar and liver-cell nuclei used as reference cells. The DNA content of large and/or mainly neuronal, and the small and/or glial cell-enriched nuclei were 6.74 +/- 0.51 and 6.21 +/- 0.30 pg, respectively, and in the cerebellum 6.10 +/- 0.28 pg per nucleus. The DNA-content values determined cytophotometrically in Feulgen-stained samples of large and small bulk isolated cortical nuclei ranged within the diploid limits, indicated by a part of a liver-cell population measured in parallel. No evidence for the large scale existence of an "extra-DNA" reported earlier [Bregnard, Knuesel and Kuenzle (1975) Histochemie 43, 59-61; Bregnard, Kuenzle and Ruch (1977) Expl Cell Res. 107, 151-157; Kuenzle, Bregnard, Hübschner and Ruch (1978) Expl Cell Res. 113, 151-160] in cortical neurons has thus been obtained.

Animals↗
PubMed1985

Growth and polyploidization of the liver of early postnatal rats treated with bleomycin.

Bleomycin was administered to 6- and 10-day-old rats and its effects were examined after a two-week survival period. Decreases in body weight and some phenotype abnormalities occurred in both age groups. Decreased relative liver weight (g/g b.w.) was observed only in animals injected on PD 10. DNA microdensitometry and microfluorometry (Feulgen reaction and Hoechst 33342 staining) revealed an increase in the frequency of intermediate DNA values in animals injected on PD6 and killed on PD20. In older animals, i.e. in rats injected on PD10 and killed on PD25, increases in 4c mononucleate and 8c binucleate cells and also a reduction of the relative number of binucleate cells appeared, as compared with age-matched controls. In both age groups, karyological abnormalities, including micronucleate cells and imbalanced DNA content in binucleate cells, were observed.

Aging↗
PubMed1985

Alpha-fetoprotein in the brain of embryonic pigs.

Alpha-fetoprotein (AFP) has been screened in the brain of pig embryos by immunocytofluorescence, crossed immunoelectrophoresis and the immunoblotting technique. Immunocytochemistry showed that the cell and tissue localization of AFP changes with age of the embryos, and/or maturation of the brain. At the earliest stages studied (embryonic days (ED) 26 to 28), the AFP in the brain prevails near the cerebral ventricles, while later (ED 35-46) AFP also appears in higher concentrations in the differentiating parenchyma of some regions of the brain (especially in deep layers of the cortical plate or the region of the differentiating hippocampus) and spinal cord. Its intracellular presence was well evident in the large neuroblasts of these regions. During the 3rd month of embryonic life AFP starts to disappear from the brain parenchyma. At all ages studied, bright fluorescence in the choroid plexus and the meningovascular apparatus is evident. In the choroid plexus as well as ependymal cells a mosaic-like pattern of fluorescence appeared, especially near its invagination into the lateral ventricles; the latter suggests transependymal transport of AFP by some cells. Single and crossed immunoelectrophoresis revealed one reacting band in the water extract of homogenate of the brain of 35-day-old pig embryos. Similarly, only one band has been stained in SDS electrophoretograms prepared from such extracts and stained by the immunoblotting technique; its molecular weight corresponded to the pig AFP. In contrast to the brain the AFP band in the liver was also found in the membrane fraction. It is concluded that maximal tissue concentrations of AFP in the pig brain are attained during the second embryonic month and that, in addition to extracellular fluids, it is also transitionally present in the cytoplasmic compartment of brain cells, especially those more advanced in maturation.

Animals↗
PubMed1979

Constancy and variability in the content of DNA in cerebellar Purkinje cell nuclei. A cytophotometric study.

A cytophotometric study of DNA content in Purkinje cells of the cerebellum of rats, cats, chicken and humans (Feulgen staining) revealed that in a certain number of cells the amount of NDA ranged between the diploid and tetraploid level (H2C cells). The incidence of H2C Purkinje cells varied among the species studied. In rats, which were studied most thoroughly, these cells amounted on average to 3%. In some rats, as well as in some cats and chickens H2C Purkinje cells were entirely absent. In the group of animals possesing H2C Purkinje cells, great interindividual differences were observed. In rats for instance, the incidence of these cells varied from 1 to 23 per cent. Topographic analyses carried out in rat and human cerebellum revealed that H2C Purkinje cells occurred more frequently in the hemispheres than in the vermis. No significant differences were found in the number of H2C Purkinje cells in healthy and Kilham-DNA-virus infected rats. Densitometric analysis of the distribution of nuclear chromatin showed that H2C Purkinje cells were richer in condensed chromatin, especially in the region of the nucleolus, which apparently contains the hyperploid surplus of DNA. It is proposed that the phenomenon of DNA hyperdiploidy arises as a result of either incomplete S-phase in some immature Purkinje cell precursors or the amplification of some DNA sequences particularly those localized in the nucleolar region.

Aged↗