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Biomedical subjects

V Mares

Publications and source records attributed to V Mares.

At least 37 records · Page 2Linked to original sources
PubMed1999

Growth and differentiation of the vascular smooth muscle and endothelial cells cultured on fluorine ion-implanted polystyrene.

The rat vascular (SMCs) and bovine endothelial cells (BECs) were cultured on conventional or fluorine ion-implanted polystyrene (5 x 10(12) and 5 x 10(14) fluorine ions/cm2). The cells grown on the implanted growth supports showed better adherence, higher volume and higher total protein content. The immunocytochemical analysis revealed that SMCs contained more of the cytoskeletal vimentin and the vascular SMC-specific alpha-actin as well as several cell adhesion-mediating molecules (vinculin, talin, alpha(v)-integrin and ICAM-1). In BECs, only the content of vimentin and talin increased, while expression of ICAM-1 was unchanged. The data suggest that cells on the ion implanted polymers could be more viable and that increased expression of some adhesion molecules mediating interactions with the host immune system is cell type-dependent.

Actins↗
PubMed1999

Different responsiveness of male and female rat aortic smooth muscle cells (SMCs) to repeated passaging in culture.

The smooth muscle cell (SMC) cultures were prepared from the aorta of male and female 8-week-old rats and used at passage 5-7 or 40-45. On day 1, low-passaged cells of both sex groups adhered to growth supports at similar numbers while after repeated passaging the adherence of female-derived cells was higher. These cells had also higher total protein content and contained more of the SMC specific alpha-actin, vimentin and alpha(v) integrins. Compared to the male type of cultures, the high passaged cells of female origin cycled at a slower rate and were undergoing massive polyploidization. Male-derived cells remained of the same morphology, ploidy and the differentiation status at all passages. Their passage response consisted mainly in faster cycling and growth to higher population densities. The data could be of importance for explanation of different incidence of hyperplastic vascular diseases in males and females.

Actins↗
PubMed1999

Ultrastructural response of the nuclear envelope (NE) of C6-glioma cells to cisplatin-induced apoptosis.

The early response of the nuclear envelope of C6-glioma cells (t < or = 24 h), treated with a cytostatic dose of cisplatin in culture (5 microg/ml) included formation of slim and deep invaginations formed by either the inner or both membranes. The invaginations made of the complete NE often extended up to the enlarged nucleoli. In some of them, nucleus-like material occurred at their cytoplasmic site suggesting its enhanced nucleus-to-cytoplasm transport. Some nuclear pores in the invagination-forming cells were covered by dome-shaped "caps" protruding into the karyoplasm. The "capped" pores were absent in the cells that were initially more damaged. At 48 to 72 h, we found a small number of large and hyperlobulated cells with some small lobules containing a rarefied chromatin and focally disintegrated NE. The lamina-free remnants of the NE with a swollen perinuclear cistern were still present at 72-96 h when the population entered the execution phase of apoptotic death.

Animals↗
PubMed1998

Modulation of porphyrin derivatives accumulation in C6 glioma cells by drugs acting on beta-adrenergic receptors. A spectrofluorometric study.

The pharmacological modulation of the uptake of porphyrin derivatives in cultured C6 glioma cells was investigated by means of spectrofluorometric analysis both in single cells and in cell homogenates. The influence of drugs acting as beta-receptor agonists or antagonists was studied in cells grown to semiconfluency. Isoproterenol (ISO), a beta-receptor agonist, enhanced the intracellular fluorescence intensity of both Photofrin and protoporphyrin IX (PpIX). A treatment with a beta-receptor antagonist I-propranolol (PRO), simultaneous with ISO, resulted in an intracellular Photofrin fluorescence signal comparable to that of the control cells, indicating the specificity of the pharmacological action. The pharmacological treatment seemed particularly effective with the aggregated species. This is suggested by the relative increase of the band at 670 nm, being greater than that in the 630 nm band in the emission spectra of Photofrin and PpIX, and by the comparison of the fluorescence intensity on cell homogenates measured both in the absence and in the presence of cetyltrimethyl-ammonium bromide as a detergent.

Animals↗
PubMed1997

Monitoring of the boron neutron-capture reaction by a simple animal model.

Sodium borocaptate (BSH, Na2Bl2HllSH), a slow neutron-capture compound, was injected into the left forebrain ventricle of 1-week-old rats (150 microg BSH/3 microl phosphate buffered saline). After 90 min, the animals were irradiated by epithermal neutrons (LVR-15 nuclear reactor in Rez near Prague, flux density 8.8 x 10(7) neutrons cm-2 s-1, 8 MW reactor power, 8.2 cGy/min) for 5, 10 or 20 min. The brains were examined histologically 8 h after irradiation. In animals irradiated for 5 to 10 min (41 and 82 cGy-Eq, respectively) lethal damage of cells was found in the external granular layer of the cerebellum and the subependymal layer of the forebrain. Irradiation for 20 min (164 cGy-Eq) caused more extensive destruction of cell populations in these regions and, in addition, dead cells appeared also in the more differentiated postmitotic compartments, namely the deeper layers of the cerebellum, layers II/III of the cerebral cortex and corpus callosum. In the forebrain periventricular layer, the extent of cell damage was declining towards the olfactory bulbs. In intact animals, as well as in those injected only with the 150 microl phosphate buffered saline, the radiation damage was low and limited only to the most sensitive dividing populations of the cerebellum and the forebrain. The study demonstrates a differentiation-dependent damage of the rat brain cells by alpha particles and presents a simple model for evaluation of the biological effectiveness of slow neutron beams constructed for neutron-capture therapy of tumors.

Animals↗
PubMed1997

Neuroimmunomodulation of natural killer (NK) cells by ergot alkaloid derivatives.

Ergot alkaloids (EAs), products of Claviceps spp., are widely used in various fields of clinical medicine (neurology, psychiatry, endocrinology). In the present work we studied the neuroimmunomodulative effect of EAs on activation of NK cells and their signalling pathways. Furthermore, the killing capability of rat NK cells in vitro was examined in the presence of glycosidic derivatives of elymoclavine, agroclavine, and liposome-encapsulated EAs. The engagement of appropriate NK cell membrane receptors by EAs cause an indirect enhancement of adenylyl cyclase system through inhibition of G-protein al,2-subunit (up to 50 % of control values). All of the tested EAs enhanced the rat NK cell-mediated cytotoxic activity in vitro, particularly against target cells of astrocyte origin (C-6 glioma). The present results argue for a possible EA immunomodulatory role of cell-mediated immunity in tumour regression processes.

Animals↗
PubMed1997

In vitro early allogeneic reaction of murine brain cortex cells.

An allogeneic reaction among brain cortex cells (mixed reaction) was demonstrated previously by H-2 alloantigen-induced uncoupling of oxidative metabolism (Kovárů Med. Biol. 58: 273, 1980). In the present study we have demonstrated that alloantigen already increased cell surface Na+,K+-ATPase activity after 100 min when the enzyme activation was highest at Mg2+/ATP ratio 4: 1. The allogeneic cell reaction was accompanied by an elevation of membrane lipid fluidity and probably also by a thermotropic lipid phase transition which might influence the membrane lipid-dependent Na+,K+-ATPase activity, while Mg2+-ATPase remained unaffected. Furthermore, the effects of proteins and peptides released into the supernatant during the allogeneic reaction were analyzed in brain cortex cells. One of the isolated active peptide fractions, FA (m.w. lower than 2.5 kD), was able to enhance Na+,K+-ATPase activity as well as to block K+-evoked O2 uptake by brain cortex cells. Thus the FA fraction simulated primary allorecognition events. The data indicate that various brain cell surface domains were influenced by a regulatory peptide fraction of the cytokine type during the early phase of allogeneic reaction. Allorecognition among brain cortex cells is directed against functionally important metabolic reactions.

Adenosine Triphosphate↗
PubMed1997

Peptide cytokines in CNS and the immune system.

This study describes the effects of cytokine peptides released into the supernatant during an early allogeneic reaction (AR) of mouse spleen lymphocytes or brain cortex cells which differ in their major histocompatibility complex (MHC). The peptides were isolated by ultrafiltration, liquid chromatography and HPLC. We found that both peptides stimulated the cell surface Na+,K+-ATPase and Ca2+-ATPase activities of quiescent spleen lymphocytes in vitro and mimicked early allogeneic cell interactions. Both brain and spleen AR peptides inhibited Concanavalin A-stimulated spleen lymphocyte proliferation, whereas 3H-TdR incorporation into DNA of the E7 neuroblastoma cell line was stimulated by these peptides. The peptide isolated from the supernatant of the allogeneic brain cell reaction inhibited phagocytosis in phorbol myristate-stimulated LA5-9/8 mouse macrophage cell line. Immunosuppressive activity of spleen AR peptide is supported by inhibition of spontaneous E rosette formation by lymphocytes. The immunosuppressive effect of isolated peptide cytokines on lectin-activated lymphocytes was comparable with the serum thymic factor (FTS, Lenfant et al. 1983). These changes demonstrate the pleiotropic cytokine actions mediated by plasma membrane of immune system and brain cells.

Animals↗
PubMed1997

Sex-dependent differences in growth and morphology of cultured vascular smooth muscle cells from newborn rats.

The morphology and proliferation of vascular smooth muscle cells (VSMC) were studied in cultures prepared from the aorta of newborn male and female Wistar rats. The doubling times (DT) of the male-derived population were 16.4+/-0.7 h and 30.0+/-2.2 h in the exponential and post-exponential growth phases, respectively. In the female donor cells, the corresponding DT values were significantly longer, i.e. 21.9+/-1.8 h and 38.0+/-2.2 h. In addition, the period of growth was shorter in the female-derived cultures. The percentage of 3H-thymidine labelled cells in male cultures was 61.0+/-3.1, 92.8+/-1.9 and 98.7+/-0.6% at 2, 27 and 52 h, respectively. In the female-derived populations, only 24.6+/-4.4, 66.1+/-3.8 and 82.8+/-2.0% of cells were labelled at the corresponding incubation intervals. As a consequence, the final population density in male cultures was 5.6 times higher. In addition, the male-derived VSMC were mainly spindle-shaped and bulgy in appearance while those from female donors were flat and polygonal which means that the cells were adhering to the growth support to a different extent. The study revealed early determination and long-term persistence of lower adhesiveness as well as higher growth potential of male VSMC, i.e. properties which may be of importance for explaining the higher incidence of vascular wall disorders in males.

Animals↗
PubMed1995

Growth related changes in sugar determinants on the surface of C6 glioma cells in culture: a cytochemical lectin-binding study.

The cell surface sugar determinants (CSSD) were examined in C6 glioma cells in cultures at different conditions of growth by peroxidase conjugates of the lectins: peanut agglutinin (PNA), Ricinus communis agglutinin (RCA), Helix pomatia agglutinin (HPA), wheat germ agglutinin (WGA), lentil agglutinin (LCA), laburnum bork agglutinin (LABA), and lotus agglutinin (TPA). It was found that the cells bound more intensively WGA, LCA, and RCA compared to PNA, HPA; the weakest staining was provided by LABA and TPA. Binding intensity for PNA significantly increased after pretreatment of the cells with neuraminidase. This indicates that a part of the beta-D-galactose residues on the surface membrane of C6 glioma cells is covered by sialic acid. The process of sialization was increased during the culturing of C6 glioma cells. Addition of cis-DDP or dBcAMP to cultures growing in medium with 10% of CS increased the number of Gal residues which are not covered by sialic acid. The expression of beta-D-galactose (Gal), N-acetyl-D-galactosamine (NAcDGal), and fucose (Fuc) residues appeared to be most responsive to changes in growth conditions and degree of cell differentiation. The expressions of N-acetyl-D-glucosamine (NAcDGlc) and mannose (Man) residues were high and seems did not depend on changing of the conditions of culturing. In C6 glioma cells cultures in which the rate of cell division, formation of the cell processes, and adhesiveness of the cells to the substratum were reduced by growing cells in MEM+, expression of beta-Gal, NAcDGal, and Fuc was considerably reduced. The decrease of expression of beta-Gal, NAcDGal, and Fuc on the surface of cell membrane was more pronounced in MEM+ with 1% of CS than in MEM+ with 10% of CS. In DbcAMP and cis-DDP treated cultures, grown in medium with 1% serum, in which cell division was inhibited without obvious changes in cell adhesiveness to the substratum, binding of PNA and HPA was increased due to higher expression of beta-Gal and NAcDGal. From these observations it was concluded that the pattern of expression of sugar residues on the cell surface varies according to the biological state of the cells and are easily affected by tissue culture conditions.

Animals↗
PubMed1995

Tissue culture loading test with storage granules from animal models of neuronal ceroid-lipofuscinosis (Batten disease): testing their lysosomal degradability by normal and Batten cells.

Storage granules (SGs) from ovine and canine models of Batten disease were found to be easily phagocytosed by four cell types studied. The cell types tested were human fibroblasts and peripheral monocytes (control and from a late infantile Batten disease patient), rat C6 cell line, and neonatal cardiomyocytes. The phagocytosed SGs elicited an increase in acid phosphatase activity which was localized in the phagolysosome. After phagocytosis SGs were followed for various times ranging from 7 to 21 days and were found to be of unchanged density (phase contrast), autofluorescence, and ultrastructural appearance. These findings point to their undergradability, or very low degree of degradability, in phagolysosomes in both normal or Batten cultured cells. The Batten disease SGs are not toxic and did not cause any adverse affect on the host cells. Either the normal clearance rate from lysosomes is too slow to be measured by this technique or subunit c accumulation in lysosomes need not result from a primary lysosomal protease defect. Subunit c may aggregate, because of the lack of some normally preventive factor, resulting in a physical barrier to the degradation of this highly apolar molecule.

Adult↗
PubMed1995

Cell kinetics of aortic smooth muscle cells in long-term cultures prepared from rats raised under conventional and SPF conditions.

Migration and proliferation of smooth muscle cells (SMC) were studied in cultures prepared from the aorta of Wistar male rats (170-200 g b.w., 8 weeks old) raised under conventional (CC) or specific pathogen-free (SPF) conditions. In primary cultures, higher movement of cells from explants was found in CC raised donors, namely in samples cultured in serum incomplete medium. In the following subcultures (passage 3-16), the growth curves were steeper and the doubling time shorter in CC type of cultures. The faster growth of SMC population from conventional donors was found to be due to a shorter cell cycle and a higher proportion of dividing cells. As a consequence, the maximum population densities were also higher in the latter type of cultures. The differences in growth, that were dependent on raising conditions, were evident for 16 passages, i.e. 7 months after explantation of cells into culture. The data suggest that breeding conditions may affect the activation of growth of SMC in blood vessels in situ.

Animals↗
PubMed1994

Inhibition of mitochondrial ATPase by dicarbopolyborate, a new enzyme inhibitor.

Polyborate anions were found to inhibit mitochondrial ATPase. Mercapto and chloro derivatives of dicarbononaborates showed full inhibition of the enzyme activity at 0.5-0.8 mM. The inhibitory effect of dodecaborates was lower. The inhibition was of competitive type with respect to ATP. The inhibition of soluble F1-ATPase indicates a direct interaction of the polyborate anion with the catalytic part of the enzyme molecule.

Animals↗
PubMed1993

Appearance of GFAP-positive cells in adult human brain cultures spontaneously decelerated in growth.

Cell cultures were derived from adult human brain biopsies [from cortical gray (cultures 9-HB-G and 33-HB-G) and white (culture 14-HB-W) and stroke-injured white matter (culture 33-HB-IW)]. The morphology and growth rate of cultured cells were examined and correlated with the presence of vimentin and glial fibrillary acidic protein (GFAP). The cultures from various brain matters differed in cell morphology and rate of growth but not in GFAP and vimentin staining. Cells of primary and rapidly proliferating cultures were GFAP-negative and vimentin-positive. Spontaneous growth deceleration occurred in culture 14-HB-W within passages 5 to 10 and in cultures 9-HB-G, 33-HB-G, and 33-HB-W within passages 17 to 20. This deceleration, as well as the successive complete growth arrest, were accompanied by an appearance of GFAP-positive cells and an elevated intensity for vimentin staining. We propose that GFAP-positive astrocytes originate from glial precursor cells that migrate from the explants and differentiate under prolonged subcultivation.

Adult↗
PubMed1993

Renewal of DNA in Purkinje cell nuclei of mouse cerebellum. A 9-month follow-up autoradiographic study.

3H thymidine was injected into pregnant mice in order to label the DNA in the dividing Purkinje cell (PC) precursors of the embryonic cerebellum. The retention of 3H-DNA was evaluated in PC nuclei of animals at the age of 25 days, 3, 6 or 9 months by light microscope autoradiography. The number of silver grains decreased in the whole nuclei by 13.6% and 19.6% in animals 6- and 9-month-old, respectively. In the nucleolar region, the loss of DNA radioactivity was more profound; the silver grain counts decreased by 22.6% and 29.1% in 6- and 9-month-old animals, respectively. No significant differences in the volume and dry mass concentration were found in the PC nuclei of 25 PD and 9 PM old animals. Therefore, the observed changes in grain density counts represent the actual measure of 3H-DNA loss, and/or "spontaneous" renewal of the DNA molecule in PC nuclei, as well as its higher expression in the nucleolar region. Furthermore, it follows from the comparison of our data with those present in the literature, that DNA synthesized in nerve cell precursors before their withdrawal from the mitotic cycle is more stable than that synthesized in postmitotic neurones. This suggests that the repair of DNA in mature neurones might be of an error-prone type.

Animals↗
PubMed1993

Renewal of DNA in Purkinje cell nuclei of mouse cerebellum. A 9-month follow-up autoradiographic study.

3H thymidine was injected into pregnant mice in order to label the DNA in the dividing Purkinje cell (PC) precursors of the embryonic cerebellum. The retention of 3H-DNA was evaluated in PC nuclei of animals at the age of 25 days, 3, 6 or 9 months by light microscope autoradiography. The number of silver grains decreased in the whole nuclei by 13.6% and 19.6% in animals 6- and 9-month-old, respectively. In the nucleolar region, the loss of DNA radioactivity was more profound; the silver grain counts decreased by 22.6% and 29.1% in 6- and 9-month-old animals, respectively. No significant differences in the volume and dry mass concentration were found in the PC nuclei of 25 PD and 9 PM old animals. Therefore, the observed changes in grain density counts represent the actual measure of 3H-DNA loss, and/or "spontaneous" renewal of the DNA molecule in PC nuclei, as well as its higher expression in the nucleolar region. Furthermore, it follows from the comparison of our data with those present in the literature, that DNA synthesized in nerve cell precursors before their withdrawal from the mitotic cycle is more stable than that synthesized in postmitotic neurones. This suggests that the repair of DNA in mature neurones might be of an error-prone type.

Aging↗