Biomedical subjects

L Vitković

Publications and source records attributed to L Vitković.

At least 19 recordsLinked to original sources

PubMed1995

HIV-1 non-specifically stimulates production of transforming growth factor-beta 1 transfer in primary astrocytes.

HIV-1 expression in monocytes/macrophages can be controlled by transforming growth factor-beta 1 (TGF-beta 1). TGF-beta 1 is present in astrocytes surrounding HIV-1-infected monocyte/macrophages in brain tissue from patients with AIDS but not from seronegative, normal individuals. We sought to determine whether or not production of TGF-beta 1 can be directly stimulated by HIV-1 in astrocytes. Astrocytes from neonatal rat cortex grown in primary culture were exposed to HIV-1 virions for 24 h. One day later, TGF-beta 1 was measured in culture supernatants by a biological assay. HIV-1 caused 1.7-2.1-fold increase in extracellular concentration of TGF-beta 1. TGF beta 1 production also was stimulated by recombinant HIV-1 proteins gp120, p66 and p24. Gp120 labeled with fluorescein was visualized inside astrocytes and its stimulatory effect was not blocked by antibodies against rat CD4. The effect was not specific to HIV-1 and its proteins, because non-opsonized Latex particles and leucine methyl ester (LME) (known to be phagocytosed and endocytosed, respectively, by astrocytes) also stimulated TGF-beta 1 production. The effect was inhibited by two inhibitors of the phago/endocytotic pathway, chloroquine and leupeptin. These results may be relevant to the neuropathogenesis of HIV-1 infection.

Animals↗

PubMed1995

Animal models recapitulate aspects of HIV/CNS disease.

Neurobehavioral and pathological data indicate that the central nervous system (CNS) becomes infected with HIV-1 soon after the virus enters the body. However, neuropathogenesis of HIV-1 infection is difficult to investigate because the brain parenchyma is not accessible to sampling during the course of AIDS. The second compartment of the CNS, cerebrospinal fluid (CSF), is accessible to sampling but how changes in the CSF relate to the changes in the parenchyma is poorly understood. Thus, knowledge of the neuropathogenesis of HIV-1 infection predominantly stems from either postmortem or in vitro studies. This raises the need for animal models of HIV infection of the CNS. Such models have been developed and are briefly reviewed here. The models faithfully recapitulate some aspects of the HIV/CNS disease. Appropriate neuropathological changes and neurobehavioral dysfunction (e.g., cognitive and motor deficits) occur in SIV-infected macaques. Central sensory electrophysiological changes and sleep disturbances occur in FIV-infected cats. Infection of the brain and behavioral changes comparable to some of the changes seen in humans occur in mice infected with a mixture of murine leukemia viruses. Genetically immunodeficient mice (e.g., SCID) accept HIV-infected human organs and or cell grafts. Evidence summarized here indicates that these HuSCID animals undergo neuropathological changes similar to those observed in brains of individuals who died with AIDS. Thus, presently available animal models provide an opportunity to investigate HIV/CNS disease, and to develop and test therapeutic interventions to prevent or cure the disease.

AIDS Dementia Complex↗

PubMed1992

Transforming growth factor-beta 1 (TGF-beta 1) expression and regulation in rat cortical astrocytes.

Transforming growth factor-beta 1 (TGF-beta 1) is a potent modulator of immune and glial cells' functions and thus, could play an important role in neuro-immune interaction. However, published reports disagree on whether or not TGF-beta 1 is expressed in normal brain. We demonstrate here the constitutive expression of TGF-beta 1 mRNA but not protein in both cerebral cortex and primary rat cortical astrocytes. Steady-state TGF-beta 1 mRNA level increased 2-fold in adult compared to neonatal cortex and during proliferation and differentiation of astrocytes in primary culture. This response was not accompanied by the appearance of detectable TGF-beta protein either in vivo or in vitro. However, both intracellular immunoreactive TGF-beta and extracellular TGF-beta 1 activity were detected upon in vitro stimulation of astrocytes with interleukin-1 (IL-1). The extracellular TGF-beta 1 increased with time of exposure to and concentration of IL-1. In contrast, the amount of TGF-beta 1 mRNA remained unchanged during stimulation of astrocytes with IL-1. These results suggest that the production of TGF-beta 1 in astrocytes is regulated at both mRNA and protein levels. The former may occur during astrocytic development, and the latter during astrocytic response to injury in association with elevation of IL-1.

Age Factors↗

PubMed1992

Central nervous system infection in a murine retrovirus-induced immunodeficiency syndrome.

Astrocyte-enriched primary glial cultures (AGC) from C57BL/6 mice were found to be highly susceptible to infection with the replication competent components of LP-BM5, consisting of the ecotropic and mink cell focus-inducing (MCF) helper murine leukemia viruses (MuLVs). The presence in infected AGC of defective LP-BM5 MuLV genome, a critical component for induction of the disease referred to as murine AIDS, was confirmed by Southern blot hybridization using a probe reactive with the p12 gag sequence of the 4.9 kb defective genome. Electron microscopic studies demonstrated C-type retrovirus particles in both astrocytes and microglial cells. In vivo studies demonstrated that the ecotropic MuLVs and the defective genome could be detected within AGC obtained form either 14-day-old mice following intraperitoneal inoculation or 7-day-old mice following intracranial inoculation. These findings suggest that: (1) the central nervous system (CNS) infection is present at an early stage in murine AIDS, (2) both astrocytes and microglial cells are possible CNS targets in which helper MuLVs replicate, and (3) these cells can harbor the defective genome that is a critical component for disease induction.

Animals↗

PubMed1991

Developmental regulation of GAP-43, glutamine synthetase and beta-actin mRNA in rat cortical astrocytes.

Steady-state levels of mRNA encoding growth-associated protein 43 (GAP-43), glutamine synthetase (GS) and beta-actin were measured during development of neonatal rat cortical astrocytes in primary culture. GAP-43 mRNA and protein decreased rapidly during the first 2 weeks and slowly thereafter. In contrast, GS mRNA increased approximately 3-fold during the first 2 weeks and reached maximum by day 15. Actin mRNA first increased up to 8 days and decreased thereafter reaching a constant amount of 15 days, similar to the initial low value. Thus, GAP-43, GS and beta-actin mRNA levels are differentially regulated during development of astrocytes in primary culture. Because the patterns of expression of astrocytic markers GS and GFAP (shown previously) in vitro and in vivo are similar to each other, primary cultures of astrocytes may be an excellent system for investigating mechanisms of developmental regulation of these genes.

Actins↗

PubMed1991

Human astrocytes stimulate HIV-1 expression in a chronically infected promonocyte clone via interleukin-6.

Human promonocyte cells chronically infected with human immunodeficiency virus type (HIV-1) (clone U1.1.5) were grown in the presence of media conditioned by human astrocytes and glioma cell lines U251 and 253. HIV-1 expression was assessed by measuring reverse transcriptase activity. All media conditioned by unstimulated and lipopolysaccharide (LPS) stimulated glial cells induced HIV-1 expression and contained detectable levels of interleukin-6 (IL-6) but not tumor necrosis factor-alpha (TNF-alpha). An antibody against IL-6, but not against TNF-alpha, reduced the induction of HIV-1 by the conditioned media in a concentration-dependent manner. The magnitude of HIV-1 induction by the conditioned media was proportional to the concentration of IL-6 in them. The data indicate that normal and transformed human astrocytes are capable of stimulating HIV-1 expression in chronically infected promonocytic cells by secreting IL-6. The results demonstrate that cytokines secreted by neural cells could play an important role in regulating HIV-1 expression in the brain.

Astrocytes↗

PubMed1990

Astrocyte-conditioned medium stimulates HIV-1 expression in a chronically infected promonocyte clone.

Human promonocytic cells chronically infected with human immunodeficiency virus-1 (HIV-1) (clone U1.1.5) were grown in the presence of media conditioned by primary rat cortical astrocytes and HIV-1 expression was assessed by measuring reverse transcriptase activity. Media conditioned by non-stimulated and lipopolysaccharide (LPS)-stimulated astrocytes induced the expression of HIV-1 2.1-fold and 4.1-fold, respectively. LPS alone, media conditioned by the uninfected parental cell line of U1.1.5 (U937), and culture media from four other cell lines, had no effect on viral expression. The magnitude of induction was time- and dose-dependent. Tumor necrosis factor alpha (TNF-alpha) was detected in LPS-stimulated astrocyte-conditioned medium and the HIV-inducing capability of the medium was neutralized, in part, by an antibody to recombinant murine TNF-alpha. These results suggest a role for astrocytes in the induction of HIV expression and thus in the pathogenesis of HIV-1 infection in brain.

Animals↗

PubMed1990

Regulation of immunoreactive GAP-43 expression in rat cortical macroglia is cell type specific.

Growth-associated protein 43 (GAP-43) is an abundant, intensely investigated membrane phosphoprotein of the nervous system (Benowitz, L.I., and A. Routtenberg. 1987. Trends Neurosci. 10:527-532; Skene, J. H. P. 1989. Annu. Rev. Neurosci. 12:127-156), with a hitherto unknown function. We have previously demonstrated that astrocytes, brain macroglial cells, contain GAP-43 (Steisslinger, H. W., V. J. Aloyo, and L. Vitković, 1987. Brain Res. 415:375-379; Vitković, L., H. W. Steisslinger, V. J. Aloyo, and M. Mersel. 1988. Proc. Natl. Acad. Sci. USA. 85:8296-8300; Vitković L., and M. Mersel. 1989. Metab. Brain Dis. 4:47-53). Results from double immunofluorescent labeling experiments presented here show that oligodendrocytes also contain GAP-43 immunoreactivity (GAP-43ir). Thus, all three macroglial cell types of the central nervous system (type I and type 2 astrocytes and oligodendrocytes) contain GAP-43. Whereas immunoreactive GAP-43 is expressed by progenitors of all macroglial cell types, the developmental regulation of its expression is cell type specific. Immunoreactive GAP-43 is downregulated in type 1 astrocytes, and constitutively expressed in both type 2 astrocytes and oligodendrocytes. These results may be relevant to potential function(s) of GAP-43.

Animals↗

PubMed1989

Growth-associated protein 43 is down-regulated in cultured astrocytes.

Growth-associated protein 43 (GAP-43: other designations--pp46, F1, B-50, p-57) is an abundant, neural, membrane-associated protein involved in synaptic plasticity and regeneration. We recently reported that GAP-43 is present in plasma membranes of cultured rat astrocytes. In the present study the level of astrocytic GAP-43 was assessed by indirect immunofluorescence labeling of cells with a specific anti-GAP-43 serum and immunoblotting of plasma membrane proteins. The results indicate that all astrocytes from 1-day-old cortex contained GAP-43 and those differentiated in culture did not. Furthermore, GAP-43 decreased dramatically during 3 weeks in culture. These results are consistent with the developmental down-regulation of GAP-43 in vivo.

Animals↗

PubMed1988

The 43-kDa neuronal growth-associated protein (GAP-43) is present in plasma membranes of rat astrocytes.

One of the neuronal growth-associated proteins, GAP-43 (molecular mass, approximately 43 kDa; pI 4.3), is abundant in growth-cone membranes and corresponds to a major protein kinase C substrate, the 46-kDa phosphoprotein (pp46), of a growth-cone-enriched subcellular fraction. This protein has the following additional designations (depending on context): B-50 (phospholipid metabolism), F1 (synaptic plasticity), and p57 (calmodulin binding). We show that a protein with the same molecular mass and isoelectric point as GAP-43, which interacts with anti-GAP-43 antibodies on immunoblots, is present in the plasma membranes of cultured neonatal rat cortical astrocytes. Double-immunofluorescence labeling of cells with a serum against glial fibrillary acidic protein and anti-GAP-43 antibody was observed. Furthermore, astrocytic protein was phosphorylated in vitro by protein kinase C and comigrated in two-dimensional PAGE with GAP-43. The data indicate that GAP-43, heretofore believed to be neuron-specific, is present in at least one class of glial cells.

Animals↗

PubMed1988

Coccus-shaped Bacillus subtilis cells are inhibited at stage 0 of sporulation.

RodA and rodB mutations cause rod-shaped Bacillus subtilis cells to become coccus-shaped when the growth temperature is increased from 30 to 45 degrees C. At 30 degrees C four rod strains sporulated as well as the genetically closely related rod+ strains. In contrast, at 45 degrees C the sporulation frequencies of rod strains decreased approximately 10(2)- to 10(4)-fold, while those of rod+ strains remained either unchanged or decreased only slightly. Temperature shift experiments and ultrastructural data indicated that coccus-shaped cells were unable to form prespore septa and were, therefore, inhibited at stage 0 of sporulation.

Bacillus subtilis↗

PubMed1988

Dexamethasone regulates glutamine synthetase expression in rat skeletal muscles.

The regulation of glutamine synthetase expression by dexamethasone was studied in rat skeletal muscles. Daily administration of dexamethasone caused striking enhancement of glutamine synthetase activity in plantaris, soleus, and diaphragm muscles. Northern blot analysis revealed that the dexamethasone-mediated increase of glutamine synthetase activity was associated with dramatically increased levels of glutamine synthetase mRNA. Both glutamine synthetase activity and mRNA levels were significantly elevated in plantaris muscle at 0.5 mg.kg-1.day-1 of dexamethasone, a dose that approximates endogenous corticosteroid levels in animals under severe stress. Quantification of changes in glutamine synthetase mRNA on the basis of total mRNA (by oligo dT hybridization) also revealed a major increase in glutamine synthetase mRNA. Dexamethasone was without effect on beta-tubulin mRNA levels, indicating that glutamine synthetase induction is not part of a global response to glucocorticoids. Dexamethasone treatment resulted in only an approximately 15% increase in glutamine synthetase activity in heart; there was no change in glutamine synthetase mRNA level in this tissue. Thus glucocorticoids regulate glutamine synthetase gene expression in rat skeletal muscles.

Animals↗

PubMed1987

Characterization of two plasma membrane proteins abundant in rat brain.

Plasma membranes were isolated from rat brain cortices and their proteins characterized by two-dimensional electrophoresis. Approximately 500 polypeptides with relative molecular weights (mol. wt.) between 20 kDa and 120 kDa and isoelectric points (pI) between 4.2 and 8.5 were visualized by silver staining. Two proteins, MP1 and MP2, comprised about 5% each of the total by mass. Their mol. wts. were 56 kDa and 43 kDa, and their pIs were 4.2 and 4.3, respectively. The two proteins were present in membranes of cultured granule neurons and cortical astrocytes and absent in liver and kidney. They were both substrates for phosphorylation by protein kinase C. MP2 is similar, if not identical, to a major phosphoprotein in growth cones, pp46 (also termed GAP-43, B-50, F1).

Animals↗

PubMed1987

Bacillus subtilis Lyt+ and Lyt- strains secrete peptidoglycan hydrolases.

The phenotypes characteristic of the pleiotropic lytic-deficient Bacillus subtilis mutants have been attributed to reductions in N-acetyl-muramoyl-L-alanine amidase (EC 3.5.1.28) and endo-beta-N-acetyl-glucosaminidase (EC 3.2.1.30). It is reported here that these peptidoglycan hydrolases are secreted. The FJ3 (lyt-1), FJ6 (lyt-2), and Ni15 (lyt-15) mutants secreted the enzymes in amounts comparable to a Lyt+ strain. Thus, the Lyt- mutants appear not be as deficient in the enzymes' synthesis as their cell-bound activities indicated. Based on the levels of cell-bound and extracellular activities measured during growth, it is suggested that the Lyt- phenotype may be due to a deficiency of the enzymes' acceptor(s) in cell walls.

Acetylglucosaminidase↗

PubMed1987

Wall turnover deficiency of Bacillus subtilis Ni15 is due to a decrease in teichoic acid.

Bacillus subtilis Ni15 is deficient in cell wall turnover. The deficiency is removed if the medium contains 0.2 M NaCl, which does not affect growth. The levels of amidase and glucosaminidase, the most likely enzymes involved in turnover, were, in stationary phase Ni15 cells, similar to those in late-exponential phase cells of a standard strain. The Ni15 enzymes were not salt sensitive. However, the Ni15 walls contained 4.7-fold less phosphorus than the walls of the standard strain. Since the phosphorus content of B. subtilis walls reflects the level of teichoic acid, it is proposed that the turnover deficiency of this strain is due to a decrease in wall teichoic acid.

Acetylglucosaminidase↗

PubMed1984

Absence of correlation between rates of cell wall turnover and autolysis shown by Bacillus subtilis mutants.

Bacillus subtilis mutants with reduced rates of cell wall autolysis reached a constant rate of wall turnover after a longer lag than the standard strain but eventually showed the same turnover rate. In reverse, a turnover-deficient mutant autolysed at a slightly higher rate than the standard strain. Consequently, there is no correlation between the rates of cell wall turnover and autolysis.

Bacillus subtilis↗

PubMed1983

Rates of peptidoglycan turnover and cell growth of Bacillus subtilis are correlated.

Peptidoglycan turnover was measured by the decrease of trichloroacetic acid-precipitable label in cells labeled with N-acetyl-D-[14C]glucosamine. The rate of turnover was reduced strongly by the inhibition of RNA or protein synthesis and weakly by the inhibition of lipid, peptidoglycan, or DNA synthesis. It increased with the growth rate (which was controlled by the concentration of oxomethylvalerate limiting the intracellular isoleucine supply) to the same degree in stringent (rel+) and isogenic relaxed (relA) strains. In these and all other strains tested, the turnover rate (k) increased with the growth rate (g) according to the equation, k = 0.70 X g1.38, even when the growth rate was systematically altered by changes in the temperature or in the composition of the medium.

Bacillus subtilis↗

PubMed1977

Purification of the extracellular protease of Bacillus licheniformis and its inhibition by bacitracin.

Sporulating cells of Bacillus licheniformis excrete three seryl proteases that are of similar size, 28,000 daltons, but of different charge at pH 6. The peptide antibiotic bactracin is released from the cells at the same time and exists, in part, as a bacitracin-protease complex that is stable throughout chromatographic procedures employed in enzyme purification. However, preextraction of crude protease with CHCl3 and subsequent gel filtration effect separation of the antibiotic and the enzyme. Three purified, bacitracin-free proteases, designated CMC I, CMC II, and CMC III and whose ratios of total activity are 1:3.7:10.3, respectively, are obtained by chromatography on carboxymethyl cellulose. The major component, CMC III, is inhibited by commercial bacitracin at near-physiological concentrations of the antibiotic.

Bacillus↗