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M A Rebello

Publications and source records attributed to M A Rebello.

36 records · Page 2Linked to original sources

The surface charge of L-A9 cells and Aedes albopictus cells infected with Marituba (Bunyaviridae) virus.

The surface charge of Marituba virus infected L-A9 cells and Aedes albopictus cells was estimated by direct measurement of their electrophoretic mobilities. Uninfected L-A9 cells and A. albopictus cells have mean electrophoretic mobilities of -1.083 microns/s X cm/V and -1.019 microns/s X cm/V, respectively. In Marituba virus infected L-A9 cells a progressive decline in the electrophoretic mobility was observed. In contrast, in Marituba virus infected A. albopictus cells the electrophoretic mobility of the cell surface was unaltered.

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Effect of high temperature on Aedes albopictus cells infected with Mayaro virus.

The multiplication of Mayaro virus in Aedes albopictus cells was drastically inhibited after incubation at 37 degrees C. The effect of short-term exposure of infected cells to high temperatures (heat shock) produced a preferential translation of the heat shock messengers when compared to the viral mRNAs. When cells were shifted back to 28 degrees C (the optimum growth temperature for Aedes albopictus cells), preferential translation of viral mRNA occurred. Although the infected cells were programmed for preferential translation of viral messengers, the thermal treatment was able to shift the translational machinery towards synthesis of heat shock proteins.

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Lactate dehydrogenase from cultured Aedes albopictus cells: kinetic and isozyme analysis.

L(+) lactate dehydrogenase (LDH) activity from cultured cells of Aedes albopictus was studied as a kinetic model of carbohydrate metabolism. Enzyme kinetics were studied in the forward (lactate as substrate) and reverse (pyruvate as substrate) reactions and the apparent Km values were obtained showing LDH higher affinity for pyruvate. The Hill coefficient values for each substrate were similar and indicate the existence of only one binding site on the enzyme. Isozyme analysis on cellulose-acetate electrophoresis presented a single band of LDH which presumably is of the LDH-5 type. The results obtained contribute to the assumption that Aedes albopictus cells have a predominance of anaerobic metabolism.

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Core transcription restores in vitro inhibition of protein synthesis induced by vaccinia virus.

When Ehrlich acistes tumor cell lysate is incubated in the presence of vaccinia core, protein synthesis is impaired. However, when the same system is coupled with viral transcription, protein synthesis is restored. The reversal of endogenous protein synthesis is inhibited by actinomycin D, suggesting that de novo RNA synthesis is required for the reversal of total protein synthesis. When the in vitro products of synthesis are analysed by polyacrylamide gel electrophoresis, two newly synthesized peptides which are not present in the noncoupled transcription-translation system are observed. These two peptides have molecular weights of 31 000 and 25 000, similar to viral early proteins.

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Marituba (Bunyaviridae) virus replication in cultured Aedes albopictus cells and in L-A9 cells.

The replication of Marituba virus (Bunyavirus genus, family Bunyaviridae), was studied in Aedes albopictus (mosquito) cells. Infection of Aedes albopictus cells with Marituba virus was characterized by an initial acute phase of infection in which large amounts of virus were produced and further by a persistent phase of infection in which virus yield was much lower. No changes in host cell DNA, RNA and protein synthesis were observed in Aedes albopictus cells infected with Marituba virus. In contrast in L-A9 (mouse fibroblasts) cells this virus shut-off the host macromolecular synthesis. During the replication of MTB virus in L-A9 cells three virus-specific proteins (G1, G2 and N) were detected. In Aedes albopictus cells, Marituba virus replicates slowly and two virus-specific proteins (G1 and N) accumulate in these cells.

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Interferon induction in mouse fibroblast L-A9 cells.

Mouse L-A9 cell interferon was induced by infection with Newcastle disease virus. Interferon production was 1.5 X 10(5) IU/10(7) cells. Interferon was partially purified by precipitation with ammonium sulphate, chromatography on CM-Sephadex and hydrophobic chromatography on octyl-agarose. The specific activity of the final preparation was 1.7 X 10(7) IU/mg protein. Treatment of L-A9 cells with 20 IU/ml interferon prior to viral infection inhibited the intracellular accumulation of reovirus-specific double-stranded RNA. Dose-response studies of the cells to interferon indicated that L-A9 cells require 10, 13 and 15 IU/ml to obtain 50% viral plaque reduction for Marituba virus, vesicular stomatitis virus and reovirus, respectively. The present results demonstrate the potential of mouse L-A9 cells as an interferon-producing system and also as a model for the study of the effect of cellular response to exogenous interferon treatment on the replication of RNA viruses.

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Autointerference of Marituba Virus (Bunyaviridae) in mouse L cells by defective interfering particles.

The growth characteristics of Marituba virus, a member of the Bunyaviridae family, were studied in L-A9 cells. Virus yield was strictly dependent on the MOI. Quantitation of infectious virus released from the cells revealed a decrease in magnitude with continued serial passage. Specificity of the Marituba virus inhibitory response was investigated in relation to interference within homologous and heterologous viral classes. Virus particles were studied by isopycnic centrifugation in sucrose gradients. Under conditions of multiple viral passages at high multiplicity, two major classes of virus particles were produced, one band at 1.19 g/ml and another at 1.16 g/ml. Particles at 1.16 g/ml were noninfectious. Our results suggest that during the replication of Marituba virus at high MOI a population of defective interfering particles is generated.

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Structural proteins of Marituba virus (Bunyaviridae).

Marituba virus was purified by rate-zonal sedimentation and equilibrium density centrifugation in sucrose gradients. The buoyant density of virus particles was 1.19 g/ml. Purified virus was dissociated and its proteins were analyzed by SDS-PAGE. Four virus polypeptides were identified and designated L, G1, G2 and N. Their average molecular weights were 190 x 10(3) [L, range (180-200) x 10(3)], 120 x 10(3) [G1, range (118-122) x 10(3)], 26 x 10(3) [G2, range (24-28) x 10(3)], and 20 x 10(3) [N, range (19-21) x 10(3)]. Polypeptides N and L are, respectively, the major and the minor viral components. G1 and G2 are glycopeptides as demonstrated by the preferential incorporation of labeled 3H-glucosamine.

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Interferon treatment of Ehrlich ascites tumor cells: effects on exogenous mRNA translation and tRNA inactivation in the cell extract.

We reported earlier that in cell extracts that were prepared from interferon-treated Ehrlich ascites tumor cells and preincubated and passed through Sephadex G-25 (S60INT), the translation of exogenous mRNA (viral and host) was impaired and the impairment could be overcome to a large extent by adding a crude tRNA preparation from Ehrlich ascites tumor cells but not from Escherichia coli. We find now that the rate of inactivation of some tRNA's (especially those specific for leucine, lysine, and serine) but not those of many others is faster in S30INT than in corresponding extracts from control cells. This increased rate of tRNA inactivation may perhaps account for the need for added RNA to overcome at least partially the impairment of translation in S30INT. The relationship of the increased rate of tRNA inactivation to the antiviral effect of interferon is unclear. So far no significant difference has been detected in the amount of tRNA needed to overcome the impairment of encephalomyocarditis virus RNA translation in S30INT between tRNA from interferon-treated cells and tRNA from control cells. Futhermore, no difference was found in the rate of inactivation in S30INT between leucine-specific tRNA's from interferon-treated and from control cells. tRNA's specific for leucine and lysine were not inactivated (unless very slowly) during incubation under out conditions in an extract from interferon-treated (or from control) cells unless the extract had been passed through Sephadex G-25 or dialyzed. The translation fo exogenous mRNA was, however, impaired in an extract from interferon-treated cells that had not been passed through Sephadex G-25. This impairment was apparently not overcome by added tRNA.

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Effect of prostaglandin A1, arsenite and aspirin on stress proteins response in mosquito cells.

The stress response of eukaryotic cells is characterized by changes in the metabolism of responding cells, most notably by increased synthesis of a group of proteins known as heat shock (HSP) proteins In this paper the effect of prostaglandin A1 (PGA1), arsenite and aspirin in Aedes albopictus cells was investigated. In cells treated with PGA1 (10 microg/ml) we observed the induction of several polypeptides with molecular masses of 87, 80, 70, 57, 29 and 23 kDa. Immunoblot analysis revealed that arsenite induces a marked synthesis of HSP70, and aspirin administered during the hyperthermic treatment caused a small increase of HSP70 synthesized.

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Mayaro virus proteins.

Mayaro virus was grown in BHK-21 cells and purified by centrifugation in a potassium-tartrate gradient (5-50%). The electron microscopy analyses of the purified virus showed an homogeneous population of enveloped particles with 69 +/- 2.3 nm in diameter. Three structural virus proteins were identified and designated p1, p2 and p3. Their average molecular weight were p1, 54 KDa; p2, 50 KDa and p3, 34 KDa. In Mayaro virus infected Aedes albopictus cells and in BHK-21 infected cells we detected six viral proteins, in which three of them are the structural virus proteins and the other three were products from processing of precursors of viral proteins, whose molecular weights are 62 KDa, 64 KDa and 110 KDa. The 34 KDa protein was the first viral protein synthesized at 5 hours post-infection in both cell lines studied.

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Studies on the replication of Mayaro virus grown in interferon treated cells.

Mayaro virus grown in interferon treated infected cells has been characterized with regard to its ability to replicate in vertebrate (TC7) and invertebrate (Aedes albopictus) cells. Virus purified from interferon treated TC7 cells adsorbs and penetrates to the same extent as the control virus. During infection, these virus particles caused inhibition of host protein synthesis and synthesized the same spectrum of viral proteins as normal virus. This population however, was apparently more sensitive to interferon treatment. Electron microscopy of TC7 cells showed the presence of numerous aberrant virus particles budding from the plasma membrane.

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Prostaglandin A1 inhibits replication of classical swine fever virus.

Prostaglandins (Pgs) have been shown to inhibit the replication of several DNA and RNA viruses. Here we report the effect of prostaglandin (PgA1) on the multiplication of a positive strand RNA virus, Classical Swine Fever Virus (CSFV) in PK15 cells. PgA1 was found to inhibit the multiplication of CSFV. At a concentration of 5 micrograms/ml, which was nontoxic to the cells, PgA1 inhibitis virus production in 99%. In PgA1 treated cells the size and number of characteristic Classical Swine Fever focus decreased in amount.

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