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R A De Torres

Publications and source records attributed to R A De Torres.

14 recordsLinked to original sources

Penicillin-binding proteins in Listeria monocytogenes.

The Penicillin-Binding Proteins (PBP) of Listeria monocytogenes 29-CCM-A: 454 (ATCC 15313) are described by the use of 125I-Penicillin X as radiotracer. The membranes of this tolerant bacilli contained at least five proteins with different affinities for the radiotracer or Dicloxacillin. The molecular weights of these proteins were estimated as 76, 74, 67, 66 and 47 KDa. Dicloxacillin induced the formation of straight filaments when present at sub-inhibitory concentrations, while Penicillin G did not induce any visible alteration in the morphology of this microorganism.

Bacterial Proteins↗

Production of Linnocuicina 819, a bacteriocin produced by Listeria innocua.

We studied the reciprocal antagonistic properties of 14 Listeria strains. Listeria innocua 29-CCM-A 819 (CIP 8011) produces a bacteriocin named Linnocuicina 819. This substance is mainly released in the exponential phase of growth and was isolated from the supernatant of TPB cultures. Thermal resistance and sensitivity to proteolytic enzymes was assayed. Linnocuicina 819 has a bactericidal mode of action producing a lytic effect.

Bacteriocins↗

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Argentina↗

Simian virus 40 deoxyribonucleic acid replication. I. Effect of cycloheximide on the replication of SV40 deoxyribonucleic acid in monkey kidney cells and in heterokaryons of SV40-transformed and susceptible cells.

Infectious deoxyribonucleic acid (DNA) was extracted from green monkey kidney (CV-1) cultures at various times after the cultures were infected with simian virus 40 (SV40) at input multiplicities of 0.01 and 0.1 plaque-forming unit (PFU) per cell. A pronounced decrease in infectious DNA was observed from 3 to 16 hr after virus infection, suggesting that structurally altered intracellular forms may have been generated early in infection. Evidence is also presented that SV40 DNA synthesis requires concurrent protein synthesis. DNA replication was studied in the presence and absence of cycloheximide in: (i) SV40-infected and uninfected cultures of CV-1 cells; (ii) cultures synchronized with 1-beta-d-arabinofuranosylcytosine (ara-C) for 24 to 30 hr prior to the addition of cycloheximide; and (iii) in heterokaryons of SV40-transformed hamster and susceptible monkey kidney cells. DNA synthesis was determined by pulse-labeling the cultures with (3)H-thymidine at various times from 24 to 46 hr after infection. In addition, the total infectious SV40 DNA was measured. Addition of cycloheximide, even after early proteins had been induced, grossly inhibited both SV40 and cellular DNA syntheses. The activities of thymidine kinase, DNA polymerase, deoxycytidylate deaminase, and thymidylate kinase were measured; these enzyme activities remained high for at least 9 hr in the presence of cycloheximide. SV40 DNA prelabeled with (3)H-thymidine before the addition of cycloheximide was also relatively stable during the time required for cycloheximide to inhibit further DNA replication.

Animals↗

Enzyme induction in green monkey kidney cultures infected with simian adenovirus.

Thymidine kinase was induced after infection of an established strain of green monkey kidney cells (CV-1) with simian adenovirus SV15. Increased levels of thymidine kinase were first observed 8 to 10 hr postinoculation (PI), and the levels increased four- to eightfold by 16 to 24 hr PI. A transient increase (1.5- to 3-fold) of deoxyribonucleic acid (DNA) polymerase activity was also observed about 18 hr PI, but the level of deoxycytidylic deaminase was not enhanced. The inductions of thymidine kinase and DNA polymerase were not obtained when protein synthesis was inhibited with 10(-5) M cycloheximide. However, the enzyme increases did take place when infected cultures were treated with 1-beta-D-arabinofuranosylcytosine (ara-C), an inhibitor of DNA synthesis and SV15 replication. The incorporation of tritium-labeled thymidine (H(3)-dT) into DNA was also stimulated 8 to 24 hr after infection with SV15.

Adenoviridae↗

Nonidentiy of some simian virus 40-induced enzymes with tumor antigen.

The complement-fixing tumor (T) antigen induced by simian virus 40 (SV40) has been prepared from SV40-infected cell cultures, from infected cell cultures treated at the time of infection with 1-beta-d-arabinofuranosylcytosine (ara-C), and from SV40-transformed cells. Upon partial purification, the T antigen exhibited the following properties: it was tightly adsorbed by calcium phosphate gel, it was precipitated by acetic acid at pH 5 or by ammonium sulfate at about 20 to 32% saturation, and it had a molecular weight greater than 250,000, as estimated by Sephadex G-200 gel chromatography. In contrast, deoxycytidylate (dCMP) deaminase, thymidylate (dTMP) kinase, and thymidine (dT) kinase were less strongly bound to calcium phosphate and were not precipitated at pH 5; these enzymes also had much lower molecular weights than the T antigen, as did dihydrofolic (FH(2)) reductase. Furthermore, higher ammonium sulfate concentrations were required to precipitate dCMP deaminase, dTMP kinase, and FH(2) reductase activities than to precipitate the T antigen. Another difference was that the T antigen was not stabilized, but dCMP deaminase, dTMP kinase, and dT kinase, were stabilized, respectively, by dCTP, dTMP, and dT or dTTP. Deoxyribonucleic acid (DNA) polymerase activity resembled the T antigen in adsorption to calcium phosphate, in precipitation by ammonium sulfate or at pH 5, and in the rate of inactivation when incubated at 38 C. However, the polymerase activity could be partly separated from the T antigen by Sephadex G-200 gel chromatography. The cell fraction containing partially purified T antigen also contained a soluble complement-fixing antigen (presumably a subunit of the viral capsid) which reacted with hyperimmune monkey sera. The latter antigen was present in very low titers or absent from cell extracts prepared from SV40-infected monkey kidney cell cultures which had been treated with ara-C at the time of infection, or from SV40-transformed mouse kidney (mKS) or hamster tumor (H-50) cells. The T antigen, however, was present in usual amounts in SV40-transformed cells or ara-C treated, infected cells.

Aminohydrolases↗

Induction of cellular deoxyribonuleic acid synthesis by simian virus 40.

The incorporation of (3)H-thymidine ((3)H-dT) into deoxyribonucleic acid (DNA) has been studied in uninfected confluent monolayer cultures of monkey kidney and mouse kidney cells, simian virus 40 (SV40)-infected cells, and in SV40-transformed mouse kidney cells. Radioautographic measurements revealed that during the period from 28 to 51 hr after productive SV40 infection of monkey kidney cultures about 80% of the cells synthesized DNA, compared to about 16% in uninfected cultures. At 28 to 43 hr after abortive SV40 infection of mouse kidney cultures, 24 to 37% of the cells synthesized DNA, compared to about 6 to 8% in uninfected cultures. The infected monkey kidney and mouse kidney cultures, respectively, incorporated about 5 to 10 times and 3 to 5 times as much (3)H-dT into DNA as did uninfected cultures. Moreover, the net DNA synthesized by SV40-infected monkey kidney cultures, estimated by colorimetric methods, substantially exceeded that of uninfected cultures. Nitrocellulose chromatography and band centrifugation experiments were performed to elucidate the kinds of DNA synthesized in the cultures. In uninfected monkey kidney cultures and at 2 to 12 hr after SV40 infection, almost all of the (3)H-dT labeled DNA sedimented more rapidly than SV40 DNA, and the radioactive DNA was denatured by heating for 12 min at 100 C (cellular DNA). Almost all of the labeled DNA obtained from abortively infected mouse kidney cultures and from SV40-transformed cells also had the properties of cellular DNA. However, approximately one-third to one-half of the labeled DNA obtained from monkey kidney cultures 28 to 51 hr after infection sedimented more slowly than cellular DNA and was not denatured by the heating (SV40 DNA). It is concluded that cellular DNA synthesis was induced during either the productive or abortive SV40 infections.

Animals↗

Virogenic properties of bromodeoxyuridine-sensitive and bromodeoxyuridine-resistant simian virus 40-transformed mouse kidney cells.

When simian virus 40 (SV40)-transformed mouse kidney cells (mKS) were grown in the presence of susceptible indicator cells, SV40 was readily recovered from: (i) 15 transformed cell lines, (ii) transformed cells subcultured 45 times over a 7-month period in medium containing antiviral serum and bromodeoxyuridine (dBU), (iii) 45 of 46 clonal lines isolated in the presence of antiviral serum, (iv) 19 of 19 secondary clones isolated from two clonal lines, and (v) dBU-resistant transformed cell lines. dBU-resistant SV40-transformed mouse kidney cell lines were selected and shown to contain the T antigen and to have normal levels of thymidylate kinase and deoxyribonucleic acid (DNA) polymerase, but to be deficient in thymidine (dT) kinase. Radioautographic and biochemical experiments demonstrated that very little (3)H-dT was incorporated into DNA of dBU-resistant cells during a 6-hr labeling period. After infection of dT kinase-deficient mKS cells with vaccinia virus, high levels of dT kinase were induced. The properties of SV40 recovered from dBU-sensitive and dBU-resistant cells were studied. SV40 recovered from transformed cells was shown to express in CV-1 cells at least six functions characteristic of parental virus: synthesis of capsid antigen, synthesis of T antigen, synthesis of viral DNA, induction of dT kinase, induction of DNA polymerase, and induction of host cell DNA synthesis. In addition, SV40 recovered from the transformed cells induced T antigen, dT kinase, deoxycytidylate deaminase, thymidylate kinase, and DNA polymerase in abortively infected mouse kidney cultures, and the virus was also capable of transforming primary cultures of mouse kidney cells.

Animals↗