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Biomedical subjects

R Díaz

Publications and source records attributed to R Díaz.

At least 91 records · Page 5Linked to original sources

Killing of Escherichia coli cells modulated by components of the stability system ParD of plasmid R1.

The proteins P10 and P12 have been shown to be gene products of a new stability system, ParD, of plasmid R1. It is now shown that an R1 miniplasmid, pAB112, carrying a trans-complementable amber mutation in the gene of the P10 protein, is lethal for the host in the absence of suppression. This lethal effect is suppressed in a supF background and also by deletions in pAB112 that affect the gene of the P12 protein. These data indicate that the P12 protein has a lethal effect on the host and that this effect is neutralized by the P10 protein. The possibility that the stabilization conferred by the ParD system could be due to a counterselection, mediated by P12, of cells that lose the plasmid at cell division, is discussed.

Escherichia coli↗

Serological response to the outer membrane lipoprotein in animal brucellosis.

The presence of antibodies to Brucella outer membrane lipoprotein was investigated in cattle and rams. Low but significant amounts of antibody were detected in sera from B. abortus-infected cattle and from B. ovis-infected rams which had developed epididymitis. Strain-19-vaccinated cattle also showed a weak albeit transient antibody response.

Animal Diseases↗

Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide.

Brucella melitensis native haptens (NH) are polysaccharides identical to the O-side chain of the smooth lipopolysaccharide (S-LPS) (E. Moreno, H. Mayer, and I. Moriyón, Infect. Immun. 55:2850-2853, 1987) which precipitate with sera from infected cattle but not from strain 19-vaccinated cattle. In the present work, NH was extracted by the hot-water method (R. Díaz, J. Toyos, M.D. Salvo, and M.L. Pardo, Ann. Rech. Vet. 12:35-39, 1981) and purified free of S-LPS and protein. Purified NH lacked the ability to coat polystyrene and sheep erythrocytes. In contrast, NH acylated with stearoyl chloride bound to both polystyrene and erythrocytes. By hemagglutination and enzyme-linked immunosorbent assay (ELISA), S-LPS and acylated NH gave similar results with blood sera from brucellosis-free, strain 19-vaccinated, and infected cattle. Moreover, a significant correlation between the results of NH ELISA and S-LPS ELISA was demonstrated with milk sera. However, in a competitive ELISA with milk sera, S-LPS in the liquid phase abrogated the binding of antibodies to acylated NH adsorbed to polystyrene, while NH in the liquid phase did not influence the binding of antibodies to polystyrene-adsorbed S-LPS. It is hypothesized that the different precipitations of NH and S-LPS with sera from infected or strain 19-vaccinated cattle are due to differences in the affinity of the antibodies produced upon vaccination or infection and in the physical state of aggregation of NH and S-LPS in aqueous solutions.

Animals↗

Identification of components of a new stability system of plasmid R1, ParD, that is close to the origin of replication of this plasmid.

We provide evidence that a mutation which derepresses an autoregulated system that is located in the vicinity of the basic replicon of R1, stabilizes the ParA- and ParB- miniplasmid of R1 pKN1562, without increasing its copy number. The system, which we have called ParD, maps inside the 1.45-kb PstI-EcoRI fragment that is adjacent to the origin of replication of the plasmid. Two proteins whose expression is coordinated are components of the system. The sequence of the PstI-EcoRI fragment was obtained. The wild-type ParD system determines in cis a basal but detectable stability.

Amino Acid Sequence↗

Immunization with Brucella melitensis Rev 1 against Brucella ovis infection of rams.

The efficacy of Brucella Melitensis Rev 1 vaccine (Rev 1) for the prophylaxis of Brucella ovis ram epididymitis was evaluated. Twenty-nine 3-month-old rams were vaccinated with 2 X 10(9) Rev 1 and 14 were revaccinated with 5 X 10(8) at 14 months of age. Six rams remained unvaccinated as a control group. All rams were challenged with 5 X 10(8) B. ovis at 21 months of age. Before being slaughtered 8 weeks later, only one vaccinated ram developed epididymitis while four of the six control rams developed testicular alterations. Genital and selected extragenital organs and lymph nodes were removed at slaughter and inoculated on selective media. B. ovis was isolated from 26.6% of the vaccinated rams, 21.4% of the revaccinated rams and 100% of control rams. Portions of epididymis, testes and vesicular glands were also used for pathological studies. More severe lesions were observed in control rams than in vaccinated ones. In conclusion, these results show that vaccination of young lambs, followed or not by revaccination, is a suitable method for the prophylaxis of B. ovis infection of rams.

Animals↗

Influence of mouse genotype on passive systemic anaphylaxis by immune complexes.

The influence of mouse genotype on passive systemic anaphylaxis (PSA) by immune complexes was studied. PSA was induced by using Brucella abortus endotoxin as the antigen and rabbit anti-Brucella endotoxin antisera. Experiments using syngeneic mice as well as mice congenic for H-2 showed that the H-2 haplotype influenced the sensitivity of mice to PSA. Among the H-2 haplotypes studied, H-2b was the most sensitive, followed by H-2k and H-2d. Experiments using passive transfer of serum as well as the complement inhibitors suramin and flufenamic acid indicated that variations in complement levels under control of H-2 may be responsible for the effects described. Cyproheptadine, a blocker of serotonin and histamine receptors, and imidazol-alpha-ketoglutarate, an inhibitor of thromboxane synthesis, inhibited PSA, indicating that platelet aggregation, possibly mediated by activated components of the complement cascade, is an important feature in the development of PSA reactions in this system. Differences between strains for protection by cyproheptadine and for the effect of complement inhibitors indicated a role of early components of the classical pathway in this model.

Anaphylaxis↗

Characterization of a Yersinia enterocolitica antigen common to enterocolitis-associated serotypes.

Yersinia enterocolitica synthesized an exocellular antigen common to the serotypes associated with enterocolitis but absent from other serotypes or from other Yersinia species. Both virulent Ca2+-dependent and avirulent Ca2+-independent isogenic pairs derived from the enterocolitis-associated serotypes synthesized the common antigen. Requirements for the synthesis of this common antigen were (i) the presence of metabolizable sugars and (ii) growth on a solid medium at 37 degrees C. The antigen was identified as a 24,000-dalton protein loosely associated with the cell surface but absent from either the cell envelope or the cytoplasmic fraction.

Animals↗

[Quantification (by nomograms) of physiopathological parameters in an experimental model of passive anaphylaxis caused by immune complexes].

Pharmacological changes in a model of passive anaphylaxis by immune complexes in mice have been studied. The results have been evaluated by nomograms to overcome the difficulties observed when using classical criteria as 50% lethal dosis or 50% infective dosis in experimental models which do not allow serial dilutions of the reagents. Mean lethal time, maximal pathogenic capacity and relative survival time can be easily determined, allowing the comparison of the immunopharmacologic protection for each administration schedule. By these criteria, we have observed a different degree of anaphylactic reaction depending on the genetic background of the hosts as well as a protection against anaphylactic shock using the antiserotoninic cyproheptadine and, to a lesser extent, by the platelet antiaggregant ticlopidine.

Anaphylaxis↗