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Biomedical subjects

A Silva

Publications and source records attributed to A Silva.

At least 91 records · Page 5Linked to original sources
PubMed1997

Glucocorticoids enhance concanavalin A-induced mitogenic response through the inhibition of nitric oxide production.

Glucocorticoids (GC) are known to inhibit mitogen-induced proliferation of T cells. In this study we show two experimental situations where the addition of GC increases lymphocyte proliferation. It has been reported by different authors that rat spleen (SPL) cells proliferate poorly after concanavalin A (Con A) activation. These poor responses have been related to the suppressor activity of macrophages. Similarly, it is known that T-cell proliferation is depressed in the presence of an excess of macrophages in the culture. Here we show that in both experimental situations, the inclusion of dexamethasone (DEX), a synthetic glucocorticoid, in the culture medium enhances the Con A-stimulated proliferation. We provide evidence that this effect is a consequence of the inhibition of nitric oxide (NO) synthesis by the hormone. Furthermore, we also demonstrate that rat SPL cells are inefficient antigen-presenting cells (APC) because of their spontaneous high production of NO. Taken together our results suggest that the effects of GC on T-cell activation may be to promote or inhibit proliferation depending on the level of endogenous NO synthesis. The possible significance of these results is briefly discussed.

Animals↗
PubMed1997

A second merozoite surface protein (MSP-4) of Plasmodium falciparum that contains an epidermal growth factor-like domain.

Merozoite surface proteins of Plasmodium falciparum play a critical role in the invasion of human erythrocytes by the malaria parasite. Here we describe the identification of a novel protein with a molecular mass of 40 kDa that is found on the merozoite surface of P. falciparum. We call this protein merozoite surface protein 4 (MSP-4). Evidence for the surface location of MSP-4 includes (i) a staining pattern that is consistent with merozoite surface location in indirect immunofluorescent studies of cultured parasites, (ii) localization of MSP-4 in the detergent phase in Triton X-114 partitioning studies, and (iii) nucleotide sequencing studies which predict the presence of an N-terminal signal sequence and a hydrophobic C-terminal sequence in the protein. Immunoprecipitation studies of biosynthetically labelled parasites with [3H] myristic acid indicated that MSP-4 is anchored on the merozoite surface by a glycosylphosphatidylinositol moiety. Of considerable interest is the presence of a single epidermal growth factor-like domain at the C terminus of the MSP-4 protein, making it the second protein with such a structure to be found on the merozoite surface.

Amino Acid Sequence↗
PubMed1997

Promoter activities in Vibrio cholerae ctx phi prophage.

Comparison of cholera toxin (CT) production directed by different gene constructs and S1 nuclease mapping revealed the presence of a ctxB-specific promoter within the ctxA coding sequence. Initiation of transcription in this region occurred in wild-type El Tor and classical biotype choleragenic vibrios. We propose that transcription from the ctxB-specific promoter and a stronger ribosomal binding site on the ctxB mRNA synergistically contribute to achieve the correct (5B:1A) subunit stoichiometry. Plasmid pB, a CT promoterless vector expressing only CTB, was used to detect promoter activity by restoration of A-subunit synthesis. Promoter activity expressed in vitro and in vivo was detected upstream of the zonula occludens toxin gene, suggesting that this factor could be produced in vivo to contribute to fluid accumulation. No promoter activity was detected in vitro and in vivo upstream from the accessory cholera enterotoxin gene.

Base Sequence↗
PubMed1997

Adherence of Vibrio cholerae to cultured differentiated human intestinal cells: an in vitro colonization model.

Choleragenic vibrios adhered to and multiplied on monolayers of the highly differentiated mucin-secreting cell line HT29-18N2. Their adherence followed first-order kinetics, was dependent on the concentration of vibrios, and was partially inhibited by lipopolysaccharide. Comparison of genetically modified vibrios showed that flagella, an active toxR gene, and the virulence cassette were not essential for initial binding. Inactivation of the hemagglutinin/protease increased binding. This highly differentiated human intestinal cell line provides a versatile new approach for studying major events occurring during intestinal colonization: adherence, multiplication, and detachment.

Bacterial Adhesion↗
PubMed1997

[Nursing staff and occupational accidents in a central supply unit].

The aims of this study are to analyse the occupational accidents that affected nursing workers in the central supply unit of the public hospitals that make part of the ERSA-2 in the city of São Paulo and also identify the work load performed by these workers. Data were obtained by verifying all accidents registers that were listed in the Worker Health Attending Service of the institutions related above and by interviewing the workers. Sample was divided in two groups. In the Group I are included sixty-one (61) nursing workers that have registered ninety seven (97) occupational accidents and in the Group II, are included forty five (45) nursing workers who have pointed seventy-eight (78) accidents during the interview. Results were evaluated by quantitative analysis and association tests have been applied, in order to verify the possible differences between the groups of workers. Thus, this study has been able of observe that the major part of affected workers are the female nursing attendants aging from twenty (20) to forty (40) years old. The occupational accidents were also analysed by considering the kind, the place and period in which the accidents occurred; the sort of objects and the injuries that caused them; the body region that has been affected in the worker and if the medical leaves has been necessary and how long it has been taken. The work load evaluated are those identified by the workers of the central supply unit as chemical, physical, biologic, mechanical and physiological loads; The psychic loads have not been mentioned. In this study, the occupational accidents occurrence shows relation with the human and the material resources as well as with the environment and also, the suggestions to minimize them have to consider these points.

Accidents, Occupational↗
PubMed1997

Transgenic expression of mouse proinsulin II prevents diabetes in nonobese diabetic mice.

IDDM in humans and in nonobese diabetic (NOD) mice is a T-cell-dependent autoimmune disease in which the beta-cells of the pancreatic islets are destroyed. Several putative beta-cell autoantigens have been identified, but insulin and its precursor, proinsulin, are the only ones that are beta-cell specific. (Pro)insulin may be a key autoantigen in IDDM. To address the role of proinsulin in the development of IDDM, we generated NOD mice transgenic for the mouse proinsulin II gene driven off a major histocompatibility complex (MHC) class II promoter to direct expression of the transgene to MHC class II bearing cells, including those in the thymus, with the aim of deleting proinsulin-reactive T-cells. The mononuclear cell infiltration of the islets (insulitis) is almost completely absent, and diabetes is prevented in these transgenic NOD mice. The mononuclear cell infiltration of the salivary glands (sialitis) and immune responses to ovalbumin (OVA) are not altered, indicating that the protective effect of the transgene is specific for islet pathology and not due to general immunosuppression. We conclude that autoimmunity to proinsulin plays a pivotal role in the development of IDDM.

Aging↗
PubMed1997

[Nurses' perception of management].

The objective of this investigation is to identify nurses perception about wards management. The collecting of the data was mode through an organized interview with sixteen nurses which works in patients wards in three hospitals of Porto Alegre. The data were examined based on the conteud analysis proposed by Bardin (1977). The results showed five categories: management conception, management characteristics, nurses activities, feelings and nursing graduation and induce us to do some reflexion about the curriculum modification, nurse-teachers improvement and the adjustment between the learning necessity and the work market. In this way is vital that professionals to look for best proficiency.

Attitude of Health Personnel↗
PubMed1996

Secretion of CTLA4Ig by an SV40 T antigen-transformed islet cell line inhibits graft rejection against the neoantigen.

In a model of transplantation rejection, we tested whether a graft manipulated to secrete an immunomodulator could protect itself from immune destruction, thus waiving the need for administration of exogenous immunosuppressants to the recipient. An insulinoma cell line, NIT, having the nonobese diabetic (NOD) genotype but also expressing the SV40 large T antigen, was transfected with CTLA4Ig in an attempt to block the CD28/B7 costimulatory pathway between antigen-presenting calls and T lymphocytes near the site of the graft. The SV40 T antigen is potent at inducing graft rejection. NIT.CTLA4Ig and control transfectants were transplanted subcutaneously into young NOD mice to determine whether CTLA4Ig secretion would abet the survival of the insulinoma graft. CTLA4Ig protein was secreted abundantly in vitro (3-5 microg/ml) and this phenotype was maintained in vivo. Tumor growth was monitored visibly, by palpation, by measuring blood glucose levels, and by death of the host from hypoglycemia caused by unregulated insulin production of the growing insulinoma. Cell growth was similar for NIT.CTLA4Ig7 and control transfectants in immunodeficient mice (nude, irradiated, or SCID mice), indicating that there was no intrinsic growth advantage of the NIT.CTLA4Ig cells. In immunocompetent NOD mice however, the survival/growth of the NIT.CTLA4Ig graft was significantly better than that of the controls. Histopathology was consistent with this finding. Donor-specific second-set grafts were acutely rejected, indicating that tolerance was not induced. CTLs were generated even when the graft secreted CTLA4Ig; there was no clear difference in in vitro immune responses generated by NIT.CTLA4Ig and control cells. We conclude that blockade of the B7 costimulation pathway by graft manipulation can contribute to transplantation success.

Abatacept↗
PubMed1996

Requirement for alpha-CaMKII in experience-dependent plasticity of the barrel cortex.

The mammalian sensory neocortex exhibits experience-dependent plasticity such that neurons modify their response properties according to changes in sensory experience. The synaptic plasticity mechanism of long-term potentiation requiring calcium-calmodulin-dependent kinase type II (CaMKII) could underlie experience-dependent plasticity. Plasticity in adult mice can be induced by changes in the patterns of tactile input to the barrel cortex. This response is strongly depressed in adult mice that lack the gene encoding alpha-CaMKII, although adolescent animals are unaffected. Thus, alpha-CaMKII is necessary either for the induction or for the expression of plasticity in adult mice.

Animals↗
PubMed1996

Expression and folding of an interleukin-2-proinsulin fusion protein and its conversion into insulin by a single step enzymatic removal of the C-peptide and the N-terminal fused sequence.

We report the expression in E. coli of a proinsulin fusion protein carrying a modified interleukin-2 N-terminal peptide linked to the N-terminus of proinsulin by a lysine residue. The key aspects investigated were: (a) the expression of the fused IL2-PI gene, (b) the folding efficiency of the insulin precursor when still carrying the N-fused peptide and (c) the selectivity of the enzymatic cleavage reaction with trypsin in order to remove simultaneously the C-peptide and the N-terminal extension. It was found that this construction expresses the chimeric proinsulin at high level (20%) as inclusion bodies; the fused protein was refolded at 100-200 micrograms/ml to yield about 80% of correctly folded proinsulin and then it was converted into insulin by prolonged reaction (5 h) with trypsin and carboxypeptidase B at a low enzyme/substrate rate (1:600). This approach is based on a single enzymatic reaction for the removal of both the N-terminal fused peptide and the C-peptide and avoids the use of toxic cyanogen bromide.

Amino Acid Sequence↗
PubMed1996

Tagging a Vibrio cholerae El Tor candidate vaccine strain by disruption of its hemagglutinin/protease gene using a novel reporter enzyme: Clostridium thermocellum endoglucanase A.

The celA gene encoding Clostridium thermocellum endoglucanase A was expressed in Vibrio cholerae on its own promoter and used to tag a candidate El Tor biotype cholera vaccine strain. Colonies of the tagged strain could be unequivocally distinguished by overlaying them with CM-cellulose indicator agar and Congo Red staining. Expression of celA did not affect growth of V. cholerae in vitro and in vivo. The celA gene was inserted in the chromosomal hap locus encoding V. cholerae hemagglutinin/protease, a putative "detachase", to create a hap- mutant that could be identified and scored by its halo of cellulolytic activity. The inactivation of hap had a positive effect on colonization in the infant mice model. The above results indicate that celA is a suitable marker gene for V. cholerae and hap is an appropriate locus for insertion of foreign DNA in vaccine development. Inactivation of hap, by increasing the duration of adherence, might decrease excretion of the resulting vaccine vector strain and thus increase its immunogenicity.

Animals↗
PubMed1996

Mechanisms underlying experience-dependent potentiation and depression of vibrissae responses in barrel cortex.

Plasticity was studied in the barrel cortex of rats and mice. Vibrissae deprivation in adult and adolescent animals caused changes in the response properties of cortical neurons to stimulation of spared and deprived vibrissae. Plasticity involved both potentiation of spared vibrissae responses and depression of deprived vibrissae responses in layer II/III of the cortex. Vibrissae response potentiation was found to require alpha CaMKII in adult cortex. Vibrissae response depression was not found to occur in adult cortex but was found in adolescent animals for the principal vibrissa. Preliminary results suggested that vibrissae response depression exhibits both hetero- and homosynaptic components.

Animals↗
PubMed1996

[Infectious endocarditis caused by Q fever. Apropos of a clinical case].

Endocarditis is a rare, but some times fatal, complication of Q fever. Its diagnosis is difficult and it is based on non-specific cardiac findings and a high title of phase I antibodies. The treatment is based on tetracyclines alone or in combination with cotrimoxazole, for long periods of time. The therapeutic efficacy is evaluated by the measurement of phase I antibodies, every three months. The relapses are frequent despite the long period of antibiotic therapy. We report what is probably the first case of Q fever prosthesis endocarditis in Portugal, as a complication following an acute episode of Q fever.

Adult↗