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Biomedical subjects

M S Azevedo

Publications and source records attributed to M S Azevedo.

At least 19 recordsLinked to original sources

Protective immunity and antibody-secreting cell responses elicited by combined oral attenuated Wa human rotavirus and intranasal Wa 2/6-VLPs with mutant Escherichia coli heat-labile toxin in gnotobiotic pigs.

Two combined rotavirus vaccination regimens were evaluated in a gnotobiotic pig model of rotavirus infection and disease and were compared to previously tested rotavirus vaccination regimens. The first (AttHRV/VLP2x) involved oral inoculation with one dose of attenuated (Att) Wa human rotavirus (HRV), followed by two intranasal (i.n.) doses of a rotavirus-like particle (2/6-VLPs) vaccine derived from Wa (VP6) and bovine RF (VP2) rotavirus strains. The 2/6-VLPs were coadministered with a mutant Escherichia coli heat-labile toxin, LT-R192G (mLT) adjuvant. For the second regimen (VLP2x/AttHRV), two i.n. doses of 2/6-VLPs+mLT were given, followed by one oral dose of attenuated Wa HRV. To compare the protective efficacy and immune responses induced by the combined vaccine regimens with individual rotavirus vaccine regimens, we included in the experiments the following vaccine groups: one oral dose of attenuated Wa HRV (AttHRV1x and Mock2x/AttHRV, respectively), three oral doses of attenuated Wa HRV (AttHRV3x), three i.n. doses of 2/6-VLPs plus mLT (VLP3x), three i.n. doses of purified double-layered inactivated Wa HRV plus mLT (InactHRV3x), mLT alone, and mock-inoculated pigs. The isotype, magnitude, and tissue distribution of antibody-secreting cells (ASCs) in the intestinal and systemic lymphoid tissues were evaluated using an enzyme-linked immunospot assay. The AttHRV/VLP2x regimen stimulated the highest mean numbers of intestinal immunoglobulin A (IgA) ASCs prechallenge among all vaccine groups. This regimen induced partial protection against virus shedding (58%) and diarrhea (44%) upon challenge of pigs with virulent Wa HRV. The reverse VLP2x/AttHRV regimen was less efficacious than the AttHRV/VLP2x regimen in inducing IgA ASC responses and protection against diarrhea (25% protection rate) but was more efficacious than VLP3x or InactHRV3x (no protection). In conclusion, the AttHRV/VLP2x vaccination regimen stimulated the strongest B-cell responses in the intestinal mucosal immune system at challenge and conferred a moderately high protection rate against rotavirus disease, indicating that priming of the mucosal inductive site at the portal of natural infection with a replicating vaccine, followed by boosting with a nonreplicating vaccine at a second mucosal inductive site, may be a highly effective approach to stimulate the mucosal immune system and induce protective immunity against various mucosal pathogens.

Adjuvants, Immunologic↗

Genotyping of group A rotavirus samples from Brazilian children by probe hybridization.

The G genotyping of 74 group A rotavirus samples was done by RNA-DNA hybridization (dot-blot) using oligonucleotide probes for the VP7 gene region of the human rotavirus serotypes/genotypes 1, 2, 3 and 4. Thirty-one samples could be genotyped by dot-blot showing the following results: G1 = 16, G4 = 6, G3 = 5, and G2 = 4. The data show circulation of genotypes G1-G4 and the predominance of G1. The knowledge of genotypes provides important information concerning rotavirus circulation in Central Brazil.

Brazil↗

Serotypes and subgroups of rotavirus isolated from children in central Brazil.

Group A rotavirus, obtained from children of Goiânia, Brazil, during 1987-1994, were analyzed for subgroup and G serotype by enzyme-linked immunosorbent assay with monoclonal antibodies. The index of serotyping obtained was 61.4% with the following proportions: G1--19.7%, G2--28.0%, G3--9.8%, G4--1.5%, and G5--2.3%. It was observed that G1 occurred from 1987 to 1989 and from 1993 to 1994, and G2 from 1990 to 1993. About 94% of the samples (85/90) could be subgrouped with the following results: 55.5% for SG II, 7.8% SG I, and 31.1% for SG non-I-non-II. Unusual relationship patterns were also detected among serotypes, subgroups, and profiles of electropherotypes in 57.0% of the samples: 20 of them were G2/SG II/"long" profile. The results suggest that variation in temporal and regional characteristics should be considered in the development of rotavirus vaccine.

Antibodies, Monoclonal↗

Superoxide anion radical generation during the oxidation of various amines by diamine oxidase.

Diamine oxidase (DAO) or histaminase is an enzyme which deaminates histamine and several aliphatic amines to their corresponding aldehydes. Hydrogen peroxide and ammonia are side products of this reaction. The purpose of the present work was to evaluate if determination of produced hydrogen peroxide reflects DAO activity or if intermediate formation of the superoxide radical could be a reason for lack of correspondence between oxygen uptake and hydrogen peroxide production at different pH. Superoxide radical formation was determined by cytochrome c reduction in the presence and absence of superoxide dismutase (SOD). Oxygen uptake was measured with an oxygen electrode and hydrogen peroxide production by a spectrophotometric method. At pH 6.6 there was no superoxide production, but at pH 7.4 there was some, and it increased markedly at pH 9.5. Oxygen uptake also increased with increasing pH, especially with histamine as substrate. These results lead us to suggest that the mechanism of action of DAO involves the intermediate generation of superoxide radicals.

Amine Oxidase (Copper-Containing)↗

Oxidative inhibition of red blood cell ATPases by glyceraldehyde.

Glyceraldehyde and other simple monosaccharides autoxidize under physiological conditions, forming dicarbonyl compounds and hydrogen peroxide via intermediate free radicals. These products may have deleterious effects on cell components. In this paper we study the effect of glyceraldehyde autoxidation on red-cell ATPase activities. The autoxidation of glyceraldehyde in imidazole-glycylglycine buffer, measured by oxygen consumption, depends on the buffer concentration and decreases in the presence of superoxide dismutase and catalase. The addition of DETAPAC inhibits the autoxidation almost completely. When human red-blood-cell membranes are incubated with glyceraldehyde, the red-blood-cell ATPase activities decrease significantly. The addition of DETAPAC, GSH and DTE (dithioerythritol) protects the enzyme from inactivation, but superoxide dismutase and catalase have no effect. Methylglyoxal (a dicarbonyl which is analogous to hydroxypyruvaldehyde derived from glyceraldehyde autoxidation) proved to have a powerful inhibitory action on ATPase activities. The addition of DTE completely protects the enzyme from inactivation, suggesting that the sulphydryl groups of the active site of the enzyme are the critical targets for dicarbonyl compounds.

Adenosine Triphosphatases↗

Decrease in blood histamine in drug-treated parkinsonian patients.

Blood histamine and serum histaminase activity were determined in a group of patients with Parkinson disease (PD) and in a group of healthy controls. Untreated PD patients present significantly higher blood histamine than normal controls. Therapy with L-dopa + cardidopa results in a significant decrease in blood histamine. This decrease becomes more accentuated when anticholinergics are added to this therapy. No significant changes were detected in plasma histaminase. The authors suggest, as a hypothesis, that the CNS contributes to blood histamine concentration and that its rise is the result of increased neuronal destruction.

Adult↗

Mechanisms of hemolysis induced by copper.

An excess of copper is the cause of hemolysis in a number of clinical conditions. Incubation of human erythrocyte (RBC) suspensions with copper (II) causes the formation of methemoglobin, lipid peroxidation and hemolysis. A new variant of the thiobarbituric acid (TBA) method, which minimizes the formation of interfering chromophores, was used to detect lipid peroxidation. Lipid peroxidation precedes hemolysis and the antioxidant vitamins C and E, which inhibit lipid peroxidation, also inhibit hemolysis. Consequently lipid peroxidation appears to be the cause of RBC destruction. Lipid peroxidation arises mostly from the oxidation of oxyhemoglobin by copper as it is inhibited in RBCs with carbon monoxyhemoglobin or methemoglobin. A direct interaction of copper with the red cell membrane seems to play only a minor role. Copper effects depend on the presence of free SH groups. Lipid peroxidation is probably initiated by activated forms of oxygen as it is increased by an inhibitor of catalase and reduced by hydroxyl radical scavengers. With higher copper concentrations hemolysis is greater: its mechanism appears different as lipid peroxidation is smaller but hemoglobin alterations, namely precipitation, are more pronounced.

Ascorbic Acid↗

The neutralization of hydroxyl radical by silibin, sorbinil and bendazac.

The compounds silibin, sorbinil and bendazac act as hydroxyl radical scavengers, when the hydroxyl radical is generated by the Fenton reaction. Hydroxyl was detected by the degradation of deoxyribose. The Authors discuss the possibility that the scavenger activities of those compounds may explain, at least in part, their therapeutic activity.

Antioxidants↗

Early increase in histamine concentration in the islets of Langerhans isolated from rats made diabetic with streptozotocin.

Sprague-Dawley rats were separated in 4 groups. G1 received streptozotocin (ST). G2 received nicotinamide (NC) followed by ST. G3 was a NC control and G4 was a citrate control. The rats were sacrificed after 28 h and the islets isolated. Histamine and histaminase were determined. In the islets there was an increase in histamine content in G1 and a smaller increase in G2. The first two groups differ significantly and also in relation to the control groups. A decrease in islet histaminase does not seem responsible for the increased histamine, since group 2 (NC + ST) which had no diabetes, had a lower activity than group 1 (ST). It is suggested that histamine liberation by ST may be related to the diabetogenic effect of this drug.

Amine Oxidase (Copper-Containing)↗

Oxygen radical generation by Maillard compounds.

Amino sugars such as galactosamine are hepatotoxic. It has been verified that toxic hepatitis induced by galactosamine is similar to that of CCl4 poisoning, and that both were inhibited by O2* scavengers. Fructosamine results from the union of glucose with the epsilon-amine of lysine. A test for fructosamine quantification is based on nitroblue tetrazolium (NBT) reduction, in which O2- is involved, the reduction being inhibited in the presence of superoxide dismutase (SOD). Given these facts, we attempted to elucidate if galactosamine and glucosamine reduce NBT and if that reduction is inhibited by SOD. This was confirmed. Subsequently, we incubated aminoacids (glycine, lysine, alanine) with glucose and galactose for 7 days and studied the action of the incubation products on NBT, using amino acids and sugars as controls. We found that NBT reduction increases proportionally to the length of incubation time of glucose/galactose with lysine, but not with other amino acids. Reduction of NBT by the Amadori compounds formed is inhibited by SOD. We suggest that oxygen radical generation by Amadori compounds must be taken into consideration as one cause of damage in diabetes of long duration.

Amino Sugars↗

Risk factors and prevalence of antibodies against hepatitis A virus (HAV) in children from day-care centers, in Goiania, Brazil.

A seroepidemiologic survey about hepatitis A virus (HAV) infection was carried out in a group comprising 310 children, ranging in age from 3 months to 9 years, from day-care centers, in Goiania, a middle sized city in the central region of Brazil. The biomarkers employed in the investigation of previous infection include total IgG and IgM anti-HAV antibodies, and for the detection of more recent infection, IgM anti-HAV antibodies were analyzed. The study was performed in 1991 and 1992. According to the results, 69.7% of the children presented total IgG/IgM anti-HAV antibodies, with 60% of the group in the age range of 1 to 3 years. Among 10 day-care centers analyzed, the prevalence of the biomarker IgM anti-HAV was 3.2%, with an uniform distribution of the cases in the group of children ranging in age from 1 to 4 years. Multivariate analysis was performed to investigate the sociodemographic factors that could influence the results. It was verified that the risk for the infection increased with the length of the attendance in the day-care centers, i.e., the risk for children with attendance of one year or more was 4.7 times higher, when compared with children with one month attendance (CI 95% 2.3-9.9). According to the results, hepatitis A is an endemic infection in day-care centers in the study area. The length of attendance in the day-care settings was demonstrated to be a risk factor for the HAV infection. Such findings suggest that if hepatitis A vaccination becomes available as a routine policy in our region, the target group should be children under one year. Moreover, those children should receive the vaccine before they start to attend the day-care centers.

Brazil↗

[Seroepidemiology of hepatitis A virus infection in street children of Goiânia-Goiás].

A cross-sectional study was performed to determine the age-prevalence of hepatitis A virus (HAV) among street youth from Goiania city--Central Brazil, and to determine if any risk factors could be identified. The surveys were conducted between 1991/1992. The street youth were 397 individuals aged 7 to 21 years old living in institutions and teenagers working at streets. Then, 313 adolescents working at streets and with family links, and 84 institutionalized ones without family were screened for antibodies against HAV. Prevalences ranged from 80.0% to 92.2% to total anti-HAV and there was not a statistically significant trend of positivity with age in this group. Univariate analysis for risk factors associated with HAV infection was performed with no statistically significant difference for adolescents working at streets and living with parents, and street youth with family links. Economic variables were not statistically associated with seropositivity, probable due to homogeneous characteristics of the group. This study disclosed that street youth have a high rate of HAV infection. These findings do not suggest differences between subgroups of populations in acquiring immunity to HVA. The public health implication and the need of screening other subgroups of population of the same city were suggested in order to discuss vaccine strategy in underdeveloped countries.

Adolescent↗

[Rotavirus and adenovirus in 0- to 5-year-old children hospitalized with or without gastroenteritis in Goiâna, GO, Brazil].

In order to detect rotavirus and adenovirus 557 feces samples from hospitalized children (0-5 years of age) were analysed from June 1987 to July 1990 in Goiânia city. Two hundred and ninety one samples were from children with diarrhoea and 266 were from children without diarrhoea. Amongst this later group, 64 samples were from children from the nursery. Two hundred and sixty one out of 557 samples were analysed by immunoelectron microscopy (IEM), polyacrylamide gel electrophoresis (SDS-PAGE) and enzymatic immunoassay for rotavirus and adenovirus (EIARA) whereas the rest (296 samples) were analysed by SDS-PAGE and EIARA. Positivity of rotavirus and adenovirus was 17.2% and 2.1% respectively. Concerning rotavirus and adenovirus there was 29.2% and 2.4% positivity within the group with diarrhoea and 4.1% and 1.5% positivity amongst children without diarrhoea (p < 0.05). Rotavirus were more prevalent amongst children which age ranged from 1 to 11 months of age. No newborn child from the nursery was positive for rotavirus. Adenovirus were detected amongst children from 1 to 3 years of age. Rotavirus circulation peak occurred between May and August (p < 0.05) and no positive case was detected from December to February. Two hundred out of 291 diarrheic samples were also studied concerning bacteria and pathogenic parasites and equal percentages (17.0%) were found for both rotavirus and pathogenic bacteria. Eighty nine samples of rotavirus were detected by SDS-PAGE and 86 of these (96.6%) belonged to the subgroup II with 13 different electrophoretic patterns. Predominance of a given electrophoretic profile was observed in each year of the study.

Acute Disease↗