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Biomedical subjects

S M Oliveira

Publications and source records attributed to S M Oliveira.

16 recordsLinked to original sources

Human immunodeficiency virus type 1 (HIV-1) genotyping in Rio de Janeiro, Brazil: assessing subtype and drug-resistance associated mutations in HIV-1 infected individuals failing highly active antiretroviral therapy.

In order to assess the human immunodeficiency virus type 1 (HIV-1) drug resistance mutation profiles and evaluate the distribution of the genetic subtypes in the state of Rio de Janeiro, Brazil, blood samples from 547 HIV-1 infected patients failing antiretroviral (ARV) therapy, were collected during the years 2002 and 2003 to perform the viral resistance genotyping at the Renageno Laboratory from Rio de Janeiro (Oswaldo Cruz Foundation). Viral resistance genotyping was performed using ViroSeq Genotyping System (Celera Diagnostic-Abbott, US). The HIV-1 subtyping based on polymerase (pol) gene sequences (protease and reverse transcriptase-RT regions) was as follows: subtype B (91.2%), subtype F (4.9%), and B/F viral recombinant forms (3.3%). The subtype C was identified in two patients (0.4%) and the recombinant CRF_02/AG virus was found infecting one patient (0.2%). The HIV-1 genotyping profile associated to the reverse transcriptase inhibitors has shown a high frequency of the M184V mutation followed by the timidine-associated mutations. The K103N mutation was the most prevalent to the non-nucleoside RT inhibitor and the resistance associated to protease inhibitor showed the minor mutations L63P, L10F/R, and A71V as the more prevalent. A large proportion of subtype B was observed in HIV-1 treated patients from Rio de Janeiro. In addition, we have identified the circulation of drug-resistant HIV-1 subtype C and are presenting the first report of the occurrence of an African recombinant CRF_02/AG virus in Rio de Janeiro, Brazil. A clear association between HIV-1 subtypes and protease resistance mutations was observed in this study. The maintenance of resistance genotyping programs for HIV-1 failing patients is important to the management of ARV therapies and to attempt and monitor the HIV-1 subtype prevalence in Brazil.

Anti-HIV Agents↗

Hypoalgesia and altered inflammatory responses in mice lacking kinin B1 receptors.

Kinins are important mediators in cardiovascular homeostasis, inflammation, and nociception. Two kinin receptors have been described, B1 and B2. The B2 receptor is constitutively expressed, and its targeted disruption leads to salt-sensitive hypertension and altered nociception. The B1 receptor is a heptahelical receptor distinct from the B2 receptor in that it is highly inducible by inflammatory mediators such as bacterial lipopolysaccharide and interleukins. To clarify its physiological function, we have generated mice with a targeted deletion of the gene for the B1 receptor. B1 receptor-deficient animals are healthy, fertile, and normotensive. In these mice, bacterial lipopolysaccharide-induced hypotension is blunted, and there is a reduced accumulation of polymorphonuclear leukocytes in inflamed tissue. Moreover, under normal noninflamed conditions, they are analgesic in behavioral tests of chemical and thermal nociception. Using whole-cell patch-clamp recordings, we show that the B1 receptor was not necessary for regulating the noxious heat sensitivity of isolated nociceptors. However, by using an in vitro preparation, we could show that functional B1 receptors are present in the spinal cord, and their activation can facilitate a nociceptive reflex. Furthermore, in B1 receptor-deficient mice, we observed a reduction in the activity-dependent facilitation (wind-up) of a nociceptive spinal reflex. Thus, the kinin B1 receptor plays an essential physiological role in the initiation of inflammatory responses and the modulation of spinal cord plasticity that underlies the central component of pain. The B1 receptor therefore represents a useful pharmacological target especially for the treatment of inflammatory disorders and pain.

Animals↗

Nitric oxide interaction with IL-10, MIP-1alpha, MCP-1 and RANTES over the in vitro granuloma formation against different Schistosoma mansoni antigenic preparations on human schistosomiasis.

Nitric oxide (NO) produced by cytokine-activated macrophages is reported to be cytotoxic against the helminth Schistosoma mansoni, although this is a controversial issue. Previous work in our laboratory identified a fraction of S. mansoni soluble adult worm antigenic preparation (SWAP), named PIII, able to elicit significant in vitro cell proliferation and at the same time lower in vitro and in vivo granuloma formation when compared either to soluble egg antigen (SEA) or to SWAP. Here we report that, in comparison to other S. mansoni antigenic preparations (SEA and SWAP), supernatants of PBMC cultivated with PIII possess higher concentrations of interleukin-10 (IL-10) and macrophage inflammatory protein (MIP-1alpha), concomitantly with lower concentrations of monocyte chemoattractant protein (MCP-1) and regulated on activation, normal T expressed and secreted (RANTES). In the particular case of NO inhibition, supernatants of PBMC cultivated with PIII present decreased IL-10 levels. Altogether, these results indicate that IL-10, MIP-1alpha, MCP-1 and RANTES are distinctively important elements in the PIII modulating role, while NO seems to be pivotal in the regulation of granulomatous responses.

Adult↗

Reduced cardiac hypertrophy and altered blood pressure control in transgenic rats with the human tissue kallikrein gene.

To evaluate the cardiovascular actions of kinins, we established a transgenic rat line harboring the human tissue kallikrein gene, TGR(hKLK1). Under the control of the zinc-inducible metallothionein promoter, the transgene was expressed in most tissues including the heart, kidney, lung, and brain, and human kallikrein was detected in the urine of transgenic animals. Transgenic rats had a lower 24-h mean arterial pressure in comparison with control rats, which was further decreased when their diet was supplemented with zinc. The day/night rhythm of blood pressure was significantly diminished in TGR(hKLK1) animals, whereas the circadian rhythms of heart rate and locomotor activity were unaffected. Induction of cardiac hypertrophy by isoproterenol treatment revealed a marked protective effect of the kallikrein transgene because the cardiac weight of TGR(hKLK1) increased significantly less, and the expression of atrial natriuretic peptide and collagen III as markers for hypertrophy and fibrosis, respectively, were less enhanced. The specific kinin-B2 receptor antagonist, icatibant, abolished this cardioprotective effect. In conclusion, the kallikrein-kinin system is an important determinant in the regulation of blood pressure and its circadian rhythmicity. It also exerts antihypertrophic and antifibrotic actions in the heart.

Angiotensin-Converting Enzyme Inhibitors↗

Rabbit kidney aminopeptidases: purification and some properties.

Aminopeptidases (EC.3.4.11...) are widely distributed in nature and have medical and biological importance due to their function in the modification and degradation of protein. Two aminopeptidases were purified from rabbit kidney homogenate by ion exchange and gel filtration chromatography columns, using aminoacyl of beta-naphthylamides and p-nitroanilides as substrates. The enzymes' homogeneity was assured by SDS-PAGE. The first enzyme (P1) has an optimum of pH 7.0, a molecular mass of 70 kDa, best catalytical efficiency for methionyl-beta-naphthylamide, is 70% inhibited by 0.5 mM Zn2+ and Co2+ ions, 3.33 mM sodium hydrocortisone succinate and 0.08 mM p-hydroxymercuribenzoate, and is little or not inhibited by EDTA, amino acids, p-nitroaniline, beta-naphthylamine, deoxicholate, bestatin and puromycin. The second enzyme (P2) has an optimum of pH 7.0, a molecular mass of 54 kDa, best catalytical efficiency for Leu-beta-naphthylamide, is inhibited by 0.5 mM ions Zn2+ (45%), 0.02 mM EDTA (94%) 0.08 mM p-hydroxymercuribenzoate (70%), 3.33 mM beta-ME (13%), 1.33 mM p-nitroaniline (40%), 1.33 mM beta-naphthylamine (17%), 1.33 mM sodium deoxicholate (96%), 3.33 mM sodium hydrocortisone succinate (60%), and is 30% activated by 0.5 mM Co2+ ions. Puromycin and bestatin are competitive inhibitors with Ki values in 10(-6) and 10(-7) M order, respectively. P1 is a methionine aminopeptidase, while P2 is a leucine aminopeptidase.

Aminopeptidases↗

Molecular cloning and functional characterization of a mouse bradykinin B1 receptor gene.

The gene encoding a putative mouse bradykinin B1 receptor was cloned from a genomic library by low stringency screening. Analysis of two isolated clones revealed a region which contains an open reading frame uninterrupted by introns and encodes a 334 amino acid protein, which exhibits seven potential transmembrane domains and is 68% identical to the human and rabbit bradykinin B1 receptors. Lipopolysaccharide-treatment induces B1 receptor transcripts in the heart, liver, and lung. Stable expression of the coding region in COS-7 cells resulted in high levels of binding sites for the specific B1 ligand des-ARG10 kallidin (Kd = 1.3 nM; Bmax = 51 fmol/mg protein). The rank order of affinity of the receptor for the agonists and antagonists was: des-Arg9BKdes-Arg9Leu8BKdes- Arg10kallidin >> Hoe-140=bradykinin. Functional coupling of the cloned receptor was demonstrated by the dose-dependent effects of des-Arg(9)BK on the extracellular acidification rate in stably transfected COS-7 cells. This effect was not produced by bradykinin and could be blocked by the B1 antagonist des-Arg9Leu8BK.

Amino Acid Sequence↗

Basic aminopeptidase from rabbit kidney: purification and partial characterization.

The aminopeptidase activity of a homogenate of rabbit kidney treated with Triton X-100 was measured using L-aminoacyl-2-naphthylamides (AA-NA). After gradient elution ion-exchange chromatography, four peaks of aminopeptidase activity were eluted. The enzyme eluted at 450 microS containing 33.5% of the activity towards Arg-NA was applied to a Superdex 75 column and presented only one protein band on 10% SDS-polyacrylamide gel electrophoresis. This enzyme has an apparent molecular mass of 78 kDa, is five-fold activated by 0.15 M NaCl and the highest Vmax/KM ratio was obtained with Arg-NA. Enzyme activity was inhibited 100% by 0.13 mM sodium p-hydroxymercuribenzoate, 20% by 0.75 mM EDTA and 100% by 0.66 mM o-phenanthroline. Puromycin and bestatin behaved like competitive inhibitors with a Ki of 0.60 mM and 5.0 microM and 5.0 microM, respectively.

2-Naphthylamine↗

ATP- and UTP-induced currents in macrophages and macrophage polykaryons.

We have investigated the currents induced by extracellular ATP (ATPo), extra-cellular UTP, and other related compounds in macrophages. At potentials of -20 to -60 mV, a typical response to ATPo puffs consists of a fast-activating inward current followed by a transient outward current. The phenomenon lasts 5-20 s, but for sustained exposure to ATP the inward current persists for up to 10 min (our longest recording time). Both currents are inhibited by Mg2+, suggesting that the phenomenon is mediated by ATP4-. The outward current can be ascribed to a Ca(2+)-dependent K+ conductance, and release of Ca2+ from intracellular stores is at least in part responsible for this current. The inward current has a reversal potential of approximately 0 mV, and it is nonspecific for monovalent cations. UTP, a nucleotide that induces an increase in the cytoplasmic concentration of free Ca2+ but does not permeabilize macrophages, and ATP-gamma-S can also induce inward and outward current similar to those described for ATP, but higher doses are required. Adenosine and AMP produce no detectable effect, whereas ADP induces a small outward current. The implications of these results to the phenomenon of ATPo-induced permeabilization of macrophage membranes to large molecules are discussed.

Adenosine↗

Toxocara spp. eggs in public squares of Sorocaba, São Paulo State, Brazil.

The visceral larva migrans (VLM) is a syndrome observed in human infection with helminth larval eggs such as the Toxocara spp. that usually infects dogs and cats. Among the risk factors involved in the occurrence of VLM, particularly important is the size of these animal populations. Sorocaba is a city with a dog population twice as large as that recommended by the World Health Organization. This fact has led to a survey of the presence of Toxocara spp. eggs in public square soils of this city. Thirty squares were selected, fifteen located in the outskirts of the city and fifteen downtown. Soil samples were collected from five distinct sites in the same area. The material was homogenized and drained and 100 g was mixed with a saturated solution of magnesium sulfate and 5% potassium iodine. The floating material was analyzed under the light microscope. Toxocara spp. eggs were found in 16 squares, nine of which were located in the outskirts of the city and seven downtown. It was concluded that Sorocaba squares present a high rate of contamination with Toxocara spp. eggs. The squares in the outskirts of the city presented a higher occurrence of these eggs in comparison with those downtown, although the difference was not statistically significant (p > 0.05).

Animals↗

[Helminth eggs and larvae in restrooms at municipal nursery schools in Sorocaba, SP, Brazil, and their frequency in children feces].

The purpose of this research was to establish a relationship between the presence of helminth eggs in the water closet elements and the frequency of these eggs in the feces of their users. 1050 kindergarten children's feces were examined by the spontaneous sedimentation method in three samples of feces, which were positive in 162 children presenting 184 eggs or helmints larvs. From the 465 water closet elements, constituted of : toilet seat, internal and external door knobs, latch, faucet handle and discharge valve, of 12 Infant Educational Centers of Sorocaba, examined by sticking some transparent adhesive tape on microscopy slides, were found 18 eggs of Ascaris lumbricoides; 1 of Enterobius vermicularis and 4 larvs of nemathoids partially deformed were found in 23 infected elements. There wasn't significance relationship between the elements of water closets and user's feces contamination.

Animals↗

[Prevalence of microbiota in the digestive tract of wild females of Lutzomyia longipalpis Lutz & Neiva, 1912) (Diptera: Psychodidae)].

We dissected the digestive tract of 245 females in pools of 35 flies forming 7 groups. These flies were Lutzomyia longipalpis originating from Lapinha Cave, Lagoa Santa, Minas Gerais. Out of the 8 species of bacteria isolated there was a predominancy of Gram negative bacterias (GNB) in the group of non-fermenters of sugar belonging to the following species: Acinetobacter lwoffii, Stenotrophomonas maltophilia, Pseudomonas putida and Flavimonas orizihabitans. The group of GNB fermenters were: Enterobacter cloacae and Klebsiella ozaenae. In the Gram positive group we isolated the genera Bacillus thuringiensis and Staphylococcus spp.

Animals↗

[Detection of transmissible forms of enteroparasites in water and vegetables consumed at schools in Sorocaba, São Paulo state, Brazil].

Water and raw vegetables consumed in ten nursery schools were evaluated for the presence of transmissible forms of enteroparasites. The water was submitted to filtration through membranes. The washed membrane water was submitted to the Faust method. The in natura and washed vegetables were washed and the water analyzed by the sedimentation method. Contamination was not detected in one school; in two schools, all the materials were contaminated; in four schools, two items were contaminated and in three, one material presented contamination. The water presented a contamination index of 0.7% contamination (Hymenolepis diminuta, Strongyloides stercoralis and Ancylostomatidae); the vegetables in natura, 3.9% (Strongyloides stercoralis, Ancylostomatidae, Ascaris lumbricoides and Giardia lamblia) and the washed samples 1.3% (Strongyloides stercoralis, Ascaris lumbricoides and Giardia lamblia). The water and vegetables are enteroparasites vehicles. The larval form predominated. The vegetables in natura presented higher contamination than those that were washed. Elimination of these forms in vegetables was not guaranteed by washing.

Brazil↗