Condições de trabalho e transtornos mentais comuns em trabalhadores da rede básica de saúde de Botucatu (SP)

Os transtornos mentais comuns (TMC) apresentam elevada prevalência em populações gerais e de trabalhadores, com consequências individuais e sociais importantes. Este estudo, transversal e descritivo, explora a relação entre demandas psicológicas, grau de controle e presença de suporte social no trabalho e prevalência de TMC em trabalhadores da rede básica de saúde de Botucatu (SP). A coleta de dados foi feita por meio de questionário autoaplicável, não identificado, com destaque para itens relativos à demanda-controle-suporte e presença de TMC (Self Reporting Questionnaire, SRQ-20). As informações foram inseridas em banco de dados construído com Excel/Office XP 2003 e a análise estatística, efetuada com o programa SAS. Constatou-se que 42,6% dos trabalhadores apresentavam TMC. A observação de associação - alta prevalência de TMC com elevado desgaste (classificação de Karasek) e baixa prevalência de TMC com baixo desgaste - indica que, no município estudado, as condições de trabalho na atenção básica constituem fator contributivo não negligenciável ao adoecimento dos trabalhadores. Revela-se a necessidade de intervenções direcionadas ao cuidado aos trabalhadores, melhoria das condições de trabalho e aumento do suporte social no trabalho.

The role of the parenchyma sheath and PCD during the development of oil cavities in Pterodon pubescens (Leguminosae-Papilionoideae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Analysis of Bothrops jararacussu venomous gland transcriptome focusing on structural and functional aspects: I - gene expression profile of highly expressed phospholipases A(2)

Snake venom glands are a rich source of bioactive molecules such as peptides, proteins and enzymes that show important pharmacological activity leading to in local and systemic effects as pain, edema, bleeding and muscle necrosis. Most studies on pharmacologically active peptides and proteins from snake venoms have been concerned with isolation and structure elucidation through methods of classical biochemistry. As an attempt to examine the transcripts expressed in the venom gland of Bothrops jararacussu and to unveil the toxicological and pharmacological potential of its products at the molecular level, we generated 549 expressed sequence tags (ESTs) from a directional cDNA library. Sequences obtained from single-pass sequencing of randomly selected cDNA clones could be identified by similarities searches on existing databases, resulting in 197 sequences with significant similarity to phospholipase A(2) (PLA(2)), of which 83.2% were Lys49-PLA(2) homologs (BOJU-1), 0.1% were basic Asp49-PLA(2)s (BOJU-II) and 0.6% were acidic Asp49-PLA(2)s (BOJU-III). Adjoining this very abundant class of proteins we found 88 transcripts codifying for putative sequences of metalloproteases, which after clustering and assembling resulted in three full-length sequences: BOJUMET-I, BOJUMET-II and BOJUMET-III; as well as 25 transcripts related to C-type lectin like protein including a full-length cDNA of a putative galactose binding C-type lectin and a cluster of eight serine-proteases transcripts including a full-length cDNA of a putative serine protease. Among the full-length sequenced clones we identified a nerve growth factor (Bj-NGF) with 92% identity with a human NGF (NGHUBM) and an acidic phospholipase A2 (BthA-I-PLA(2)) displaying 85-93% identity with other snake venom toxins. Genetic distance among PLA(2)s from Bothrops species were evaluated by phylogenetic analysis. Furthermore, analysis of full-length putative Lys49-PLA(2) through molecular modeling showed conserved structural domains, allowing the characterization of those proteins as group II PLA(2)s. The constructed cDNA library provides molecular clones harboring sequences that can be used to probe directly the genetic material from gland venom of other snake species. Expression of complete cDNAs or their modified derivatives will be useful for elucidation of the structure-function relationships of these toxins and peptides of biotechnological interest. (C) 2004 Elsevier SAS. All rights reserved.

Structural insights for fatty acid binding in a Lys49-phospholipase A(2): crystal structure of myotoxin II from Bothrops molojeni complexed with stearic acid

The crystal structure of dimeric Lys49-phospholipase A2 myotoxin-II from Bothrops moojeni (MjTX-II) co-crystallized with stearic acid (C18H36O2) has been determined at a resolution of 1.8 angstrom. The electron density maps permitted the unambiguous inclusion of six stearic acid molecules in the refinement. Two stearic acid molecules could be located in the substrate-binding cleft of each monomer in positions, which favor the interaction of their carboxyl groups with active site residues. The way of binding of stearic acids to this Lys49-PLA(2)s is analogous to phospholipids and transition state analogues to catalytically active PLA(2)s. Two additional stearic acid molecules were located at the dimer interface region, defining a hitherto unidentified acyl-binding site on the protein surface. The strictly conserved Lys122 for Lys49-PLA(2)s may play a fundamental role for stabilization of legend-protein complex. The comparison of MjTX-II/satiric acid complex with other Lys-PLA(2)s structures whose putative fatty acids were located at their active site is also analysed. Molecular details of the stearic acid/protein interactions provide insights to binding in croup I/II PLA(2)s and to the possible interactions of Lys49-PLA(2)s with target membranes. (c) 2004 Elsevier SAS. All rights reserved.

cDNA sequence and molecular modeling of a nerve growth factor from Bothrops jararacussu venomous gland

The complete nucleotide sequence of a nerve growth factor precursor from Bothrops jararacussu snake (Bj-NGF) was determined by DNA sequencing of a clone from cDNA library prepared from the poly(A) + RNA of the venom gland of B.jararacussu. cDNA encoding Bj-NGF precursor contained 723 bp in length, which encoded a prepro-NGF molecule with 241 amino acid residues. The mature Bj-NGF molecule was composed of I 18 amino acid residues with theoretical pI and molecular weight of 8.31 and 13,537, respectively. Its amino acid sequence showed 97%, 96%, 93%, 86%, 78%, 74%, 76%, 76% and 55% sequential similarities with NGFs from Crotalus durissus terrificus, Agkistrodon halys pallas, Daboia (Vipera) russelli russelli, Bungarus multicinctus, Naja sp., mouse, human, bovine and cat, respectively. Phylogenetic analyses based on the amino acid sequences of 15 NGFs separate the Elapidae family (Naja and Bungarus) from those Crotalidae snakes (Bothrops, Crotalus and Agkistrodon). The three-dimensional structure of mature Bj-NGF was modeled based on the crystal structure of the human NGF. The model reveals that the core of NGF, formed by a pair of P-sheets, is highly conserved and the major mutations are both at the three beta-hairpin loops and at the reverse turn. (C) 2002 Societe francaise de biochimie et biologic moleculaire/Editions scientifiques et medicales Elsevier SAS. All rights reserved.

Molecular characterization of BjussuSP-I, a new thrombin-like enzyme with procoagulant and kallikrein-like activity isolated from Bothrops jararacussu snake venom

A thrombin-like enzyme, named BjussuSP-I, isolated from Bothrops jararacussu snake venom, is an acidic single-chain glycoprotein with M-r = 61,000, pI similar to 3.8 and 6% sugar. BjussuSP-I shows high proteolytic activity upon synthetic substrates, such as S-2238 and S-2288. It also shows procoagulant and kallikrein-like activity, but is unable to act on platelets and plasmin. These activities are inhibited by specific inhibitors of this class of enzymes. The complete cDNA sequence of BjussuSP-I with 696 bp encodes open reading frames of 232 amino acid residues, which conserve the common domains of thrombin-like serine proteases. BjussuSP-I shows a high structural homology with other thrombin-like enzymes from snake venoms where common amino acid residues are identified as those corresponding to the catalytic site and subsites S1, S2 and S3 already reported. In this study, we also demonstrated the importance of N-linked glycans, to improve thrombin-like activity of BjussuSP-I toxin. (c) 2007 Elsevier Masson SAS. All rights reserved.

Structural and functional properties of Bp-LAAO, a new L-amino acid oxidase isolated from Bothrops pauloensis snake venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identification of continuous interaction sites in PLA(2)-based protein complexes by peptide arrays

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular cloning and biochemical characterization of a myotoxin inhibitor from Bothrops alternatus snake plasma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The use of carbon and nitrogen stable isotopes for the detection of poultry offal meal in meat-type quail feeds

The objective of the present study was to trace the inclusion of poultry offal meal (POM) in the diet of meat-type quails reared for a long period using the technique of stable isotopes. A number of 320 quails were randomly distributed into eight treatments: vegetable diet (T1), and a diet containing 8% POM were fed until the end of the experimental period (T2) or replaced by the vegetable diet on day 42 (T3), 56 (T4), 70 (T5), 84 (T6), 98 (T7), and 112 (T8). Breast muscle samples were collected from four birds randomly selected per treatment every 14 days. The obtained isotope results were submitted to multivariate analysis of variance (MANOVA) with the aid of the GLM procedure of statistical SAS program. Treatments were different from T1 when birds were sacrificed at least two weeks after the diet was changed. T2 results were different from T1 in all evaluated periods. It was concluded that it is possible to trace poultry offal meal inclusion in a strictly vegetable diet after the diet was changed for at least 14 days.

Production of pro- and anti-inflammatory cytokines by monocytes from patients with paracoccidioidomycosis

Monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of granulomatous diseases. We investigated the production of pro- and anti-inflammatory cytokines by monocytes from patients with active paracoccidioidomycosis. Peripheral blood monocytes from 37 patients and 29 healthy controls were cultivated with or without 10 mug/ml of lipopolysaccharide (LPS) for 18 h at 37 degreesC, and the cytokine levels were determined in the culture supernatants by enzyme immunoassay. The results showed that the endogenous levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), IL-6, IL-8, IL-10 and transforming growth factor beta detected in the supernatant of patient monocytes cultivated without stimulus were significantly higher than those produced by healthy controls. These data demonstrated that monocytes from patients with active paracoccidioidomycosis produce high levels of cytokines with both inflammatory and anti-inflammatory activities. However, patient monocytes produced significantly lower TNF-alpha and IL-6 levels in response to LPS when compared to normal subjects, suggesting an impairment in their capacity to produce these cytokines after LPS stimulation. Concentrations of IL-1beta, IL-8 and IL-10 in cultures stimulated with LPS were higher in patients than in controls. These results suggest that an imbalance in the production of pro- and anti-inflammatory cytokines might be associated with the pathogenesis of paracoccidioidomycosis. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

High expression of human monocyte iNOS mRNA induced by Paracoccidioides brasiliensis is not associated with increase in NO production

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A synthetic peptide selectively kills only virulent Paracoccidioides brasiliensis yeasts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Seed predation and fruit damage of Solanum lycocarpum (Solanaceae) by rodents in the cerrado of central Brazil

Although neotropical savannas and grasslands, collectively referred to as cerrado, are rich in seed-eating species of rodents, little is known about seed predation and its determinants in this habitat. In this study, we investigated seed predation and damage to fruits of the widespread shrub Solarium lycocarpurn. In addition, the influence of two possible determinants (distance from the parental plant and total crop size) on the feeding behaviour of Oryzorrys scotti (Rodentia, Sigmodontinae) was also examined. O. scotti were captured more frequently close to the shrubs or on shrub crops, indicating that these rodents were attracted to the shrubs and that seed predation was probably distance-dependent. Moreover, the proportion of damaged fruit on the plant decreased as the total crop size increased; consequently, more productive plants were attacked proportionally less by rodents. This pattern of fruit damage may reflect predator satiation caused by the consumption of a large amount of pulp. Alternatively, secondary metabolites in S. lycocarpum fruits may reduce the pulp consumption per feeding event, thereby limiting the number of fruits damaged. (c) 2006 Elsevier Masson SAS. All rights reserved.

Seed dispersal and spatial distribution of Attalea geraensis (Arecaceae) in two remnants of Cerrado in Southeastern Brazil

The seed dispersal system of Attalea geraensis (Arecaceae), an acaulescent palm, was investigated during one year in two Cerrado fragments in the state of São Paulo, southeastern Brazil. A. geraensis had inflorescences and infrutescences throughout the year. Two scatter-hoarding rodents (the spiny rat, Clyomys bishopi and agoutis, Dasyptocta azarae) were identified as seed predators/dispersers, able to move seeds up to 30 in from the palms, although most of the fruits (57.5%) were dispersed less than 2 in. The removal rates were high and after 20 days, 97.2% of the fruits were removed. Fruit fate was not related to fruit mass, length and diameter. The application of Morisita's index showed a more clumped distribution of adults in the smaller fragment, probably because of the absence of agoutis. Higher seed removal by rodents in the large Cerrado remnant may decrease seed predation by beetles. (C) 2007 Elsevier Masson SAS. All rights reserved.