Efeitos hemodinâmicos e metabólicos imediatos determinados pelas soluções de cloreto de sódio a 7,5% e de sua associação ao dextran 70 a 6% na reanimação do choque hemorrágico: estudo experimental em cães

JUSTIFICATIVA E OBJETIVOS: O dextran associado à solução hipertônica de cloreto sódio a 7,5% apresenta efeitos hemodinâmicos benéficos no controle prolongado da reanimação no choque hemorrágico. O objetivo deste estudo foi verificar se a associação do dextran à solução de cloreto de sódio a 7,5% apresentaria vantagens na avaliação imediata dos parâmetros hemodinâmicos e metabólicos na reanimação em modelo de choque hemorrágico controlado em cães. MÉTODO: Foram estudados 16 cães submetidos à hemorragia controlada até que a pressão arterial média atingisse 40 mmHg e permanecesse assim por até 30 minutos. Estes foram divididos em G1, com administração de NaCI a 7,5%, e G2, com administração NaCI a 7,5% combinada com dextran 70 a 6%, no volume de 4 mL.kg-1, durante três minutos. Foram avaliados os parâmetros hemodinâmicos e metabólicos. Consideraram-se quatro momentos: M1 - 10 minutos após o preparo cirúrgico, M2 - obtido na metade da fase de choque, M3 - obtido dois minutos após o final da administração das soluções, M4 - 30 minutos após o início da reanimação. RESULTADOS: Após a reanimação, não houve diferença significativa dos valores da FC, PAM, PCP e IRVS. O G2 apresentou valores maiores do IC em M4. Os valores da SvO2 foram menores no G1, final do experimento. A C(a-v)O2 foi maior no G1 nos momentos M3 e M4. Os valores do VO2 aumentaram nos dois grupos em M4 e os valores do lactato plasmático aumentaram progressivamente até M3 e diminuíram em M4. Houve aumento dos valores do Na plasmático e redução do hematócrito nos dois grupos. CONCLUSÕES: O G2 mostrou melhor desempenho hemodinâmico principalmente após 30 minutos do início da reanimação. Observou-se, também, maior expansão plasmática e melhor perfusão tecidual na associação do dextran ao NaCl a 7,5%.

Spatial distribution of arboviral mosquito vectors (Diptera, Culicidae) in Vale do Ribeira in the South-eastern Brazilian Atlantic Forest

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

Botulismo de origem alimentar

O botulismo alimentar ocorre pela ingestão de toxinas pré-formadas pelo Clostridium botulinum, consideradas as mais potentes dentre as toxinas conhecidas. Caracteriza-se como uma doença de extrema gravidade, de evolução aguda, provoca distúrbios digestivos e neurológicos, em conseqüência à ingestão de diversos tipos de alimentos. As conservas caseiras estão entre os alimentos que oferecem maior risco à população consumidora. Os produtos de origem animal são frequentemente associados aos surtos da doença, destacando-se os embutidos, tais como salsichas, salames, presuntos e patês. Derivados do leite e enlatados, bem como produtos fermentados, são passíveis de provocar a intoxicação. As outras formas naturais da doença são botulismo por feridas e botulismo infantil, normalmente associado ao consumo de mel contendo esporos do Clostridium botulinum.

Intoxicação por veneno de sapo em um canino

O sapo do gênero Bufo possui nas suas glândulas paratóides uma secreção mucóide contendo toxinas como bufaginas e Bufotoxinas, que são esteróides cardiogênicos. Os cães podem atacar os sapos, entrando em contato com o veneno por meio das mucosas. Um canino, da raça Bulldog Francês, foi encaminhado ao Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (UFRGS) para a necropsia com histórico de provável intoxicação por veneno de sapo. Na necropsia o canino apresentava pulmões aumentados de volume, avermelhados e com edema, e rins de coloração vermelho-escura. As alterações microscópicas indicaram congestão, hemorragia e edema pulmonar. Nos rins, no baço e nos linfonodos foi observada congestão. As análises toxicológicas para os venenos de rotina foram negativas. Porém, a investigação do veneno de sapo a partir de cromatografia por camada delgada e gasosa demonstrou resultado positivo, revelando ser esta a causa da morte do canino.

Caso para diagnóstico

A paracoccidioidomicose é enfermidade potencialmente grave podendo levar ao óbito. Lesões cutâneas específicas são freqüentes e propícias ao diagnóstico precoce. A disseminação para órgãos do sistema monocítico-macrofágico corresponde à forma aguda-subaguda da enfermidade. Derrame pleural e ascite, raros na paracoccidioidomicose, no presente caso foram conseqüentes à pancreatite aguda hemorrágica. Nos casos graves a anfotericina B e o suporte hospitalar adequado são decisivos na recuperação.

Tratamento da hiperidrose palmar com onabotulinumtoxinA veiculada por iontoforese ou fonoforese: relato de casos

Hiperidrose palmar idiopática é doença frequente, que tem grande impacto na qualidade de vida dos pacientes, e seu tratamento definitivo (simpatectomia) associa-se a risco cirúrgico e efeitos adversos. Fármacos, como onabotulinumtoxinA, podem ser veiculados percutaneamente por iontoforese ou fonoforese e contribuir no tratamento da hiperidrose. Os autores apresentam quatro casos em que houve melhora objetiva e subjetiva da sudorese após dez sessões consecutivas de iontoforese ou fonoforese, sem evidências de efeitos adversos. Os resultados clínicos mantiveram-se por 16 semanas de observação após a interrupção do tratamento. Técnicas de veiculação percutânea de medicamentos devem ser percebidas como opções nos tratamentos dermatológicos.

Análise de dados de gastroenterite hemorrágica canina para identificar fatores de risco por regressão logística

No presente estudo, ajustou-se um modelo de regressão logística para prever a probabilidade de óbito de cães acometidos por gastroenterite hemorrágica. O modelo Logístico é recomendado para variáveis-resposta dicotômicas em estudo de Coorte. Registraram-se 176 animais censitariamente atendidos com gastroenterite hemorrágica em quatro clínicas veterinárias da cidade de Lavras, sul de Minas Gerais, entre os anos de 1992 e 1999. Após terem sido selecionadas por meio do teste de Pearson ou teste exato de Fisher, ajustou-se o modelo considerando-se as variáveis sexo, idade, diárias de internação e número de atendimentos. A estimação dos parâmetros foi feita pelo método da máxima verossimilhança. Conclui-se que quando os cães acometidos por gastroenterite hemorrágica são atendidos apenas uma vez, aqueles com idade superior a 6 meses possuem 15,45 vezes mais chances de morrerem (P<0,05) do que aqueles com menos de 6 meses de idade. Quando os animais que apresentam a enfermidade possuem mais de 6 meses de idade, a chance de morrerem, se forem atendidos apenas uma vez, é 20,251 vezes maior (P<0,05) do que se recebessem de 2 a 7 atendimentos.

Purification, sequencing and structural characterization of the phospholipase A(1) from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae)

The biochemical and functional characterization of wasp venom toxins is an important prerequisite for the development of new tools both for the therapy of the toxic reactions due to envenomation caused by multiple stinging accidents and also for the diagnosis and therapy of allergic reactions caused by this type of venom. PLA(1) was purified from the venom of the neotropical social wasp Polybia paulista by using molecular exclusion and cation exchange chromatographies; its amino acid sequence was determined by using automated Edman degradation and compared to the sequences of other vespid venom PLA(1)'s. The enzyme exists as a 33,961.40 da protein, which was identified as a lipase of the GX class, liprotein lipase superfamily, pancreatic lipases (ab20.3) homologous family and RP2 sub-group of phospholipase. P. paulista PLA(1) is 53-82% identical to the phospholipases from wasp species from Northern Hemisphere. The use restrained-based modeling permitted to describe the 3-D structure of the enzyme, revealing that its molecule presents 23% alpha-helix, 28% beta-sheet and 49% coil. The protein structure has the alpha/beta fold common to many lipases; the core consists of a tightly packed beta-sheet constituted of six-stranded parallel and one anti-parallel beta-strand, surrounded by four alpha-helices. P. paulista PLA(1) exhibits direct hemolytic action against washed red blood cells with activity similar to the Cobra cardiotoxin from Naja naja atra. In addition to this, PLA(1) was immunoreactive to specific IgE from the sera of P. paulista-sensitive patients. (c) 2007 Elsevier Ltd. All rights reserved.

Molecular determinants of binding of a wasp toxin (PMTXs) and its analogs in the Na+ channels proteins

The structural specificity of alpha-PMTX, a novel peptide toxin derived from wasp venom has been studied on the neuromuscular synapse in the walking leg of the lobster. alpha-PMTX is known to induce repetitive action potentials in the presynaptic axon due to sodium channel inactivation. We synthesized 29 analogs of alpha-PMTX by substituting one or two amino acids and compared threshold concentrations of these mutant toxins for inducing repetitive action potentials. In 13 amino acid residues of alpha-PMTX, Arg-1, Lys-3 and Lys-12 regulate the toxic activity because substitution of these basic amino acid residues with other amino acid residues greatly changed the potency. Determining the structure-activity relationships of PMTXs will help clarifying the molecular mechanism of sodium channel inactivation. (C) 2000 Elsevier B.V. Ireland Ltd. All rights reserved.

Correlations between differences in amino-terminal sequences and different hemolytic activity of sticholysins

Sticholysins I and II (St I and St II) are cytolysins produced by the sea anemone Stichodactyla helianthus. In spite of their 93% sequence homology, St II is more hemolytic against human erythrocytes than St 1. In order to establish the possible causes of this difference, we studied the hemolytic activity of synthetic peptides containing sequences from the N-termini of both proteins. The results demonstrated that the differences in hemolytic activity of the toxins could be ascribed at least partly to differences in their N-termini. (c) 2007 Elsevier Ltd. All rights reserved.

A new class of neurotoxin from wasp venom slows inactivation of sodium current

The effects of alpha-pompilidotoxin (alpha-PMTX), a new neurotoxin isolated from the venom of a solitary wasp, were studied on the neuromuscular synapses in lobster walking leg and the rat trigeminal ganglion (TG) neurons. Paired intracellular recordings from the presynaptic axon terminals and the innervating lobster leg muscles revealed that alpha-PMTX induced long bursts of action potentials in the presynaptic axon, which resulted in facilitated excitatory and inhibitory synaptic transmission. The action or alpha-PMTX was distinct from that of other known facilitatory presynaptic toxins, including sea anemone toxins and alpha-scorpion toxins, which modify the fast inactivation of Na+ current. We further characterized the action of alpha-PMTX on Na+ channels by whole-cell recordings from rat trigeminal neurons. We found that alpha-PMTX stowed the Na+ channels inactivation process without changing the peak current-voltage relationship or the activation time course of tetrodotoxin (TTX)-sensitive Na+ currents, and that alpha-PMTX had voltage-dependent effects on the rate of recovery from Na+ current inactivation and deactivating tail currents. The results suggest that alpha-PMTX slows or blocks conformational changes required for fast inactivation of the Na+ channels on the extracellular surface. The simple structure of alpha-PMTX, consisting of 13 amino acids, would be advantageous for understanding the functional architecture of Na+ channel protein.

ANTAGONISM OF THE MYOTOXIC EFFECTS OF BOTHROPS-JARARACUSSU VENOM AND BOTHROPSTOXIN BY POLYANIONS

The effects of heparin and other polyanions on the myotoxicity of Bothrops jararacussu venom and purified bothropstoxin (BthTX) were investigated. The release rate of creatine kinase (CK) from isolated extensor digitorum longus muscle and the plasma CK activity of mice were used to quantify the results. The myotoxic effects of B. jararacussu venom or BthTX were inhibited by preincubation of these agents with one of the following: a heterogeneous heparin preparation (designated 'heparin'), low mol. wt heparin (H-4500) or dextran sulfates (DS-8000 and DS-500,000). Non-sulfated dextran (D-40,000) and two chondroitin sulfates were ineffective. The antimyotoxic effects of the polyanions are ascribed to their forming inactive acid-base complexes with the basic myotoxins of Bothrops venoms. Gel-filtration experiments in Sephadex provided direct evidence for complex formation between heparin and BthTX. Intravenous (i.v.) administration of H-4500 or DS-8000 opposed the increase in plasma CK activity induced by a subsequent i.m. injection of venom or BthTX. In contrast, pretreatment with i.v. heparin or DS-500,000 enhanced the venom-induced increase in plasma CK activity. This effect was not observed (1) when the animals were treated with a polyvalent antivenom, which inhibits the coagulation and local stasis induced by Bothrops venoms, and (2) when BthTX, which has no thrombotic or hemorrhagic properties, was the myotoxic agent. The potentiation of the venom-induced increase in plasma CK activity by heparin and DS-500,000 is ascribed to improved washout of the CK released from damaged fibers, because of the anticoagulant properties of the drugs.

Dissociation of enzymatic and pharmacological properties of piratoxins-I and -III, two myotoxic phospholipases A(2) from Bothrops pirajai snake venom

Piratoxins (PrTX) I and III are phospholipases A(2) (PLA(2)s) or PLA(2) homologue myotoxins isolated from Bothrops pirajai snake venom, which also induce myonecrosis, bactericidal activity against Escherichia coli, disruption of artificial membranes, and edema. PrTX-III is a catalytically active hemolytic and anticoagulant Asp49 PLA(2), while PrTX-I is a Lys49 PLA, homologue, which is catalytically inactive on artificial substrates, but promotes blockade of neuromuscular transmission. Chemical modifications of His, Lys, Tyr, and Trp residues of PrTX-I and PrTX-III were performed, together with cleavage of the N-terminal octapeptide by CNBr and inhibition by heparin and EDTA. The lethality, bactericidal activity, myotoxicity, neuromuscular effect, edema inducing effect, catalytic and anticoagulant activities, and the liposome-disruptive activity of the modified toxins were evaluated. A complex pattern of functional differences between the modified and native toxins was observed. However, in general, chemical modifications that significantly affected the diverse pharmacological effects of the toxins did not influence catalytic or membrane disrupting activities. Analysis of structural changes by circular dichroism spectroscopy demonstrated significant changes in the secondary structure only in the case of N-terminal octapeptide cleavage. These data indicate that PrTX-I and PrTX-III possess regions other than the catalytic site, which determine their toxic and pharmacological activities. (C) 2001 Academic Press.

Evaluation of survival of Staphylococcus aureus and Clostridium botulinum in charqui meats

Charqui meats were prepared in laboratory conditions in order to carry out experiments to observe the possibility of development of enterotoxigenic Staphylococcus aureus and Clostridium bottilinum proteolytic type B spores and their toxins. Results demonstrated that the harsh processing conditions, high salt concentration, relative high temperature, a, values, inhibited the growth of both bacteria. Under our experimental conditions, S. aureus would survive throughout the sequence of salting steps i.e. brine followed by rock salting and the sunshine drying step. However, at final a(w) value of 0.70-0.75 would create conditions to inhibit its development. The other experiment revealed that C. botulinum spores germination also was impaired because of these low a(w) values. Under these conditions, charqui meats revealed to be safe products in relation to toxins from both enterotoxigenic S. aureus and C. botulinum. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

A rapid procedure for the isolation of the Lys-49 myotoxin II from Bothrops moojeni (caissaca) venom: Biochemical characterization, crystallization, myotoxic and edematogenic activity

A, M. Soares, V, M, Rodrigues, M. I. Homsi-Brandeburgo, M. H. Toyama, F, R, Lombardi, K. Arni and J. R, Giglio. A rapid procedure for the isolation of the Lys-49 myotoxin II from Bothrops moojeni (caissaca) venom: Biochemical characterization, crystallization, myotoxic and edematogenic activity. Toxicon 36, 503-514, 1998.-Bothrops moojeni snake venom was fractionated on a CM-Sepharose column which was previously equilibrated with 0.05 M ammonium bicarbonate buffer at pH 8.0 and subsequently eluted with an ammonium bicarbonate concentration gradient from 0.05 to 0.5 M at constant pH (8.0) and temperature (25 degrees C). The fraction which eluted last (M-VI) showed, after direct lyophilization, a single band by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE, indicating an approximate M,. of 14 000 and 77 000, in the presence and absence of dithiothreitol, respectively. Its amino acid composition revealed a high level of hydrophobic and basic amino acids as well as 13 half-cystine residues. Its isoelectric point and extinction coefficient (E-1.0cm(1.0mg/ml) at 278 nm and pH 7.0) were 8.2 and 1.170, respectively. M-VI was devoid of phospholipase A(2) (PLA(2)) activity on egg yolk, as well as of hemorrhagic, anticoagulant and coagulant activities, but could induce drastic necrosis on skeletal muscle fibres as well as rapid and transient edema on the rat paw. Its N-terminal sequence: SLFELGKMILQETGKNPAKSYGVYGCNCGVGGRGKPKDATDRCCYVHKCCYK.... revealed high homology with other Lys 49 PLA(2)-like myotoxins from other bothropic venoms. Orthorhombic crystals of M-VI? which diffracted to a maximal resolution of 1.6 Angstrom. were obtained and indicated the presence of a dimer in the asymmetrical unit. (C) 1998 Elsevier B.V. Ltd. All rights reserved.