Coral snake venoms: mode of action and pathophysiology of experimental envenomation

Coral snakes, the New World Elapidae, are included in the genera Micniroides and Micrurus. The genus Mlcrurus comprises nearly all coral snake species and those which are responsible for human snake-bite accidents. The following generalizations concerning the effects induced by their venoms, and their venom-properties can be made. Coral snake venoms are neurotoxic, producing loss of muscle strenght and death by respiratory paralysis. Local edema and necrosis are not induced nor blood coagulation or hemorrhages. Proteolysis activity is absent or of very low grade. They display phospholipase A2 activity. Nephrotoxic effects are not evoked. The main toxins from elapid venoms are postsynaptic and presynaptic neurotoxins and cardiotoxins. Phospholipases A2 endowed with myonecrotic or cardiotoxin-like properties are important toxic components from some elapid venoms. The mode of action of Micrurus frontalis, M. lemniscatus, M. corallinus and M. fulvius venoms has been investigated in isolated muscle preparations and is here discussed. It is shown that while M. frontalis and M. lemniscatus venoms must contain only neurotoxins that act at the cholinergic end-plate receptor (postsynaptic neurotoxins), M. corallinus venom also inhibits evoked acetylcholine release by the motor nerve endings (presynaptic neurotoxin-like effect) and M. fulvius induces muscle fiber membrane depolarization (cardiotoxin-like effect). The effects produced by M. corallinus and M. fulvius venoms in vivo in dogs and M. frontalis venom in dogs and monkeys are also reported.

In vitro differential activity of phospholipases and acid proteinases of clinical isolates of Candida

INTRODUCTION: Candida yeasts are commensals; however, if the balance of normal flora is disrupted or the immune defenses are compromised, Candida species can cause disease manifestations. Several attributes contribute to the virulence and pathogenicity of Candida, including the production of extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate the in vitro activity of phospholipases and acid proteinases in clinical isolates of Candida spp. METHODS: Eighty-two isolates from hospitalized patients collected from various sites of origin were analyzed. Phospholipase production was performed in egg yolk medium and the production of proteinase was verified in a medium containing bovine serum albumin. The study was performed in triplicate. RESULTS: Fifty-six (68.3%) of isolates tested were phospholipase positive and 16 (44.4%) were positive for proteinase activity. C. tropicalis was the species with the highest number of positive isolates for phospholipase (91.7%). Statistically significant differences were observed in relation to production of phospholipases among species (p<0,0001) and among the strains from different sites of origin (p=0.014). Regarding the production of acid protease, the isolates of C. parapsilosis tested presented a larger number of producers (69.2%). Among the species analyzed, the percentage of protease producing isolates did not differ statistically (χ2=1.9 p=0.5901 (χ2=1.9 p=0.5901). CONCLUSIONS: The majority of C. non-albicans and all C. albicans isolates were great producers of hydrolytic enzymes and, consequently, might be able to cause infection under favorable conditions.

Comparison of recombinant A2-ELISA with rKE16 dipstick and direct agglutination tests for diagnosis of visceral leishmaniasis in dogs in Northwestern Iran

INTRODUCTION: Various methods are used for the diagnosis of visceral leishmaniasis (VL), such as microscopic examination, culture and inoculation of laboratory animals; however, serological assays are commonly used for the detection of antibodies in serum samples with a wide range of specificity and sensitivity. METHODS: The purpose of this study was to compare three serological methods, including rA2-ELISA, the recombinant KE16 (rKE16) dipstick test and the direct agglutination test (DAT), for the detection of antibodies against VL antigens. The assays utilized 350 statistically based random serum samples from domestic dogs with clinical symptoms as well as samples from asymptomatic and healthy dogs from rural and urban areas of the Meshkinshahr district, northwestern Iran. RESULTS: Samples were assessed, and the following positive rates were obtained: 11.5% by rKE16, 26.9% by DAT and 49.8% by ELISA. The sensitivity among symptomatic dogs was 32.4% with rKE16, 100% with DAT and 52.9% with ELISA. Conversely, rA2-ELISA was less specific for asymptomatic dogs, at 46.5%, compared with DAT, at 88.9%. CONCLUSIONS : This study recommends rA2-ELISA as a parallel assay combined with DAT to detect VL infection among dogs. Further evaluations should be performed to develop an inexpensive and reliable serologic test for the detection of Leishmania infantum among infected dogs.

Multifactorial control of water and saline intake: role of a2-adrenoceptors

Water and saline intake is controlled by several mechanisms activated during dehydration. Some mechanisms, such as the production of angiotensin II and unloading of cardiovascular receptors, activate both behaviors, while others, such as the increase in blood osmolality or sodium concentration, activate water, but inhibit saline intake. Aldosterone probably activates only saline intake. Clonidine, an a2-adrenergic agonist, inhibits water and saline intake induced by these mechanisms. One model to describe the interactions between these multiple mechanisms is a wire-block diagram, where the brain circuit that controls each intake is represented by a summing point of its respective inhibiting and activating factors. The a2-adrenoceptors constitute an inhibitory factor common to both summing points

Distribution of the a2, a3, and a5 nicotinic acetylcholine receptor subunits in the chick brain

Nicotinic acetylcholine receptors (nAChRs) are ionotropic receptors comprised of a and ß subunits. These receptors are widely distributed in the central nervous system, and previous studies have revealed specific patterns of localization for some nAChR subunits in the vertebrate brain. In the present study we used immunohistochemical methods and monoclonal antibodies to localize the a2, a3, and a5 nAChR subunits in the chick mesencephalon and diencephalon. We observed a differential distribution of these three subunits in the chick brain, and showed that the somata and neuropil of many central structures contain the a5 nAChR subunit. The a2 and a3 subunits, on the other hand, exhibited a more restricted distribution than a5 and other subunits previously studied, namely a7, a8 and ß2. The patterns of distribution of the different nAChR subunits suggest that neurons in many brain structures may contain several subtypes of nAChRs and that in a few regions one particular subtype may determine the cholinergic nicotinic responses

cDNA and deduced primary structure of basic phospholipase A2 with neurotoxic activity from the venom secretion of the Crotalus durissus collilineatus rattlesnake

To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR), we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49), from the venom of Crotalus durissus collilineatus (Cdc PLA2). The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC). The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World’s N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World.

Estudo das latências e amplitudes dos potenciais evocados auditivos de média latência em indivíduos audiologicamente normais

Estudo de coorte contemporânea com corte transversal. O Potencial Evocado Auditivo de Média Latência (PEAML) é gerado entre 10 e 80ms e possui múltiplos geradores, com maior contribuição da região tálamo-cortical. O estabelecimento de critérios de normalidade para os valores de latência e amplitude é necessário para uso clínico. OBJETIVOS: Analisar a latência e amplitude do PEAML em indivíduos sem alterações audiológicas, e verificar a confiabilidade da amplitude Pa-Nb. MATERIAL E MÉTODO: Foram coletados os PEAML de 25 indivíduos durante o ano de 2005 e analisados os componentes Na, Pa, Nb para cada orelha testada (A1 e A2), e posicionamento de eletrodo (C3 e C4). RESULTADOS: Observou-se diferença estatisticamente significante entre os valores médios de latência para C3A1 e C4A1 com relação aos componentes Na e Pa, não sendo encontrada esta diferença para o componente Nb e valores médios das amplitudes Na-Pa e Pa-Nb. CONCLUSÃO: Foram estabelecidos os valores das médias e desvios padrão para os parâmetros latência e amplitude dos componentes Na, Pa, Nb, e Na-Pa e Pa-Nb, nas condições C3A1, C4A1, C3A2, C4A2, proporcionando os parâmetros para a análise e interpretação deste potencial.

Avaliação da função cognitiva da aprendizagem em crianças com distúrbios obstrutivos do sono

Os distúrbios obstrutivos do sono são relativamente freqüentes na população pediátrica, porém o impacto da perda do sono na aprendizagem e função cognitiva não está bem estabelecido. OBJETIVO: Avaliar se pacientes com distúrbios obstrutivos do sono apresentam alteração de aprendizagem, memória e atenção. CASUÍSTICA E MÉTODO: Foram avaliadas 81 crianças de 6 a 12 anos de idade, divididas em 3 grupos: grupo SAHOS (n=24), grupo Ronco Primário (n=37) e grupo Controle (n=20), através de testes de aprendizagem (Teste de Rey) e cognitivos (Dígito, Código, Cancelamento de Letras e Símbolos). Todas as crianças realizaram polissonografia. RESULTADOS: O grupo SAHOS (n=24) e o grupo Ronco Primário (n=37) apresentaram diferença estatisticamente significante nas variáveis A1 (p=0,001) do Teste de Rey quando comparados ao grupo controle. O grupo Ronco Primário apresentou ainda diferenças nas variáveis A2, A4, AT e A6 do Teste de Rey (p=0,020; p=0,05; p=0,004; p=0,05, respectivamente) em relação ao grupo controle (n=20). CONCLUSÃO: Crianças com distúrbios obstrutivos do sono apresentam piores resultados no teste de aprendizagem e memória (Teste de Rey), principalmente o grupo RP, quando este é comparado ao grupo SAHOS. Os testes de atenção apresentam resultados semelhantes entre os grupos.

Volumes de substratos comerciais, solo e composto orgânico afetando a formação de mudas de maracujazeiro-amarelo em diferentes ambientes de cultivo

Este trabalho teve como objetivo avaliar o crescimento de mudas de maracujazeiro- amarelo em três diferentes tamanhos de recipientes, em três condições de cultivo protegido, utilizando seis diferentes substratos. Foram empregados três ambientes de cultivo: (A1) estufa em arco, coberta de filme de polietileno de 150 μm, abertura zenital e tela termorrefletora de 50% sob o filme; (A2) viveiro agrícola de tela de monofilamento, com 50% de sombra; e (A3) viveiro agrícola de tela termorrefletora, com 50% de sombra. Foram testados três volumes (V1 = xx cm³ -sacolas de polietileno de 7,5 x 11,5 cm; V2 = xx cm33 - sacolas de polietileno 10,0 x 16,5 cm; e V3 = xx cm33 -sacolas de polietileno de 15,0 x 21,5 cm) e seis substratos (S1 = solo; S2 = Plantmax®; S3 = vermiculita; S4 = fibra de coco; S5 = fibra de coco chips; e S6 = Organosuper®, composto orgânico comercial). O delineamento experimental foi inteiramente casualizado, em esquema de parcelas subsubdivididas, em que os ambientes foram as parcelas, os recipientes de diferentes volumes as subparcelas e os substratos as subsubparcelas. Aos 50 dias após a semeadura, foram medidos a altura das plantas, o comprimento da raiz, a massa de matéria seca da parte aérea e das raízes. A partir das massas de matéria seca determinaram-se a relação massa de matéria seca de parte aérea e raiz e massa de matéria seca total. A estufa foi o melhor ambiente quando se utilizou o recipiente de XX cm33 o qual proporcionou mudas maiores, com maior biomassa seca aérea, radicular e total. A vermiculita foi o melhor substrato, porém o solo adubado é uma alternativa menos dispendiosa para a região.

Deslizamento de rodados de tratores de pneus em diferentes operações agrícolas

Nos tratores agrícolas, o deslizamento dos rodados ocorre por diversos fatores, entre eles o esforço de tração necessário para deslocar determinado implemento e o tipo de superfície que está em contato com a banda de rodagem dos pneus. Este trabalho tem o objetivo de avaliar o deslizamento do rodado de pneus de um trator em diferentes sistemas de preparo do solo na cultura do milho, no município de Bambuí, Estado de Minas Gerais. O experimento foi conduzido a campo, na safrinha outonal de 2011, na Fazenda Varginha, do Instituto Federal Minas Gerais - campus Bambuí, em um Latossolo Vermelho distroférrico típico. O experimento foi montado em esquema de parcelas sub-subdivididas, tendo, nas parcelas, as operações, nas subparcelas as marchas e, nas sub-subparcelas, as rotações, no delineamento em blocos casualizados, com três repetições, totalizando 81 unidades experimentais. Os fatores e níveis estudados foram (PC) preparo convencional, (CM) cultivo mínimo e (PD) plantio direto. Para cada tratamento, foram utilizadas três marchas (A2, A3 e B1), em três rotações (1600, 1900 e 2200 rpm). Pelos resultados, conclui-se que, no preparo convencional, o maior índice de deslizamento ocorre na operação de semeadura; no cultivo mínimo, na operação de escarificação e, no plantio direto, na operação de semeadura.

The measurement by serological means of the impact of the Hong Kong/68 influenza virus on a population

In order to obtain evidence on the size of the impact of the Hong Kong/68 variant of influenza A2 virus on the population of São Paulo, Brazil, serum samples taken in 1967 before this variant appeared and during successive years after it appeared were examined for their antibody content. Haemagglutination-inhibition tests performed on a total of 2726 serum samples from adults showed a sharp decrease in 1969 of the proportion of sera without antibody to the Hong Kong/68 variant and a corresponding mercase in the proportion with high titres. It was concluded that about three-quarters of the adult population became infected at some time after the variant appeared, the majority in the first year of prevalence.

A eficácia do antiveneno botrópico-crotálico na neutralização das principais atividades do veneno de Bothrops jararacussu

A mionecrose é um dos efeitos causados pelo veneno de Bothrops jararacussu. Uma miotoxina com homologia estrutural à fosfolipase A2 (PLA2), mas sem atividade enzimática, foi isolada desse veneno. O veneno de Crotalus durissus terrificus apresenta também atividade miotóxica, que vem sendo atribuída à crotoxina e à PLA2 (crotoxina B), o componente básico do complexo crotoxina. O veneno de Bothrops jararacussu apresenta três proteínas, que têm identidade imunológica com a PLA2 da crotoxina. O presente trabalho comparou a eficiência dos antivenenos polivalentes comerciais produzidos pelo Instituto Butantan - o antiveneno botrópico (AB) e o antiveneno botrópico-crotálico (AB/C) - na neutralização das atividades letal, hemorrágica, coagulante e miotóxica do veneno de B. jararacussu. Os dois antivenenos neutralizaram de maneira semelhante a atividade hemorrágica, mas o AB/C foi três vezes mais potente que o AB em neutralizar a ação miotóxica e duas vezes mais potente na neutralização da letalidade e na ação coagulante do veneno de B. jararacussu. Os dados sugerem que a utilização do AB/C pode ser vantajosa no tratamento de pacientes picados por serpentes dessa espécie

Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming

Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was not effective in neutralizing the desfibrinating and edema-forming activities of B. atrox venom and the hemorrhagic and coagulant actions of both venoms. Heparin diminished the effectiveness of the ABS in the neutralization of the hemorrhagic and edema-forming activities of the B. atrox venom. However, heparin in the 6 IU dose was capable of neutralize the edema-forming of the B. erythromelas and increase the effectiveness of the ABS. Heparin also neutralized the phospholipasic A2 activity of B. atrox (14.3%) and B. erythromelas (28.0%) venoms. For B. erythromelas venom, the associated treatment, heparin and ABS, was more effective in the neutralization of its lethal activity.

Antifungal susceptibilities of clinical and environmental isolates of Cryptococcus neoformans in goiânia city, Goiás, Brasil

We evaluated the antifungal activities of amphotericin B, fluconazole, itraconazole and voriconazole in 70 Cryptococcus neoformans strains obtained from cerebrospinal fluid from AIDS patients and 40 C. neoformans strains isolated from the environment. Four clinical isolates were identified as C. neoformans var. gattii. The susceptibility test was done using a broth microdilution method according to NCCLS M27-A2. Range minimal inhibitory concentrations (MICs) for C. neoformans clinical isolates were 0.06-1.0 µg/mL for amphotericin B, 0.125-8 µg/mL for fluconazole, 0.03-0.5 µg/mL for itraconazole and 0.03-0.25 µg/mL for voriconazole. C. neoformans environmental isolates showed range MICs 0.015-0.125 µg/mL, 0.25-2.0 µg/mL, 0.007-0.125 µg/mL and 0.03-0.25 µg/mL for amphotericin B, fluconazole, itraconazole and voriconazole respectively. The MICs results obtained from clinical and environmental isolates showed similar pattern of susceptibility and no resistance has been found in our isolates.

ANTIFUNGAL ACTIVITY OF Cymbopogon nardus (L.) Rendle (CITRONELLA) AGAINST Microsporum canis FROM ANIMALS AND HOME ENVIRONMENT

Dermatophytosis is a common zoonosis in urban centers. Dogs and cats have played an important role as its disseminators. Environmental decontamination is essential for the prevention of its propagation to humans and animals. However, sanitizers or disinfectants with antifungal activity, currently available, have high toxicity. The present study evaluated the in vitro effects of an extract of citronella (Cymbopogon nardus) on 31 Microsporum canis isolates from animals and home environments. Susceptibility tests were performed based on document M38-A2 (2008) of the Clinical and Laboratory Standards Institute with modifications for natural products. Although susceptibility variation was observed between the fungus tested, the concentrations that inhibited the growth of 50 and 90% of the microorganisms were low (19.5 and 78 µg/mL, respectively). Thus, this citronella extract showed potent fungistatic and fungicide activities against M. canis isolated from animals and home environments. Therefore, it could be an alternative for dermatophytosis prophylaxis in the home environment.