Análise citogenética clássica, molecular e ultraestrutural em escorpiões da família Buthidae: um enfoque evolutivo

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

Distribuição Geográfica dos Escorpiões no município de Presidente Prudente - SP nos anos de 2012 e 2013

No Brasil os acidentes por picada de escorpião são considerados um problema para saúde pública, uma vez que este agravo pode levar ao óbito caso não haja um tratamento adequado. Assim, no município de Presidente Prudente/SP o órgão responsável pelo combate aos escorpiões é o Centro de Controle de Zoonoses (CCZ), que tem por finalidade notificar a aparição de escorpiões no município, promovendo portanto, as devidas informações à população com finalidade de evitar acidentes. Desse modo, com o uso dos Sistemas de Informações Geográficas (SIGs) aplicados à técnicas de estatística espacial, foi possível elaborar mapas de distribuição das notificações de escorpiões referente aos anos de 2012 e 2013, por meio dos métodos da geocodificação de endereços; estimador de intensidade de Kernel; índice de Moran função (LISA); quantidade de notificações por setor censitário e interpolação por vizinho mais próximo. Como resultado desta análise, o trabalho fornece informações ao CCZ sobre a distribuição de tal fenômeno para que fosse possível um controle direcionado os locais mais infestados, economizando tempo e recursos financeiros, visando o bem estar da população

Análise citogenética clássica, molecular e ultraestrutural em escorpiões da família Buthidae: um enfoque evolutivo

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

Estudo peptídico e determinação do perfil de metabólitos de venenos de escorpiões da família buthidae: Tityus serrulatus, Tityus bahiensis e Tityus obscurus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise do transcriptoma das glândulas venenosas do escorpião Tityus stigmurus

In Brazil, accidents with scorpions are considered of medical importance, not only by the high incidence, but also for the potentiality of the venom from some species in determining severe clinical conditions. Tityus stigmurus is a widely distributed scorpion species in Northeastern Brazil and known to cause severe human envenomations, inducing pain, hyposthesia, edema, erythema, paresthesia, headaches and vomiting. The present study uses a transcriptomic approach to characterize the molecular repertoire from the non-stimulated venom gland of Tityus stigmurus scorpion. A cDNA library was constructed and 540 clones were sequenced and grouped into 37 clusters, with more than one EST (expressed sequence tag) and 116 singlets. Forty-one percent of ESTs belong to recognized toxin-coding sequences, with antimicrobial toxins (AMP-like) the most abundant transcripts, followed by alfa KTx- like, beta KTx-like, beta NaTx-like and alfa NaTx-like. Our analysis indicated that 34% include other possible venom molecules , whose transcripts correspond to anionic peptides, hypothetical secreted peptides, metalloproteinases, cystein-rich peptides and lectins. Fifteen percent of ESTs are similar to cellular transcripts. Sequences without good matches corresponded to 11%. This investigation provides the first global view of cDNAs from Tityus stigmurus. This approach enables characterization of a large number of venom gland component molecules, which belong either to known or atypical types of venom peptides and proteins from the Buthidae family

Aplicação de nanopartículas de quitosana com potencial de adjuvante na produção de soro contra o venneno do escorpião Tityus serrulatus

In Brazil, several species of scorpions are known to cause accidents which can lead to death, which are mainly belonging to the genus Tityus. The scorpion Tityus serrulatus is the main responsible for more severe cases. Anti-scorpion serums are routinely produced by various institutions, despite their effectiveness, quality and action depends on how quickly treatment is started. Studies have been developed in the search for appropriate technologies to encapsulate and release recombinant or natives proteins capable of inducing antibody production. In this context, chitosan copolymer which can be obtained from the partial deacetylation of chitin or in some microorganisms and it is biocompatible and biodegradable has been widely used for this purpose. This study aimed to search for a system release from chitosan nanoparticles for peptide / protein of the venom of the scorpion T. serrulatus, able to provide a new model of immunization in animals, in order to obtain a potential novel polyclonal serum, anti-venom T. serrulatus. The chitosan nanoparticles were prepared by ionic gelation with polyanion tripolyphosphate (TPP). After standardizing the concentrations of TPP and chitosan was evaluated the efficiency of incorporation of bovine serum albumin (BSA) and scorpion venom, showed particle size compatible with the intended purpose. The particles showed adequate size around 200nm. The crosslinking was confirmed by absorption spectroscopy in the infrared. After verified the high encapsulation efficiency (EE) for acid bicinconínico method (BCA) protein assay and the particle size distribution, the success of the technique was proven and the potential for in vivo application of nanoparticles. The experimental animals were vaccinated and the antibodies measured by ELISA

Uma abordagem peptidômica do venedo do escorpião Tityus serrulatus

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

Aspectos epidemiológicos e clínicos do escorpionismo na região de Santarém, Estado do Pará, Brasil

Este trabalho é um estudo prospectivo e descritivo dos aspectos epidemiológicos e clínicos de 72 envenenamentos por escorpiões admitidos no Hospital Municipal de Santarém, Estado do Pará, Brasil, entre fevereiro de 2000 a fevereiro de 2001. Trouxeram o animal 8,3% das vítimas, os quais foram identificados como T. cambridgei. O sexo masculino foi acometido em 83,3%. A idade das vítimas e o tempo para o socorro médico foram respectivamente de 33,6±18,3 anos e 4,6±3,2 horas em média. Os membros superiores foram acometidos em 51,5% dos casos. As manifestações locais estiveram presentes em 91,7% e as sistêmicas em 98,6% dos envenenamentos. Entre os sintomas locais encontramos: parestesia em 79,2%, dor em 52,8%, e edema em 26,4% dos casos. Nas manifestações sistêmicas predominou as queixas neurológicas em 97,2% das vítimas, sendo o sintoma de sensação de "choque elétrico" pelo corpo (88,9%) o mais freqüente. No exame neurológico os sinais mais encontrados foram: mioclonias (93,0%), dismetria (86,1%), disartria (80,6%) e ataxia de marcha (70,8%). Classificou-se como moderados 76,4% dos envenenamentos, sem nenhum caso grave. Deixaram de realizar a soroterapia 32,7% dos casos moderados, por ausência de soro específico no momento do atendimento. O escorpionismo da região de Santarém mostra um comportamento clínico regional diferente daqueles descritos no Brasil e de outros locais da Amazônia e, apresenta uma clínica predominantemente neurológica, ainda não descrita na literatura brasileira.

Agressividade relacionada ao contexto alimentar em escorpião-amarelo Tityus Serrulatus (Lutz & Mello, 1922)(Scorpiones, Buthidae)

Scorpions are among the oldest living groups of animals, and are present in almost every continent. Although many studies are made about the toxins present in their venom, little is known about their behavior. The aim of this paper is to investigate the expression of agressiveness in the brazilian yellow scorpion Tityus serrulatus, specifically the relation between agressiveness, motor activity and feeding condition. The study was divided in two parts. First, we used 16 adult individuals, placed in two terrarium, to establish an ethogram. Each group was observed for 36h, and the behaviors displayed were described and categorized as agonistic or non-agonistic. In the second part of the experiment, we used 32 adult animals in three different nutritional states: Feeding (still ingesting food), Sated (1 to 4 days since last meal) and Deprived (14 to 25 days since last meal). The individuals were paired, each pair placed in a terrarium and observed for 30min. Behaviors displayed were timed in seconds. Our results show that only Feeding individuals displayed agonism towards others, and no relation was observed between motor activity and feeding conditions

Análise de agrupamentos para o reconhecimento de padrões de infestação de aracnídeos em zonas urbanas

Pós-graduação em Biociências - FCLAS

Síntese e caracterização de pequenos peptídeos lineares do veneno de Tityus serrulatus (Buthidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação funcional e estrutural de um novo peptídeo antimicrobiano do escorpião Tityus stigmurus

In Brazil, there is a high incidence of venomous animals. Among them, scorpions are highlighted by their medical importance, and for being their venom a source of several molecules with biological and pharmacological activity not yet fully understood, including several bioactive peptides. Antimicrobial peptides (AMPs) are components of the immune system in prokaryotes and eukaryotes, used in the first line of defense against microorganisms. In the present study, we characterized the first PAM previously identified through transcriptome of the venom gland of the scorpion Tityus stigmurus, named Stigmurin. The characteristics of Stigmurin were investigated by computational modeling and construction of dendrogram. In vitro tests investigated the antibacterial, antifungal, haemolytic and cytotoxic effects of crude venom and Stigmurin. In addition, the structural characteristics of Stigmurin were investigated by circular dochroism in water, 2, 2 , 2- trifluoethanol (TFE) and sodium dodecyl sulfate (SDS) and the models were refined by molecular dynamics simulations. The results showed that the selected sequence encodes a mature protein of 17 amino acid residues and the dendrogram reveals a case of convergent evolution. The crude venom showed no antimicrobial activity, however, Stigmurin exhibited a broad spectrum of antibacterial activity, with minimal inhibitory concentrations (MIC) ranging from 31.25 and 250 µg/mL for different strains, while the hemolytic activity at these concentrations was low. In cytotoxicity studies, the crude venom was unable to reduce cell viability in VERO E6 cells; in contrast, its activity in SiHa cells was significantly higher, corresponding to IC50 of 3.6 µg/mL. For Stigmurin the concentration sable to decrease cell viability of Vero E6 and SiHa cells in 50% were 275.67 µg/mL and 212.54 µg/mL, respectively. The dichroism spectra revealed the conformational flexibility, with predominating extended and β–sheet structures, as well as a remark able renaturation ability. The results suggest that Stigmurin could be considered as a potential antiinfective drug

Análise fitoquímica e avaliação das atividades anti-inflamatória, antipeçonhenta e citotóxica de extratos aquosos de Aspidosperma pyrifolium e Ipomoea asarifolia

Envenomation caused by venomous animals, mainly scorpions and snakes, are a serious matter of public health. Tityus serrulatus is considered the most venomous scorpion in South America because of the high level of toxicity of its venom. It is responsible for causing serious accidents, mainly with kids. The species Bothrops jararaca is a serpent that has in its venom a complex mixture of enzyme, peptides and other molecules. The toxins of the venom of B. jararaca induce local and systemic inflammatory responses. The treatment chosen to serious cases of envenomation is the intravenous administration of the specific antivenom. However, the treatment is not always accessible to those residents in rural areas, so that they use medicinal plant extracts as the treatment. In this context, aqueous extracts, fractions and isolated compounds of Aspidosperma pyrifolium (pereiro) and Ipomoea asarifolia (salsa, salsa-brava), used in popular medicine, were studied in this research to evaluate the anti-inflammatory activity in the peritonitis models induced by carrageenan and peritonitis induced by the venom of the T. serrulatus (VTs), and in the local oedema model and inflammatory infiltrate induced by the venom of the B. jararaca, administrated intravenously. The results of the assays of cytotoxicity, using the MTT, showed that the aqueous extracts from the plant species presented low toxicity to the cells that came from the fibroblast of the mouse embryo (3T3).The chemical analysis of the extracts by High Performance Liquid Chromatography revealed the presence of the rutin flavonoid, in A. pyrifoliu, and rutin, clorogenic acid and caffeic acid, in I. asarifolia. Concerning the pharmacological evaluation, the results showed that the pre-treatment using aqueous extracts and fractions reduced the total leukocyte migration to the abdominal cavity in the peritonitis model caused by the carrageenan and in the peritonitis model induced by the T. serulatus venom. Yet, these groups presented anti-oedematous activity, in the local oedema model caused by the venom of the B. jararaca, and reduced the inflammatory infiltrate to the muscle. The serum (anti-arachnid and anti-bothropic) specific to each venom acted inhibiting the inflammatory action of the venoms and were used as control. The compounds identified in the extracts were also tested and, similar to the plant extracts, showed meaningful anti-inflammatory effects, in the tested doses. Thus, these results are indicating the potential anti-inflammatory activity of the plants studied. This is the first research that evaluated the possible biological effects of the A. pyrifolium and I. asarifolia, showing the biological potential that these species have.

Efeito anticonvulsivante de frações isoladas da peçonha da formiga Dinoponera quadriceps (Formicidae: Ponerinae)

A epilepsia é uma patologia crônica do sistema nervoso central que afeta cerca de 65 milhões de indivíduos no mundo. Aproximadamente 30% desses indivíduos desenvolvem crises convulsivas que persistem apesar do tratamento monitorado com drogas antiepiléptica s. Assim, há uma evidente necessidade do desenvolvimento de novos fármacos antiepilépticos e as peçonhas podem ser uma excelente fonte de modelos. Nesse contexto, enquanto já vários estudos sobre peçonhas de serpentes, escorpiões e aranhas, pouco se sabe s obre as peçonhas de formigas. Estudos prévios do nosso laboratório demonstraram que a peçonha desnaturada da formiga Dinoponera quadríceps protegeu camundongos de crises convulsivas e morte induzidas por bicuculina (BIC). Nesse contexto, o objetivo desse t rabalho foi investigar o potencial anticonvulsivante de frações isoladas da peçonha de D. quadríceps em crises convulsivas induzidas pela BIC, bem como uma análise dos efeitos dessas frações no comportamento natural dos camundongos no campo aberto. Os anim ais foram divididos em grupos, os quais receberam injeções (1 mg/ml i.c.v.) de seis frações distintas e tiveram seu comportamento geral observado no campo aberto durante 3 0 min. No segundo experimento, o s animais receberam as mesmas frações 20 min antes da administração de bicuculina (10 mg/ml). Em seguida, foi analisado o comportamento motor convulsivo desses animais durante 30 minutos no campo aberto. No primeiro experimento, não foram observadas alterações comportamentais. Já no segundo experimento, a ad ministração prévia de DqTx1, DqTx3, DqTx4 e DqTx6 aumentou a latência para o desenvolvimento de crises tônico - clônicas. Além disso, todas as frações, exceto DqTx5, aumentaram a latência para a morte dos animais. Ainda, os melhores resultados foram obtidos com a fração DqTx6, que protegeu 62,5% dos animais testados contra o desenvolvimento de crises tônico - clônicas e 100% dos animais contra a morte.