Orientador: José Paulo Cabral de Vasconcellos / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-28T03:59:37Z (GMT). No. of bitstreams: 1 Teixeira_MyrianKatiaIser_M.pdf: 2342582 bytes, checksum: 2e51b6261b602fa9f018d1bd96ed680a (MD5) Previous issue date: 2015 / Resumo: Objetivo: Avaliar o efeito e a segurança da toxina botulínica tipo A quando aplicada na região do músculo elevador da pálpebra superior, para a indução de ptose palpebral protetora em gatos domésticos. Métodos: Neste estudo do tipo longitudinal, série de casos com intervenção, um total de 10 gatos foram submetidos à quimiodenervação do músculo elevador da pálpebra superior esquerdo, através da aplicação transpalpebral de 10 unidades de toxina botulínica do tipo A. Alterações sistêmicas, mobilidade ocular, função visual, pressão intraocular, o aparecimento, grau e duração da ptose foram avaliados antes da aplicação, diariamente, durante os sete primeiros dias e, posteriormente, nos dias 14, 21 e 28 após a aplicação. A mensuração da fenda palpebral foi realizada também no olho contralateral que funcionou como controle. Resultados: O início do efeito clínico foi observado entre os dias 1 e 4 após a aplicação; a ptose máxima foi observada entre o quinto e o sétimo dia e a duração média de ação da toxina foi de 21 dias. O tempo máximo para recuperação da ptose foi de 28 dias. A porcentagem média de redução da fenda palpebral foi de 39,66% (16,55% ¿ 59,64%). A análise qualitativa demonstrou que duas gatas (20%) apresentaram cobertura corneal maior que 50%, sete gatas (70%) obtiveram cobertura corneal entre 25 e 49% e uma gata (10%) mostrou cobertura corneal menor que 25%. Os valores da pressão intraocular permaneceram dentro dos limites de normalidade. A toxina botulínica não causou efeitos adversos nos gatos desse estudo. Conclusão: O uso de toxina botulínica tipo A no músculo elevador da pálpebra superior foi seguro e promoveu ptose palpebral temporária parcial nos gatos desse experimento / Abstract: Purpose: To evaluate the effect and safety and of botulinum toxin A for the induction of palpebral ptosis in felines. Methods: In this prospective interventional study, a total of 10 cats underwent transpalpebral chemodenervation of levator palpebral superioris with 10 units of botulinum toxin type A in the left eye. The systemic changes, ocular mobility, visual function, intraocular pressure, and the onset, degree and duration of ptosis were evaluated before application, on a daily basis during the first seven days and on days 14, 21 and 28 after application.The palpebral edge of the contralateral eye was also measured. Results: A clinical effect was observed beginning between the first and fourth days after botulinum toxin A administration. The extent of ptosis was maximal between the fifth and seventh days after administration, and ptosis was observed for a mean duration of 21 days. The maximum time for recovery of ptosis was 28 days. The palpebral edge was reduced by an average of 39.6% (16.55% - 59.64%). Qualitative analysis showed that two cats (20%) had greater than 50% coverage corneal, seven cats (70%) achieved corneal coverage between 25 and 49% and one cat (10%) showed corneal coverage less than 25%. Intraocular pressure values were within normal limits. Botulinum toxin did not cause undesirable effects. Conclusions: The use of botulinum toxin A in the levator palpebrae superioris muscle was safe and provided transient, partial palpebral ptosis in all of the studied cats / Mestrado / Ciencias Biomedicas / Mestra em Ciências Médicas

A Bothrops jararaca é a serpente de maior importância médica no Brasil. Vários estudos foram realizados com o objetivo de caracterizar os componentes do veneno de serpentes, entretanto, a base molecular dos genomas das serpentes é pouco conhecida. Assim, foi realizado o sequenciamento e montagem do genoma da serpente Bothrops jararaca. Foram construídas bibliotecas tipo shotgun e mate-pair para realização de corridas de sequenciamento usando a tecnologia Illumina e sequências complementares foram obtidas em equipamento PACBIO RS II. Uma biblioteca de BACs também foi construída e 768 pools de 12 BACs foram sequenciados. Um grande conjunto de segmentos genômicos foi obtido e foi possível identificar genes de várias toxinas, entre elas SVMPs, SVSPs, BPPs, CRISPs e VEGF. Ainda foi possível depreender o contexto genômico de muitos destes genes e identificamos os principais elementos repetitivos genômicos. Estes achados são relevantes para o entendimento da função e evolução do sistema venenífero e podem servir de base para outros estudos futuramente. / The pit viper Bothrops jararaca is the most medically important snake in Brazil. Several studies were conducted in order to characterize the components of snake venom. However, the molecular basis of snake genomes is poorly known. Hence, it was carried out the sequencing and assembly of the Bothrops jararaca snake genome. Shotgun and mate-pair libraries were constructed to perform sequencing runs using Illumina technology and complementary sequences were obtained in PACBIO RS II equipment. A BAC library was also constructed and 768 pools of 12 BACs were sequenced. A large number of genomic segments was obtained. It was possible to identify genes of several toxins, including SVMPs, SVSPs, BPPs, CRISPs and VEGF. In addition, it was possible to infer the genomic context related to most of these genes and identify the main genomic repetitive elements. These findings are relevant for understanding the function and evolution of the venom system and it provides the basis for further studies.

A Qualificação de Performance (QP) é requisito das Boas Práticas de Fabricação e consiste de uma documentação de todas as atividades envolvidas para produzir um produto específico, neste caso, a toxina diftérica. O objetivo é garantir que as especificações do produto atendam aos resultados das análises de rotina. O Instituto Butantan introduziu um biorreator para ser utilizado nesta produção e a QP foi considerada e desenvolvida. Análises da temperatura e pressão foram realizadas durante três ciclos de esterilização. Sensores de temperatura e indicadores biológicos foram previamente distribuídos no equipamento considerando pontos críticos pré-observados. Os resultados da curva de temperatura e pressão validaram o equipamento. Três ciclos de crescimento de C. diphtheriae foram realizados e monitorados. Os resultados da DO ou turbidez medidos tanto por um espectrofotometro convencional como por um sensor de biomassa on line mostraram títulos da toxina já a partir de 48 horas (teste de floculação). A toxina foi caracterizada bioquimicamente. / Performance qualification (PQ) is requisite of the Good Manufacturing Practices and consists on complete documentation of all activities involved to produce a specific product, in this case, the diphtheria toxin. The goal is to ascertain all product specifications that were determined by its routine analysis. Instituto Butantan introduced a bioreactor to be use in the diphtheria toxin production and the PQ must be considered and developed. Analysis of the temperature and the pressure were performed during three sterilization cycles. Temperature sensors and biological indicators where previously distributed in the equipment, considering critical pre-observed points. The temperature and pressure curve results validated the equipment. Three C. diphtheriae growth cycles were performed and monitored. The optical density (OD) or turbidity results measured either by one conventional spectrophotometer and/or by one on line biomass sensor showed toxin titers after 48 hours of the harvesting (flocculation test). The toxin was biochemically characterized.

A espécie Bothrops atrox é responsável por grande parte dos acidentes no estado do Amazonas. No entanto, ainda são poucos os estudos sobre as toxinas que compõem o veneno dessa serpente, os mecanismos envolvidos na sintomatologia dos acidentes, bem como de formas de inibição. Os soros antiveneno utilizados atualmente com o objetivo de neutralizar as atividades sistêmicas e locais dos venenos mostraram reatividade cruzada entre componentes dos venenos de serpentes do gênero Bofhrops (MOURA DA SILVA et al., 1990). No entanto alguns componentes do veneno não apresentaram essa reatividade (SILES-VILARROEL et al.., 1974), mostrando a existência de toxinas espécie específicas.Os venenos de serpentes estão sujeitos a grandes variações induzidas por diversos aspectos ontogenéticos e influencia do habitat. Assim, essas variações podem gerar toxinas espécie específicas cujos mecanismos de ação ainda são desconhecidos e que os anticorpos presentes nos antivenenos disponíveis não sejam capazes de reconhecer e neutralizar eficientemente. O gênero Bothrops possui espécies extremamente variáveis, algumas de difícil classificação taxonõmica, e novas espécies têm sido descobertas recentemente. Como contribuição para o acúmulo de informações a respeito das diferentes composições do veneno do gênero Bothrops, e para o conhecimento de toxinas já isoladas e as ainda não isoladas na espécie tipo Bothrops atrox, foi construída uma biblioteca de cDNA da glândula de veneno.Os dados obtidos são importantes para a elaboração de um painel da expressão gênica dessa espécie e permitirá a identificação de toxinas que podem ser comuns ou não ao veneno de outras espécies do gênero. Aliado a isso, esses dados permitirão a correlação com o estudo proteõmico, e possivelmente fornecerão subsídios para a melhoria da terapêutica empregada no tratamento dos casos envenamento na região. / The species Bothrops atrox is responsible for most accidents in state of Amazonas. However, there are few studies on toxins that make up the venom of this snake, mechanisms involved in symptomatology of accidents, as well as inhibition forms. Sera antivenom currently used in order to neutralize the systemic and local activities of the venoms showed cross-reactivity between components of the venom of Bothrops (MOURA DA SILVA et al., 1990). However some components of the venom showed no reactivity (SILES-VILARROEL et al., 1974), indicating the existence of species-specific toxins. Snake venoms are subject to large variations induced by several aspects and influences of ontogenetic habitat. Thus, these variations can produce toxins whose mechanisms of species-specific action are still unknown and antibodies present in available antivenoms maybe not are capable to recognize and neutralize some toxins efficiently. Bothrops species have highly variable, some of difficult taxonomic classification and new species have been discovered recently. As a contribution to gain of information about of different compositions of Bothrops venom and to knowledge of toxins not yet isolated and the isolated in Bothrops atrox type species we constructed a venom gland cDNA library . The data obtained are important for the development of gene expression panel of this species and enable the identification of toxins common or not in the venom of other species. These data will allow correlation with the proteomic study, and possibly provide input for improving the therapeutic used in the treatment of accidents cases in the region.

MENVSC / Department of Ecology and Resource Management / See the attached abstract below

Aflatoxin is a poisonous mold that has spread around the world, posing a threat to food security and the agricultural economy. A total of 4,5 million people are in danger of being exposed to aflatoxins on a long-term basis around the world. Acute toxicity is caused by consuming significant amounts of toxins in a short period of time, which can lead to death in the worst cases, whereas chronic toxicity is caused by consuming small quantities over a longer period of time, which can lead to low birth weight, immunosuppression, restricted growth in children, and liver cancer in the worst cases. The occurrence of aflatoxins in West Africa was recognized, studied, and investigated in this paper based on a literature review. The findings demonstrate that large levels of aflatoxins have been found in West African raw materials and food and the human body. Children under the age of five, pregnant women, and breastfeeding mothers are the most vulnerable to aflatoxins. Furthermore, the findings reveal that the majority of the population is unaware of aflatoxins and its health implications. Inadequate governmental systems, low societal development, lack of access to health care, a low educated population, climate variability and climate change, high levels of illiteracy, and a lack of laboratories are only a few of the many obstacles that the region has in limiting aflatoxins concentrations. Sorting procedures, the use of tarpaulins, and seminars have all helped raise awareness and knowledge and reduce contamination and consumption of aflatoxin-contaminated foods. This study argues that a multi-sectoral strategy is needed to promote food security and local education. Increased limitations and regulations and higher standards may be able to help limit aflatoxin contamination and exposure.

With cardiovascular disease, diabetes mellitus, and neurodegenerative conditions on the rise, understanding their pathogenesis is paramount to tackling this public health crisis. Current research indicates that the primary cause of these diseases is mitochondrial dysfunction in the affected patients. While genetics plays a role in these conditions, lifestyle choices and exposure to toxins also significantly contribute to their development. Unfortunately, early-stage diagnosis can be difficult due to overlapping symptoms with other diseases. Developing innovative therapies that can prevent or reverse the deterioration of metabolic dysfunctions is critical to establishing early intervention. My research focused on investigating molecular targets linked with Friedrich's Ataxia, an inherited metabolic disorder, through conducting functional in-vitro studies using human-derived cell samples, as well as developing inventive animal models created via Xenopus laevis tadpoles to evaluate the effects of environmental stressors. My investigations have uncovered promising treatment options that improve mitochondrial function, mitigate oxidative stress, and elucidate critical mechanisms involved in environmentally induced disruptions to mitochondria. / Doctor of Philosophy / Metabolic dysfunction is a widespread health issue that affects millions of individuals each day. Its associated disorders, such as cardiovascular disease, diabetes, and neurodegenerative conditions, are rising due to various factors ranging from genetic predispositions to environmental and lifestyle-related risks. Therefore, there's an urgent need to identify this disorder early on and develop innovative treatment options. Considering this growing public health concern, it has become imperative to establish new methods for detecting metabolic dysfunction at its nascent stage while also exploring potential therapeutic interventions. Our research utilized cells derived from affected patients and animal models in devising novel approaches toward understanding the molecular mechanisms underpinning metabolic dysfunction. Our findings revealed several pathways and molecular targets contributing significantly to this condition, which could effectively be leveraged to develop targeted therapeutic strategies to combat its effects. Expanding our knowledge base will enable us to stay updated with emerging insights on treating metabolic dysfunction effectively while substantially improving patient outcomes.

No description available.

Buruli ulcer disease (BUD) remains a ‘mysterious disease’ due to the unknown mode of M. ulcerans transmission and pathogenesis. To understand these, it is important to determine the reservoir of the organism in its natural environments, and stress response and interactions of M. ulcerans in its natural niche and during infection of a host. The major virulence factor of M. ulcerans is mycolactone, a lipid cytotoxin that is encoded on a giant plasmid pMUM001. Genetic analysis suggests that plasmid pMUM001 was acquired by M. ulcerans during evolution from its progenitor, M. marinum. Coincidental evolution of virulence hypothesis suggests that many microbes evolve to acquire traits to outcompete or overcome biotic and abiotic forces during their normal life cycle in the outside-host environment, which can confer virulence during infection of a human host. Hence in this study, we exposed M. ulcerans to selective abiotic forces such as UV, and dynamic oxygen and temperature conditions to determine their effect on M. ulcerans growth, and mycolactone and global gene expression. We also studied the role of mycolactone in determining polymicrobial interaction of M. ulcerans in its natural aquatic habitat by exposing mycolactone coated and uncoated slides in M. ulcerans endemic and non-endemic aquatic locations and determining differences in microbial community composition between them. Further, we studied quorum quenching ability of mycolactone against an opportunistic pathogen, S. aureus. The results obtained showed that exposure of M. ulcerans to abiotic stresses such as higher temperature and lower than optimal oxygen conditions modulate its global and mycolactone gene expression. Further, we also showed that mycolactone can impact overall microbial community structure in a polymicrobial environment in its natural, aquatic habitat. Mycolactone also effected virulence and quorum sensing in an opportunistic pathogen, S. aureus, without inhibiting its growth. These findings are important as they provide insight toward potential reservoirs or environmental niches which may harbor M. ulcerans and inform new potential mechanisms of pathogenesis. Further, our novel research of synergistic or antagonistic interactions within the complex polymicrobial communities colonizing skin and aquatic habitats is a powerful approach in determining M. ulcerans colonization efficiency, resiliency, and transmission mechanisms.

No description available.