abstract: ABSTRACT The purpose of this study was to investigate physical activity (PA) program characteristics preferred by low-income childbearing age Latinas and the relationship with the participants’ personal characteristics, cultural values, and acculturation. This was an exploratory study guided by the Preferences and Health Behavior Model (PaHBM), developed by this investigator. Recruitment occurred at three sites; two sites were located in Phoenix, AZ and one site was located in Houston, TX. Non pregnant Latinas between 18 to 35 years old were included (N=275). Latinas were excluded if they were pregnant, incarcerated, physically or mentally disabled, or had chronic diseases (e.g. cardiovascular disease). Quantitative data were collected using the Predictors and Preferences of Physical Activity Research Intervention Participation in an Underserved Latina Community Questionnaire, developed by this author, along with the Brief Acculturation Rating Scale for Mexican Americans, and the Mexican American Cultural Values Scale. The hypotheses were tested utilizing Chi-square, Pearson correlation and logistic regression. Annual family income, parity, country of origin, BMI and acculturation were the personal characteristics significantly associated with preferred PA program by this group of Latinas. Latino women were heterogeneous in their preferences. In general, sixty percent endorsed dancing as the type of activity preferred, 20% preferred PA education and 20% preferred walking. Major differences were found between the types of activity the Latino women were currently participating in compared to their preferred type of activity. Of the 124 who reported to be walking/jogging, almost half (49) preferred dancing, 22 preferred PA education and only 12 preferred walking. The study findings add to the existing knowledge by looking at factors that should be considered when developing PA interventions as well as when prescribing or recommending PA to this population. These results demonstrate the need to identify the preferred PA program characteristics of Latinas prior to developing interventions. Failure to know the patient’s preferred PA program characteristics may result in prescribing or recommending an undesired activity and decrease participation in PA interventions. RESUMEN El propósito de este estudio fue identificar las características preferidas en un programa de actividad física por una comunidad de mujeres Latinas de bajos recursos económicos y en edad fértil, así como la relación de esas características con sus propias características personales, sus valores culturales y su adaptación a la comunidad Anglosajona. Este fue un estudio exploratorio guiado por el “Modelo Preferencias y Comportamiento Saludables” (PaHBM), por sus siglas en Ingles, desarrollado por esta investigadora. El reclutamiento de las Latinas ocurrió en tres sedes: Una en Houston, TX y dos en Phoenix, AZ. Las mujeres Latinas fueron incluidas si tenían entre 18 y 35 años de edad. Se excluyeron mujeres que estaban embarazadas, estuvieran encarceladas, físicamente o mentalmente incapacitadas o que sufrieran alguna enfermedad crónica. Los datos cuantitativos fueron recolectados a través de una encuesta llamada “Predictores y preferencias de participación en un programa investigativo de actividad física”, desarrollada por la autora de este estudio, además utilizando la escala breve de aculturación para Mejicanos Americanos y la escala de valores culturales en Mejicanos Americanos. Las hipótesis fueron probadas utilizando el Chi-cuadrado, la correlación de Pearson, y la regresión lógica. Las características personales más asociadas con las características del programa preferido fueron el salario anual de la familia, el número de hijos, el país de origen, y el índice de masa corporal. En general, 60% prefirieron bailar, 20% clases de actividad física y 20% caminar. Mayores diferencias se encontraron en el tipo de actividad en las que las mujeres Latinas estaban participando, comparado con lo que ellas preferían. De 124 participantes que estaban caminando o trotando, 49 Latinas (39%) preferían bailar, 22 Latinas (17%) preferían clases de actividad física y solo 12 Latinas (10%) prefirieron caminar. Estos resultados demuestran la necesidad de identificar las características del programa de actividad física antes de crear dicho programa. Estos resultados son una adición a los conocimientos existentes, en los que se identificaron factores que deben ser considerados cuando se planea un programa así como cuando se prescribe o se recomienda actividad física a esta población. Sera un fracaso no conocer las preferencias de una paciente para mantenerse físicamente activa porque puede resultar en la prescripción o recomendación de actividades que la paciente no desea y esto se traducirá en reducción de la participación en programas de actividad física. / Dissertation/Thesis / Doctoral Dissertation Nursing and Healthcare Innovation 2016

abstract: This body of research sought to explore relationships between parenting practices, physical activity resources, and Hispanic children’s physical activity. Guided by the Family Ecological Model (FEM) and the Ecological Model of Physical Activity (EMPA) this study examined the influence of parents on children’s physical activity through an integrative review. A cross sectional study was conducted to investigate potential relationships between parental perception safety at school, gender, and children’s physical activity. A cross sectional study was also utilized to examine potential correlations between parenting practices, physical activity resources, and children’s physical activity. Parental role modeling of physical activity and parental support for physical activity emerged as parenting practices that have considerable potential to impact children’s physical activity. Gender differences among children’s physical activity were also a key finding of this study with boys participating in more physical activity than boys. While quality of physical activity resources did not have significant associations with parenting practices or children’s physical activity, more research is needed to determine how resources for physical activity may impact parenting practices, and children’s physical activity. / Dissertation/Thesis / Doctoral Dissertation Nursing and Healthcare Innovation 2017

This aim of this thesis evolved following a review of the literature investigating the factors which influence children’s participation in outdoor play. The review was conducted in light of theory recommending that when seeking to promote physical activity, considering context-specific behaviours and behaviour-specific determinants can enhance the effectiveness of interventions. An initial focus on outdoor play was warranted given the capacity for promotion of physical activity during leisure-time, concerns that children’s independent time outdoors is becoming increasingly restricted, and limited research focus on this domain of physical activity. The synthesised quantitative and qualitative evidence indicated that independent mobility, parental perceptions of safety and the availability of other children to play with were important factors related to outdoor play. However, the review also demonstrated that current understanding of how, where and with whom children spend their leisure-time is limited, and that traditional notions of children’s outdoor time may need to be re-evaluated. These deficiencies were in part due to the complexity of defining and measuring children’s outdoor play. The contributions of different indoor and outdoor leisure-time contexts towards total daily moderate to vigorous physical activity (MVPA) was identified as a particular knowledge gap. The aim of this thesis was therefore to develop greater understanding of the indoor and outdoor contexts of children’s leisure-time physical activity. A novel approach to context-specific physical activity research was devised. This method incorporated use of accelerometry to record physical activity intensity with high resolution, Global Positions System (GPS) receivers to automatically record indoor or outdoor location, and diary data to provide complementary contextual detail. Rather than utilise a domain label such as outdoor play, this method sought to combine measurement tools to not only objectively record physical activity intensity, but also build a picture of the context of this activity using combinations of contextual attributes. Children at the transition between primary and secondary school were the focus of the research due to the changes in independence which occur at approximately this age (10-13 years). The research consisted of three studies presented across three chapters. Chapter Five used data collected between 2006 and 2008 from children aged 10–11 years from Bristol involved in the Personal and Environmental Associations with Children's Health (PEACH) project. Given the association of outdoor play with independent mobility and the availability of other children, the chapter quantified who children spent their time with when indoors or outdoors after school, and measured associations with MVPA. Using a newer GPS receiver, Chapter Six aimed to assess the feasibility of using GPS data to differentiate indoor and outdoor location, and establish a cut-point for use in free-living individuals. Chapter Seven then used this GPS method in combination with accelerometry and diary data provided by children aged 11-13 years from Edinburgh. Owing to concerns that children’s unstructured outdoor time is restricted by parents in favour of adult organised sport and clubs, the chapter aimed to record the profile of children’s physical activity. This was achieved by recording whether indoor and outdoor leisure-time physical activity was structured or unstructured, and exploring relationships between periods spent in these contexts and total daily MVPA. Chapter Six demonstrated that using the signal-to-noise ratio from GPS data is an accurate tool for differentiating indoor and outdoor location, with 96.8% of all ten-second epochs correctly classified. Together the findings of Chapters Five and Seven suggest that children obtain their physical activity in multiple contexts and that no single context appears to fulfil the recommendation of 60 minutes of MVPA per day. Chapter Five showed that children spent most of the after school period with parents or alone, especially when indoors. However when participants were outdoors with other children, multivariate regression analyses indicated that these periods were most strongly associated with MVPA. Complementing these findings, Chapter Seven revealed that in a relatively active and affluent sample, participants accumulated most of their MVPA in school-time or unstructured leisure-time contexts (both indoors and outdoors). The results revealed that these active children spent more than one hour in unstructured outdoor leisure-time contexts each day. However, associations with MVPA were weaker than expected, and whilst being outdoors was favourable compared to being indoors, it was apparent that there is scope to maximise MVPA further when children are outdoors. The median contributions of structured leisure-time contexts to daily MVPA were minimal regardless of indoor or outdoor location. Deconstructing leisure-time according to contextual attributes recorded by a combination of measurement tools proved to be an informative approach for understanding variation in children’s MVPA. Taken together the findings of the thesis indicate potential for leisure-time to contribute greater volumes of MVPA. The results emphasise the importance of children being outdoors, the value of unstructured forms of physical activity and the necessity for children to spend time with their peers. It is clear from these studies that indoor time is also a vital source of MVPA. The work presented in this thesis makes a valuable contribution to our understanding of how children spend their leisure-time and how this relates to physical activity. Further research is required to explore the many other contextual attributes of children’s leisure-time, so that indoor and outdoor environments can be manipulated as part of multi-component interventions that promote physical activity as effectively as possible.

The worldwide prevalence of obesity is reaching epidemic proportions in both adults and children and leading to increased risk of non-communicable diseases (WHO, 2004), including coronary heart disease, circulatory disease, cancer and type 2 diabetes. In the UK, increasing levels of obesity may relate to a decrease in sport and physical activity participation as only 39% of males and 32% of females are sufficiently active to meet stated targets for physical activity (Townsend et al., 2012). In Britain, approximately 8% of the population originate from ethnic minorities that includes a high proportion of individuals from Arabic countries, with a significantly greater odds ratio for a sedentary leisure-time physical activity pattern than people from other countries. The thesis contains three studies which identify the patterns of physical activity in Arabic males (men and boys) in Liverpool, and explores the barriers and motivations to adopting physically active lifestyles, before addressing the feasibility of an intervention to enhance levels of physical activity. Study 1 examined the patterns of physical activity in 62 Arabic men and 65 boys during 7 consecutive days of continuous accelerometry recording. Men and boys were more active during weekdays than weekend days. Although boys were more active than men, they did not perform sufficient minutes of moderate to vigorous activity (60 min per day) to reach recommendations of MVPA. The men, however completed 190 mins per week of activity in the moderate intensity category and therefore satisfied Government guidelines. Study 2 investigated the barriers and motivators in becoming physically active by using focus group semi-structured interview techniques, followed by transcription and content analysis. The findings of the study indicated that physical activity plays a significant role in the individual’s health and that Arabic males perceive several benefits of physical activity for the individual, such as self-confidence, mental health and improved physical condition. There was a mixed interpretation and understanding of physical activity in these groups, with barriers to becoming more active cited as lack of time and socio-cultural barriers of not being accustomed to being physically activity. Facilitators, that encouraged participants to become physically active, included religion and enjoyment. Study 3 used a mixed methods approach to investigate the feasibility of an awareness raising intervention to increasing the levels of physical activity in Arabic males who owned exergames at home. An intervention group of men and boys were provided with physical activity guidelines. Changes in their physical activity levels were measured (using accelerometry) 4 weeks after receiving the guidelines and compared with a control group. The intervention provoked more light activity, moderate and MVPA activity in the men but no reduction in their sedentary behaviour. In boys, light and moderate activity increased, sedentary behavior decreased, but there was no significant difference in MVPA levels. Semi-structured interviews showed that the men found physical activity guidelines alone were not sufficient to motivate them to change their physical activity levels, but the boys found the provision of these useful. Conclusion. This study demonstrated that Arabic men met recommendations for physical activity, whereas boys did not. Barriers to the adoption of physical activity revolved mainly around a lack of understanding of physical activity and guidelines. The intervention strategy was regarded as family-focused and entertaining but not useful in promoting sustainable change in physical activity levels.

Orientador: Antonia Cecília Zacagnini Amaral / Banca: Elizabeti Yuriko Muto / Banca: Otto Bismarck Fazzano Gadig / Resumo: Este trabalho tem como propósito indicar a atividade alimentar diária de Antigonia capros, Synagrops spinosus e Urophycis mystacea (Teleostei). Essas espécies apresentam hábitos demersais e são muito freqüentes na plataforma continental externa e talude superior das regiões Sudeste e Sul do Brasil. Para a identificação de padrões de atividade alimentar diária, foram analisados resultados de coletas realizadas em diferentes horários ao longo do dia, agrupados em cinco períodos: amanhecer, manhã, tarde, entardecer e anoitecer. Em cada período foram identificados estômagos em diferentes graus de repleção e de digestão. Após essa análise, foi observado que cada espécie apresentou características específicas na captura de alimento. Nas três espécies foi verificado que houve aumento na atividade alimentar nos períodos do dia com menor intensidade luminosa. Nestes períodos ocorreu maior consumo de animais bentônicos. Constatou-se também que nos horários de maior e menor atividade, houve variação entre o consumo de organismos bentônicos e pelágicos / Abstract: The aim of this study was to investigate the daily feeding activity of Antigonia capros, Synagrops spinosus and Urophycis mystacea (Teleostei). These species present demersal behavior and are very frequent in Brazilian South and Southeast outer continental shelf and the continental slope. We carried out collections in different times of the day clustered in five periods: dawn, morning, afternoon, nightfall and night, in order to identify the patterns of daily feeding activity. In each period the different levels of digestion and repletion of the material in the stomachs were identified. Through this analysis we oberved that each species presented specific food capturing characteristics. An increase in the feeding activity was observed in the less lightened periods. In these periods there was higher consumption of benthonic animals. We have also found out a relation between the periods of higher and lower activity and the consumption of benthonic and pelagic organisms / Mestre

Made available in DSpace on 2014-06-11T19:27:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-09Bitstream added on 2014-06-13T18:31:20Z : No. of bitstreams: 1 pavan_fr_me_arafcf.pdf: 465333 bytes, checksum: a257ef7f9c510b42345a93dc770ca838 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O Mycobacterium tuberculosis, principal agente da tuberculose (TB), é responsável pela morte anual de dois a três milhões de pessoas no mundo e por prejuízos econômicos globais de aproximadamente 12 bilhões de dólares ao ano. Estima-se que 1/3 da população mundial esteja infectada com o bacilo na forma latente. Apesar disso, nenhuma nova droga específica contra o M. tuberculosis foi desenvolvida desde 1960. O presente trabalho objetivou a investigação do potencial anti-M. tuberculosis intra e extracelular juntamente com a citotoxicidade de 66 compostos envolvendo diferentes classes de ligantes como tiossemicarbazonas, semicarbazonas, hidrazonas, diiminas, fosfinas e bases de Schiff, juntamente com dois diferentes metais (vanádio e rutênio) formando diferentes e inéditos complexos. Para as diferentes análises biológicas in vitro, 3 técnicas já padronizadas foram utilizadas para a detecção da Concentração Inibitória Mínima (CIM), da Citotoxicidade (IC50) e da Atividade Intracelular. Dos 66 compostos analisados neste trabalho, 7 complexos contendo o rutênio, cis-[RuCl2(NO)(BPA)] (G1), cis- [Ru(pic)(dppe)2]PF6 (G6), [Ru(pic)(dppb)(bipy)]PF6 (G7), [Ru(pic)(dppb)(Mebipy)] PF6 (G8), [Ru(pic)(dppb)(Cl-bipy)]PF6 (G9), [Ru(pic)(dppb)(fen)] (G12), cis-[RuCl2(dppb)(bipy)] (G14), foram qualificados como potenciais agentes anti- TB, porque apresentaram atividade inibitória melhor do que algumas drogas comumente utilizadas no tratamento da tuberculose, baixa citotoxicidade e alta atividade inibitória intracelular. / The Mycobacterium tuberculosis, the major agent of tuberculosis (TB), is responsible for approximately 2-3 million deaths annually, with a global economic injury of approximately $12 billion per year. It is estimated that 1/3 of the worldwide population are infected with the latent form bacilli. Although, no new specific drug against M. tuberculosis was developed since 1960. The objective of this study was the investigation of intra and extracellular anti-M. tuberculosis activity and cytotoxicity of 66 compounds involving different class of ligants as thiosemicarbazones, semicarbazones, hydrazones, diimines, phosphines and Schiff bases with two different metals (vanadium and ruthenium) resulting different and unknown complexes. For the different biologicals in vitro analyses, three standardized techniques had been used for the detection of the Minimal Inhibitory Concentration (MIC), Cytotoxicity (IC50) and Intracellular Activity. Of 66 compounds analyzed in this study, 7 complexes containing the ruthenium, cis-[RuCl2(NO)(BPA)] (G1), cis-[Ru(pic)(dppe)2]PF6 (G6), [Ru(pic)(dppb)(bipy)]PF6 (G7), [Ru(pic)(dppb)(Me-bipy)]PF6 (G8), [Ru(pic)(dppb)(Cl-bipy)]PF6 (G9), [Ru(pic)(dppb)(fen)] (G12), cis- [RuCl2(dppb)(bipy)] (G14), were qualified as potential anti-TB agents, because they presented inhibitory activity better than some drugs commonly used in the TB treatment, low cytotoxicity and high intracellular inhibitory activity.

Made available in DSpace on 2014-06-11T19:28:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-05-14Bitstream added on 2014-06-13T20:36:46Z : No. of bitstreams: 1 zanetti_mc_me_rcla.pdf: 772213 bytes, checksum: 7896b5d7cf2e3d207bcbbc502ba1f503 (MD5) / Atualmente, nossa sociedade vem sofrendo uma transformação sem precedentes, no qual, a população, de uma maneira geral, tem se deparado com inúmeros distúrbios, sejam eles, de ordem: social, psicológica e biológica. Esses distúrbios têm refletido em uma menor qualidade de vida e saúde de nossa população, bem como, aumento nos níveis de estresse, diminuição na qualidade de sono, entre outros. Por outro lado, o exercício físico, e a Educação Física escolar vem sendo apontados como importantes agentes promotores de saúde. Pensando nisso, procuramos investigar a influência destas atividades, sobre a estrutura do sono de 58 alunas do sexo feminino, com idades entre 11 e 12 anos (l11.47), da cidade de Limeira SP. Para a coleta dos dados foram aplicados três questionários, no qual, o questionário 1, foi composto por perguntas relativas: à estrutura de sono; problemas de saúde; uso de medicamentos; relacionamento familiar e com amigos; hábitos pessoais; entre outros; o questionário 2 (PAQ-C), foi utilizado para medir o nível de atividade física regular e classificar as adolescentes em ativas e sedentárias; o questionário 3, foi estruturado e utilizado para verificar a percepção subjetiva de qualidade de sono. Posteriormente, aplicamos o teste Qui-quadrado para verificar se havia diferença entre as respostas apresentadas pelas alunas ativas e sedentárias para as questões referentes ao questionário 1, e a questão relativa à percepção subjetiva de qualidade de sono (questionário 3). Adicionalmente, empregamos a análise residual nas questões, onde foi encontrada diferença significativa. Este teste foi utilizado para verificar o comportamento das variáveis e categorias, bem como, determinar as variáveis que mais contribuíram para a diferença encontrada. O teste t para amostras independentes foi aplicado para verificar se havia diferença... / Currently, our society has been suffering a transformation without precedents, in which, the population, in a general way, if has come across with innumerable disorders, is they, social, psychological and biological order. These disorders have reflected in a lesser life quality and health of our population, increase in the levels of stress, reduction in the sleep quality, among others. On the other hand, the physical exercise, and the Physical Education in school have been pointed as important promotional agents of health. Thinking about this, we decided to investigate the influence of these activities, on the structure of the sleep of 58 girls, with ages between 11 and 12 years (l11.47), of the city of Limeira - SP. For the collection of the data three questionnaires had been applied, in which, questionnaire 1, were composed for questions about: sleep structure; health problems; medicine use; relationship in the family and with friends; personal habits; among others; questionnaire 2 (PAQ-C), was used to measure the level of regular physical activity and to classify the adolescents in sedentary or active; questionnaire 3, was structuralized and used to verify the subjective perception of the sleep quality. After, we apply the Qui-square test to verify if it had difference between the answers presented for the active and sedentary girls for the referring questions to questionnaire 1, and the relative question to the subjective perception of the sleep quality (questionnaire 3). Additionally, we use the residual analysis in the questions, where difference was found significant. This test was used to verify the behavior of the variable and categories, and to determine the variable that had more contributed for the joined difference. Test t for independent samples was applied to verify if it had difference between the schedule to sleep and to wake up, and the daily averages... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:24:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-25Bitstream added on 2014-06-13T20:11:56Z : No. of bitstreams: 1 cefali_lc_me_arafcf.pdf: 635100 bytes, checksum: 20424d01dd8e9cc7f5c20d87bb20eec3 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O licopeno é um carotenóide com poderosa ação antioxidante, encontrado em maior quantidade no tomate, utilizado no combate a inúmeras doenças como vários tipos de câncer, dentre eles o de próstata, e doenças cardiovasculares. Sua ação tópica é pouco explorada e, por isso, a proposta de ser utilizado no combate à aceleração do envelhecimento cutâneo, incorporado em uma emulsão. Um estudo preliminar foi realizado para a escolha do tipo de tomate que apresenta maior concentração de licopeno e/ou maior atividade antioxidante. O método de quantificação do fitoquímico foi validado e, em função dos testes de avaliação antioxidante in vitro, o tomate do tipo salada foi o escolhido para o desenvolvimento do fitocosmético. Além disso, o extrato foi submetido à avaliação da citotoxicidade in vitro que constatou a não ocorrência de toxicidade em fibroblastos e macrófagos. Assim, o cosmético foi desenvolvido contendo fase oleosa constituída de derivados de karité e submetido a testes para avaliar sua estabilidade, identificar as características da emulsão, avaliar seu comportamento reológico e sua atividade cosmética pelos testes de permeação cutânea e verificando a eficácia quanto à ação antioxidante do licopeno incorporado na emulsão. Foi possível observar que o fitocosmético é estável, de acordo com as condições experimentais realizadas neste estudo, permite a retenção do licopeno na epiderme e na derme e apresenta atividade antioxidante, sendo promissor para o combate à aceleração do envelhecimento cutâneo. / Lycopene, a carotenoid with high antioxidant activity, is found in greater quantities in tomatoes used to combat many diseases such as various types of cancers, including of the prostate, and cardiovascular diseases. Its topical action is not explored and therefore the proposal to be used to combat aging, incorporated into an emulsion. A preliminary study was conducted to evaluate which tomato had higher concentration of lycopene and/or higher antioxidant activity. The method of quantification of the phytochemical was validated and the lycopene extract was evaluated by testing in vitro antioxidant activity. Tomato salad was chosen for the development of the phytocosmetic. Furthermore, this extract was subjected to the evaluation of cytotoxicity in vitro activity and it was found that the non-occurrence of toxicity in fibroblasts and macrophages. Thus was developed a product containing oily phase consists only derived about shea which was submitted to tests to assess its stability, to identify the characteristics of the emulsion, to evaluate the rheological behavior and cosmetic activity carried out tests such as skin permeation and checking the effectiveness in the antioxidant action of lycopene incorporated into emulsion. Through the results it was possible to observe that the phytocosmetic is stable, allows the retention of lycopene in the epidermis and dermis, shows antioxidant activity and it’s promising to combat the acceleration of skin aging

Made available in DSpace on 2014-06-11T19:24:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-06-29Bitstream added on 2014-06-13T18:20:27Z : No. of bitstreams: 1 barros_lc_me_botfm.pdf: 1332337 bytes, checksum: 59f455ce412e913486df8681ce6b0e01 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / A giroxina isolada do veneno de Crotalus durissus terrificus apresenta atividades coagulante e neurotóxica, caracterizada pelo “rolamento em barril”. É uma serinoprotease do tipo trombina-símile que tem a capacidade de converter o fibrinogênio em fibrina. Visando a atenuação destas atividades, a irradiação com 60Co aparece como uma importante ferramenta. O presente estudo teve por objetivo isolar e purificar a giroxina e avaliar o efeito da irradiação com 60Co sobre suas atividades coagulante e tóxica. O isolamento da giroxina envolveu duas etapas cromatográficas: gel filtração em coluna Sephadex G-75 e afinidade em coluna Benzamidina-Sepharose 6B. O alto grau de pureza foi confirmado por RP-HPLC C2/C18 e pela análise eletroforética, que revelou uma massa molecular de aproximadamente 30 kDa. A giroxina nativa catalisou a hidrólise dos substratos cromogênicos S-2238 e S-2288, demonstrando tratar-se de uma serinoprotease pertencente à subclasse das enzimas trombina-símile, estável em diferentes pHs (5,5 a 8,5), sensível aos metais Mn2+ e Cu2+ e aos inibidores de serinoprotease PMSF e benzamidina. Apresentou melhor atividade coagulante sobre o plasma humano entre os pHs 6,0 e 7,4. A irradiação da giroxina nas doses de 0,5; 1,0 e 2,0 kGy anulou completamente suas atividades coagulante e tóxica. Os ensaios de toxicidade in vivo mostraram apenas alterações comportamentais sem demonstrar o rolamento em barril. Este fato sugere que as toxinas purificadas são mais sensíveis à irradiação, pois não há proteção mútua entre as proteínas presentes no veneno total. A giroxina nativa também não causou o bloqueio da contração neuromuscular in vitro sugerindo que a sua ação não tem efeito sobre o sistema nervoso periférico nas concentrações utilizadas / Gyroxin isolated from Crotalus durissus terrificus venom presents coagulant and neurotoxic activities. It belongs to the thrombin-like enzyme group capable of converting fibrinogen into fibrin. To reduce these toxic activities, the irradiation with Cobalt-60 appears to be an important tool. The present study was carried out in order to isolate and purify the gyroxin and evaluate the effects of irradiation with Cobalt-60 on coagulant and toxic activities. The gyroxin isolation consisted of two chromatographic steps: gel filtration (Sephadex G-75) and affinity (Benzamidine-Sepharose 6B). The high purity level of gyroxin was confirmed by RP-HPLC C2/C18 and electrophoretic analysis that showed a molecular weight of 30 kDa. The native gyroxin hydrolyzed the chromogenic substrates S-2238 and S-2288, indicating that this enzyme is a serine proteinase that belongs to the group of thrombin-like enzymes, stable when submitted to pHs from 5.5 to 8.5 and inhibited by Mn2+, Cu2+, PMSF and benzamidine. It was capable of coagulating human plasma at pH 6.0 and 7.4. The gyroxin irradiated at 0.5, 1.0 and 2.0 kGy doses neutralized the coagulant and toxic activities. The in vivo toxic study showed only behavioral alterations with no barrel rotation. This fact suggests that purified toxins are more sensitive to the irradiation because they e mutual protection with the other proteins present in the total venom. The native gyroxin was not able to block in vitro neuromuscular contraction, suggesting that the action of gyroxin, in the concentration used in this study, has no effect on the peripheral nervous system

Made available in DSpace on 2014-10-09T12:49:21Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:02:30Z (GMT). No. of bitstreams: 1 09994.pdf: 3905779 bytes, checksum: 7cdc8d8ae9585729a6a818ed202c59c8 (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP