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No. of bitstreams: 1 guiraldi_lm_me_bot.pdf: 5832304 bytes, checksum: abf9a0919d7f06699fed2f2c09bad1b1 (MD5) Previous issue date: 2016-07-22 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A família Trypanosomatidae inclui agentes etiológicos responsáveis por ocasionar doenças em humanos e demais animais, englobando dezenas de protozoários pertencentes a diferentes gêneros, destacando-se os protozoários representantes dos gêneros Leishmania, causadores das leishmanioses e Trypanosoma, causador de tripanosomíase. Enquanto a transmissão das leishmanioses ocorre devido à picada de insetos fêmeas de flebotomíneos pertencentes ao gênero Phlebotomus no Velho Mundo e Lutzomyia no Novo Mundo, a transmissão da tripanossomíase depende da espécie de tripanosoma envolvida na infecção, podendo ocorrer pela picada de insetos adultos do gênero Glossina; pelas fezes de triatomíneos, sobretudo do gênero Triatoma e mecanicamente por moscas hematófagas, principalmente do gênero Stomoxys. Além dos humanos, animais domésticos e silvestres podem atuar como reservatórios de tripanosomatídeos, incluindo os primatas não humanos, os quais estão associados com o ciclo enzoótico das leishmanioses e tripanosomíase, sendo que a infecção natural por protozoários do gênero Trypanosoma e Leishmania em mamíferos selvagens é comum na natureza. O objetivo do trabalho foi pesquisar tripanosomatídeos em primatas de cativeiro procedentes do Parque Zoológico Municipal de Bauru – SP por meio de anticorpos contra o parasito e pela amplificação do DNA do protozoário. Para tanto, foram coletadas amostras de sangue de 39 primatas. A técnica molecular de Reação em Cadeia da Polimerase (PCR) foi aplicada na pesquisa de Leishmania braziliensis, Leishmania amazonensis e Leishmania infantum, utilizando-se primers espécie-específicos, direcionados à região do kDNA dos parasitos, sendo todos os animais negativos. Porém, com a utilização de primers para a região ribossomal (ITS-1), específicos para a família Trypanosomatidae, 37 dos 39 (94,9%) animais foram positivos para Trypanosoma spp. Realizou-se também a prova sorológica de Reação de Imunofluorescência Indireta (RIFI) para Leishmania braziliensis, Leishmania infantum e Leishmania amazonensis, revelando um animal da espécie Erythrocebus patas (macaco-pata) positivo para Leishmania braziliensis com título 160. Os resultados demonstram que os primatas do zoológico estão susceptíveis à infecção por tripanosomatídeos, alertando-se para a necessidade de busca ativa de vetores flebotomíneos e triatomíneos nos recintos e arredores, como medida preventiva para o risco de infecção aos funcionários e visitantes. / The family Trypanosomatidae includes etiological agents responsible for causing diseases in humans and other animals. There are dozens of protozoa belonging to different genres, especially protozoa representatives of Leishmania genre, which causes leishmaniasis and Trypanosoma, responsible for trypanosomiasis. While the transmission of leishmaniasis is due to the bites of female sand flies belonging to the genus Phlebotomus in the Old World and Lutzomyia in the New World, the transmission of trypanosomiasis depends on the species of trypanosome involved in the infection. It may occurs by the bite of adult insects of the genus Glossina or by the feces of insects, especially by the genre Triatoma and mechanically by bloodsucking flies, especially the Stomoxys genre. In addition to humans, domestic and wild animals can act as trypanosomatids reservoirs, including non-human primates, which are associated with the enzootic cycle of leishmaniasis and trypanosomiasis; the natural infection by Trypanosoma and Leishmania protozoa in wild mammals is common in nature. The aim of the study was to investigate trypanosomatids in captive primates coming from Municipal Zoological Park of Bauru by antibodies against the parasite and the amplification of the DNA of the parasite. For this, blood samples were collected from 39 primates. The molecular technique of Polymerase Chain Reaction (PCR) was applied in research for Leishmania braziliensis, Leishmania amazonensis and Leishmania infantum using species-specific primers directed to the region of kDNA parasites; all animals were negative by this technique. However, with the use of primers for the ribosomal region (ITS-1) specific to Trypanosomatidae family, 37 of 39 (94.9%) animals were positive for Trypanosoma spp. It was also conducted serological evidence by the Immunofluorescente Antibody Test (IFAT) for Leishmania braziliensis, Leishmania infantum e Leishmania amazonensis, revealing one (01) animal Erythrocebus patas (patas monkey) positive for Leishmania braziliensis, with titer 160. The results show that primates from Municipal Zoological Park of Bauru are susceptible to infection by trypanosomatids, alerting the need for active search for sandfly vectors and triatomines in the grounds and surrounding areas, as a preventive measure for the risk of infection for staff and visitors. / FAPESP: 2014/12187-0

Orientador: Paulo Maria Ferreira de Araujo / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-05T10:50:34Z (GMT). No. of bitstreams: 1 Felippe_PauloAnselmoNunes_M.pdf: 1330148 bytes, checksum: d967293882138043d69f9b910c8a220a (MD5) Previous issue date: 2005 / Resumo: Com o objetivo de verificar a existência de anticorpos reativos aos antígenos virais da dengue e da febre amarela no sangue de macacos prego (Cebus apella) cativos no Brasil, procedeu-se a coleta de sangue de 227 animais, oriundos de 17 cidades, (concentradas nas regiões sul e sudeste) de 4 estados do Brasil, no período compreendido entre os anos de 2000 e 2001. Para tanto realizamos o teste de inibição da hemaglutinação e também padronizamos um ELISA indireto utilizando um conjugado comercial. O teste de Inibição da Hemaglutinação não detectou nenhuma reatividade dos soros estudados frente aos antígenos da dengue (DENI, DEN II). Encontramos, no teste de ELISA, uma reatividade de cerca de 97 % das amostras ao DEN I; 68% ao DII, 74% ao DEN III e 81% a febre amarela (FA). Não observamos diferenças estatisticamente significativas de reatividade entre machos e fêmeas, porém a observamos entre animais adultos e velhos para DEN I e FA. Os soros previamente tratados com a extração pela acetona, utilizada no teste de inibição da hemaglutinação apresentaram uma significativa perda de reatividade quando testados de forma pareada com amostras não tratadas ao ELISA indireto. Os resultados encontrados não são compatíveis com a epidemiologia da dengue e da febre amarela no Brasil, uma vez que primatas oriundos de dois estados da federação onde sabidamente não havia a transmissão por ocasião da coleta apresentaram uma reatividade importante, o que sugere a existência de anticorpos naturais reagentes aos antígenos virais pesquisados e aponta no sentido de que estes possam ter alguma importância na resistência destes primatas a estas enfermidades / Abstract: With the objective to verify the existence of reactive antibodies to viral antigens to dengue and yellow fever in the blood of capuccin monkeys (Cebus apella) captive in Brazil, it was proceeded collection from blood of 227 deriving animals of 17 cities (concentrate in south and southeast regions) of 4 states of the federacy in the understood period enters the years of 2000 and 2001. For in such a way we carry through the test of Hemaglutination Inhibition and also we standardize an indirect ELISA using one commercial conjugate. The test of Hemaglutination Inhibition did not detect reactivity of the studied seruns front to studied antigens of the dengue (DENI, DEN II). We find, in the test of ELISA, a reactivity of about 97 % of the samples to DEN I; 68% to the DII, 74% to DEN III and 81% the yellow fever (FA). We do not observe statistical significant differences of reactivity between males and females, however we observe it between adult and old animals for DEN I and FA. The seruns previously treated with the extraction for acetone, used in the test of Hemaglutination Inhibition had presented a significant loss of reactivity when tested bodily with samples not treated to the indirect ELISA. The results are not compatible with the epidemiology of the dengue and of the yellow fever in Brazil, a time that deriving primates of two states of the federacy where knew did not have the transmission for occasion of the collection had presented an important reactivity, what it suggests the existence of reacting natural antibodies to viral antigens searched and points in the direction of that these can have some importance in the resistance of these primates to these diseases / Mestrado / Imunologia / Mestre em Genética e Biologia Molecular

Philosophiae Doctor - PhD / In this century most major medical advances have resulted in part from research on animals and non-human primates such as the African green monkey and therefore often serve as a critical link between basic research and human clinical application. Due to its close evolutionary relationship to humans, the African green monkey is known to be an excellent and most sought after models for studies of human cardiovascular disease (CVD). While the human genome project and some others related to model organisms are very well advanced or even complete, little sequence information has been acquired for the African green monkey. Given the importance of this species in biomedical research generally and CVD specifically, and the fundamental significance of sequence data, it is critical that this paucity of genome information concerning this specific animal model be addressed in order to better define the molecular basis and to further understand the mechanism of cholesterol metabolism in this species which will also contribute immensely to primatology. There is a growing interest in the role of genetic polymorphisms in predicting susceptibility to disease and responsiveness to drug interventions. Since plasma lipid abnormalities are risk factors for coronary atherosclerosis, determination of these plasma lipid concentrations, especially for genes involved in lipid transport and metabolism may be influenced by genetic variations. In this study, the African green monkey was used as a model to evaluate the effect of niacin on plasma lipids and reverse cholesterol transport by examine gene expression and the influence of several polymorphisms found in genes that are involved in cholesterol metabolism in humans. A survey of genetic variation spanning ten prioritised “candidate” genes was conducted, all of which are known to produce proteins that play key roles in the reverse cholesterol pathway (RCT), and in the homeostatic regulation of blood lipid profiles related to cardiovascular health and disease. everse transcription polymerase chain reaction (RT-PCR) was used to evaluate mRNA expression of those “candidate” genes. Twenty two coincident singlenucleotide polymorphisms (cSNPs), reported to play a vital role in RCT, were genotyped within these genes. This study’s findings implicate a subset of six of the twenty two genetic variants, spanning five “candidate” genes. To assess possible involvement of these prioritised “candidate” genes and their polymorphisms, biochemical analyses of known risk factors of coronary artery disease such as HDL-C and LDL-C were conducted. Eight healthy African green monkeys were entered in this study of which four were treated with niacin at an escalating dosage. Their mean lipid-lowering response following drug therapy was analysed, compared to those with the same genotype in a control group. Niacin treatment was associated with a considerable reduction in LDL-Cholesterol, up-regulation of HDL synthesis, and increase of apo A-1 levels. Gene expression had minimal effect on niacin treatment, except CYP7A1 which was down-regulated at the same time when considerable change in HDL-C, LDL-C and apoA-1 levels was observed. The presence of CYP7A1:Asn233Ser polymorphism may have played a critical role in metabolising niacin and influencing the up-regulation of HDL-C synthesis in the African green monkey. Although cholesterol lowering alone may explain the anti-atherosclerotic effect of niacin on HDL-C, in this study, gene expression data also shed some light in supporting the hypothesis that genetic variants may influence the expression of genes involved in RCT, which may also have played a role in the anti-atherosclerotic effect of the drug.

FAULT LINE examines the fragile humanity connected to the themes of sexuality, violence, addiction, family dynamics, and death. The book is not broken into sections; rather, as poems build upon one another to explore a narrative arc, FAULT LINE tracks a single speaker’s experience from girlhood to the verge of independent womanhood. The speaker employs formal structures such as the prose poem, sestina, and particularly the list poem to examine the fluidity of inner experience and also the culture at large while challenging the narrow definitions of femininity and masculinity. FAULT LINE works to not only address the question of blame but also the literal breaks in lines of poetry. By looking at a single speaker’s struggle, the book, like life, is both humorous and horrifying.

Testing is an important step in the software development process in order to increase the reliability of the software. There are a number of different methods available to test software that use different approaches to find errors, all with different requirements and possible results. In this thesis we have performed a series of tests on our own mobile application developed for the Android platform. The thesis starts with a theory section in which most of the important terms for software testing are described. Afterwards our own application and test cases are presented. The results of our tests along with our experiences are reviewed and compared to existing studies and literature in the field of testing. The test cases have helped us find a number of faults in our source code that we had not found before. We have discovered that automated testing for Android is a field where there are a lot of good tools, although these are not often used in practice. We believe the app development process could be improved greatly by regularly putting the software through automated testing systems.

Orientador: Lucilene Silva Ruiz e Resende / Resumo: Realizaram-se estudos imuno-hematológicos em 19 macacos sul- americanos, sendo 9 Sapajus sp. (macacos-prego) e 10 Alouatta sp. (macacos bugios). O Teste de Antiglobulina Direta (TAD) não revelou moléculas IgG de imunoglobulina e C3d do complemento, semelhantes às humanas, nas membranas dos eritrócitos dos animais. A Pesquisa de Anticorpos Irregulares (PAI) mostrou-se positiva nos soros de 18 animais adultos sendo negativa no soro do único filhote do estudo, um Sapajus sp. de 2 meses de idade. A Prova de Compatibilidade (PC) foi negativa dentro de cada gênero, revelando-se positiva entre os 2 gêneros de macacos e, destes, com humanos. O filhote de macaco-prego, entretanto, mostrou PC compatível, tanto com o gênero Alouatta sp., quanto com humanos. / Abstract: An immunohematological study was carried out in 19 South-American monkeys corresponding to 9 Sapajus sp. (Capuchin monkeys) and 10 Alouatta sp. (Howler monkeys). The Direct Antiglobulin Test (DAT) didn’t show humanlike molecules of IgG-type immunoglobulin and C3d complement fraction on the animal’s erythrocyte membranes. Serum Irregular Antibodies Screening (IAS) was found in 18 adult animals but not in the only studied cub, which was a 2 month-old Capuchin monkey. The Compatibility Test (CT) was negative among members of a same genre of animals being positive among Sapajus sp. and Alouatta sp., and among the monkey’s genres and humans. The Capuchin monkey cub, however, showed a negative CT with both, Howler monkeys and humans. / Mestre

Cortical injury, such as that following stroke, is one of the leading causes of long-term disabilities world-wide. While some neuroprotective agents given within hours of stroke can reduce damage, there are currently no neurorestorative therapeutics that can enhance long-term recovery. To address this, we tested Mesenchymal Stem Cell (MSC) derived Extracellular Vesicles (EVs) as a treatment for cortical injury in rhesus monkeys (Macaca mulatta). Monkeys treated with EVs 24 hours after injury and again at 14 days after injury recovered more completely and more rapidly than monkeys given a vehicle control. However, the cellular changes associated with enhanced recovery remained unknown. In this dissertation, it was hypothesized that EVs modulated cells within the brain to enhance recovery after cortical injury. To explore this hypothesis, three specific aims were tested. Aim 1: To determine the effects of EVs on microglial reactivity. Since EVs in this study were derived from MSCs, it was hypothesized that they would have an immunomodulatory effect. Using immunohistochemistry, image analyses, and 3-D reconstruction, we showed that microglia shifted from reactive, damaging phenotypes towards homeostatic, surveilling functions in EV-treated monkeys. These effects correlated with reduced time to recovery, suggesting that reduced microglial reactivity enhanced recovery. Aim 2: To assess the effects of EVs on myelination. Because MSCs have regenerative effects, it was hypothesized that these MSC-derived EVs would improve neurorestoration. Using immunohistochemistry, qRT-PCR, Spectral Confocal Reflectance microscopy, and ELISA, we assessed myelination after cortical injury with and without EV treatment. EVs limited oligodendrocyte damage and increased densities of mature oligodendrocytes to enhance myelin maintenance. These effects correlated with improved recovery, suggesting the importance of myelination in recovery after cortical injury. Aim 3: To assess the neuroprotective role of EVs on infarct volumes. While it was hypothesized that EVs would reduce the densities of inflammatory cells (astrocytes, macrophages/microglia, T-cells), hemosiderin accumulation, and infarct volume, we found that EVs did not alter these endpoints. Collectively, our results suggest that EVs modulated microglia and oligodendrocytes to promote neurorestoration. Overall, these findings demonstrate the therapeutic potential of EVs for neurorestoration after cortical injury.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第17819号 / 医博第3817号 / 新制||医||999(附属図書館) / 30634 / 京都大学大学院医学研究科医学専攻 / (主査)教授 金子 武嗣, 教授 大森 治紀, 教授 福山 秀直 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医科学) / 甲第18185号 / 医科博第50号 / 新制||医科||4(附属図書館) / 31043 / 京都大学大学院医学研究科医科学専攻 / (主査)教授 朝長 啓造, 教授 松岡 雅雄, 教授 小柳 義夫 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Rehabilitation-release is a form of species reintroduction where sick, injured, or rescued animals are rehabilitated before release back into the wild. Published research on rehabilitation releases of rehabilitant non-human primates is limited, and released troop mortality rates are generally high or difficult to determine. The objective of this study was to add to the limited scientific literature on primate rehabilitation and release by investigating factors affecting survival rates and welfare of a rehabilitant troop of vervet monkeys (Chlorocebus pygerythrus rufoviridis) released in Malawi in 2016, using pre-existing datasets from the Lilongwe Wildlife Trust. I hypothesized that 1) higher social rank, more complete forest strata use, close proximity to troop members, and frequent predator vigilance would be associated with greater survival, and 2) rank stability/ group cohesion will be strong post-release, activity budgets will show low levels of stress-related behaviors, and behavioral diversity will increase post-release, suggesting welfare improvements. The Lilongwe Wildlife Trust troop had a survival rate of 36%, which is comparable to other vervet releases. Using a combination of linear modeling, survival analysis, and preliminary social network analysis, I found that being a juvenile, being more highly ranked, and being in close proximity to others was significantly associated with lower risk of death – but these results were not consistent and should be considered with caution. Contrary to predictions, forest strata use did not differ greatly across individuals despite differences in survival. Interestingly, the troop’s mean hourly count of predator vigilance decreased post-release, but this did not influence individual survival. In support of my predictions, the troop’s dominance hierarchy appeared stable post-release, group cohesion was strong, and activity budgets showed low levels of stress-related behaviors. However, mean behavioral diversity across individuals decreased post-release, contrary to predictions. These findings suggest that vervet dominance hierarchy, age, and social proximity may influence post-release survival with higher ranking individuals, juveniles, and highly socially connected individuals more likely to survive. Juveniles may be more ecologically adaptable than adults and so better able to survive in a new habitat. Lower ranked individuals, as well as those with low social connectedness, may be more disconnected from the troop while traveling or foraging, placing them at a higher risk of predation but more research is needed to confirm this. Decreased behavioral diversity post-release may have been caused by an increase in foraging and troop movement and generalized behavior categorization may have limited the accuracy of behavioral diversity measurements. Future studies that wish to use behavioral diversity to assess welfare should use highly specific ethograms to capture unique behaviors. Release troops may also benefit from pre-release feeding regimes, such as platform feeders, that encourage more complete canopy use as well as more time at the release site prior to the start of the rainy season. Predator-awareness training is highly recommended to strengthen anti-predator behaviors, especially if the troop has any wild individuals. Finally, the Lilongwe Wildlife Trust’s extensive pre- and post-release monitoring provides vital insight into the troop’s social dynamics, behavioral repertories, and overall survival. Other rehabilitation centers should follow this strategy, since all newly monitored and reported releases will add valuable information to the development of the vervet monkey rehabilitation and release program.