Submitted by Edisangela Bastos (edisangela@ufpa.br) on 2014-01-27T17:38:58Z No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_ExpressaoGenicaViabilidade.pdf: 1020011 bytes, checksum: 6d7d985ee1bbd74238e1030e0549c524 (MD5) / Approved for entry into archive by Ana Rosa Silva(arosa@ufpa.br) on 2014-01-29T14:04:09Z (GMT) No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_ExpressaoGenicaViabilidade.pdf: 1020011 bytes, checksum: 6d7d985ee1bbd74238e1030e0549c524 (MD5) / Made available in DSpace on 2014-01-29T14:04:09Z (GMT). No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_ExpressaoGenicaViabilidade.pdf: 1020011 bytes, checksum: 6d7d985ee1bbd74238e1030e0549c524 (MD5) Previous issue date: 2011 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo desse trabalho foi analisar a ativação e o crescimento de folículos préantrais de Sapajus apella, submetidos a um sistema de cultivo in vitro em curto-prazo. Nesse sentido, dois experimentos foram realizados neste trabalho. Experimento I: exposição à fresco e à crioprotetores de biopsias de fragmentos do córtex ovariano. Ambos os fragmentos ovarianos foram submetidos à extração total de RNA e síntese de cDNA. Após a amplificação do cDNA por PCR em tempo-real (RT-PCR), os software GeNorm, bestkeeper e Normfinder foram usados para avaliar a estabilidade dos genes GAPDH (glyceraldehyde-2-phosphate dehydrogenase), HPRT1 (hypoxanthine phosphoribosyltransferase 1) e TBP (TATA-binding protein). Experimento II: o tecido do córtex ovariano de quatro fêmeas foi oletado e dividido em nove pedaços de 1 mm³. Um fragmento ovariano (grupo controle) foi imediatamente dividido em dois pedaços, os quais foram destinados para análise da viabilidade ou por RT-PCR. Os 8 fragmentos restantes foram individualmente cultivados in vitro em um meio constituído de TCM suplementado com 100 ng/mL EGF (T1), com adição de 10 μM de BME (T2), 100 ng/mL de BMP4 (T3), 25 IU de PMSG (T4), 10 μM de BME e 100 ng/mL de BMP4 (T5), 10 μM de BME, 25 IU de PMSG (T6), 100 ng/mL de BMP4, 25 IU de PMSG (T7) ou 10 μM de BME, 100 ng/mL de BMP4, 25 IU de PMSG (T8). Os resultados demonstraram que no tecido do córtex ovariano de S. apella, os genes HPRT1 e TBP foram os mais apropriados como genes de referência, podendo ser usados como parâmetro para normalizar dados em estudos futuros. Ao contrário, o GAPDH se apresentou como menos estável dos genes de referencia testados. Após o cultivo in vitro, todos os tratamentos alcançaram percentuais similares da viabilidade de folículos pré-antrais. O tecido ovariano cultivado na presença de GF+BME/BMP4/PMSG resultou no aumento da taxa de ativação e crescimento folicular, assim como no aumento da expressão de AMH, BMP15 e GDF9, genes conhecidos como indicadores específicos de desenvolvimento folicular. Dessa forma, o HPRT1 e TBP são os genes de referência mais estáveis, na exposição à crioprotetores, a fresco e no o cultivo de tecidos de córtex ovariano de S. apella. Os folículos pré-antrais são capazes de desenvolverem-se in vitro quando cultivados em meio suplementado com PMSG, BME e BMP4. A viabilidade folicular, entretanto, permaneceu independentemente do meio de cultivo in vitro e o uso de fatores de crescimento, como marcadores de desenvolvimento folicular, foi crucial para identificar o melhor meio de cultivo in vitro. / The aim of the present study was to investigate the stability of three reference genes in the ovarian tissue of capuchin monkeys (Sapajus apella) and to develop a short-term in vitro culture system for the activation and growth of preantral follicles from capuchin monkeys. To this end two experiment were conducted as follow. Experiment I: Fresh and cryoprotectant exposed ovarian biopsies were used. Both fresh and exposed ovarian tissues were subjected to total RNA extraction and synthesis of cDNA. After amplification of cDNA by real-time PCR, the GeNorm, Bestkeeper and Normfinder software were used to evaluate the stability of glyceraldehyde-2-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and TATA-binding protein (TBP). Experiment II: Ovarian tissue from four healthy mature females were collected and divided into nine ovarian cortical pieces of 1 mm³. One ovarian fragment (control) was immediately divided in two pieces, which were subjected to viability analysis or qRT-PCR. The remaining 8 fragments were individually cultured in vitro in a medium consisting of TCM supplemented with 100 ng/mL EGF (T1), either or not added with 10 μM BME (T2), 100 ng/mL BMP4 (T3), 25 IU PMSG (T4), 10 μM BME and 100 ng/mL BMP4 (T5), 10 μM BME, 25 IU PMSG (T6), 100 ng/mL BMP4, 25 IU PMSG (T7) or 10 μM BME, 100 ng/mL BMP4, 25 IU PMSG (T8). Results demonstrated that, in the ovarian tissue from capuchin monkeys, HPRT1 and TBP were the most suitable reference genes and thus could be used as parameters to normalize data in future studies. In contrast, GAPDH appeared as the least stable gene among the tested reference genes. After in vitro culture, all treatments resulted in similar percentages of viable preantral follicles. Ovarian tissue cultured in the presence of EGF + BME/BMP4/PMSG resulted in an increased rate of follicular activation and growth, as well as in the up regulation of AMH, BMP15 and GDF9, specific markers of follicular development. In conclusion, HPRT1 and TBP are the most stable reference genes in fresh and cryoprotectant exposed ovarian tissue from capuchin monkeys. Preantral follicles are able to develop in vitro when cultured in a medium supplemented with PMSG, BME and BMP4. Follicular viability, however, was maintained independently on the culture medium. The use of growth factors as markers of follicular development was crucial to identify the best culture medium.

Submitted by Edisangela Bastos (edisangela@ufpa.br) on 2014-01-29T20:29:36Z No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_ExpressaoGenicaViabilidade.pdf: 1672759 bytes, checksum: f731371d76d549439735b8deb95e258f (MD5) / Approved for entry into archive by Ana Rosa Silva(arosa@ufpa.br) on 2014-02-03T16:27:20Z (GMT) No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_ExpressaoGenicaViabilidade.pdf: 1672759 bytes, checksum: f731371d76d549439735b8deb95e258f (MD5) / Made available in DSpace on 2014-02-03T16:27:20Z (GMT). No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_ExpressaoGenicaViabilidade.pdf: 1672759 bytes, checksum: f731371d76d549439735b8deb95e258f (MD5) Previous issue date: 2012 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo do presente estudo foi desenvolver um protocolo de congelação para a preservação de folículos pré-antrais de Sapajus apella (macaco-prego). Para este fim, fragmentos ovarianos foram expostos à diferentes soluções crioprotetoras, adicionadas ou não por antioxidantes (selênio e trolox), congelados e cultivados in vitro por 24/horas. Análises morfológicas, ultraestruturais, de viabilidade e estresse oxidativo foram desenvolvidas. A coleta do material foi realizada no Centro Nacional de primatas (CENP) e nove macacos-prego maduras e saudáveis foram usadas. Biopsias ovarianas de 1 mm³ foram coletadas por laparoscopia exploratória. Os folículos coletados foram classificados de acordo com sua fase de desenvolvimento em primordial, primário ou secundário. A viabilidade folicular foi observada através da utilização de marcadores fluorescentes (Iodeto de propídeo e Hoechst) qRT-PCR foi usado para avaliar a expressão de hormônios e fatores de crescimento. O TEAC foi usado para mensurar o estresse oxidativo no tecido. Os resultados mostraram que a solução congelação contendo trolox não afetou a morfologia folicular e expressão gênica. A criopreservação resultou em elevadas taxas de viabilidade folicular quando o trolox estava presente na solução, porém a expressão de genes codificando BMP4 e KL foi negativamente afetada. Nossos achados mostraram um efeito favorável da adição do trolox à solução de congelação. Entretanto, a viabilidade folicular e expressão gênica foram afetados após cultivo in vitro. / The aim of the present study was to develop a freezing protocol for the preservation of preantral follicles from Sapajus apella (capuchin monkeys). To this end, ovarian fragments were exposed to different cryoprotectant solutions, added or not by antioxidants (selenium or trolox), frozen and in vitro cultived by 24 hours. Morphology, ultrastructure, viability, oxidative stress and mocelular analyses were performed. The collection site was in the Primates Nacional Center (CENP) and nine healthy mature female capuchin monkeys were used. Ovarian biopsies of 1 mm3 were collected by laparoscopy. The follicles were classified accordingly to their developmental phase in primordial, primary or secondary. Follicular viability scored using fluorescent markers (propidium iodide and Hoechst). qRT-PCR was used to evaluate the expression of hormones and growth factors. TEAC was used to measure oxidative stress in the tissue. In cryoprotectant solution containing trolox did not affected follicular morphology and gene expression. Cryopreservation resulted in higher rates of follicular viability when trolox was present in the solution, but expression of genes encoding BMP4 and KL was negatively affected. Our findings show a favorable effect of adding trolox to a cryopreservation solution. However, follicular viability and gene expression was affected after in vitro culture.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-07-03T11:58:02Z No. of bitstreams: 1 barbaramarunrocha.pdf: 2524124 bytes, checksum: 60e8676bc9d19488d92267243d731e7b (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-08-08T12:24:09Z (GMT) No. of bitstreams: 1 barbaramarunrocha.pdf: 2524124 bytes, checksum: 60e8676bc9d19488d92267243d731e7b (MD5) / Made available in DSpace on 2017-08-08T12:24:09Z (GMT). No. of bitstreams: 1 barbaramarunrocha.pdf: 2524124 bytes, checksum: 60e8676bc9d19488d92267243d731e7b (MD5) Previous issue date: 2014-02-28 / O presente trabalho teve como objetivo o estudo taxonômico de nematóides parasitos de três espécies de primatas Neotropicais, Callithrix penicillata (Geoffroy, 1812), Sapajus apella (Linnaeus, 1758) e Allouatta guariba (Humboldt, 1812), doados mortos à Universidade Federal de Juiz de Fora, pelo IBAMA/JF. Foram necropsiados 26 espécimes de primatas, sendo 22 C. penicillata, 2 S. apella e 2 A. guariba. Os helmintos foram coletados, quantificados e tratados de acordo com técnicas helmintológicas usuais. Para o estudo taxonômico, foram feitas imagens e desenhos, além da morfometria dos mesmos espécimes. Foram feitas análises de distribuição, intensidade, densidade e abundância das comunidades parasitárias. Foram coletados 10416 nematóides, pertencendo a quatro diferentes gêneros, com uma diferença significativa na quantidade de helmintos de C. penicillata (1391), S. apella (4590) e A. guariba (4435). Foram registrados Primasubulura jacchi em C. penicillata, Trypanoxyuris (Trypanoxyuris) minutus em A. guariba e S. apella, Parabronema (=Squamanema) bonnei em A. guariba, além do registro de uma nova espécie de Onchocercinae nos pulmões de um espécime de A. guariba, Courduriella sp..O registro de T. (T.) minutus em S. apella representa o primeiro relato deste parasito em hospedeiro deste gênero na América do Sul e com a coleta de Parabronema bonnei, foi possível fazer uma redescrição da espécie que não possui detalhamento de características morfológicas e morfométrica disponíveis na literatura. / The present work aimed to the taxonomic study of nematode parasites of three species of Neotropical primates, donated, already dead, the Federal University of Juiz de Fora, from IBAMA/JF. They are Callithrix penicillata (Geoffroy, 1812), Sapajus apella (Linnaeus, 1758) and Allouatta guariba (Humboldt, 1812). Twenty-six specimens of primates were necropsied, being twenty-two of C. penicillata, two S. apella and two A. guariba. All colected helminths were quantified and treated in accordance with usual helminthological techniques. For the taxonomic study, pictures and drawings were made, for morphometry of the same specimens. Distribution analysis, intensity, density and abundance of parasite communities were made. 10416 nematodes were collected belonging to four different genera, with a significant difference in the number of nematodes in C. penicillata (1391) and S. apella and A. guariba monkey (9025). Primasubulura jacchi were recorded in C. penicillata, Trypanoxyuris (Trypanoxyuris) minutus in A. guariba and S. apella e Parabronema (= Squamanema) bonnei in A. guariba, other than registration of a new species of Onchocercinae, lungs of a specimen A.guariba monkey, named Courduriella sp. The record of T. (T.) minutus in S. apella is the first report of this parasite in this host genus in South America and the collecting Parabronema bonnei was possible to make a redescription of the species that lacks detailed morphological and morphometric characteristics available in the literature.

This thesis examines the relationship between individual differences in learning, personality, and social success in two groups of brown capuchin monkeys (Sapajus sp.) housed at the “Living Links Centre for Human Evolution” at Edinburgh Zoo, UK. Being able to learn quickly and efficiently likely helps primates achieve social success (defined here in terms of centrality within a social network), such as acquiring knowledge of others or learning social skills. Therefore, individuals that are better at learning were predicted to have greater social success than other group members. This prediction, however, contrasts with hypotheses generated from two other disciplines at the individual level: 1) the study of behavioural innovation, and 2) the study of individual differences, i.e. “personality”. In terms of behavioural innovation, better learners should have less social success than other group members because they are expected to rely more on problem-solving, rather than physical combativeness or status, to gain access to socioecological resources. In terms of personality, learning should have little or no direct relationship with social success because other individual differences, like sociability and fearfulness, should mediate primates’ social decision making. This thesis investigates each of these hypotheses. Personality was assessed in 127 capuchins from 7 international sites using the Hominoid Personality Questionnaire, and then validated at Living Links (LL) using behavioural codings; this was the first-ever description of personality structure in brown capuchins. Brown capuchins have five personality dimensions: Assertiveness, Openness, Sociability, Neuroticism, and Attentiveness. Ratings were consistent across observers, and predicted relevant behaviours among the LL capuchins over a year later (e.g. scores on Sociability predicted time spent in close proximity to others). “Social success” in the LL capuchins was assessed in terms of centrality in spatial proximity networks. Individual scores on social network centrality were significantly correlated with scores derived from a Principal Components Analysis of eight affiliative and agonistic behaviours among the LL capuchins, indicating that spatial proximity is a reliable measure of the quality of subjects’ social embeddedness within their groups. Social rank and two personality traits (Assertiveness and Sociability) were positively related to network centrality, while another personality trait (Neuroticism) was negatively related to centrality. Sociability was a significant predictor of network centrality even after controlling for social rank and the other personality traits, highlighting the importance of this personality trait in shaping the social success of capuchins beyond that of basic social rules (e.g. kinship, sex, and rank). Individual learning was assessed in the LL capuchins by administering two operant tasks to subjects under conditions of free choice participation. In Task 1, thirteen monkeys participated, and eight individuals met learning criteria (i.e. >80% trials correct over 3 consecutive sessions). In Task 2, fifteen monkeys participated, and five individuals met learning criteria; the monkeys that learned this second task were also among those individuals that learned Task 1. For monkeys that regularly participated in both tasks (i.e. >50% of sessions), their average performances (i.e. % trials correct) were significantly correlated with individual scores on Assertiveness, but not the other four personality traits, or individual differences in attention span during testing, the percent of sessions subjects participated during testing, the amount of scrounging events subjects directed towards others within their social group, or the percent of observation time subjects spent feeding within their main indoor/outdoor enclosures. In terms of social success, relatively better learners had lower social rank and network centrality compared to relatively poor learners. Also, compared to poorer learners, better learners were generally less likely to direct affiliative acts (e.g. grooming, food sharing, coalitionary support) to other group members. Controlling for Assertiveness (i.e. the only variable related to individual differences in subjects’ average learning performance), individual differences in learning performance were no longer significantly related to social rank, network centrality, or the amount of affiliative acts subjects initiated with others. Collectively, such findings contrast the hypothesis that better learners should (concurrently) be more socially successful than poorer learners, and instead are more reflective of hypotheses pertaining to behavioural innovation and/or the study of individual differences. Social rank and certain traits of personality (Assertiveness, Openness, Neuroticism, and Sociability) appear to interact with capuchins’ patterns of social interaction, and one personality trait (Assertiveness) may mediate how individual differences in learning are associated with differences in social success.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-05-15T11:45:34Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_CriopreservacaoSemenMacaco.PDF: 1133952 bytes, checksum: fac97a88b90276c4785ddf98ac2b835c (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-05-17T12:51:30Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_CriopreservacaoSemenMacaco.PDF: 1133952 bytes, checksum: fac97a88b90276c4785ddf98ac2b835c (MD5) / Made available in DSpace on 2017-05-17T12:51:30Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_CriopreservacaoSemenMacaco.PDF: 1133952 bytes, checksum: fac97a88b90276c4785ddf98ac2b835c (MD5) Previous issue date: 2015-02-24 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / O objetivo principal do presente estudo foi estabelecer um protocolo eficiente de criopreservação seminal em Sapajus apella para a manutenção da viabilidade espermática. Para isso, foi comparado o desempenho dos diluidores TES-TRIS, ACIN e ACP-118® para escolher do melhor diluidor durante a dissolução do coágulo seminal e resfriamento, para posteriormente ser estabelecido a melhor concentração de glicerol (3; 5 e 7%) a ser adicionado no diluidor, bem como avaliar a suplementação do antioxidante catalase (10 µg e 50 µg) no meio de congelação seminal, no intuito de melhorar os parâmetros espermáticos. Foram utilizados seis machos adultos de S. apella oriundos do Centro Nacional de Primatas. As coletas do sêmen foram realizadas por eletroejaculação com probe retal, após a contenção química dos animais com Cloridrato de Ketamina e Cloridrato de Xilazina. Na primeira fase do projeto, o sêmen obtido foi diluído em TES-TRIS, ACIN e ACP®-118, mantido em banho-maria a 37 °C. Após dissolução do coágulo, o sêmen foi resfriado em geladeira a 4°C por 90 minutos, e avaliadas após 28h quanto a motilidade, o vigor, e o percentual de integridade de membrana antes e após o resfriamento. ACP®-118 foi o melhor diluidor para preservar a motilidade dos espermatozoides e integridade de membrana plasmática após 28 horas de incubação. A partir desse resultado, foi realizada a criopreservação avaliando diferentes concentrações de glicerol (3, 5 e 7%). Os espermatozoides criopreservados a 3% de glicerol apresentaram melhores resultados. Na segunda fase do projeto, foi realizada a criopreservação do sêmen de S. apella em diluidor ACP®-118 suplementado ou não com antioxidante catalase (10µg/mL e 50µg/mL). Todos os tratamentos foram eficazes na manutenção de parâmetros espermáticos, sendo possível a recuperação da motilidade pós-descongelamento. O tratamento catalase 50 foi o melhor de manutenção do vigor durante após resfriamento, e na integridade de membrana plasmática após a descongelação espermática. Em conclusão, ACP-118® pode ser usado de forma eficiente como diluidor para a criopreservação de sêmen S. apella adicionado de 3% de glicerol, além de que a adição do antioxidante catalase mostrou efeito benéfico durante esse processo. / The main objective of the present study was to establish an efficient semen cryopreservation protocol in Sapajus apella for maintaining sperm viability. For this, we compared the performance of TES-TRIS, CWS and ACP-118® extenders during the seminal coagulum dissolution and cooling. Then, in order to improve the sperm parameters, we also determined the glycerol concentration (3; 5 and 7%) as well as to evaluate the effect of catalase antioxidant (10 µg and 50 µg). Therefore, six adult males of S. apella from the National Primate Center were used. Semen was collected by electro-ejaculation with rectal probe after chemical restraint of animals with ketamine and xylazine hydrochloride. In the first phase of the project, the semen obtained was diluted in TES- TRIS, CWS and ACP®-118 extenders, kept in a water bath at 37 °C. After coagulum dissolution, the semen was cooled in the refrigerator at 4 °C for 90 minutes and evaluated after 28h for motility, vigor and plasma membrane integrity percentage before and after the cooling. ACP®-118 was the best extender to preserve the motility sperm and plasma membrane integrity after 28 hours of incubation. Based on these results, cryopreservation was performed by evaluation of different concentrations of glycerol (3, 5, and 7%). The cryopreserved sperm to 3% glycerol had better results. In the second project phase was performed S. apella semen cryopreservation in ACP®-118 extender supplemented or not with catalase antioxidant (10 µg/mL and 50 µg /mL). All treatments were effective in maintaining sperm parameters and was possible to recovery the post-thaw motility. The catalase 50 treatment was the best for maintenance of the vigor after cooling and plasma membrane integrity in post-thaw sperm. So, we concluded that ACP-118® extender can be efficiently used for S. apella semen cryopreservation added 3% glycerol, moreover, the addition of the catalase antioxidant showed beneficial effect during this process.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-03-27T14:24:45Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoSeriadaPerfil.pdf: 1895325 bytes, checksum: a44c51bf5d6c8db53d565476c04edbf1 (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-04-10T16:32:36Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoSeriadaPerfil.pdf: 1895325 bytes, checksum: a44c51bf5d6c8db53d565476c04edbf1 (MD5) / Made available in DSpace on 2017-04-10T16:32:36Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoSeriadaPerfil.pdf: 1895325 bytes, checksum: a44c51bf5d6c8db53d565476c04edbf1 (MD5) Previous issue date: 2017-02-23 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / Estudo de um sistema de veiculação de quimioterápicos, denominado LDE, com composição lipídica semelhantes com as lipoproteínas de baixa densidade naturais do corpo. As LDE apresentam vantagens sobre as formas químicas comerciais, pois são capazes de se concentrar nos tecidos neoplásicos após injeção na corrente circulatória, podendo assim direcionar-se aos tumores. As LDE podem ser utilizadas como “portadoras” do quimioterápico paclitaxel (PTX) para possível redução da toxicidade e aumento da sua ação terapêutica. A utilização de primatas não humanos como modelos experimentais in vivo é de grande importância em trabalhos de aplicação à saúde humana, devido as suas semelhanças anatômicas, bioquímicas e filogenéticas com os primatas humanos, gerando resultados que podem ser interpretados de forma mais próxima e segura para os fenômenos em seres humanos. O projeto objetivou avaliar a toxicidade crônica das nanopartículas associadas com o quimioterápico Paclitaxel (LDE-PTX) em indivíduos da espécie Sapajus apella, a partir da determinação de parâmetros hematológicos e bioquímicos, e suas possíveis alterações. Durante a pesquisa foram utilizados 15 animais, divididos em grupos: Controle negativo (CN); Experimental (EXP1 e EXP2) onde os animais receberam a LDE-PTX por via intravenosa em duas doses diferentes de 175 mg⁄m2 e 250 mg⁄m2 respectivamente; e o Controle positivo (CP1 e CP2) onde os animais receberam por via intravenosa o fármaco na forma comercial nas mesmas doses utilizadas no grupo experimental, respectivamente. Os primatas foram acompanhados durante 6 ciclos de quimioterapia, com intervalo de 3 semanas. A análise hematológica e bioquímica foi realizada a cada ciclo através dos valores do eritrograma, leucograma, plaquetogrma, fosfatase alcalina, proteína total, albumina e globulina, bilirrubina total e frações, glicemia, amilase e lipase sérica. A análise dos eletrólitos sódio e potássio, foi realizada no soro dos animais nos dias das coletas de materiais. Os dados foram expressos em média ± desvio padrão e submetidos à análise de variância ANOVA, com pós-teste de Bonferroni com significância para p<0,05, através do BioEstat®5.3. Os resultados obtidos demonstraram vantagens da utilização de LDE-PTX, já que os teste hematológicos demonstram que houve uma menor toxicidade em todos os ciclos quimioterápicos e a não alteração da maioria dos parâmetros bioquímicos, demonstram que a toxicidade do fármaco testado associado a LDE apresentam menor efeito tóxico do que sua versão comercial. Conclui-se com as análises dos resultados que a toxicidade hematológica e bioquímica foi menor no tratamento com o PTX associado à LDE do que o tratamento do PTX na sua forma comercial. / Study of a chemotherapy delivery system, called LDE, with lipid composition similar to the natural low density lipoproteins of the body, denominated by the acronym LDL. LDEs have advantages over commercial chemical forms, since it is able to concentrate in the neoplastic tissues after injection into the circulatory chain, thus being able to target the tumors. LDE can be used as a "carrier" of paclitaxel (PTX) for possible reduction of toxicity and increase of its therapeutic action. The use of non-human primates as in vivo experimental models are of great importance in human health applications due to their anatomical, biochemical and phylogenetic similarities with human primates, generating results that can be interpreted more closely and safely to The phenomena in humans. The aim of the project was to evaluate the chronic toxicity of nanoparticles associated with to chemotherapy Paclitaxel (LDE-PTX) in individuals of the Sapajus apella species, based on the determination of hematological and biochemical parameters and their possible alterations. During the research 15 animals were used, divided into groups: Negative control (CN); Experimental (EXP1 and EXP2) where the animals received LDE-PTX intravenously two different doses of 175 mg / m2 and 250 mg / m2 respectively; and positive control (CP1 and CP2) where the animals intravenously received the drug in commercial form at the same doses used in the experimental group, respectively. Primates were accompanied for 6 cycles of chemotherapy, with interval of 3 weeks. Hematological and biochemical analysis was performed at each cycle through erythrogram and leukogram, alkaline phosphatase, total protein, albumin and globulin, total bilirubin and fractions, glycemia, amylase and serum lipase. E of the sodium and potassium eletrolytes were carried out in the serum of the animals during the collection days. Data were expressed as mean ± standard deviation and submitted to analysis of variance ANOVA, with Bonferroni post-test with significance for p <0.05, through BioEstat®5.3. The obtained results demonstrated advantages of the use of LDE-PTX, since the hematological tests demonstrate that there was a lower toxicity in all the chemotherapeutic cycles and the non-alteration of the majority of the biochemical parameters, demonstrate that the toxicity of the tested drug associated to LDE present smaller Effect toxic than its commercial version. It was concluded from the analysis of the results that hematological and biochemical toxicity was lower in treatment with PTX associated with LDE than treatment of PTX in its commercial form.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-05-15T17:31:45Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_ExpressaoGenicaViabilidade.PDF: 2527584 bytes, checksum: a92c30bb13a081c6f66b18e729cf04d0 (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-05-19T14:44:20Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_ExpressaoGenicaViabilidade.PDF: 2527584 bytes, checksum: a92c30bb13a081c6f66b18e729cf04d0 (MD5) / Made available in DSpace on 2017-05-19T14:44:20Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_ExpressaoGenicaViabilidade.PDF: 2527584 bytes, checksum: a92c30bb13a081c6f66b18e729cf04d0 (MD5) Previous issue date: 2012-05-25 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A vitrificação é uma biotécnica que tem se mostrando cada vez mais promissora em várias espécies, dentre elas ruminantes domésticos (como ovelhas, cabras e vacas), e em primatas não humanos (PNH) (como o cynomologus e rhesus), e consiste na redução ultra-rápida da temperatura mediante a presença de altas concentrações de agentes crioprotetores (ACP’s) em nitrogenio liquido. Desta forma o objetivo deste estudo foi o desenvolvimento de uma metodologia de criopreservação por vitrificação de Folículos Ovarianos Pré-antrais (FOPA’s) em Sapajus apella. Para tanto, foram utilizadas femêas (n=9) adultas de Sapajus apella pertencentes ao plantel do Centro Nacional de Primatas (CENP). Foram realizadas coletas de córtex de tecido ovariano por meio de laparotomia exploratória de modo a não inviabilizar o animal reprodutivamente. Os fragmentos expostos a 8 tratamentos diferentes: Etilenoglicol (EG) 40% + Sacarose (Sac) 0,5 M dissolvidos em TCM-199 adicionados ou não de Selenio (2,5, 5 ou 10 ng/ml) ou Trolox (25, 50 ou 100 mM). Após a exposição aos agentes crioprotetores foi analisada a viabilidade folicular antes e após a vitrificação, a partir da morfologia folicular, expressão dos genes Hsp70, Erp29, Erp60 e Sod1 através da análise por Reação em Cadeia da Polimerase (PCR), além de realizar a dosagem de estresse oxidativo atravéz da TEAC (do inglês Total Equivalent Antioxidant Activity). As análises mostraram que a vitrificação permitiu a manutenção da viabilidade folícular mediante a exposição previa a elevadas concentrações de ACP’s, principalmente quando suplementados com trolox a 50 μM (que resultou em elevadas taxas de sobrevivência folicular) e aumento na expressão da enzima antioxidante Sod1. Enquanto na ausência do mesmo foi observado o aumento nas taxas de folículos degenerados e com vacuolização citoplasmática, além da redução na expressão da enzima antioxidante Sod1 e elevação da expressão da chaperona Erp29. / Vitrification is a biotech that has been increasingly showing promise in several species, among them domestic ruminants (such as sheep, goats and cows) and in nonhuman primates (NHP) (as cynomologus and rhesus), and consists in reducing ultra-rapid temperature by the presence of high concentrations of cryoprotective agents (PCA's) in liquid nitrogen. Thus the aim of this study was to develop a methodology for cryopreservation by vitrification of preantral follicles (PF's) in Sapajus apella. For this purpose, we used females (n = 9) adult Sapajus apella squad belonging to the National Primate Center (CENP). Samples were taken from the ovarian cortex by laparotomy so as not to destabilize the animals reproducibly. The fragments exposed to 8 different treatments: Ethylene glycol (EG) + 40% Sucrose (Sac) dissolved in 0.5 M TCM-199, added to Selenium (2,5, 5 or 10 ng / ml) or Trolox (25, 50 or 100 mM). Following exposure to cryoprotective agents was analyzed follicular viability before and after vitrification, from the follicular morphology, expression of Hsp70 genes, Erp29, Erp60 SOD1 and through the analysis by Polymerase Chain Reaction (PCR), in addition to performing measurement of oxidative stress thru the TEAC (English Total Equivalent Antioxidant Activity). The analyzes showed that vitrification allowed the maintenance of follicular viability by previous exposure to concentrations of ACP's alevadas, especially when supplemented with 50 mM trolox (which resulted in high follicular survival rates) and increased expression of the antioxidant enzyme SOD1. While in the absence of it was observed an increase in rates of follicles degenerate and vacuolated, and reduced expression of the antioxidant enzyme SOD1 and increased expression of chaperone Erp29.