Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-06-22T19:22:17Z No. of bitstreams: 1 TES_DANIEL_RODRIGO_MARINOWIC_COMPLETO.pdf: 16202491 bytes, checksum: 5bb7aaf0de3a700a163c7437ecf2adef (MD5) / Made available in DSpace on 2016-06-22T19:22:17Z (GMT). No. of bitstreams: 1 TES_DANIEL_RODRIGO_MARINOWIC_COMPLETO.pdf: 16202491 bytes, checksum: 5bb7aaf0de3a700a163c7437ecf2adef (MD5) Previous issue date: 2016-03-03 / Changes in the cerebral cortex development are presented as a group of distinct defects with a not well-defined pathogenesis. The focal cortical dysplasia (FCD) is one of the most frequent form of cortical development malformation, that encompasses multiple types of changes both in the cortical architecture and cytological abnormalities. It is an underlying pathology of a significant proportion of partial epilepsy refractory to drug treatment. Especially the limited number of cases and the lack of suitable experimental models rarely document the mechanisms involved with the genesis of FCD. The scarse predictive preclinical models that can be used to study the pathophysiology and to translate the therapeutic discovery from animal models to human use, strengthens the need to study the brain development from cells originated from patients that harbor these central nervous systems diseases. The generation of iPSC cells and the differentiation into specific cells and tissues will provide important testing and an unique ability to study the development and progress of CNS diseases. The objective of this study was to establish a cellular model of focal cortical dysplasia through the generation of induced pluripotent stem cells (iPSC) from fibroblasts derived from affected patients. Human fibroblasts were obtained from skin biopsies from two patients and cultured up to the fifth passage. Immunofluorescence analysis of AKT/mTOR signaling pathways was performed in the dysplastic brain tissue. iPSC cells were generated from fibroblasts through transfection with a viral vector containing the genes OCT4, KLF4, SOX2, and C-MYC and characterized by immunohistochemistry. Cell migration co-cultured with fibroblast and iPSC was investigated. Morphological and molecular characterization of neurodifferentiated iPSC were performed by immunohistochemistry and PI3K/ATK/mTOR signaling pathway analysis. Both patients were diagnosed with FCD type IIb. The AKT and mTOR phosphorylated and non-phosphorylated was higher in brain sections from patient 01. iPSC clones from patients and controls were generated from skin fibroblasts. Both iPSC from patients and controls showed polarization and morphology similar to nerve cells. In the cell migration assay, fibroblasts derived from FCD migrate with greater intensity within 24 hours (p<0,0001) and 48 hours (p<0,001), and iPSC cells did not present difference in cell migration. During neurodifferentiation, iPSC cells from patients with FCD showed lower values of 4EBP-1, ?-catenin, CIAP-1, CIAP-2 and PI3K, and higher values of MCL 1 gene expression. Changes in cell migration in adult tissue, uncontrolled cell proliferation, cell adhesion protein deficiency, and alterations in the expression of genes responsible for apoptosis and PI3K pathway that are implicated with an complete and well successful CNS formation. Alterations in some of these were detected in cells and iPSC from patients with FCD and may be related to the ethiopathology of the disease. / As altera??es do desenvolvimento do c?rtex cerebral apresentam-se como um grupo de malforma??es com uma distin??o e patog?nse ainda n?o bem definidas. A displasia cortical focal (DCF) ? uma das formas mais frequentes de malforma??es do desenvolvimento cortical, sendo a patologia subjacente a uma parcela significativa de epilepsias parciais refrat?rias ao tratamento medicamentoso. A displasia cortical focal engloba m?ltiplos tipos de altera??es tanto na arquitetura cortical quanto em anormalidades citol?gicas. Palmini e colaboradores classificaram as displasias corticais focais de acordo com observa??es na subst?ncia branca e na arquitetura da camada cortical. Os mecanismos envolvidos na g?nese da DCF s?o pouco investigados, principalmente pelo n?mero limitado de casos e a falta de modelos experimentais adequados e suas causas provavelmente est?o relacionadas a muta??es som?ticas. A falta de modelos pr?-cl?nicos preditivos que possam ser utilizados no estudo da fisiopatologia e o hist?rico enfraquecido quando se trata em traduzir a descoberta terap?utica de modelos animais para o uso humano, fortalece a necessidade de estudar o desenvolvimento cerebral a partir de c?lulas originadas do pr?prio paciente. A gera??o de c?lulas iPSC e diferencia??o tecidual espec?fica de c?lulas de pacientes acometidos por doen?as neurol?gicas relevantes possui um valor inestim?vel para a realiza??o de testes al?m de fornecerem uma capacidade adicional e ?nica para estudar o desenvolvimento inicial e a progress?o das patologias associadas ao SNC. O objetivo do presente trabalho ? estabelecer um modelo celular de Displasia Cortical Focal atrav?s da gera??o de c?lulas-tronco pluripotentes induzidas (iPSC) a partir de fibroblastos de pacientes afetados. Fibroblastos humanos foram obtidos de bi?psias de pele de dois pacientes com DCF e foram cultivados at? a quinta passagem. Foi realizada an?lise imunopatol?gica e das vias de sinaliza??o AKT/mTOR no tecido cerebral displ?sico. C?lulas iPSC foram geradas a partir dos fibroblastos atrav?s da exposi??o a vetores virais contendo os genes OCT4, KLF4, SOX2, e C-MYC e caracterizadas por imunohistoqu?mica. Foi realizado ensaio de migra??o celular dos fibroblastos (24h, 48h e 72h) e das iPSC (3 dias e 7 dias). As c?lulas iPSC foram neurodiferenciadas e analisadas nos per?odos de 14 dias, 22 dias e 35 dias. Nesses per?odos, foram realizadas an?lises morfol?gicas, imunohistoqu?mica (polariza??o) e moleculares (genes relacionados a via PI3K/ATK/mTOR). Ambos os pacientes foram diagnosticados com DCF tipo IIb. O paciente 01 apresentou valores maiores que o paciente 02 em rela??o ? ?rea marcada das vias AKT e mTOR, tanto na via fosforilada como n?o fosforilada no tecido cerebral, com diferen?as estatisticamente significativas. Clones iPSC dos pacientes e controles foram gerados e caracterizados a partir dos fibroblastos de pele. Tanto as iPSC dos pacientes quanto do controle apresentaram morfologia e polariza??o semelhante a c?lulas nervosas ap?s protocolo de neurodiferencia??o. No ensaio de migra??o celular, fibroblastos com DCF migraram com mais intensidade em 24 horas (p<0,0001) e 48 horas (p<0,001). As c?lulas iPSC n?o apresentaram diferen?a no potencial de migra??o celular nos per?odos analisados. Durante o protocolo de neurodiferencia??o, as c?lulas iPSC dos pacientes com DCF apresentaram valores menores de express?o dos genes 4EBP-1, ?-Catenina, CIAP-1, CIAP-2 e PI3K, e valores mais elevados da express?o de MCL 1. Altera??es na migra??o celular no tecido adulto e em processos como de prolifera??o celular acentuada, defici?ncia de prote?na de ades?o celular, altera??o na express?o de genes respons?veis pelo controle de apoptose e altera??o na via PI3K respons?vel no sistema nervoso central pela sobreviv?ncia celular, controle de apoptose, migra??o neuronal, desenvolvimento morfol?gico dos neur?nios e forma??o das neurotransmiss?es, puderam ser evidenciadas nas c?lulas dos pacientes com DCF em rela??o aos pacientes controle e podem estar relacionadas com a forma??o do c?rebro com displasia.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-08-03T18:08:58Z No. of bitstreams: 1 DIS_RAFAEL_LISBOA_LOPES_COMPLETO.pdf: 9085477 bytes, checksum: b4223f63e2f669559a5830cd87644a78 (MD5) / Made available in DSpace on 2016-08-03T18:08:58Z (GMT). No. of bitstreams: 1 DIS_RAFAEL_LISBOA_LOPES_COMPLETO.pdf: 9085477 bytes, checksum: b4223f63e2f669559a5830cd87644a78 (MD5) Previous issue date: 2016-03-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Hsp70 (Heat shock protein 70 kDa) is a chaperone protein which has intra- and extracellular functions. In the early studies, only intracellular functions were operated, as to aid in folding, prevent aggregation and protein refolding. In 80?s a study has shown that this protein can be exported into the extracellular environment exerting effects on other cells, such as the immune system. Our group are exploring the role of this protein in key immune response cells such as macrophages and dendritic cells. Present study noted that the Hsp70 of Mycobacterium tuberculosis can polarize macrophages to a M2 phenotype and this response is dependent on IL-10. Peritoneal and bone marrow derived macrophages were treated with DnaK (Hsp70 prokaryotic homolog) from M. tuberculosis for 24 hours. After this period there was an increase of M2 classic phenotype markers, such as FIZZ1, YM1, CD206, IL-10 and Arg1. On the other hand, there was a decrease in expression of IL-6, MCP-1, TNF-?, NO, MHC class II and CD86 (M1 markers), and these cells are able to promote tumor growth in allogeneic model. It was also observed with KO cells or blocking IL-10R that this modulation is dependent on IL-10. In addition, this cellular modulation is not restricted to macrophages; such protein acting on dendritic cells by modulating them with an immature phenotype, as has been shown in prior works of our group (Borges et al, 2010;. Motta et al., 2007 ). However, it is not yet known what specific region of the protein is essential for that purpose and whether it is restricted to that homologues, DnaK from M. tuberculosis. Therefore, another objective was to express and purify in the same system their counterparts Hspa1a (murine Hsp70) and HSPA1A (human Hsp70) and subsequently testing them in parallel in immunological experimental models. Work is currently in progress with regard to purification of Hspa1a and initial dendritic cells activation state tests were conducted. It is believed that whether there is an effect with DnaK homologues, this effect is less obvious, according to previous results. / A Hsp70 (Heat shock protein 70 kDa) ? uma prote?na de choque t?rmico de 70 kDa que possui fun??es intra e extracelulares. Nos estudos iniciais, somente as fun??es intracelulares eram exploradas, como o aux?lio no dobramento, impedir a agrega??o e o redobramento prot?ico H? d?cadas mostrou-se que essa prote?na pode ser exportada para o ambiente extracelular e exercer efeitos sobre outras c?lulas, como as do sistema imune. O nosso grupo v?m explorando o papel dessa prote?na em c?lulas chave da resposta imune, como macr?fagos e c?lulas dendr?ticas. O presente trabalho observou que a Hsp70 de Mycobacterium tuberculosis pode polarizar macr?fagos a um fen?tipo M2 e que essa resposta ? dependente de IL-10. Macr?fagos peritoneais e derivados de medula ?ssea foram tratados com DnaK (hom?logo procari?tico da Hsp70) de Mycobacterium tuberculosis por 24h. Ap?s esse per?odo observou-se o aumento de marcadores cl?ssicos de fen?tipo M2, como: FIZZ1, YM1, CD206, IL-10, Arg1. Por outro lado, houve uma diminui??o na express?o de IL-6, MCP-1, TNF-?, NO, MHC de classe II e CD86, al?m de essas c?lulas serem capazes de promover o crescimento tumoral em modelo alog?nico. Observou-se tamb?m, com c?lulas KO ou bloqueando o IL-10R que essa modula??o ? dependente de IL-10. Al?m disso, essa modula??o celular n?o ? restrita aos macr?fagos, podendo essa prote?na agir sobre c?lulas dendr?ticas, modulando-as a um fen?tipo imaturo, como j? mostrado em trabalho anteriores do grupo (1,2). Por?m, n?o se sabe ainda qual regi?o espec?fica da prote?na ? essencial para esse efeito e se ele ? restrito a esse hom?lo, DnaK de M. tuberculosis. Portanto, outro objetivo do trabalho foi expressar e purificar no mesmo sistema os seus hom?logos Hspa1a (Hsp70 murina) e HSPA1A (Hsp70 humana) e posteriormente, test?-las em paralelo em modelos experimentais imunol?gicos. O trabalho est? em andamento atualmente em rela??o a purifica??o da Hspa1a e os primeiros testes de estado de ativa??o de c?lulas dendr?ticas foram realizados. Acredita-se que se h? um efeito dos hom?logos de DnaK, esse efeito ? menos evidente, segundo resultados pr?vios.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-10-24T15:48:35Z No. of bitstreams: 1 DIS_CALINO_FERREIRA_PACHECO_NETO_COMPLETO.pdf: 463906 bytes, checksum: b8097cd7556014c152e145a0681d517a (MD5) / Made available in DSpace on 2016-10-24T15:48:36Z (GMT). No. of bitstreams: 1 DIS_CALINO_FERREIRA_PACHECO_NETO_COMPLETO.pdf: 463906 bytes, checksum: b8097cd7556014c152e145a0681d517a (MD5) Previous issue date: 2014-03-31 / Rhizoctonia solani is a pathogenic ubiquitous fungus in Solanum tuberosum cultures, causing severe losses in production. Plant resistance can be induced by molecules that promote natural plant defense system. In the present study, the efficiency of the inducers Acibenzolar-Smethyl (BION?), Ceratocystis fimbriata extract and Xanthomonas axonopodis extract against the pathogenic fungus Rhizoctonia solani in potato plants was evaluated. Plants treated with the inducer XTH were resistant to the pathogen at 21 dpi. The biotic inducer XTH was more efficient than Bion? and CTS on promoting plant resistance. The highest activity of POX was observed at 9 dpi on the XTH treatment. Inoculation of R. solani caused a progressive increase in the PPO activity in XTH- and control-plants. Both inducers CTS and Bion? caused a reduction on PPO activity over time. Levels of phenolics compounds and flavonoids were similar among all treatments. Plants were more susceptible to R. solani when induced with CTS or Bion?. XTH did not show a negative effect on plant resistance. XTH presented a good potential as inducer, delaying disease progression and promoting plant resistance against R. solani. / Apesar da batata ser um dos alimentos mais consumidos do mundo, a sua produ??o apresenta s?rias dificuldades devido a doen?as causadas por fungos, bact?rias e v?rus, reduzindo a produtividade e aumentando os custos de produ??o. Assim, este trabalho teve por objetivo analisar os mecanismos envolvidos na defesa cruzada de Solanum tuberosum cv. ?gata, atrav?s a a??o de indutores bi?ticos no metabolismo de fenilpropan?ides. Plantas jovens de S. tuberosum mantidas em casa de vegeta??o foram aspergidas com indutores de defesa: extratos autoclavados de Xanthomonas axonopodis e de Ceratocystis fimbriata, al?m do indutor sint?tico Bion?. Ap?s cinco dias do tratamento, as plantas foram desafiadas com o fungo patog?nico Rhizoctonia solani, sendo analisados o progresso da doen?a e os par?metros bioqu?micos: compostos fen?licos totais, flavon?ides quercet?nicos e atividade das enzimas polifenoloxidase e peroxidases. As plantas tratadas com o indutor XTH foram resistentes ao pat?geno at? 21 dpi. O indutor bi?tico XTH foi mais eficiente que o Bion? e CTS na promo??o da resist?ncia vegetal. A atividade mais elevada da POX foi observada em 9 dpi com o tratamento XTH. A inocula??o de R. solani causou o aumento progressivo da atividade da PPO nas plantas dos tratamentos XTH e controle. Os indutores CTS e Bion? causaram a diminui??o da atividade de PPO ao longo do experimento. Os n?veis dos compostos fen?licos e dos flavonoides foram similares entre todos os tratamentos. As plantas foram mais suscet?veis ao pat?geno R. solani quando aspergidas previamente com CTS ou Bion?. O XTH apresenta um bom potencial de utiliza??o como indutor de resist?ncia, retardando o progresso da doen?a e promovendo a resist?ncia contra R. solani.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2017-03-17T15:08:38Z No. of bitstreams: 1 DIS_JOSE_CRISTIANO_MENGUE_MODEL_COMPLETO.pdf: 3495211 bytes, checksum: 700bfa91afac93bfa629684755fcc8ce (MD5) / Made available in DSpace on 2017-03-17T15:08:38Z (GMT). No. of bitstreams: 1 DIS_JOSE_CRISTIANO_MENGUE_MODEL_COMPLETO.pdf: 3495211 bytes, checksum: 700bfa91afac93bfa629684755fcc8ce (MD5) Previous issue date: 2017-01-24 / The surface passivation is an important step in solar cell manufacturing since it intends to fix the surface defects. In order to develop p-type crystalline silicon solar cells, solar grade, with surface passivation provided by titanium dioxide film, atmospheric pressure chemical vapor deposition (APCVD) and electron beam deposition (E-Beam) were carried out. For the films deposited by E-Beam, the thickness of TiO2, which has produced the most efficient solar cell, was of 80 nm. It was observed that how thicker the film, higher was the internal quantum efficiency (IQE) for short wavelengths, indicating a surface passivation that changes according to the thickness. The cells in which the film deposition on the front face was performed by APCVD were as efficient in to reduce the reflection as those with films deposited by E-Beam, although the first technique did not produce films with high homogeneity with regard to thickness. When the both techniques to deposit films on the back surface were compared, it was observed that the better results were obtained with APCVD films. Regarding the thickness of the films obtained by APCVD, it was not observed difference between the electrical characteristics of solar cells with thin and thick films. The most efficient solar cell produced in this work used TiO2 films obtained by chemical vapor deposition on both sides, reaching the efficiency of 15.6% and short-circuit current density of 34.9 mA/cm?. / A passiva??o ? uma importante etapa no processo de produ??o de c?lulas solares, pois visa corrigir os defeitos de superf?cie. Com o objetivo de desenvolver c?lulas solares de sil?cio cristalino tipo p, grau solar, com BSF (back surface field ? campo retrodifusor) seletivo e com passiva??o da superf?cie proporcionada por filme de di?xido de tit?nio, realizou-se deposi??o qu?mica em fase vapor a press?o atmosf?rica (APCVD) e por canh?o de el?trons (E-Beam). Para os filmes depositados por E-Beam, verificou-se que a espessura de TiO2 que produziu a c?lula solar mais eficiente foi de 80 nm. Observou-se que quanto maior a espessura do filme, mais elevada foi a efici?ncia qu?ntica interna (EQI) para comprimentos de onda curtos, indicando uma passiva??o de superf?cie vari?vel com a espessura. As c?lulas em que a deposi??o de filme na face frontal foi realizada via APCVD se mostraram t?o eficientes na redu??o da reflex?o quanto as que receberam filme por E-Beam, embora a primeira t?cnica n?o produziu filmes de elevada homogeneidade no que se refere a espessura. Ao comparar as duas t?cnicas utilizadas para deposi??o de filme na face posterior, verificou-se que os melhores resultados foram obtidos com filmes depositados por APCVD. No que se refere a espessura do filme posterior obtido por APCVD, n?o se observou diferen?a entre as caracter?sticas el?tricas de c?lulas solares com filmes finos e espessos. A c?lula solar mais eficiente produzida neste trabalho utilizou filmes de TiO2 obtidos por deposi??o qu?mica em fase vapor em ambas as faces, atingindo 15,6 % de efici?ncia e 34,9 mA/cm? de densidade de corrente de curto-circuito.

Submitted by PPG Medicina e Ci?ncias da Sa?de (medicina-pg@pucrs.br) on 2018-09-05T18:11:52Z No. of bitstreams: 1 ALINE_ZAPARTE.pdf: 2153343 bytes, checksum: 3c21541f64c5540c0571cbd6752b2141 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-09-11T11:13:44Z (GMT) No. of bitstreams: 1 ALINE_ZAPARTE.pdf: 2153343 bytes, checksum: 3c21541f64c5540c0571cbd6752b2141 (MD5) / Made available in DSpace on 2018-09-11T11:21:30Z (GMT). No. of bitstreams: 1 ALINE_ZAPARTE.pdf: 2153343 bytes, checksum: 3c21541f64c5540c0571cbd6752b2141 (MD5) Previous issue date: 2018-04-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Esophageal cancer, in general, is diagnosed at an advanced stage, which leads to impairment in the therapies employed, resulting in a high mortality rate. It is classified into two subtypes: adenocarcinoma and squamous cell carcinoma; both differ in histology, etiology and epidemiology. Purinergic signaling uses nucleotides and nucleosides in the extracellular medium as signaling molecules, and has been linked to different types of carcinomas. These molecules bind to G-protein coupled adenosine receptors, characterized as P1 (A1, A2A, A2B and A3), to P2X ionotropic receptors (P2X1-P2X7), and to P2Y receptors (P2Y 1, 2, 4, 6, 11, 12, 13 14), coupled to G protein. Given the evidence described in the literature and the lack of data correlating purinergic signaling with esophageal cancer, we analyzed the role of P2Y2 (P2Y2R) and P2Y12 (P2Y12R) receptors in the processes of proliferation, colony formation capacity, migration, adhesion, enzymatic activity of the ectonucleotidases and signaling pathways after the nucleotide stimulation. For this, we used the Kyse- 30 and Kyse-450 cells, representative of squamous cell carcinoma, and the OE-33 line, representative of adenocarcinoma. Also, we verified the expression of P2Y2R in biopsies of patients with squamous cell carcinoma and adenocarcinoma, compared to non-neoplastic tissues. We observed that the biopsy specimens express the P2Y2 receptor, but at different labeling intensities. The cell lines expressing P2Y2 and P2Y12R, have different responses to the stimulus with the nucleotides ADP, ATP and UTP, but the blockade of these receptors leads to a decrease in proliferation, polyclonal population formation, adhesion and migration. Regarding the pathways related to the action of P2Y2R, we verified the activation of ERK1 / 2 and Akt at different times after the stimulation with ATP and UTP. The data presented in this study demonstrate that the modulation of purinergic receptors P2Y2 and P2Y12 may become a promising tool for achieving efficacy in the treatment of esophageal cancer. / O c?ncer de es?fago, em geral, ? diagnosticado em est?gio avan?ado, o que leva a um preju?zo nas terapias empregadas, resultando em uma alta taxa de mortalidade. ? dividido em dois subtipos: adenocarcinoma e carcinoma de c?lulas escamosas; os dois subtipos diferem quanto a histologia, etiologia e epidemiologia. A sinaliza??o purin?rgica utiliza nucleot?deos e nucleos?deos no meio extracelular como mol?culas sinalizadoras e j? foi relacionada com diferentes tipos de carcinomas. Essas mol?culas ligam-se a receptores para adenosina acoplados a prote?na G, caracterizados como P1 (A1, A2A, A2B e A3), a receptores ionotr?picos P2X (P2X1- P2X7), e ainda a receptores P2Y (P2Y1, 2, 4, 6, 11, 12,13 14), acoplados a prote?na G. Diante das evid?ncias descritas na literatura e a falta de dados correlacionando a sinaliza??o purin?rgica com c?ncer de es?fago, analisamos o papel dos receptores P2Y2 (P2Y2R) e P2Y12 (P2Y12R) nos processos de prolifera??o, capacidade de forma??o de col?nias, migra??o, ades?o e atividade enzim?tica das ectonucleotidases e vias de sinaliza??o envolvidas, ap?s o est?mulo com nucleot?deos. Para isso, utilizamos as c?lulas Kyse-30 e Kyse-450, representativas de carcinoma de c?lulas escamosas, e a linhagem OE-33, representativa de adenocarcinoma. Tamb?m, verificamos a express?o do P2Y2R em bi?psias de pacientes com carcinoma de c?lulas escamosas e adenocarcinoma, comparadas com tecidos n?o neopl?sicos. Observamos que as amostras de bi?psia expressam o receptor P2Y2, por?m em diferentes intensidades de marca??o. As linhagens celulares expressam P2Y2 e P2Y12R, possuem diferentes respostas frente ao est?mulo com os nucleot?deos ADP, ATP e UTP, por?m o bloqueio desses receptores leva ? diminui??o da prolifera??o, forma??o de popula??o policlonal, ades?o e migra??o. Quanto ?s vias relacionadas ? a??o do P2Y2R, verificamos a ativa??o de ERK1/2 e Akt em diferentes tempos ap?s o est?mulo com ATP e UTP. Os dados apresentados neste estudo demonstram que a modula??o de receptores purin?rgicos P2Y2 e P2Y12, pode tornar-se uma ferramenta promissora para alcan?ar efic?cia no tratamento do c?ncer de es?fago.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-11-22T20:18:42Z No. of bitstreams: 1 CleonilsonMafraBarbosa_TESE.pdf: 4055728 bytes, checksum: 4de55338df2c07716d2c555f19dcc09a (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-11-22T23:43:24Z (GMT) No. of bitstreams: 1 CleonilsonMafraBarbosa_TESE.pdf: 4055728 bytes, checksum: 4de55338df2c07716d2c555f19dcc09a (MD5) / Made available in DSpace on 2017-11-22T23:43:24Z (GMT). No. of bitstreams: 1 CleonilsonMafraBarbosa_TESE.pdf: 4055728 bytes, checksum: 4de55338df2c07716d2c555f19dcc09a (MD5) Previous issue date: 2017-07-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Esse trabalho prop?s uma rota alternativa de s?ntese de catalisadores para rea??es de oxida??o do metanol e etanol a serem aplicados em c?lulas a combust?vel, sendo estes a base de ?xidos e precursores mistos de ni?bio e t?ntalo puros e dopados com cobalto, cobre e n?quel, obtidos a partir do mineral de base, a columbita/tantalita. Inicialmente, foi efetivado um planejamento experimental da purifica??o total deste min?rio, na sequ?ncia, foi realizada a dopagem usando um percentual de 10%, em massa. Os tratamentos t?rmicos foram realizados em tr?s diferentes temperaturas (110, 400 e 600 ?C). Na sequ?ncia, foi efetivada a s?ntese do precursor oxalato, que foi utilizado, por conseguinte, na s?ntese dos precursores dopados onde estes passaram pelos mesmos procedimentos dos ?xidos. O min?rio puro e tratado foi caracterizado por fluoresc?ncia de raios X (FRX) e difra??o de raios X (DRX) que mostraram a sua total purifica??o. O precursor foi avaliado atrav?s das an?lises de difra??o de raios X (DRX), espectroscopia de infravermelho (IV), an?lises t?rmicas (TG/DTG/DSC e DTA) e a microscopia eletr?nica de varredura (MEV); apresentando part?culas inferiores a 0,2 micrometros, um alto valor de perda de massa (76,6 %) e uma estrutura porosa de formas irregulares. Os catalisadores puros e dopados foram submetidos ?s an?lises por difra??o de raios X (DRX), espectroscopia de fotoel?trons excitados por raios X (XPS), microscopia eletr?nica de varredura (MEV) e microscopia eletr?nica de transmiss?o (MET), que apresentaram fortes ind?cios de propriedades catal?ticas para a oxida??o devido a sua r?pida redu??o. Na caracteriza??o el?trica, estes catalisadores foram avaliados pela t?cnica da voltametria de pulso diferencial (DPV) atrav?s de sensores em rea??o para a oxida??o dos ?lcoois. As an?lises demonstraram que estes materiais s?o ?timos condutores, porque aumentaram a passagem de corrente el?trica do eletrodo de trabalho em at? duas ordens de grandeza superior ao eletrodo de ouro. Os melhores desempenhos para as rea??es foram observados principalmente com o dopante de cobre, seguido por o n?quel, o puro e depois o de cobalto, considerando ainda que os materiais obtidos possuem caracter?sticas apropriadas para aplica??o em eletrodos de c?lulas a combust?vel. / This paper proposes an alternative route for producing catalysts for methanol and ethanol oxidation reactions to be applied on fuel cells. Those catalysts are based on oxides and precursors of mixed niobium and tantalum materials in their pure and doped (with Co, Cu or Ni) forms. These materials are obtained from columbite/tantalite, which is the base mineral for Nb, Ta. At first, an experimental planning for the complete purification of the mineral was performed. After purification, 10%wt. doping with each of the metals, and thermal treatment at three different temperatures (110,400 and 600?C) was carried out. Un-doped purified oxides were then subject to complexation process followed by metal addition (doping) and thermal treatment. Purified and thermally treated mineral was characterized by X-Ray Fluorescence (XRF) and X-Ray Diffraction and complete purification was attained. Complex precursors were evaluated on the basis of XRD, Infra-Red Spectroscopy (IR), thermal behavior (TG/DSC and DTA) and morphology (Scanning Electron Microscopy) and presented particle sizes under 0.2 ?m, elevated weight loss (76.6%) and a porous structure of irregular shape. Pure and doped catalysts were characterized on XRD, XPS (X-Ray Excited Photon Spectroscopy), SEM and TEM (Transmission Electron Microscopy) basis, presenting indication of catalytic properties interesting for oxidation reactions, such as quick surface reduction. Electrical evaluation of the catalysts was performed according to Differentia Pulse Voltammetry (DPV) with the use of micro sensors during alcohol oxidation reactions. These analyses indicated the excellent conducting characteristics of the materials as electric current flow was increased in two orders of magnitudes in comparison to gold electrodes. The best catalytic behaviors were observed when dopping was performed with copper, followed by nickel, without and cobalt dopant addition. Therefore, the synthesized materials presented characteristics that indicate their suitability for use as fuel cells electrodes.

Made available in DSpace on 2014-12-17T14:05:23Z (GMT). No. of bitstreams: 1 JoanaCMT_TESE.pdf: 5399741 bytes, checksum: 9af0d84203a9976314d4eb5bba7e4067 (MD5) Previous issue date: 2013-03-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Human mesenchymal stem cells (MSC) are powerful sources for cell therapy in regenerative medicine. The long time cultivation can result in replicative senescence or can be related to the emergence of chromosomal alterations responsible for the acquisition of tumorigenesis features in vitro. In this study, for the first time, the expression profile of MSC with a paracentric chromosomal inversion (MSC/inv) was compared to normal karyotype (MSC/n) in early and late passages. Furthermore, we compared the transcriptome of each MSC in early passages with late passages. MSC used in this study were obtained from the umbilical vein of three donors, two MSC/n and one MSC/inv. After their cryopreservation, they have been expanded in vitro until reached senescence. Total RNA was extracted using the RNeasy mini kit (Qiagen) and marked with the GeneChip ? 3 IVT Express Kit (Affymetrix Inc.). Subsequently, the fragmented aRNA was hybridized on the microarranjo Affymetrix Human Genome U133 Plus 2.0 arrays (Affymetrix Inc.). The statistical analysis of differential gene expression was performed between groups MSC by the Partek Genomic Suite software, version 6.4 (Partek Inc.). Was considered statistically significant differences in expression to p-value Bonferroni correction &#706;.01. Only signals with fold change &#707; 3.0 were included in the list of differentially expressed. Differences in gene expression data obtained from microarrays were confirmed by Real Time RT-PCR. For the interpretation of biological expression data were used: IPA (Ingenuity Systems) for analysis enrichment functions, the STRING 9.0 for construction of network interactions; Cytoscape 2.8 to the network visualization and analysis bottlenecks with the aid of the GraphPad Prism 5.0 software. BiNGO Cytoscape pluggin was used to access overrepresentation of Gene Ontology categories in Biological Networks. The comparison between senescent and young at each group of MSC has shown that there is a difference in the expression parttern, being higher in the senescent MSC/inv group. The results also showed difference in expression profiles between the MSC/inv versus MSC/n, being greater when they are senescent. New networks were identified for genes related to the response of two of MSC over cultivation time. Were also identified genes that can coordinate functional categories over represented at networks, such as CXCL12, SFRP1, xvi EGF, SPP1, MMP1 e THBS1. The biological interpretation of these data suggests that the population of MSC/inv has different constitutional characteristics, related to their potential for differentiation, proliferation and response to stimuli, responsible for a distinct process of replicative senescence in MSC/inv compared to MSC/n. The genes identified in this study are candidates for biomarkers of cellular senescence in MSC, but their functional relevance in this process should be evaluated in additional in vitro and/or in vivo assays / C?lulas-tronco mesenquimais humanas (CTMH) s?o muito ?teis na terapia celular. O longo per?odo de cultivo pode resultar em senesc?ncia replicativa ou estar relacionado com o aparecimento de altera??es cromoss?micas respons?veis pela aquisi??o de um car?ter tumorig?nico in vitro . Neste estudo, foi comparado o transcriptoma de CTMH jovens e senescentes obtidas de diferentes doadores. Al?m disso, pela primeira vez, o perfil de express?o de CTMH com uma invers?o cromoss?mica parac?ntrica (CTMH/inv) foi comparado ao de CTMH que possuem cari?tipo normal (CTMH/n) em passagens jovens e senescentes de cultivo in vitro . As CTMH utilizadas neste estudo foram isoladas da veia do cord?o umbilical de tr?s dadores, dois CTMH/n e de um CTMH/inv. Ap?s a criopreserva??o, elas foram expandidas in vitro at? alcan?arem a senesc?ncia. O RNA total foi extra?do utilizando o RNeasy mini kit (Qiagen), marcado, purificado e fragmentado com o ? 3 GeneChip IVT expresso Kit (Affymetrix, Inc.). Subsequentemente, o RNA fragmentado foi hibridado no microarranjo Affymetrix Human Genome U133 Plus 2.0 (Affymetrix, Inc.). A an?lise estat?stica da express?o diferencial foi realizada usando o Partek Suite Software Genomic, vers?o 6.4 (Partek, Inc.). Foram consideradas estatisticamente significativas as diferen?as na express?o com valor de P &#706;0.01 corrigido com Bonferroni. Apenas os sinais com fold change &#707;3.0 foram inclu?dos na lista de diferencialmente expressos. Diferen?as na express?o g?nica observadas no estudo dos microarranjos foram confirmadas por resultados de RT-PCR em tempo real. Para a interpreta??o biol?gica dos dados foram utilizados: IPA (Ingenuity Systems) para an?lise de enriquecimento de fun??es; STRING 9,0 para a constru??o de redes de intera??es; Cytoscape 2,8 para a visualiza??o das redes e an?lises de gargalos com o aux?lio do software GraphPad Prism 5.0. O pluggin BiNGO do Cytoscape foi utilizado para avaliar a representa??o de categorias funcionais no Gene Ontology nas redes biol?gicas. A compara??o entre senescentes e jovens em cada grupo de CTMH mostrou que h? uma diferen?a no perfil de express?o, sendo maior nas senescentes do grupo CTMH/inv. Os resultados tamb?m mostraram que h? diferen?a nos perfis de express?o entre as CTMH/inv e CTMH/n, sendo maior a diferen?a quando as c?lulas est?o senescentes. Novas xiv redes foram identificadas para genes relacionados com a resposta ao longo do tempo de cultivo nos dois grupos de CTMH. Foram identificados genes que podem coordenar fun??es importantes mais enriquecidas nas redes, como por exemplo, CXCL12, SFRP1, EGF, SPP1, MMP1 e THBS1. A interpreta??o biol?gica destes dados sugere que a popula??o de c?lulas CTMH/inv tem diferentes caracter?sticas constitucionais, relacionadas com o seu potencial de prolifera??o, diferencia??o e resposta a est?mulos, respons?veis por um processo de senesc?ncia replicativa em CTMH/inv distinto das CTMH/n. Os genes identificados neste estudo s?o candidatos a marcadores da senesc?ncia celular em CTMH, mas a sua relev?ncia funcional neste processo deve ser testada em experi?ncias adicionais in vitro e/ou in vivo

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-10-11T22:29:37Z No. of bitstreams: 1 ClarissaFaveroDemeda_TESE.pdf: 3254557 bytes, checksum: 71f116b43b1323319edbd07fa141e478 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-10-17T23:03:20Z (GMT) No. of bitstreams: 1 ClarissaFaveroDemeda_TESE.pdf: 3254557 bytes, checksum: 71f116b43b1323319edbd07fa141e478 (MD5) / Made available in DSpace on 2016-10-17T23:03:20Z (GMT). No. of bitstreams: 1 ClarissaFaveroDemeda_TESE.pdf: 3254557 bytes, checksum: 71f116b43b1323319edbd07fa141e478 (MD5) Previous issue date: 2016-02-17 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Os exossomos s?o respons?veis pela comunica??o c?lula-c?lula e podem influenciar na progress?o tumoral, met?stase e efic?cia terap?utica. Dentre as c?lulas capazes de secretar exossomos est?o as c?lulas tumorais e as c?lulas imunes. Sabe-se que a presen?a das c?lulas imunes ? importante para erradicar os tumores. No entanto, achados recentes demonstram que a inflama??o pode promover o crescimento tumoral. Os macr?fagos associados a tumores (TAMs) s?o conhecidos por apresentarem diferentes subtipos, M1 e M2, capazes de secretarem exossomos. O presente estudo se prop?s a observar o comportamento dos exossomos derivados dos TAMs, dos subtipos 1 e 2, frente a cultura de c?lulas humanas SCC-25, HSC-3 e SAS derivadas de CE de l?ngua, por meio da an?lise da capacidade de invas?o, prolifera??o e viabilidade das c?lulas tumorais na presen?a dos exossomos. Observou-se que as microves?culas derivadas dos TAMs apresentam positividade para CD63, caracterizando-as como exossomos. Os exossomos dos TAMs do subtipo M2 foram os ?nicos a apresentarem marca??o para TGF-?, quando em compara??o com os exossomos M1, THP1 e das linhagens celulares de CE, sugerindo que os exossomos M2 podem ser respons?veis pela express?o de TGF-? nas c?lulas tumorais, uma vez que s?o internalizados. Nos ensaios de migra??o, observou-se que as c?lulas SCC-25 em presen?a de meio de cultura DMEM F/12, apresentaram maior capacidade de invas?o frente aos exossomos M2 (p?0,001), para concentra??o de 0,1 ?g/ml. Para as c?lulas HSC-3 e SAS, n?o foi observada rela??o estatisticamente significante entre a presen?a de exossomos cultivados juntamente com as c?lulas tumorais e a capacidade de invas?o celular (p>0,05). Quando os exossomos foram colocados no compartimento inferior do transwell, as c?lulas HSC-3 em presen?a dos exossomos M2 (1,0 ?g/ml) apresentaram maior capacidade de invas?o (p?0,001). O teste de viabilidade demonstrou que as c?lulas HSC-3 tornam-se mais vi?veis frente ? presen?a dos exossomos M2 (p?0,001) na concentra??o de 50 ?g/ml. Para as c?lulas SCC-25, o resultado foi o mesmo (p?0,05). A imunofluoresc?ncia demonstrou a internaliza??o dos exossomos nas linhagens celulares estudadas. Os achados sugerem que a presen?a de exossomos M2, frente ?s culturas de c?lulas de CE de l?ngua, pode ser um campo de pesquisa importante para futuros estudos com terapias-alvo. / Squamous cell carcinoma (SCC) of the oral tongue is one of the most common malignant lesions in the oral cavity and is characterized by presenting a locally invasive and aggressive behavior. The exosomes are responsible for cell-cell communication and may influence tumor progression, metastasis and therapeutic efficacy. Among the cells that can secrete exosomes are tumor cells and immune cells. It is known that the presence of immune cells is important to eradicate tumors. However, recent findings suggest that inflammation may promote tumor growth. The tumor associated macrophages (TAMs) are known to have subtypes, M1 and M2, that secrete exosomes. This study?s goal was to observe the behavior of derivatives TAMs exosomes, subtypes 1 and 2, against human cell culture SCC-25, HSC-3 and SAS derived from SCC of oral tongue, through the analysis of invasiveness, proliferation and viability of tumor cells in the presence of exosomes. It was observed that the microvesicles derived from TAMs are positive for CD63, characterizing them as exosomes. The exosomes of the M2 subtype TAMs were the only ones to present TGF-? marking, as compared with M1 exosomes, THP1 and SCC cell lines, suggesting that M2 exosomes may be responsible for TGF-? expression in tumor cells. In the migration tests, it was found that the SCC-25 cells in the presence of culture medium DMEM F / 12 showed higher invasion capacity in the presence of M2 exosomes (p?0,001) to a concentration of 0.1 ?g/ml. For HSC-3 and SAS cells, there was no significant statistical relationship between the presence of exosomes and invasiveness (p> 0.05) for the exosomes derived from TAMs. When the exosomes were placed in the lower compartment of the transwell, the HSC-3 cells in the presence of M2 exosomes (1.0 ?g/ml) had higher invasiveness (p?0,001). The viability test showed that HSC-3 cells became more viable in the presence of M2 exosomes (p?0.001) at a concentration of 50 ?g/ml. For SCC-25 cells, the result was the same (p?0.05). Immunofluorescence showed the internalization of exosomes in the cell lines studied. These findings suggest that the presence of M2 exosomes in the SCC of the oral tongue cell cultures may be an important research field for future studies of targeted therapies.

Submitted by Raniere Barreto (raniere.barros@ufvjm.edu.br) on 2018-05-10T17:58:30Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) alcides_alves_freitas.pdf: 1707403 bytes, checksum: f9233629066db73fd877b2885aa875bd (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-05-14T14:47:32Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) alcides_alves_freitas.pdf: 1707403 bytes, checksum: f9233629066db73fd877b2885aa875bd (MD5) / Made available in DSpace on 2018-05-14T14:47:32Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) alcides_alves_freitas.pdf: 1707403 bytes, checksum: f9233629066db73fd877b2885aa875bd (MD5) Previous issue date: 2017 / O boldo da terra (Plectranthus barbatus) ? popularmente utilizado para o tratamento de dist?rbios gastrintestinais e para doen?as hep?ticas. Devido ? exist?ncia de um grande n?mero de esp?cies dispon?veis para pesquisa e estudos farmacol?gicos, o estudo dessa planta torna-se importante para o conhecimento t?cnico-cient?fico, especialmente com a finalidade do desenvolvimento de novos f?rmacos. Com isto, o objetivo desse trabalho foi detectar prote?nas ativas de Plectranthus barbatus (boldo da terra) e avaliar a atividade biol?gica em c?lulas A549 e RAW264.7. As amostras dos procedimentos de extra??o das folhas e caule do P. barbatus foram submetidas ? quantifica??o de prote?na. Foi detectado em gel de poliacrilamida SDS-PAGE 12% prote?nas com peso molecular em torno de 30kDa e 94kDa o que ? descrito na literatura como lectinas e lipoxigenases. Os extratos foram caracterizados por cromatografia l?quida de alta efici?ncia com picos aparentes em 16 e 27 minutos. N?o foi detectada atividade de inibi??o da tripsina. Os resultados dos testes biol?gicos em cultura de c?lulas demonstraram que o extrato purificado de inibidores de protease n?o alterou a viabilidade celular de ambas as linhagens, no entanto, foi capaz de inibir a produ??o de ?xido n?trico na concentra??o de 10 ?g/ml para folha e caule e 100 ?g/ml para folha. Este trabalho demonstra pela primeira vez a extra??o de prote?nas em folhas e caule de Plectranthus barbatus e a atividade dessa mol?cula em cultura celular. Esse extrato n?o alterou a viabilidade celular de ambas as linhagens celulares, podendo ser caracterizados como n?o citot?xico nas concentra??es testadas. Conclui-se, portanto, que embora as folhas, caules e flores do Plectranthus barbatus seja utilizado amplamente pela popula??o esse trabalho demonstrou a detec??o de lectina e lipoxigenase at? agora desconhecidos nessa esp?cie em estudo. / Disserta??o (Mestrado Profissional) ? Programa de P?s-Gradua??o em Tecnologia, Sa?de e Sociedade, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2017. / The boldo da terra (Plectranthus barbatus) is popularly used for the treatment of gastrointestinal disorders and for liver diseases. Due to the existence of a large number of species available for research and pharmacological studies, the study of this plant becomes important for technical-scientific knowledge, especially for the purpose of developing new drugs. With this, the objective of this work was to detect active proteins of Plectranthus barbatus and to evaluate the biological activity in cells A549 and RAW264.7. Samples of P. barbatus leaf and stem extraction procedures were submitted to protein quantification. SDS-PAGE was detected in 12% proteins with molecular weight around 30kDa and 94kDa which is described in the literature as lectins and lipoxygenases. The extracts were characterized by high performance liquid chromatography with apparent peaks at 16 and 27 minutes. No trypsin inhibition activity was detected. The results of the biological tests in cell culture demonstrated that the purified protease inhibitor extract did not alter the cell viability of both strains, however, it was able to inhibit the production of 10 ?g / ml nitric oxide to leaf and And 100 ?g / ml for leaf. This work demonstrates for the first time the extraction of proteins in leaves and stem of Plectranthus barbatus and the activity of this molecule in cell culture. This extract did not alter the cellular viability of both cell lines and could be characterized as non-cytotoxic at the concentrations tested. It was concluded, therefore, that although the leaves, stems and flowers of Plectranthus barbatus were used extensively by the population, this work demonstrated the detection of lectin and lipoxygenase hitherto unknown in this species under study.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Biologia Celular e do Desenvolvimento, Florianópolis, 2015. / Made available in DSpace on 2016-05-24T17:30:08Z (GMT). No. of bitstreams: 1 337730.pdf: 3042114 bytes, checksum: b1db87538875d11b0c2804170ae64711 (MD5) Previous issue date: 2015 / Elevados níveis do aminoácido homocisteína (Hcy) são caracterizados como uma desordem metabólica denominada - hiperhomocisteinemia, a qual está envolvida com a ocorrência de anomalias congênitas. No sistema nervoso central (SNC) especificamente, esta condição está associada ao dano ao DNA, citotoxicidade neuronal, estresse oxidativo e prejuízo na síntese de neurotransmissores. Neste estudo, foi avaliado o efeito da dose alta de Hcy sobre a expressão das proteínas do ciclo celular, diferenciação e sobrevivência neural. Adicionalmente, investigamos o efeito da Hcy sobre a ultraestrutura das células do mesencéfalo em embriões de Gallus domesticus. Para tal, os embriões foram tratados com 48 horas de incubação (E2) onde foram organizados dois grupos experimentais: controle - 50 µl salina e Hcy - 20 µmol Hcy/50 µl salina. Os embriões foram analisados em duas idades - E6 e E10. Através da técnica de imuno-histoquímica, verificou-se uma redução significativa da proliferação celular na camada ependimária do mesencéfalo nos embriões em E6. A avaliação da expressão das demais proteínas envolvidas no ciclo celular como p53, p21, ciclina E e PCNA foi realizada pela combinação das técnicas de citometria de fluxo e imuno-histoquímica. O tratamento com Hcy induziu um aumento na expressão da proteína p53, e uma diminuição na expressão das proteínas p21 e ciclina E. Para as proteínas envolvidas no processo apoptótico, como BCL2 e BAK, não foram encontradas diferenças significativas nas duas idades analisadas. Na análise da diferenciação neural encontramos um aumento significativo na expressão da proteína GFAP em E6, o que caracterizamos como um processo de resposta celular denominado gliose reativa. Porém, esta resposta foi normalizada nos embriões na idade E10. Para a análise de sobrevivência neural, houve uma redução na expressão de BDNF e GDNF nos embriões expostos à Hcy na idade de E6. Por fim, através da técnica de microscopia eletrônica de transmissão, a análise ultraestrutural das células revelou alterações relacionadas ao espaço perinuclear e a formação anômala de membrana nos embriões expostos à Hcy nas idades de E6 e E10. Os resultados demonstram que, embora a Hcy não tenha induzido anomalias congênitas aparentes no SNC, o tratamento comprometeu os processos de proliferação celular, diferenciação glial e sobrevivência neural em G. domesticus.<br> / Abstract : High levels of amino acid homocysteine (Hcy) are characterized as a metabolic disorder named hyperhomocysteinemia, which is related to the occurrence of congenital anomalies. Specifically in the nervous system (CNS), this condition is associated with DNA damage, neuronal cytotoxicity, oxidative stress and impaired neurotransmitter synthesis. This study evaluated the effect of Hcy on the expression of cell cycle proteins, neuronal differentiation and survival. Additionally we investigated the effect of Hcy on the ultrastructure of midbrain in embryos aged E6 and E10. Thus, fertilized eggs of Gallus domesticus were treated at 48 hours of incubation (E2). Two experimental groups were organized: Control - 50 µl saline and Hcy - 20 µmol Hcy/50 µl saline. Embryos were analyzed at two embryonic ages - E6 and E10. Immunohistochemistry showed a significant reduction in expression cell proliferation in the ependymal layer of the midbrain at E6. Evaluation of expression of the other proteins involved in cell cycle such as p53, p21, cyclin E and PCNA were performed by the combination of flow cytometry and immunohistochemistry Hcy treatment induced an increase in p53 expression, and a decrease in the expression of the proteins p21 and cyclin E. Proteins involved in apoptosis, as BCL2 and BAK, not showed significant differences in both ages examined. The analysis of neural differentiation found a significant increase in protein expression GFAP at E6, which characterized as a process of cellular response termed reactive gliosis, but this response was standardized at E10. The quantification of the proteins neuronal survival BDNF and GDNF were quantified by flow cytometry and a significant reduction was found in embryos exposed to Hcy at E6, and this pattern is reversed at E10. Finally, by technique of transmission electron microscopy, the ultrastructural analysis of the cells showed changes related to the perinuclear space and the anomalous formation of membrane in embryos exposed to Hcy at E6 and E10. The results demonstrated that although Hcy did not induce congenital anomalies in the CNS, the treatment impaired the cellular processes of proliferation, differentiation glial and neuronal survival in G. domesticus.