Objetivo: Determinar potenciales factores de riesgo para diabetes mellitus gestacional (DMG). Métodos: Estudio de casos y controles realizado en el Hospital Alberto Sabogal mediante recolección de historias clínicas del 2009 a 2014. Se define como caso las gestantes con diagnóstico de DMG mediante una prueba de tolerancia oral a la glucosa (PTOG), previa glucosa en ayunas anormal y control a la gestante sin valores indicativos de DMG. Las variables de interés fueron paridad, antecedente de cesáreas, abortos y recién nacido con mayor peso. Modelos de regresión logística fueron calculados para estimar odd ratios (OR) e intervalos de confianza al 95% (IC95%). Resultados: Se incluyeron 84 casos y 336 controles. En el modelo multivariado, la multiparidad incrementó el riesgo de DMG (OR= 3,54; IC95% 1,55 – 8,14). También, antecedente de abortos, a partir del segundo aborto (OR= 3,40, IC95% 1,55 – 7,44) y cesáreas previas (1 cesárea OR= 3,5 IC95% 1,89 – 6,47 y 2+ cesáreas OR=8,35 IC95% 3,50 – 19,95). La multiparidad, dos o más abortos y mayor número de cesáreas son factores de riesgo para DMG. / Objectives: To identify risk factors for gestational diabetes mellitus (GDM). Methods: A case-control study was performed in Alberto Sabogal Hospital, collecting medical records from 2009 to 2014. A case was defined as a pregnant women diagnosed with GDM by an oral glucose tolerance test (OGTT) after an abnormal fasting glucose and control was defined as a pregnant women without GDM indicative values. The study outcome was GDM. The variables of interest were multiparity, previous cesarean section, abortions, newborn with the greatest weight. Logistic regression were used to calculate the odds ratio (OR) and a confidence interval of 95% (IC95%). Results: 84 cases and 336 controls were included. In the multivariate model, multiparity increased risk of GDM (OR= 3.54, 95% CI 1.55 to 8.14). As well history of abortions, from the second abortion (OR= 3.40, 95% CI 1.55 to 7.44) and previous cesarean section are also related (cesarean section 1 OR= 3.5 95% CI 1.89 to 6.47 and 2+ cesarean OR= 8.35 95% CI 3.50 to 19.95). Multiparity, two or more abortions, a biggest number of cesarean sections are GDM risk factors.

Grief and causal attribution are two of the most commonly observed reactions to early miscarriage, yet little is known about these reactions or whether a relationship exists between them. This exploratory and descriptive correlational study examined the maternal grief intensities, the causal attributions, and the relationship between them in a convenience sample of 15 women who spontaneously aborted at 16 weeks' or less gestation. Women responded to both a written questionnaire and a semi-structured Interview at 6 to 10 weeks post-miscarriage. Their responses Indicated both current and retrospective reactions to their miscarriages. Responses were analysed using nonparametric statistics and content analysis. Maternal grief Intensities were found to vary widely at the time of the miscarriage, but all decreased significantly 6 to 10 weeks later. All women reacted to their miscarriage with attribution-seeking behaviors. The explanations most women formed were comprised of more than one causal attribution. Attributions were observed to have four distinct characteristics. Causal attributions were found to be either philosophical or physically oriented; to be organic, non-specific or maternal/self-blaming In origin; to be either dominant or non-dominant, and/or to refer to causalities immediate or prior to the physical event. At the time of the miscarriage a positive correlation between grief Intensity and maternal/self-blaming attributions and between grief Intensity and philosophical attributions was found. These relationships were not observed 6 to 10 weeks later. A positive correlation was found between grief intensity and attributions to maternal emotions at both the time of the miscarriage and 6 to 10 weeks later. / Applied Science, Faculty of / Nursing, School of / Graduate

Introdução: Alterações no ambiente fetal conferem um risco aumentado para doenças crônicas como obesidade, doença cardiovascular, hipertensão arterial e diabetes tipo 2. As evidências sugerem que a restrição de crescimento intrauterino (RCIU) pode programar de forma persistente as preferências alimentares, e acredita-se que esse tipo de alteração comportamental, pode explicar, pelo menos em parte, o aumento do risco para essas doenças em indivíduos que sofreram RCIU. Portanto, torna-se importante entender os fatores associados e mecanismos envolvidos nesse comportamento. O objetivo deste trabalho foi investigar o efeito da RCIU no comportamento alimentar em animais e humanos, assim como os possíveis mecanismos envolvidos na sua programação. Métodos: Ratas Sprague Dawley prenhes foram randomizadas para o grupo controle (Adlib), que recebeu dieta padrão ad libitum ou grupo restrição 50% (FR), que recebeu 50% do consumo habitual de genitoras alimentadas ad libitum. As dietas foram oferecidas a partir do dia 10 de gestação até o dia 21 de lactação. Em até 24h após o nascimento, foi realizada a adoção cruzada formando os grupos: Adlib_Adlib, FR_Adlib, FR_FR, Adlib_FR. O consumo de ração padrão foi comparado entre todos os grupos. A preferência alimentar, a preferência condicionada por lugar induzida por alimento palatável, assim como a fosforilação da enzima tirosina hidroxilase e os níveis de receptores D2 no núcleo acumbens foram comparados entre os grupos de interesse (Adlib_Adlib e FR_Adlib). Nos humanos, 75 jovens, classificados quanto à RCIU, participaram de avaliação antropométrica, bioquímica e de comportamento alimentar (teste de escolha alimentar, no qual todos recebiam um valor monetário para compra de um lanche, e Dutch Eating Behaviour Questionnaire, DEBQ). Dados de neuroimagem funcional em repouso entre regiões relacionadas à recompensa de 28 indivíduos foram processados e analisados, de um total de 43 exames realizados. Resultados: No estudo experimental, viu-se que o consumo de ração padrão não foi diferente entre os grupos. Ratos restritos apresentaram preferência pela dieta palatável, mas menor condicionamento de preferência ao lugar associado ao alimento palatável. A fosforilação da tirosina hidroxilase no núcleo acumbens foi maior nestes animais no estado basal, mas após exposição ao doce essa diferença entre os grupos permaneceu apenas nos machos. A RCIU também se associou a menores níveis de receptores D2 no núcleo acumbens. No estudo clínico, encontrou-se que a menor razão de crescimento fetal (indicativo de maior RCIU) e alto índice de massa corporal predizem um estilo alimentar restritivo visto pelo DEBQ. Pessoas nascidas com RCIU também usaram menor quantidade do um recurso financeiro oferecido no teste de escolha alimentar após um período de jejum. Os dados de neuroimagem funcional sugerem que os indivíduos restritos apresentam um padrão de conectividade em repouso alterado entre o córtex orbito-frontal, o estriado ventral/dorsal e a amígdala. Conclusão: A RCIU esteve associada com preferência por alimentos palatáveis e alterações no sistema dopaminérgico no estudo experimental e alterações da conectividade em repouso entre áreas do sistema mesocorticolímbico no estudo clínico. As alterações observadas no sistema dopaminérgico dos animais restritos indicam que esse sistema estaria envolvido na programação da preferência alimentar nesses indivíduos. Além disso, o padrão de conectividade em repouso observado nos indivíduos restritos sugere que alterações em determinadas regiões do sistema de recompensa poderiam estar associadas com mudanças no comportamento alimentar. As alterações neurocomportamentais observadas confirmam a existência de programação fetal do comportamento alimentar pela RCIU, apontando modificações persistentes no sistema de recompensa do cérebro, o que pode ser visto como um fator de risco para o desenvolvimento de obesidade e suas comorbidades. / Introduction: Fetal environment changes can lead to adaptations that are associated with increased risk for obesity, cardiovascular disease, hypertension and diabetes in adult life. Evidence suggests that intrauterine growth restriction (IUGR) can persistently program the subject’s preference for palatable foods. It is believed that feeding behavior alterations can explain, at least in part, the increased risk for chronic diseases in IUGR individuals. Therefore, it becomes important to understand the factors and mechanisms involved in this behavior. The aim of this study was to explore how IUGR affects feeding behavior of animals and humans, as well as to verify the potential mechanisms related to this behavioral programming. Methods: Time-mated pregnant Sprague-Dawley rats were randomly allocated to Control (receiving standard chow ad libitum) or 50% food restricted (FR), receiving 50% of the ad libitum-fed dam’s habitual intake. These diets were provided from day 10 of pregnancy throughout day 21 of lactation. Within 24 hours after birth, pups were crossfostered, forming four groups: Adlib_Adlib, FR_Adlib, FR_FR, Adlib_FR. Standard chow consumption was compared between all groups. Food preference, conditioned place preference to a palatable diet, and the nucleus accumbens tyrosine hydroxylase phosphorylation and D2 receptor levels were analyzed focusing on two groups of interest (Adlib_Adlib and FR_Adlib). In humans, 75 youths were classified regarding IUGR and had anthropometric data, biochemical data, and feeding behavior (food choice task, in which everyone received a monetary value to purchase a snack, and Dutch Eating Behaviour Questionnaire) assessed. Forty three neuroimaging exams were performed and resting state functional connectivity between brain regions related to reward of 28 individuals were processed and analyzed. Results: In the experimental study, standard chow consumption was not different between groups. IUGR adult rats had increased preference for palatable food, but showed less conditioned place preference to a palatable diet compared to controls. At baseline, the accumbal tyrosine hydroxylase phosphorylation was increased in IUGR rats compared to controls. After sweet food exposure, the difference between groups remained only in males. Accumbal D2 receptors levels were decreased in IUGR rats. In the clinical study, it was found that low birth weight ratio (indicative of higher IUGR) and high body mass index predict a restrained eating style as seen by the DEBQ. IUGR individuals used a smaller quantity of a financial resource offered in the food choice task after a fasting period. Resting state functional connectivity data suggest that IUGR individuals had an altered pattern of connectivity between the orbitofrontal cortex, the ventral/dorsal striatum and the amygdala. Conclusion: IUGR was associated with a preference for palatable foods and alterations in the dopaminergic system in the experimental study, as well as changes in the resting state functional connectivity between regions of the mesocorticolimbic pathway in the clinical study. Alterations in the mesolimbic dopaminergic system observed in IUGR rats indicate an important role in the programming of food preferences. Moreover, the IUGR pattern of brain connectivity observed suggests that alterations in certain regions involved in reward processing and evaluation could be associated with changes in eating behavior. Neurobehavioral changes observed confirmed the existence of a fetal programming of feeding behavior associated with IUGR, pointing out to persistent modifications in the brain reward system, which can be seen as a risk factor for the development of obesity and its comorbidities.

Orientadores: Ricardo Barini, Jose Guilherme Cecatti / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-07T09:44:20Z (GMT). No. of bitstreams: 1 Silva_LucianaVivas_M.pdf: 2243768 bytes, checksum: 779bfadbc13a9b63606a5548362e4609 (MD5) Previous issue date: 2006 / Mestrado / Tocoginecologia / Mestre em Tocoginecologia

Submitted by Samira Ramos (samira.ramos@unifenas.br) on 2018-04-25T21:05:10Z No. of bitstreams: 1 TAIS APARECIDA PATROCINIO.pdf: 726977 bytes, checksum: 8cef7ddcd6970bf3d2ccd912eb038060 (MD5) / Made available in DSpace on 2018-04-25T21:05:10Z (GMT). No. of bitstreams: 1 TAIS APARECIDA PATROCINIO.pdf: 726977 bytes, checksum: 8cef7ddcd6970bf3d2ccd912eb038060 (MD5) Previous issue date: 2017-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG / This study evaluated the effect of AR33 (patent-pending formula), a resveratrol analogue, in the culture of in vitro fertilized embryos. Cumulus-oocyte complexes (COCs) recovered from bovine ovaries collected at the slaughterhouse were matured in vitro for 24 h and fertilized in vitro for 20 h, both at 38.8 °C under 5% CO2 in air and high humidity. Probably partially nude zygotes were randomly distributed in two experiments. Experiment 1: 0 (control, n = 347), 0.1 μM (n = 337), 0.5 μM (n = 277) and 2.5 μM AR33 (n = 343) with 2.5% fetal bovine serum (FBS) and experiment 2: 2.5 μM AR33 (n = 381), 0.5 μM resveratrol (n = 381), both with 2.5% SFB and 0 (control, n = 341) with 10% FBS. The base medium for all treatments was SOFaa and incubation conditions were 38.8 °C under 5% CO2 in air and high humidity. Half of the culture medium was fed on days 3 and 5 after fertilization. The cleavage rate was evaluated on day 3 and the blastocyst rate (B1) on days 7 and 8 post-fertilization. At day 8, the blastocysts were fixed and subsequently submitted to analysis of the number of cells and apoptotic index. Cleavage and blastocyst rates were analyzed by logistic regression models (Proc Logistic), and the number of cells and apoptotic index by mixed linear models (Proc Mixed) using the SAS statistical package. In experiment 1, the cleavage rate (P <0.05) was higher at 2.5 μM (69.0 ± 4.4%) than at 0, 0.1 and 0.5 μM AR33 (62.1 ± 2.0%, 60.7 ± 5.9% and 56.7 ± 5.8%, respectively). At day 7, the Bl rate was similar (P> 0.05) between 0.1, 0.5 and 2.5 μM (18.1 ± 5.4%, 17.5 ± 2.9% and 19.4 ± 3.3%, respectively) and all were higher (P <0.05) 05) at 0 μM AR33 (12.4 ± 2.5%). At day 8, only 0.1 and 2.5 μM (21.0 ± 5.0% and 24.6 ± 3.3%) were higher than 0 μM AR33 (15.2 ± 2.5%). There was no difference (P> 0.05) between treatments regarding total cell number (TC) and internal cell mass (MCI); However, the apoptotic index in CT and MCI was higher for 0 and 2.5 μM (11.36 and 9.89%, 20.52 and 15.85%) than in 0.1 and 0.5 μM AR33 (4.66 and 4.82%, 8.47 and 10.92%). In the experiment 2 the cleavage rate (P <0.05) was higher in the control (80.8 ± 3.4%) than in the treatment with 0.5 μM resveratrol (76.4 ± 3.6%), but similar to 2.5 μM AR33 (76.9 ± 1.2%). There were no differences (P> 0.05) for the Bl rate on days 7 and 8. The apoptotic index in the CT and MCI was higher in the control (8.9 and 14.9%, respectively) than in the 2.5 μM AR33 and 0.5 μM resveratrol (6.4 and 5.5%, 11.1 and 8.8%, respectively). In conclusion, resveratrol and its synthetic analogue tested in this study improve bovine embryonic development in culture medium supplemented with 2.5% FBS under 5% CO2 in air. / Este estudo avaliou o efeito de AR33 (fórmula com patente-pendente), um análogo de resveratrol, no cultivo de embriões fecundados in vitro. Complexos cumulus-oócitos (COCs) recuperados de ovários bovinos coletados no matadouro, foram maturados in vitro durante 24 h e fertilizados in vitro por 20 h, ambos em 38.8 °C sob 5% de CO2 em ar e alta umidade. Prováveis zigotos parcialmente desnudos foram distribuídos aleatoriamente em dois experimentos. Experimento 1: 0 (controle, n=347), 0.1 µM (n=337), 0.5 µM (n=277) e 2.5 µM de AR33 (n=343) com 2,5% de soro fetal bovino (SFB), e experimento 2: 2.5 µM de AR33 (n=381), 0.5 µM de resveratrol (n=381), ambos com 2,5% SFB e 0 (controle, n=341) com 10% SFB. O meio base para todos os tratamentos foi SOFaa e as condições de incubação foram de 38.8 °C sob 5% de CO2 em ar e alta umidade. Metade do meio de cultura foi renovado (feeding) nos dias 3 e 5 após a fertilização. A taxa de clivagem foi avaliada no dia 3 e a taxa de blastocisto (Bl) nos dias 7 e 8 pós-fecundação. No dia 8, os blastocistos foram fixados e posteriormente submetidos a análise do número de células e índice apoptótico. As taxas de clivagem e de blastocistos foram analisadas por modelos de regressão logística (Proc Logistic), e o número de células e índice apoptótico por modelos lineares mistos (Proc Mixed) usando o pacote estatístico SAS. No experimento 1, a taxa de clivagem (P<0.05) foi maior para 2.5 µM (69.0±4.4%) do que para 0, 0.1 e 0.5 µM de AR33 (62.1±2.0%, 60.7±5.9% e 56.7±5.8%, respectivamente). No dia 7, a taxa de Bl foi semelhante (P>0.05) entre 0.1, 0.5 e 2.5 µM (18.1±5.4%, 17.5±2.9% e 19.4±3.3%, respectivamente) e todos eles foram superiores (P<0,05) à 0 µM AR33 (12.4±2.5%). No dia 8, apenas 0.1 e 2.5 µM (21.0±5.0% e 24.6±3.3%) foram maiores do que 0 µM AR33 (15.2±2.5%). Não houve diferença (P>0,05) entre tratamentos quanto ao número de células totais (CT) e da massa celular interna (MCI); contudo, o índice apoptótico nas CT e na MCI foram maiores para 0 e 2.5 µM (11.36 e 9.89%; 20.52 e 15.85%) do que em 0.1 e 0.5 µM AR33 (4.66 e 4.82%; 8.47 e 10.92%). No experimento 2 a taxa de clivagem (P<0,05) foi maior no controle (80.8±3.4%) do que no tratamento com 0.5 µM resveratrol (76.4±3.6%), e este último semelhante à 2.5 µM AR33 (76.9±1.2%). Não houve diferenças (P>0,05) para a taxa de Bl nos dias 7 e 8. O índice apoptótico nas CT e MCI foi maior no controle (8.9 e 14.9%, respectivamente) do que para 2.5 µM AR33 e 0.5 µM resveratrol (6.4 e 5.5%; 11.1 e 8.8%, respectivamente). Em conclusão, o resveratrol e o seu análogo sintético testado neste estudo melhoram o desenvolvimento embrionário bovino em meio de cultura suplementado com 2,5% SFB sob 5% de CO2 em ar.

Orientador: Leandro Gustavo de Oliveira / Resumo: Introdução: O Lúpus Eritematoso Sistêmico (LES) é uma doença sistêmica de caráter autoimune, que acomete mulheres em idade reprodutiva, sendo que a inter-relação entre a doença e a gestação determina importantes desfechos maternos e perinatais. Objetivos: Descrever os desfechos maternos e perinatais em gestações de pacientes portadoras de LES e avaliar o impacto da nefrite lúpica sobre os resultados encontrados. Métodos: Este é um estudo observacional descritivo desenvolvido para avaliar as inter-relações entre gestação e lúpus eritematoso sistêmico em pacientes atendidas na Maternidade do Hospital das Clínicas da Faculdade de Medicina de Botucatu – HCFMB. O período de estudo foi de janeiro de 2010 a agosto de 2019. Resultados: Foram avaliadas 38 gestações em 31 pacientes com LES. A média das idades foi de 27,4 + 6 anos. A média das idades gestacionais ao nascimento foi de 36 + 3 semanas. As principais intercorrências observadas foram: anemia (39,4%), nefrite lúpica (29%) e hipertensão arterial crônica (10,5%). Hidroxicloroquina foi utilizada em 47,4% das gestações. Em 51,4% das pacientes houve necessidade de antecipação do parto e em 13,1% houve piora da função renal. A incidência de pré-eclâmpsia foi de 19,4%. Prematuridade ocorreu em 20% dos casos e restrição de crescimento fetal, em 19,4%. Nefrite lúpica determinou maior ocorrência de flare (p<0,05) e maior necessidade de antecipação do parto (p< 0,05). Conclusão: O presente estudo permitiu avaliar a inter-relação entre L... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Systemic Lupus Erythematosus is an autoimmune systemic disease that affects women of reproductive age, and the interrelation between disease and pregnancy determines important maternal and perinatal outcomes. Objectives: To describe maternal and perinatal outcomes in pregnancies of patients with SLE and to evaluate the impact of lupus nephritis on the results found. Methods: Descriptive observational study developed to assess the interrelation between pregnancy and systemic lupus erythematosus in patients attended at the Maternity of the Clinics Hospital from Botucatu Medical School – HCFMB. The study period corresponded to January 2010 until August 2019. Results: Thirty-eight pregnancies were evaluated in 31 patients with SLE. Their average age was 27.4 + 6.0 years. The average gestational age at birth was 36 + 3 weeks. The main clinical complications observed were anemia (39.4%), lupus nephritis (29%) and chronic hypertension (10.5%). In 51,4% of the patients, it was necessary to anticipate delivery and in 13.1%, there was worsening of the renal function. Prematurity occurred in 20% of cases and FGR in 19,4%. Hydroxicloroquine was used in 47,4% of the pregnancies. Lupus nephritis determined a higher occurrence of flare (p<0.05) and a greater need to anticipate delivery (p<0.05). Conclusion: The present study made it possible to relate the main clinical characteristics and the maternal and perinatal outcomes from the pregnant women with SLE treated at the prena... (Complete abstract click electronic access below) / Mestre

Neural stem (progenitor) cells (NPCs) from fetal tissue are an ideal transplantable cell source. They divide rapidly, are able to generate cells of all three neural lineages and do not divide uncontrolled once transplanted into a host organism. To obtain large quantities of cells for transplantation strategies and to eliminate primary cell contaminations, long periods of in vitro cultivation are necessary. Mouse NPCs are a crucial tool for further investigations of neural stem cells because they make the employment of transgenic animals in vivo and cells in vitro possible. So far only short-term expanded fetal mouse NPCs have been shown to generate dopaminergic neurons and it is not clear whether this was due to differentiation or a result of increased survival of primary dopaminergic neurons. The aims of the thesis were to characterize mouse fetal NPCs, to establish the long-term expansion of fetal mouse NPCs and the generation of dopaminergic neurons in long-term expanded fetal mouse NPCs, to investigate the signaling mechanisms involved in the differentiation of mouse fetal NPCs towards the dopaminergic phenotype and to compare short and long-term expanded NPCs. Long-term expanded fetal mesencephalic NPCs could be grown under suspension and adherent culture conditions and showed self- renewing capacity as well as markers typical for NPCs. They could be differentiated into the three major cell types of the nervous system, but suspension NPCs had a larger potential to generate neurons than adherently grown NPCs. Signaling cascades involved in this process were p38 and Erk1/2 mediated. Long-term expanded NPCs did not have the potential to generate neuronal sub-types. Importantly, they did not generate dopaminergic neurons. Mouse fetal NPCs from three different developmental stages (E10, E12, and E14) were employed but were not able to differentiate into dopaminergic neurons using factors known to stimulate in vitro dopaminergic specification. When cultivated in vitro for short periods, fetal mesencephalic NPCs were able to generate dopaminergic neurons. By eliminating all primary Th- positive neurons, FACS-sorting of NPCs proved a de novo generation of dopaminergic neurons, because after cultivation and differentiation of Th- depleted cell solutions dopaminergic neurons were present in the culture. However, these newly generated neurons failed to incorporate BrdU, making a generation without cell division from precursors probable. The precursor population of short cultures differed from long-term expanded cultures suggesting an ‘aging’ effect of in vitro conditions. IL-1 was a potent inducer of the dopaminergic neuronal phenotype in short-term expanded in vitro cultures and was expressed in vitro as well as in vivo at E14. Several important conclusions concerning fetal mouse stem cell behavior could be drawn from the results of this work: Firstly, the results showed for the first time that in fetal mouse mesencephalic NPCs dopaminergic neurons differentiate from precursors without cell division, therefore consuming those progenitors. Therein fetal mouse NPCs differ significantly from rat and human NPCs or respond differently to the same in vitro conditions that need to be optimized for fetal mouse NPCs. Secondly, less committed precursors find appropriate conditions to proliferate but not to generate the more committed DA precursors that are able to generate dopaminergic neurons. The hallmarks of stem cells, self-renewal and multipotentiality, seem to be part of a delicate balance, that, when unsettled, goes in favor of one side without the possibility of returning to the previous status. Further research should focus on two coherent issues: the isolation of more pure populations of progenitors and the more precise characterization of progenitor populations to find out which in vitro conditions need to be provided to keep the balance between proliferation and differentiation potential. The knowledge gained about stem cells this way would help establish cell sources for transplantation strategies. / Stammzellen sind ein wichtiges Werkzeug für regenerative Therapien im Bereich der neurodegenerativen Erkrankungen wie der Parkinson’schen Erkrankung. Ein besonderer Vorteil von Stammzellen gegenüber dem bereits zur Transplantation verwendeten Primärgewebe, ist ihre Fähigkeit zur fortlaufenden Zellteilung, so dass ausreichende Mengen zur Transplantation zur Verfügung stehen. Der Vorteil von fetalen neuralen Stammzellen (fNSZ) ist ihre genomische Stabilität, die dazu führt, dass bei Transplantationen keine Tumore entstehen. Dennoch ist der Großteil ihrer Eigenschaften und Potentiale noch unbekannt und die optimalen Wachstumsbedingungen für eine lange in vitro Kultur und optimale Differenzierung in dopaminerge Neuronen müssen erforscht werden, um bessere Transplantate herzustellen. Insbesondere Stammzellen der Maus sind für die Forschung von immenser Wichtigkeit, da sie die Arbeit mit transgenen Tieren ermöglichen. Die Zielsetzungen dieser Arbeit waren die Charakterisierung der fNSZ der Maus, die Langzeitexpansion und die anschließende Differenzierung in dopaminerge Neurone. Die Signalkaskaden der frühen Differenzierung und die Unterschiede von kurz- und langzeitkultivierten Stammzellen wurden untersucht. Es konnte gezeigt werden, dass fNSZ der Maus nach Langzeitkultivierung in alle Zelltypen des zentralen Nervensystems, also Neuronen und Glia differenzieren und die dabei aktivierten Signalkaskaden p38 und Erk1/2 vermittelt sind. Das Differenzierungspotential zu neuronalen Subtypen (also auch zu dopaminergen Nervenzellen) verloren diese fetalen Stammzellen unter Kulturbedingungen schnell. Das steht im Gegensatz zu fetalen Stammzellen aus Ratte oder dem Menschen, die auch nach langer Kultivierung ihr dopaminerge Potential erhalten. Nur nach Kurzzeitkultivierung waren dopaminerge Neurone nachzuweisen, die jedoch nicht durch Zellteilung aus Vorläuferzellen hervorgegangen waren. Die Eliminierung aller primären Neurone aus der Mittelhirnisolation durch FACS-sorting von Th-Gfp transgenen Mäusen bewies die de novo Generation der dopaminergen Neurone aus Vorläuferzellen ohne Zellteilung während der Kultivierung der Stammzellen. Diese Ergebnisse zeigten, dass in fetalen mesenzephalen NSZ der Maus dopaminerge Neurone von spezialisierten Vorläuferzellen differenzieren, wodurch diese der Kultur verloren gehen. Weniger spezialisierte Vorläuferzellen finden Bedingungen, die ihre Kultivierung ermöglichen, sind aber nicht in der Lage, spezifischere Vorläuferzellen zu bilden. Die Markenzeichen von Stammzellen, Selbsterneuerung (durch Zellteilung) und das Potential, die Zelltypen des Nervensystems zu generieren, scheinen fein balancierte Zustände zu sein, die bei einer Störung nicht wiederherzustellen sind. Die Ergebnisse dieses Projektes sind von großer Bedeutung für die Forschung zur Zellersatztherapie der Parkinson’schen Erkrankung, deren ultimatives Ziel es ist, eine sichere und verlässlich expandierbare Zellquelle zu etablieren, die fähig ist, in dopaminerge Neurone zu differenzieren. Solche Stammzellen würden Bemühungen um Transplantationsstrategien für neurodegenerative Erkrankungen unterstützen und vorantreiben.

Orientador: Lígia Maria Suppo de Souza Rugolo / Resumo: INTRODUÇÃO: O efeito da idade gestacional e do crescimento fetal nos imunocomponentes do leite materno ainda é pouco conhecido. OBJETIVO: Determinar a quantidade de citocinas no colostro em função da idade gestacional e do crescimento fetal. MÉTODO: Estudo transversal, envolvendo mães de recém-nascidos prematuros (PT) e de termo (T), nascidos na Maternidade da Faculdade de Medicina de Botucatu – UNESP, em 2014-2015. Critério de inclusão: gestação única; ausência de diabetes materno, ou uso de medicações/drogas ilícitas, sorologias negativas; recém-nascido com peso adequado (AIG) ou pequeno para idade gestacional (PIG) e sem malformação. Excluídos: não obtenção do colostro, mastite, uso de medicamento pela puérpera. Foram estudados 4 grupos: PT-PIG (n=18), PT-AIG (n=42), T-PIG (n=45), T-AIG (controle, n=42). No colostro coletado entre 24-72 horas pós-nascimento, foram dosadas as citocinas (IL-1β, IL-6, IL-8, IL-10, IL-12 e TNF-α) por citometria de fluxo. Na comparação entre grupos utilizou-se o Qui-quadrado ou teste Exato de Fisher, ANOVA, e Correlação de Pearson. RESULTADOS: As características maternas foram semelhantes nos 4 grupos. A idade gestacional média foi 34 semanas nos prematuros e 39 semanas nos termos. Os níveis de citocinas do colostro não diferiram entre os grupos de termo e pretermo. O grupo T-PIG apresentou maior quantidade de citocinas comparado aos demais. Nos 4 grupos houve correlação entre as citocinas, especialmente nos T-PIG. CONCLUSÃO: A quantidade de c... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: INTRODUCTION: The effect of gestational age and foetal growth in the imunno componentes of human milk is not well stablished. OBJECTIVE: To determine cytokines levels in colostrum according to gestational age and foetal growth. METHOD: Cross-sectional study with mothers of term (T) and preterm (PT) infants, with birth weight appropriate-for-gestational age (AGA) and small-for-gestational age (SGA), born at the Maternity of Botucatu School of Medicine- UNESP, during 2014 -2015. Inclusion criteria were: single gestation; absence of diabetes mellitus; no maternal use of medications or illicit drugs; negative maternal serology; and newborn without malformation. Mothers with mastitis, or use of medication, or failure to obtain colostrum, were excluded. Four groups were studied: PT-SGA (n=18), PT-AGA (n=42), T-SGA (n=45), and T-AGA (control, n=42). Colostrum was collected between 24-72 hours after birth, and cytokines (IL-1β, IL-6, IL-8, IL-10, IL-12 and TNF-α) were measured by flow cytometry. Chi-square or Fisher's exact test, ANOVA and Pearson's correlation were used to evaluate differences among the groups. RESULTS: Maternal characteristics did not differ between groups. The mean gestational age was 34 and 39 weeks for preterm and term groups. Cytokines levels in the colostrum were not different between term and preterm groups. However cytokines levels were significantly higher in colostrum from mothers of T-SGA. A positive correlation between cytokines was found in all groups, ... (Complete abstract click electronic access below) / Mestre

Prenatal alcohol exposure (PAE) alters limb development that may lead to structural and functional abnormalities of the limb reported in children diagnosed with Fetal Alcohol Spectrum Disorder. To determine whether PAE alters the central representation of the forelimb we used the rodent barrel cortex as our model system where it was possible to visualize and quantitatively measure the size of the forepaw representation in the forepaw barrel subfield (FBS) in first somatosensory cortex. In the present study, we examined the effects of PAE on pattern and size of the forepaw and forepaw representation in FBS in neonatal rats at gestational day 32 that corresponds to postnatal day 9. Pregnant Sprague-Dawley rats were chronically intubated with binge doses of ethanol (6 g/kg) from gestational day 1 through gestational day 20. The offspring of the ethanol treated dams comprised the ethanol (EtOH) group. The effect of PAE on the EtOH group was compared with a nutritional-controlled pairfed (PF) group and a normal chowfed (CF) group. The ventral (glabrous) surface area of the forepaw digits, length of digit 2 through digit 5, and the corresponding glabrous forepaw digit representations in the FBS were measured and compared between treatment groups. In rats exposed to in utero alcohol, the sizes of the overall glabrous forepaw and forepaw digits were significantly reduced in EtOH pups compared to CF and PF pups; overall glabrous forepaw area was 11% smaller than CF controls. Glabrous digit lengths were also smaller in EtOH rats compared to CF controls and significantly smaller in digit 2 through digit 4. The glabrous digit representation in FBS was 18% smaller in the EtOH group when compared to the CF treatment. However, PAE did not produce malformations in the forepaw or alter the pattern of the forepaw representation in FBS; instead, PAE significantly reduced both body and brain weights compared to controls. Unexpectedly, little or no correlation was observed between the size of the glabrous forepaw compared to the size of the glabrous forepaw representation in the FBS for any of the treatment groups. The present findings of PAE-related alterations in sensory periphery and the central cortical representation may underlie deficits in sensorimotor integration reported among children with Fetal Alcohol Spectrum Disorder.

Indiana University-Purdue University Indianapolis (IUPUI) / Fetal alcohol spectrum disorder is a disease caused by prenatal alcohol exposure. It is characterized by craniofacial abnormalities, growth retardation, central nervous system defects, learning disabilities and a variety of other minor defects. Even though it affects 2-5% of individuals born every year, very little is known about the mechanisms that cause it. The zebrafish (Danio rerio) presents as an interesting and efficient model for studying this disease. This study provides some insight into the mechanisms underlying observed FASD phenotypes and, more specifically, the transcription factor elf3, which is downregulated in response to ethanol exposure during early embryonic development. Here we show a number of elf3 target genes that are downregulated during early development in response to ethanol exposure. We also give some insight into the expression pattern of elf3 in relation to zygotic genome activation. Translation blocking morpholino oligonucleotides were used to implicate Elf3 in epiboly movements during gastrulation and zebrafish tail development. Taken together these results help to strengthen the zebrafish as a model for FASD in addition to giving greater insight into both the expression pattern and role of Elf3 during development.