Submitted by Karem Dantas (karem.c.dantas@gmail.com) on 2018-10-08T13:33:57Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Teorema de Pick.pdf: 18162866 bytes, checksum: b5924b5d514e4978acc23fddace5a5cb (MD5) / Approved for entry into archive by Marcos Roberto Gomes (mrobertosg@gmail.com) on 2018-10-08T14:49:39Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Teorema de Pick.pdf: 18162866 bytes, checksum: b5924b5d514e4978acc23fddace5a5cb (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-10-09T17:15:54Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Teorema de Pick.pdf: 18162866 bytes, checksum: b5924b5d514e4978acc23fddace5a5cb (MD5) / Made available in DSpace on 2018-10-09T17:15:54Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Teorema de Pick.pdf: 18162866 bytes, checksum: b5924b5d514e4978acc23fddace5a5cb (MD5) Previous issue date: 2018-06-20 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The purpose of this project is Pick’s Theorem, which is about of the calculation of simple polygons with vertices in the points of mesh grid at the plane. The theorem let us to calculate the area using counting, analyzing the points in the edge and the interior of the polygon at the mesh grid. We going to talk about a little bit of Georg Alexander Pick history, beyond the necessary observations to understand the theorem’s demonstration and complexity. At last, we going to show you Pick’s Theorem activities, in the Geoboard and on the GeoGebra software as didactic resources for classroom. / O foco deste trabalho é o Teorema de Pick. Esse teorema se refere ao cálculo de áreas de polígonos simples com vértices nos pontos de uma malha quadriculada no plano, o teorema permite calcular a área usando contagem, analisando os pontos do bordo e do interior do polígono em uma malha quadriculada. Apresentaremos um pouco da história de Georg Alexander Pick, além de observações necessárias para se compreender a demonstração do teorema e sua extensão. Finalmente mostramos atividades com o teorema de Pick no Geoplano e no software GeoGebra como recursos didáticos para a sala de aula.

Cholesterol is essential for many life processes, including correct development, fluidity of cell membranes, production of steroid hormones and bile acids, and is a major component of myelin. Smith-Lemli-Opitz Syndrome (SLOS) and Niemann-Pick disease type C (NPC) are devastating diseases and both involve dysregulation of cholesterol homeostasis. SLOS is caused by a defect in 7-dehydrocholesterol reductase (DHCR7), resulting in increased levels of 7-dehydrocholesterol (7DHC) and decreased levels of cholesterol. On the other hand, NPC is caused by defects in NPC1 or NPC2. These genes encode two lysosomal proteins that are responsible for the transport of cholesterol and other lipids out of lysosomes. Consequently, defects in these proteins results in accumulation of unesterified cholesterol within late endosomes/lysosomes. The severity ranges of both disorders are broad, and no or limited therapeutic options are available. This thesis aimed to establish the incidence and mechanisms behind SLOS and NPC in order to aid development of novel therapeutic interventions. Using a bioinformatics approach, <b>Chapter 2</b> showed that the estimated incidences of classical SLOS and NPC were similar to clinical reports. However, the analysis suggested that a late onset form of NPC1 could be more prevalent. <b>Chapter 3</b> investigated the behavioural phenotype of the SLOS Dhcr7^{T93M/&Delta;3-5} mouse model and it was found that some parallels could be drawn to the behaviour observed in SLOS patients. It also highlighted some defects in neuro-anatomy that could potentially explain certain cognitive defects. <b>Chapter 4</b> explored the suggestion of oxidative stress in the Dhcr7^{T93M/&Delta;3-5} mouse model. However, this study did not support a role for oxidative stress in this model. Genetic insights were generated via an RNAseq study on SLOS and NPC patient fibroblasts, which suggested that the WNT signalling pathway could play a role in the pathogenesis of SLOS. This pathway was also highlighted when the cells were treated with miglustat, the only approved therapy for NPC. However, the main pathway apparently affected by this drug in both SLOS and NPC was cell proliferation.

Niemann-Pick type C2 (NPC2) protein is a soluble protein that binds unesterified cholesterol. The protein helps transport unesterified cholesterol in tandem with the membrane protein Niemann-Pick type C1 (NPC1). Defects in either of proteins can cause Niemann-Pick type C disease (NPC), which results in the accumulation of unesterified cholesterol and lipids in the late endosome and lysosome. NPC is an autosomal recessive lysosomal storage disease affecting 0.35~2.20 per 100,000 people. Here we present the structural analysis of the human NPC2 glycoprotein, including expression, purification, functional analysis, homology modeling, and crystallographic studies. Human NPC2 was expressed from baculovirus-infected Trichoplusia ni (Tn5) insect cells. The construct contained a hexahistidine purification epitope tag, and the protein was purified using Nickel affinity column chromatography. The purified protein was used in binding studies with dehydroergosterol (DHE), showing that human NPC2 was functional. Using the structure of bovine NPC2, we made a homology model and mapped the human mutations onto the model. Some modeled proteins, such as the V30M and S67P variants, are unclear as to how they lead to disease, thus a structure of the human protein would be informative. Crystallization screens of human NPC2 were performed and led to crystals with a needle-like morphology, which diffracted to 4Å resolution. The structure of human NPC2 will be useful for understanding the mechanism of cholesterol binding and trafficking in cells, and to better understand the human metabolic disease NPC.

Two-neutron pick up reactions have been performed on targets of 149Sm and radioactive 151Sm using 18 MeV protons. The outgoing tritons from the 151Sm target were analyzed with a magnetic spectrograph at 16 angles between 6 degrees and 8 degrees. Unlike the two-neutron transfer data on neighbouring even-even targets, the angular distributions indicated l=o transitions to many levels in the final nucleus. Partial angular distributions for the 149Sm (p, t) 147Sm reaction were obtained, but showed only one strong l=o transition populating the 147Sm ground state. In addition, spectra from the 152Sm (p, t) reaction were measured at several angles in order to provide normalization to previous results. / Thesis / Master of Science (MSc)

A hipótese do presente estudo sugere que a baixa fertilidade de vacas Holandesas (Bos taurus) repetidoras de serviço (RS) está relacionada à baixa qualidade oocitária que, por sua vez, é associada ao quadro de resistência periférica a insulina (RPI). Ainda, a indução de lactação (IL) em vacas Holandesas (Bos taurus) RS pode reverter o quadro de RPI e, consequentemente, melhorar a qualidade oocitária e a produção in vitro de embriões (PIVE). Para testar a hipótese proposta, este estudo objetivou avaliar o efeito da fase da lactação e da gestação [Exp. 1], o efeito da secagem de RS [Exp. 2] e o efeito da IL [Exp. 3] sobre a RPI, a qualidade oocitária e a PIVE em vacas da raça Holandesa (Bos taurus). Nos três estudos foram realizados testes de tolerância à glicose (TTG) para avaliar a RPI através do perfil hormonal sérico de insulina e glicose. Além disso, avaliou-se o perfil bioquímico sérico e folicular e a qualidade oocitária através da PIVE. Verificou-se que, em resposta à infusão de 0,3mg glicose/kg PV, as vacas RS no final da lactação secretaram 53% mais insulina e captaram 40% menos de glicose quando comparadas as vacas no terço inicial de lactação (Exp 1). Esses achados são indicativos do estabelecimento do quadro de RPI nas vacas RS em lactação. Durante o período seco, as vacas RS secretaram 96% mais insulina e captaram 56% menos glicose que as vacas no terço inicial da lactação e as vacas RS em lactação, respectivamente (Exp. 2). Ainda, as vacas com lactação induzida secretaram 11% menos insulina e captaram a mesma quantidade de glicose que vacas paridas em fases semelhantes de lactação (Exp 3), demonstrando que o protocolo de IL foi eficiente em alterar o perfil metabólico, revertendo o quadro de RPI presente nas vacas RS. Nos Exp. 1, 2 e 3 foram verificadas maiores concentrações plasmática de triglicérides (TG; P &lt; 0,05), colesterol total (COL; P < 0,05) e LDL (P < 0,05) no soro sanguíneo em vacas RS quando comparadas com vacas no terço inicial de lactação. Durante o período seco (Exp. 2 e 3), observou-se incremento desses metabólitos, destacando aumento na concentração de TG (P < 0,05), COL (P < 0,05) e LDL (P < 0,05) plasmático em vacas secas quando comparadas as vacas em lactação [início e final (RS) da lactação]. No liquido folicular foram observadas variações no perfil bioquímico para COL e TG. Nos Exp. 1, 2 e 3, verificou-se que vacas RS possuem maior concentração de TG (P < 0,05) e COL (P < 0,05) no fluido folicular do que vacas no terço inicial de lactação. Contrariando a hipótese inicialmente proposta, as vacas RS em lactação e as vacas secas apresentaram maior taxa de blastocisto (P < 0,05) e número de blastocistos por OPU (P < 0,05) que as vacas no terço inicial de lactação (Exp. 1, 2 e 3). Através do perfil de insulina circulante em resposta ao TTG foi possível demonstrar o estabelecimento do quadro de RPI em vacas RS (P < 0,05). Além disso, constatou-se agravamento da RPI em vacas secas (P < 0,05). Esse quadro foi associado ao aumento do escore de condição corporal (P < 0,05) e do peso vivo (kg; P < 0,05) nas vacas RS. Em conclusão, não foi verificada associação negativa entre RPI, qualidade oocitária e PIVE em vacas Holandesas (Bos taurus) RS. Apesar da indução de lactação em vacas Holandesas (Bos taurus) RS alterar o metabolismo e diminuir o quadro de RPI, não foi verificado efetivo positivo na qualidade oocitária e na PIVE. / The hypothesis of the present study suggests that low fertility of repeat breeders (RB) Holstein (Bos taurus) cows is related to low oocyte quality, which is associated with peripheral insulin resistance (PIR). Also, induction of lactation (IL) in RB Holstein (Bos taurus) cows can revert PIR and, consequently, improve oocyte quality and in vitro embryo production (IVEP). In order to test the proposed hypothesis, the objective of this study was to evaluate the effect of phase of lactation and gestation [Exp. 1], effect of drying RB [Exp. 2] and effect of IL [Exp. 3] on PIR, oocyte quality and IVEP of Holstein (Bos taurus) cows. In all three studies, glucose tolerance tests (GTT) were performed to evaluate PIR through the serum hormonal insulin and glucose profile. In addition, we evaluated the serum and follicular biochemical profile and oocyte quality through IVEP. It was verified that, in response to a 0.3mg glucose/kg of body weight (BW), RB cows at the end of lactation secreted 53% more insulin and captured 40% less glucose when compared to cows in the initial third of their lactation (Exp. 1). These findings are indicative of the establishment of PIR in RB lactating cows. During the dry period, RB cows secreted 96% more insulin and captured 56% less glucose than cows in the initial third of their lactation and RB lactating cows, respectively (Exp. 2). Also, cows with induced lactation secreted 11% less insulin and captured the same amount of glucose than calved cows in similar lactation phase (Exp. 3), demonstrating that the IL protocol was efficient to alter the metabolic profile, reverting PIR present in RB cows. In Exp. 1, 2 and 3 higher plasmatic concentrations of triglycerides (TG; P<0.05), total cholesterol (COL; P<0.05) and LDL (P<0.05) were verified in the blood serum in RB cows when compared to cows in the initial third of their lactation. During the dry period (Exp. 2 and 3), we observed the increment of these metabolites, and a notable elevation of the plasmatic TG (P < 0.05), COL (P < 0.05) and LDL (P < 0.05) in dry cows when compared to lactating cows [beginning and end (RB) of lactation]. In the follicular fluid, it was possible to observe variations in the biochemical profile for COL and TG. In Exp. 1, 2 and 3, it was verified that RB cows have higher concentration of TG (P < 0.05) and COL (P < 0.05) in the follicular fluid than cows in the initial third of their lactation. Contrary to the initially proposed hypothesis, RB lactating cows and dry cows presented higher blastocyst rate (P<0.05) than cows in the initial third of lactation (Exp. 1, 2 and 3). Through the circulating insulin profile in response to the GTT it was possible to demonstrate the establishment of PIR in RB cows (P<0.05). Also, it was observed worsening of the PIR in dry cows (P<0.05). This condition was associated with an increase in body condition score (P<0.05) and BW (kg; P<0.05) in RB cows. In conclusion, no negative association between PIR, oocyte quality and IVEP was observed in RB Holstein (Bos taurus) cows. Although induction of lactation in RB Holstein (Bos taurus) cows altered the metabolism and reduced PIR, no positive effect was observed in oocyte quality and IVEP.

A hipótese do presente estudo sugere que a baixa fertilidade de vacas Holandesas (Bos taurus) repetidoras de serviço (RS) está relacionada à baixa qualidade oocitária que, por sua vez, é associada ao quadro de resistência periférica a insulina (RPI). Ainda, a indução de lactação (IL) em vacas Holandesas (Bos taurus) RS pode reverter o quadro de RPI e, consequentemente, melhorar a qualidade oocitária e a produção in vitro de embriões (PIVE). Para testar a hipótese proposta, este estudo objetivou avaliar o efeito da fase da lactação e da gestação [Exp. 1], o efeito da secagem de RS [Exp. 2] e o efeito da IL [Exp. 3] sobre a RPI, a qualidade oocitária e a PIVE em vacas da raça Holandesa (Bos taurus). Nos três estudos foram realizados testes de tolerância à glicose (TTG) para avaliar a RPI através do perfil hormonal sérico de insulina e glicose. Além disso, avaliou-se o perfil bioquímico sérico e folicular e a qualidade oocitária através da PIVE. Verificou-se que, em resposta à infusão de 0,3mg glicose/kg PV, as vacas RS no final da lactação secretaram 53% mais insulina e captaram 40% menos de glicose quando comparadas as vacas no terço inicial de lactação (Exp 1). Esses achados são indicativos do estabelecimento do quadro de RPI nas vacas RS em lactação. Durante o período seco, as vacas RS secretaram 96% mais insulina e captaram 56% menos glicose que as vacas no terço inicial da lactação e as vacas RS em lactação, respectivamente (Exp. 2). Ainda, as vacas com lactação induzida secretaram 11% menos insulina e captaram a mesma quantidade de glicose que vacas paridas em fases semelhantes de lactação (Exp 3), demonstrando que o protocolo de IL foi eficiente em alterar o perfil metabólico, revertendo o quadro de RPI presente nas vacas RS. Nos Exp. 1, 2 e 3 foram verificadas maiores concentrações plasmática de triglicérides (TG; P &lt; 0,05), colesterol total (COL; P < 0,05) e LDL (P < 0,05) no soro sanguíneo em vacas RS quando comparadas com vacas no terço inicial de lactação. Durante o período seco (Exp. 2 e 3), observou-se incremento desses metabólitos, destacando aumento na concentração de TG (P < 0,05), COL (P < 0,05) e LDL (P < 0,05) plasmático em vacas secas quando comparadas as vacas em lactação [início e final (RS) da lactação]. No liquido folicular foram observadas variações no perfil bioquímico para COL e TG. Nos Exp. 1, 2 e 3, verificou-se que vacas RS possuem maior concentração de TG (P < 0,05) e COL (P < 0,05) no fluido folicular do que vacas no terço inicial de lactação. Contrariando a hipótese inicialmente proposta, as vacas RS em lactação e as vacas secas apresentaram maior taxa de blastocisto (P < 0,05) e número de blastocistos por OPU (P < 0,05) que as vacas no terço inicial de lactação (Exp. 1, 2 e 3). Através do perfil de insulina circulante em resposta ao TTG foi possível demonstrar o estabelecimento do quadro de RPI em vacas RS (P < 0,05). Além disso, constatou-se agravamento da RPI em vacas secas (P < 0,05). Esse quadro foi associado ao aumento do escore de condição corporal (P < 0,05) e do peso vivo (kg; P < 0,05) nas vacas RS. Em conclusão, não foi verificada associação negativa entre RPI, qualidade oocitária e PIVE em vacas Holandesas (Bos taurus) RS. Apesar da indução de lactação em vacas Holandesas (Bos taurus) RS alterar o metabolismo e diminuir o quadro de RPI, não foi verificado efetivo positivo na qualidade oocitária e na PIVE. / The hypothesis of the present study suggests that low fertility of repeat breeders (RB) Holstein (Bos taurus) cows is related to low oocyte quality, which is associated with peripheral insulin resistance (PIR). Also, induction of lactation (IL) in RB Holstein (Bos taurus) cows can revert PIR and, consequently, improve oocyte quality and in vitro embryo production (IVEP). In order to test the proposed hypothesis, the objective of this study was to evaluate the effect of phase of lactation and gestation [Exp. 1], effect of drying RB [Exp. 2] and effect of IL [Exp. 3] on PIR, oocyte quality and IVEP of Holstein (Bos taurus) cows. In all three studies, glucose tolerance tests (GTT) were performed to evaluate PIR through the serum hormonal insulin and glucose profile. In addition, we evaluated the serum and follicular biochemical profile and oocyte quality through IVEP. It was verified that, in response to a 0.3mg glucose/kg of body weight (BW), RB cows at the end of lactation secreted 53% more insulin and captured 40% less glucose when compared to cows in the initial third of their lactation (Exp. 1). These findings are indicative of the establishment of PIR in RB lactating cows. During the dry period, RB cows secreted 96% more insulin and captured 56% less glucose than cows in the initial third of their lactation and RB lactating cows, respectively (Exp. 2). Also, cows with induced lactation secreted 11% less insulin and captured the same amount of glucose than calved cows in similar lactation phase (Exp. 3), demonstrating that the IL protocol was efficient to alter the metabolic profile, reverting PIR present in RB cows. In Exp. 1, 2 and 3 higher plasmatic concentrations of triglycerides (TG; P<0.05), total cholesterol (COL; P<0.05) and LDL (P<0.05) were verified in the blood serum in RB cows when compared to cows in the initial third of their lactation. During the dry period (Exp. 2 and 3), we observed the increment of these metabolites, and a notable elevation of the plasmatic TG (P < 0.05), COL (P < 0.05) and LDL (P < 0.05) in dry cows when compared to lactating cows [beginning and end (RB) of lactation]. In the follicular fluid, it was possible to observe variations in the biochemical profile for COL and TG. In Exp. 1, 2 and 3, it was verified that RB cows have higher concentration of TG (P < 0.05) and COL (P < 0.05) in the follicular fluid than cows in the initial third of their lactation. Contrary to the initially proposed hypothesis, RB lactating cows and dry cows presented higher blastocyst rate (P<0.05) than cows in the initial third of lactation (Exp. 1, 2 and 3). Through the circulating insulin profile in response to the GTT it was possible to demonstrate the establishment of PIR in RB cows (P<0.05). Also, it was observed worsening of the PIR in dry cows (P<0.05). This condition was associated with an increase in body condition score (P<0.05) and BW (kg; P<0.05) in RB cows. In conclusion, no negative association between PIR, oocyte quality and IVEP was observed in RB Holstein (Bos taurus) cows. Although induction of lactation in RB Holstein (Bos taurus) cows altered the metabolism and reduced PIR, no positive effect was observed in oocyte quality and IVEP.

In dieser Arbeit beleuchten wir die subzelluläre Lokalisation von Flotillin unter Einfluss von Cholesterin. Wir zeigen, dass die zelluläre Lokalisation abhängig vom Cholesterinniveau der Zelle ist. Eine Cholesterindepletion bringt Flotillin in die Plasmamembran, sowie umgekehrt eine Überversorgung mit Cholesterin Flotillin in cholesterinhaltige, endosomale Strukturen führt. Dabei ist die Umverteilung abhängig von der Integrität des Zytoskeletts. Außerdem zeigt die vorliegende Arbeit, dass die Umverteilung von Flotillin von der Plasmamembran hin zu endosomalen, intrazellulären Kompartimenten abhängig vom Vorhandensein von zwei putativen Cholesterinbindungs-/Interaktionsdomänen ist. Aus den gewonnenen Daten ergeben sich weiterhin Hinweise, dass Cholesterin an Flotillin gebunden in das späte Endosomen transportiert wird. In weiterführenden Versuchen unserer Gruppe zeigte sich, dass die exosomale Freisetzung von Cholesterin bei ansteigenden zellulären Cholesterinkonzentrationen erhöht wird und dass die exosomale Cholesterinfreisetzung von Flotillin abhängig ist. Die Daten deuten auf eine mögliche Rolle von Flotillin und Exosomen bei der zellulären Cholesterinhomöostase hin.

Bakgrund: En organisations olika avdelningar är ständigt i behov av rätt information vid rätt tillfälle. Information i realtid är av stor vikt för en distributör då deras överlevnad är beroende av kunden. Plockprocessen i ett lager hos ett företag utgör en stor kostnad och är en viktig aktivitet att förbättra för att bli mer lönsam och effektivare. Syfte: Syftet med studien är att skapa ett beslutsunderlag gällande implementering av ett digitalt verktyg för en manuell plockprocess samt ekonomisk aspekt och förändringsarbete. Det generella beslutsunderlaget ska kunna tillämpas av företag som agerar grossister inom distribution och som vill digitalisera plockprocessen för att få information i realtid. Metod: Studien omfattas av en kvalitativ forskningsmetod där fallstudie har utförts. Forskarna har tillämpat jämförande studie gällande det digitala verktygen. Empiri har samlats in genom snöbollsurval och använt strukturerade och semi- strukturerade intervjuer samt utfört observationer. Resultat: Studien har analyserat tre digitala verktyg utifrån Hall Mibas förutsättningar. Studien har analyserat de lämpliga verktygen utifrån ett finansiellt perspektiv med hjälp av en Pay-back kalkyl samt testat resultatet med en känslighetsanalys. En sammanställning av en möjlig implementeringsprocess har förklaras med hjälp av ASAP Roadmap och forskarna har utformat en checklista för företag att följa genom implementeringens gång.

O objetivo do primeiro estudo foi avaliar a produção in vitro de embriões (PIVE) em vacas holandesas não lactantes submetidas a aspiração oocitária (OPU) posteriormente ao protocolo de superestimulação folicular similar ao descrito por Nivet et al. (2012) em comparação à realização da OPU em dia aleatório do ciclo estral. Para tal, vacas holandesas não lactantes e não gestantes foram distribuídas aleatoriamente em delineamento tipo crossover em Controle (n = 35), em que as vacas não foram tratadas com FSH, mas submetidas a uma sessão de aspiração em dia aleatório do ciclo estral; ou p-FSH (n = 35), em que, 36 horas após a OPU para sincronização da onda folicular, as vacas foram tratadas com p-FSH por 3 dias e 44 horas após, submetidas a sessões de OPU. O número total de complexos cumulusoócito (CCO) recuperados e o número de oócitos viáveis foram semelhantes entre os grupos controle e p-FSH. Além disso, não houve aumento na proporção de CCO viáveis (CCO viáveis / CCO total recuperado). Da mesma forma, não se detectaram diferenças no número de embriões / sessão de OPU e taxa de blastocistos. O protocolo de superestimulação folicular não melhorou a PIVE em vacas holandesas não lactantes. O experimento 2 testou a hipótese de que vacas leiteiras de alta produção se tornam cada vez mais resistentes à insulina com o avançar da lactação, e consequentemente, a qualidade do oócito é comprometida. Foram utilizadas vacas holandesas em 50 (51,5 ± 3,7; n = 30), 100 (102,3 ± 9,4; n = 30) e 150 (154,5 ± 18,9; n = 30) dias em lactação (DEL). Durante o teste de tolerância à glicose (TTG), não houve diferença entre grupos para qualquer variável relacionada à glicose circulante. No entanto, medidas de insulina circulante foram diferentes em vacas aos 150 DEL em comparação com 50 ou 100 DEL, tais como: maior insulina basal, pico, &Delta; máx de insulina e AUC 5-60. Porém, não houve diferença entre os grupos para o número ou percentagem de oócitos viáveis. Assim, as vacas desenvolveram resistência à insulina com o aumento do DEL. No entanto, o aumento da resistência à insulina não foi associado com alteração detectável na qualidade dos oócitos aspirados de folículos pequenos e médios. O experimento 3 foi para avaliar se o aumento de insulina circulante durante os períodos de pré e pós desvio folicular aumenta o desenvolvimento inicial e final, do folículo, bem como do corpo lúteo (CL). Além disso, por induzir a ovulação de um folículo maior, o CL resultante de vacas com alta insulina circulante também é maior e mais esteroidogênico, refletindo em maiores concentrações circulantes de progesterona (P4). O delineamento experimental utilizado foi o quadrado latino em arranjo fatorial 2x2, em quatro grupos experimentais: 1) CC = água pré e pós desvio folicular (n = 16); 2) CP = água e propilenoglicol (PPG) pré e pós desvio folicular, respectivamente (n = 16); 3) PC = PPG e água pré e pós desvio folicular, respectivamente (n = 16) e 4) PP = PPG pré e pós desvio folicular (n = 16). O aumento agudo e transitório, durante os períodos de pré e pós desvio não aumentou o desenvolvimento folicular, luteal e concentrações plasmáticas de P4. / The aim of the first study was to evaluate the in vitro embryo production (IVEP) in nonlactating Holstein cows subjected to ovum pick-up (OPU) after ovarian superstimulation with a protocol similar to that described by Nivet et al. (2012) in comparison with OPU at a random day of the estrous cycle. Nonlactating Holstein cows were randomly assigned in a crossover design to: Control (n = 35) in which cows were not treated with p-FSH, but subjected to OPU at a random day of the estrous cycle; or p-FSH (n = 35), in which, 36 hours after OPU to synchronize follicle wave, the cows were treated with p-FSH for 3 days and 44 hours later, subjected to OPU sessions. The total number of cumulus-oocyte complex (COC) recovered and the number of viable oocytes were similar between control and p-FSH groups. In addition, there was no increase in the proportion of viable COC (viable COC / overall COC recovered). Likewise, we detected no differences in the number of embryos / OPU session and blastocyst rate. Follicle superstimulation protocol with p-FSH did not improve IVEP in nonlactating Holstein cows. Experiment 2 tested the hypothesis that high-producing dairy cows become increasingly resistant to insulin with advancing lactation, and consequently oocyte quality is compromised. We used Holstein cows at 50 (51.5 ± 3.7; n = 30), 100 (102.3 ± 9.4; n = 30) and 150 (n = 30 154.5 ± 18.9) days in milk (DIM). During the glucose tolerance test (GTT), there was no difference between groups for any variable related to circulating glucose. However, circulating insulin measurements such as basal insulin, peak insulin, &Delta; max and AUC 5-60 were higher for cows at 150 DIM. Nevertheless, there was no difference between groups for the number or percentage of viable oocytes. Therefore, although cows developed insulin resistance with increasing DIM, this has not been associated with detectable change in the quality of oocytes aspirated from small and medium follicles. The third experiment assessed whether the increase in circulating insulin during periods of pre- and post-follicle deviation increases the initial and final follicle size and corpus luteum (CL) volume. Moreover, by inducing ovulation of greater follicles, resulting in greater CL, cows with high circulating insulin also have higher circulating progesterone (P4). The experimental design was a Latin square in a 2x2 factorial arrangement in four groups: 1) CC = water pre and post follicle deviation (n = 16); 2) CP = water pre and propylene glycol (PPG) post follicle deviation (n = 16); 3) PC = PPG and water pre and post follicle deviation, respectively (n = 16), 4) PP = PPG pre and post follicle deviation (n = 16). Acute and transient circulating insulin increase during periods of pre and post follicle deviation has not affected follicle development, luteal volume or plasma concentrations of P4.

Niemann-Pick tipo C (NPC) é uma doença de depósito lisossomal, ocasionada por alterações no tráfico de colesterol não esterificado, decorrente de alterações bialélicas nos genes NPC1 ou NPC2, ambos definindo uma doença autossômica recessiva, progressiva e irreversível, caracterizada por manifestações viscerais, neurológicas e psiquiátricas, não necessariamente combinadas. Com o propósito de descrever as características fenotípicas e genotípicas de pacientes com NPC, objetivou-se relatar dados demográficos, formas clínicas classificadas por idade, sinais e sintomas neurológicos e psiquiátricos, achados de ressonância magnética (RM) de encéfalo e ultrassonografia de abdome, assim como teste de filipin, mutações observadas e o tratamento com N-butyldeoxynojirimycin (Miglustat). De uma casuística de 12 pacientes atendidos entre 2000-2014, por revisão de prontuários, no Ambulatório de Neurogenética do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto, havia 7 mulheres e 5 homens, idade média de 20 anos (entre 2 e 42), sendo dez pacientes da etnia branca e dois mulatos, procedentes de 3 estados brasileiros: São Paulo, Mato Grosso do Sul e Minas Gerais. As formas clínicas identificadas foram infantil, juvenil e adulto. A idade do primeiro sintoma neurológico ocorreu entre 1 e 27 anos (media 9,5). Dentre os achados viscerais, dois pacientes encontravam-se assintomáticos, os demais apresentaram icterícia prolongada/colestase, hepatomegalia e esplenomegalia. Todos os pacientes apresentaram em diferentes momentos da evolução manifestações neuropsiquiátricas, tais como: paralisia do olhar vertical, ataxia/quedas, epilepsia, mioclonias, distonia, disartria, disfagia, fraqueza muscular, espasticidade, declínio cognitivo/demência, sintomas psicóticos, atraso escolar, distúrbio de comportamento, cataplexia gelástica e hipotonia neonatal. A idade de diagnóstico variou de 0 a 41 anos, com uma média de 14,5 anos. O tempo entre a idade do primeiro sintoma neurológico e o diagnóstico da doença variou de 0 a 14 anos, tempo médio de 5,3 anos. O teste de Filipin demonstrou seis resultados positivos e seis variantes. A RM de encéfalo apresentou três diferentes tipos de alteração: atrofia cerebral em 6 casos, atrofia cerebelar e desmielinização em 7. A ultrassonografia de Abdome resultou em três alterações: hepatomegalia em 8, esplenomegalia em 10 e hepatoesplenomegalia em 8. O resultado do teste genético molecular em 11 pacientes evidenciou alterações no gene NPC1 e uma paciente não possuía o resultado. A mutação c.3104C>T foi a mais frequente, em oito pacientes; c.3548G>A, de significado incerto, em um paciente, e, as demais mutações encontradas: c.3493G>A, c.3019C>G. O tratamento com N-butyl deoxynojirimycin (Miglustat) foi realizado por todos os pacientes, o tempo entre o diagnóstico e o início da medicação variou de 0 a 9 anos, média de 2,9 anos. Concluímos que o registro da doença NPC deve ser feito através de uma coleta de dados detalhada e contínua, pois sua heterogeneidade fenotípica e genotípica sugerem um número subestimado de casos, não só por sua raridade, mas também pelo desconhecimento da doença, já que há poucos grupos estudados e publicados. O seu reconhecimento precoce, associado ao adequado manejo clínico, podem retardar a progressão implacável da doença e aumentar a expectativa de vida dos pacientes. / Niemann-Pick type C (NPC) is a lysosomal storage disease caused by abnormal unesterified cholesterol trafficking, resulting from biallelic changes in NPC1 or NPC2 genes, both defining an autosomal recessive progressive and irreversible disease characterized by visceral, neurological and psychiatric manifestations, not necessarily combined. In order to describe the phenotypic and genotypic characteristics of patients with NPC, this work aimed to report demographic data, clinical forms classified by age, neurological and psychiatric signs and symptoms, brain magnetic resonance imaging (MRI) and abdominal ultrasound findings, Filipin test, the gene mutations, and the treatment with N-butyldeoxynojirimycin (Miglustat). A series of 12 patients were studied, treated between 2000-2014, by review of medical records of the Neurogenetics Clinic at the Hospital of Clinics, Ribeirão Preto Medical School, Brazil. There were 7 women and 5 men, mean age 20 years (from 2 to 42); 10 caucasian and 2 mulattos, coming from three Brazilian states: São Paulo, Mato Grosso do Sul and Minas Gerais. Infantile, Juvenile and adult clinical forms were identified. The age of the first neurological or psichiatric symptoms occurred between 1 and 27 years (mean 9.5). Among the visceral findings, two patients were asymptomatic, and the others had prolonged jaundice / cholestasis, hepatomegaly and splenomegaly. All patients had at different times of evolution symptoms, such as paralysis of vertical gaze, ataxia/falls, epilepsy, myoclonus, dystonia, dysarthria, dysphagia, muscle weakness, spasticity, cognitive decline/dementia, psychotic symptoms, school delay, disorders behavior, gelastic cataplexy and neonatal hypotonia. Age at diagnosis ranged from 0 to 41 years, with a mean of 14.5 years. The interval between the first signs of the disease and the onset of treatment ranged from zero to 14 years, with an average of 5.3 years. Filipin test resulted six positive and six variant form. The MRI scans showed three different types of changes: brain atrophy in 6 cases, cerebellar atrophy in 7 and demyelination in 7. Abdominal ultrasound revealed 8 patients with hepatomegaly, 10 with splenomegaly and 8 hepatosplenomegaly. The results of the molecular genetic testing on 11 patients showed changes in NPC1 gene and a patient did not have the result. Mutation c.3104C>T was more frequent in eight patients; c.3548G>A, of uncertain significance in a patient, and other mutations found: c.3493G>A and c.3019C>G. Treatment with N-butyl deoxynojirimycin (Miglustat) was carried by all patients; the time between diagnosis and beginning of the treatment ranged from 0 to 9, with an average of 2.9 years. We conclude that NPC disease registry should be done through a collection of detailed and continuous data because their phenotypic and genotypic heterogeneity suggest an underestimated number of cases, not only for its rarity but also by unawareness about the disease, and the fact that there are few published studies. The early recognition, coupled with appropriate clinical management, may slow the progression of the disease and increase life expectancy of patients.