Introdução: Diabetes Mellitus é a alteração metabólica mais comum na gestação, com prevalência variável de acordo com a população e métodos diagnósticos. Diabetes monogenética ou Diabetes do adulto de início no jovem (Maturity-Onset Diabetes of the Young - MODY) consiste em um subtipo ocasionado por defeito primário na secreção de insulina determinado por herança autossômica dominante, sendo responsável por aproximadamente 1 a 2% dos casos. O diagnóstico genético de gestantes e neonatos acometidos pode propiciar manejo específico, possibilitando melhor prognóstico evolutivo da doença em curto e longo prazo. Objetivos: Analisar alterações moleculares do gene GCK em gestantes com diagnóstico de Diabetes Mellitus e seus neonatos. Metodologia: Estudo transversal, com amostragem consecutiva, sendo incluídas gestantes diabéticas em seguimento no Ambulatório de Endocrinopatias em Obstetrícia do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, no período de agosto de 2013 a dezembro de 2015. Amostras sanguíneas maternas e de cordão umbilical foram colhidas no momento do parto e enviadas para extração de DNA e reação em cadeia de polimerase (PCR) para identificação de alterações moleculares no gene GCK. Resultados: Foram encontrados duas mutações que geraram dois polimorfismos no gene da glucoquinase, respectivamente em 13 e em sete pacientes; e uma mutação deletéria que cursou com diagnóstico de MODY GCK em dois pacientes (uma amostra materna e uma de neonato), totalizando alterações em 10,6% dos pacientes. Este diagnóstico permitirá intervenções profiláticas e terapêuticas, interferindo beneficamente na evolução natural da doença. / ntroduction: Diabetes mellitus is the most common metabolic disorder in pregnancy, with prevalence varying according to the population and diagnostic methods. Monogenetic diabetes or Maturity - Onset Diabetes of the Young (MODY) consists of a subtype caused by primary defect in insulin secretion, which is determined by autosomal dominant inheritance and it is responsible for approximately 1-2 % of cases. Genetic diagnosis of affected pregnant women and newborns can provide specific management enabling better evolutionary prognosis of the disease in short and long term. Objectives: To analyze the molecular changes in the GCK gene in pregnant women diagnosed with Diabetes Mellitus and their neonates. Methods: Cross-sectional study with consecutive sampling, that included diabetic pregnant women in follow-up at the Clinical Hospital of the Ribeirão Preto Medical School, University of São Paulo, from August 2014 to December 2015. Maternal and umbilical cord blood samples was collected at time of delivery and sent for DNA extraction and polymerase chain reaction (PCR) to identify molecular changes in the GCK gene. Results: We found mutations that generated two different polymorphisms in the glucokinase gene, one in 13 patients, and the other in seven patients; and a deleterious mutation that was diagnosed with MODY GCK in two patients (a maternal sample and in a neonate), resulting in change in 10,6% of the patients. This diagnosis will determine prophylactic and therapeutic interventions interfering beneficially in the natural evolution of the disease.

Abstract Dietary vegetables and fruit may play a significant role in atherosclerosis. We investigated the effects of a high intake of vegetables, berries, and citrus fruit along with a diet low in total and saturated fat on plasma concentrations of lipids, lipoprotein(a), antioxidants, oxidised LDL (OxLDL), folate, homocysteine, and on serum paraoxonase-1 activity. We also determined whether gene polymorphisms affect diet response of plasma homocysteine and serum paraoxonase-1 activity. Thirty-seven healthy females consumed two diets (low and high vegetable diets) in a controlled crossover intervention. The plasma measurements were determined at the baseline and at the end of diet periods. The average plasma concentrations of total, LDL, and HDL cholesterol were 5.0 mmol/l, 2.8 mmol/l, and 1.7 mmol/l, respectively, on the low vegetable diet, and decreased by 8%, 8%,and 5%, respectively, in response to the high vegetable diet. The high vegetable diet increased the plasma concentrations of alpha-carotene, beta-carotene, lutein-zeaxanthin, beta-cryptoxanthin, and vitamin C by 133%, 134%, 107%, 65%, and 25%, respectively, compared with the low vegetable diet. There were no differences in the plasma concentrations of OxLDL between the low and high vegetable diets. The mean serum paraoxonase-1 activity was lower at the end of the high vegetable diet (226 U/l) than at the end of the low vegetable diet (240 U/l). Subjects having a genotype with high baseline paraoxonase-1 activity showed the most extensive reduction in their serum enzyme activities. The high vegetable diet enhanced the serum and erythrocyte folate concentrations by 78% and 14%, respectively, and reduced the plasma homocysteine by 13% compared with the low vegetable diet. The dietary treatment was effective even among subjects homozygous for C677T mutation in methylenetetrahydrofolate reductase gene, who are susceptible to high homocysteine levels. In conclusion, a high intake of vegetables, berries, and citrus fruit resulted in reduced plasma total and LDL cholesterol concentrations and enhanced plasma antioxidant levels. The high vegetable diet also effectively increased blood folate concentrations and reduced plasma homocysteine concentration.

Abstract Chlamydia pneumoniae is an obligatory intracellular pathogen that causes upper and lower respiratory tract infections. Like other Chlamydial species, also C. pneumoniae has a tendency to cause persistent infections, which have been associated with different cardiovascular, neurological, and respiratory diseases. In addition, a few studies have reported an association between C. pneumoniae seropositivity and an elevated body mass index (BMI), and it has been shown that C. pneumoniae is capable of infecting preadipocytes and adipocytes. The main aims of this study were to study if certain gene polymorphisms regulate the serum levels of innate immunity and inflammation proteins, and if the polymorphisms are associated with markers of C. pneumoniae infection; to compare different methods in detection of C pneumoniae in atherosclerotic tissue; and to study if serum levels of chlamydial LPS (cLPS) are associated with BMI. The serum levels of inflammatory and innate immunity markers, namely interleukin 6 (IL-6), C-reactive protein (CRP), LPS-binding protein (LBP), and soluble CD14, in apparently healthy individuals were found to correlate with each other and possibly be regulated by the polymorphisms of genes important in inflammation and innate immunity. Especially the serum LBP levels may be regulated by the LBP (rs2232618) and toll-like receptor 4 (rs4986790) polymorphisms. The IL-6 (rs1800795) polymorphism was found to be associated with C. pneumoniae antibody positivity. C. pneumoniae DNA and cLPS could be found from atherosclerotic tissue. A new, cLPS enzyme immunoassay method was developed in this study, and it might provide a standardized, commercial method for the detection of chlamydia in tissue samples, if the sensitivity of the method could be increased e.g. by testing multiple pieces of tissue. In situ hybridization method was found to be complicated by technical problems and the repeatability of polymerase chain reaction was poor. C. pneumoniae IgG positivity and elevated serum cLPS and CRP levels were associated with an elevated BMI. There was also a strong association between cLPS levels and inflammation as measured by CRP levels. The lack of association between serum total endotoxin activity and BMI implies that the association between infection and an elevated BMI may be specific to certain pathogens.

Abstract Respiratory syncytial virus (RSV) is one of the most common pathogens for early childhood respiratory tract infections. Most children are infected by RSV, approximately 1% of infants require hospital care. RSV infections are often accompanied by otitis media (OM). Surfactant proteins A and D (SP-A and SP-D) are involved in lung function and innate immunity. The proteins are capable of recognizing surface patterns in pathogens and function as a defense before the acquired immunity is developed. The Toll-like receptor (TLR) family is associated to several pathogens. The role of the TLR-family is to recognize pathogens and activate the immune defense. The aim of the thesis was to investigate the genetic association of RSV and OM infections in infants. A candidate gene approach was used study SP-A, SP-D and TLR4 in severe RSV bronchiolitis. Association between SP-A and OM was also studied. A case-control study setup was used for all studies. 1700 samples were collected for the studies. The results revealed genetic association between SP-A gene variation and severe RSV infections. SP-A allele 1A3 was overrepresented in RSV infants, the allele 1A was present more often in the control population. SP-D allele Met11 and genotype Met/Met were predisposing to severe RSV infections. The TLR4 gene did not show direct association with severe RSV. However, we showed for the first time difference in association in two separate epidemics. In the OM study, an association was shown between SP-A gene variations and otitis media in children. The present results have brought new information about innate defense and the genetic variations and associations involved. The results will help understand the mechanisms of innate defense and predisposition to infections. The results also present possibilities to investigate and develop new treatment strategies.

La sécrétion de vasopressine (AVP) et de corticolibérine (CRH) déclenchent la sécrétion d'ACTH via leurs récepteurs V1B et CRF1, puis celle des catécholamines et des corticostéroïdes, résultant dans les effets physiologiques induits par le stress. Trois mutations résultant de variants non-synonymes affectant le récepteur V1B (K65N, R364H et G191R) ont été associées chez l'homme à des troubles psychiatriques tels que la phobie, l'hyperactivité et le comportement suicidaire. Nous avons étudiés les propriétés pharmacologiques de ces variants V1B qui pourraient être la cause de ces pathologies. Après construction de chacun des plasmides codant pour chaque variant, nous les avons exprimés dans des cellules HEK293 et avons établi les caractéristiques de chaque récepteur telles que son affinité de liaison, ses couplages à la PLC, à la MAP kinase et la mobilisation du Ca2+. Nous avons observé que le variant R191 avait un couplage à la PLC et à la MAP kinases fortement augmenté (+50%), alors que les autres variants montraient un baisse de ces couplages. Les différences n'étant pas dûes à des constantes d'activation (Kact) pour l'AVP différents. L'association avec les β-arrestines n'est pas équivalente pour tous les variants, le R191 montrant une forte association aux β-arrestines 1 et 2 alors que le variant N65 ne s'associe plus. Ces données indiquent que le déficit en ERK-phosphorylée peut être dû à la fois à une baisse de la production d'Inositol-P et de DAG via la voie Gq et à une baisse de signalisation directe via les β-arrestines. Nous avons également examiné les propriétés d'internalisation de ces variants en utilisant des lignées cellulaires stables exprimant chacun des récepteurs couplés à une EGFP. Nous avons observé, par microscopie confocale, que la cinétique et le taux d'internalisation étaient similaires pour tous les variants. Par contre, la voie du traffic intracellulaire est modifiée pour certains. Les variants possédant la mutation C terminale H364 (H364 et N65/H364) sont préférentiellement adressés à la voie de dégradation lysosomiale alors que les autres utilisent la voie classique de recyclage endosomial comme le récepteur sauvage, ainsi que le révèlent les marquages spécifiques de ces compartiments. En conclusion, les résultats obtenus durant cette thèse révèlent que les variants du récepteurs V1B trouvés chez des patients atteints de troubles affectifs ont des propriétés pharmacologiques de couplage et de trafic intracellulaire modifiées. Ces mutations pourraient participer à une plus grande vulnérabilité des patients au stress. Ainsi, elles pourraient être recherchées, à titre de marqueurs génétiques, chez des patients considérés comme à risque.Cette thèse translationnelle a été financée par un contrat doctoral CHRU/UM1. / The release of vasopressin (AVP) and corticoliberin (CRH) triggers the release of ACTH by the mean of V1B and CRF1 receptors, releasing catecholamines and corticosteroids and regulating stress. Genetic modifications of either element may produce severe consequences. Three non-synonymous variants of the V1B receptor gene were found in patients with phobia, hyperactivity or suicidal conducts (respectively K65N, R364H and G191R). To explore the pharmacological properties of these V1B variants, we generated and expressed in HEK293 cells cDNA constructs of each variant. We evaluated ligand affinity, PLC coupling, MAP kinase activation, Ca2+ mobilization. The capacity of R191 variant to accumulate inositol phosphates and to activate MAP kinase under AVP activation was higher as compared to N65 and H364 variants. However, all variants exhibited the same Kact for AVP. V1B variants were not similar in β-arrestin interaction either, with N65 variant impaired for this function, indicating that the deficit in ERK phosphorylation measured previously may result of 2 additional mechanisms, one due to a lower IP/DAG production with impaired coupling to Gq, and the other due to a decrease in β-arrestin interaction. Finally, we compared internalization properties using stable cell lines expressing EGFP–tagged V1B variants and confocal microscopy. First, we observed no difference neither in the internalization kinetics nor in amplitude (70% of total receptors internalized). However, H364 variant showed a different trafficking as compared to K65/G191/R364 (“wt”) and N65, with less cytoplasmic recycling endosomes and more lysosomial addressing as revealed by Lamp1 co-labelling.Altogether, these data reveal that V1B receptor variants in phobic and suicidal patients display modified coupling and/or desensitization processes. These mutations could participate to a predisposition to depression and could be used as diagnosis genetic markers in risky patients.M Manière's Ph.D Scholarship was granted on a translational CHRU/UM1.

Bien que le Tacrolimus soit un immunosuppresseur largement prescrit en transplantation rénale, ses effets néphrotoxiques limitent son utilisation. En effet, le Tacrolimus contribue au développement de lésions de fibrose interstitielle rénale et d’atrophie tubulaire à plus ou moins brève échéance. Parmi les mécanismes qui interviennent dans le processus de fibrogenèse, la transition épithélio-mésenchymateuse (TEM) a été évoquée. Dans ce processus, une cellule épithéliale polarisée perd l’expression de certains de ses marqueurs épithéliaux (E-cadhérine, cytokératine, β-caténine membranaire...) au profit d’un phénotype mésenchymateux (expression de marqueurs mésenchymateux tels que vimentine ou translocation nucléaire de la β-caténine, sécrétion de composants de la matrice extracellulaire). Ainsi, afin d’identifier des déterminants moléculaires de la néphrotoxicité induite par le Tacrolimus et d’évaluer la contribution du processus de TEM, plusieurs approches in vitro et in vivo ont été combinées.Tout d’abord, un modèle de culture primaire de cellules tubulaires proximales rénales (CTP), cibles privilégiées des xénobiotiques au niveau rénal a été développé à partir de pièces opératoires. Ce modèle a été caractérisé : analyse de la stabilité sur 5 passages, de l’expression de marqueurs épithéliaux proximaux et mésenchymateux, et de la résistance trans-épithéliale... De plus, la comparaison de la capacité métabolique des échantillons de tissu rénal sain à différents modèles cellulaires (HEK293, HK-2, CTP) nous a permis de montrer que les CTP est le modèle le plus pertinent. De plus, l’exposition de cellules rénales au Tacrolimus induit une modification du phénotype des cellules.De plus, le développement d’un modèle murin de néphrotoxicité induite par le Tacrolimus a été réalisé (exposition durant 28 jours à une dose de 1 mg/kg/j de Tacrolimus par implantation sous-cutanée de pompes Alzet® ou par injection intra-péritonéale). Les données histologiques et d’expression génique montrent que les reins de souris traitées par Tacrolimus présentent des zones localisées d’expression des marqueurs mésenchymateux (vimentine) et de fibrose (collagène, α-SMA, miR-21). Par ailleurs, la variabilité interindividuelle des effets néphrotoxiques du Tacrolimus étant potentiellement associée à la présence de polymorphismes génétiques (SNP), les ADN de patients transplantés rénaux et de leur greffon ont été génotypés pour des SNP (i) affectant les CYP3A5 et ABCB1, intervenant dans la prise en charge du Tacrolimus, (ii) affectant la cavéoline-1, impliquée dans le processus de fibrose. Nos résultats montrent que deux SNP affectant le donneur (CYP3A5 6986A>G et ABCB1 3435C>T) sont significativement associés à une plus faible expression des marqueurs de TEM (expression de novo de la vimentine et translocation nucléaire de la β-caténine) et à un nombre moins important de lésions de fibrose rénale sur les biopsies de greffons à 3 mois post-greffe. Enfin, les patients porteurs d’un greffon de génotype CAV1 rs4730751AA ont une perte de fonction rénale plus rapide. Ces patients semblent développer plus fréquemment des lésions de fibrose. Dans le cadre de la transplantation rénale, ces résultats suggèrent que certains SNP du donneur influencent la néphrotoxicité du Tacrolimus et que son métabolisme in situ est un élément clé dans la compréhension de la fibrogenèse du greffon.Au total, les résultats obtenus nous ont permis d’identifier des facteurs individuels de vulnérabilité à la toxicité du Tacrolimus. De telles données, utilisées comme outil prédictif d’une néphrotoxicité plus ou moins importante, aideraient à un meilleur choix de traitement immunosuppresseur. A terme, sur le plan médico-économique, notre étude pourrait permettre d’améliorer la prise en charge de la néphrotoxicité des immunosuppresseurs, et ainsi réduire les coûts de traitement liés aux pertes de greffons induites par la toxicité du Tacrolimus. / Although widely prescribed in kidney transplantation, Tacrolimus use is limited by its nephrotoxic effects. Indeed, Tacrolimus contributes to the development of renal interstitial fibrosis lesions and tubular atrophy with a large variability between patients. Among the mechanisms involved in fibrogenesis, the epithelial-mesenchymal transition (EMT) has been proposed. EMT is a dynamic process by which a polarized epithelial cell loses its epithelial markers (E-cadherin, cytokeratin, membrane β-catenin...) and acquires a mesenchymal phenotype (de novo expression of vimentin or nuclear translocation of β-catenin, secretion of extracellular matrix components). Thus, to identify molecular determinants of Tacrolimus-induced nephrotoxicity and to evaluate the contribution of EMT, several in vitro and in vivo approaches were combined.First, a model of primary culture of renal proximal tubular cells (PT cells), the main target of xenobiotics in kidney, has been developed and characterized: phenotypic stability, functional properties, expression of proximal and mesenchymal markers and transepithelial resistance. In addition, the comparison of the metabolic capacity of the healthy renal tissue samples to different cell models (HEK293, HK-2 CTP) has revealed that PT cells is the most appropriate model. Furthermore, renal cells exposure to Tacrolimus induced a modification of the cell phenotype.Moreover, the development of a murine model of Tacrolimus-induced nephrotoxicity has been performed (28 days-exposure at 1 mg/kg/day by subcutaneous implantation of Alzet® pumps or by intra-peritoneal injection). Histological and gene expression data indicated that kidney of Tacrolimus-treated mice exhibited localized expression of mesenchymal markers (vimentin) and fibrosis areas (collagen, α-SMA, miR-21).Furthermore, as the interindividual variability of Tacrolimus nephrotoxic effects is potentially associated with genetic polymorphisms (SNPs), renal transplant recipients and their corresponding graft were genotyped for (i) CYP3A5 and ABCB1 SNPs, involved in Tacrolimus cellular processing, (ii) caveolin-1 SNP, involved in fibrosis. Our results showed that two SNPs affecting the donor (CYP3A5 6986A> G and ABCB1 3435C> T) were significantly associated with a lower expression of EMT markers (vimentine de novo expression and nuclear translocation of β-catenin) together with less fibrosis lesions evaluated on renal graft biopsies performed at 3 months post-transplant. Finally, patients with a CAV1 rs4730751AA graft displayed a more severe renal function decrease. These patients also developed more frequently fibrotic lesions. In the context of renal transplantation, these results suggest that some donor SNPs modulate Tacrolimus-induced nephrotoxicity and that its in situ metabolism is a key element in the graft fibrogenesis understanding.Overall, these data allowed us to identify some molecular determinants of Tacrolimus-induced nephrotoxicity. Early identification of patients at high risk of Tacrolimus renal toxicity represents one of the most important and future challenges in kidney transplantation to tailor treatment before the development of irreversible lesions. Although preliminary, our data suggest that the genetic make-up of donors as well as the early detection of nephrotoxicity markers such as mesenchymal markers, may improve to the medical management of renal transplant patients.

Orientadores: Ricardo Della Coletta, Ana Lucia Carrinho Ayroza Rangel / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-26T13:40:24Z (GMT). No. of bitstreams: 1 Moreira_HelenaraSalvatiBertolossi_D.pdf: 1239702 bytes, checksum: 4e6973ffe9eab3d15e1180f4b76d52d2 (MD5) Previous issue date: 2014 / Resumo: As fissuras lábio-palatinas não-sindrômicas (FL/PNS) representam as malformações faciais mais comuns em seres humanos e são causadas por uma combinação de fatores genéticos e ambientais sob um modelo de herança multifatorial. O objetivo deste estudo foi caracterizar o perfil epidemiológico (características clínicas, demográficas e ambientais) dos indivíduos afetados por FL/PNS residentes na região oeste do estado do Paraná na busca por fatores de risco para o desenvolvimento das FL/PNS e também verificar a associação de polimorfismos em genes que codificam enzimas do sistema do reparo do DNA na susceptibilidade das FL/PNS. Para a primeira parte do estudo, foram coletadas 194 amostras de saliva de pacientes com FL/PNS e seus pais, sendo que os mesmos foram entrevistados com um questionário, contendo perguntas relacionadas aos pacientes com fissura e aos seus genitores, visando levantar dados referentes aos aspectos ambientais relacionados às fissuras. A segunda parte do estudo envolveu a análise dos polimorfismos rs1136410 no gene ADPRT, rs1052133 no gene OGG1, rs1800734 no gene MLH1, rs1130409 no gene APEX1, rs861539 no gene XRCC3, rs1801321 no gene RAD51, rs25487, rs25489, rs3213245 e rs1799782 no gene XRCC1 e rs13181 e rs1799793 no gene ERCC2 em 223 trios (pai, mãe e paciente com fissura) pelo teste de desequilíbrio de transmissão (TDT). As amostras utilizadas para a investigação de polimorfismo foram provenientes de 4 Centros diferentes de tratamento da FL/P, composta por 93 trios de Minas Gerais, 74 do Paraná, 34 da Bahia e 22 da Paraíba. Entre os pacientes avaliados, um predomínio de homens, leucodermas e afetados por fissuras lábio-palatinas (FLP) foi observado. Entre as alterações sistêmicas, as otorrinolaringológicas foram significantemente mais prevalentes em fissuras palatinas (FP) em comparação com as fissuras labiais (FL; p=0,013). Mais de 80% das mães dos pacientes com FL/PNS reportaram que não fizeram uso de suplementos vitamínicos durante o primeiro trimestre de gestação. Entre os 12 polimorfismos avaliados, rs3213245 no gene XRCC1 demonstrou uma significante associação com FL (p=0,03) e rs13181 do gene ERCC2 demonstrou tendência de associação com FLP (p=0,06). O haplótipo formado pelos polimorfismos rs25487, rs25489, rs321345 e rs1799782 no gene XRCC1 também foi significantemente associado as FLs (p=0,02). Os resultados deste estudo revelaram o perfil epidemiológico dos pacientes com FL/PNS atendidos no oeste do estado do Paraná e demonstram que os polimorfismos nos genes XRCC1 e ERCC2, que codificam enzimas associadas ao sistema e reparo do DNA, podem estar associados à suscetibilidade ao desenvolvimento das FL/PNS / Abstract: Nonsyndromic cleft lip and palate (NSCL/P) represents the most common facial malformation in humans, and they are caused by a combination of genetic and environmental factors under a multifactorial model of inheritance. The objective of this study was to characterize the epidemiologic profile (clinical, demographical and environmental features) of individuals affected by the NSCL/P in the west of the Paraná state searchingfor risk factors for the development of NSCL/P, and to verify the association of polymorphisms in genes that encode enzymes of the DNA repair system in the susceptibility of the NSCL/P. For the first part of the study, 194 patients with NSCL/P and their parents were interviewed with a questionnaire containing questions associated with the patients with cleft and their parents, addressing points of environmental aspects related to the clefts only in the western of the city of Parana. The genetic study involved the analysis of the polymorphisms rs1136410 in the gene ADPRT, rs1052133 in the gene OGG1, rs1800734 in the gene MLH1, rs1130409 in the gene APEX1, rs861539 in the gene XRCC3, rs1801321 in the gene RAD51, rs25487, rs25489, rs3213245 and rs1799782 in the gene XRCC1 and rs13181 and rs1799793 in the gene ERCC2 in 223 trios (father, mother and the patient with cleft) by the disequilibrium of transmission test (TDT). The sample to the investigation of the polymorphism were from four centers of FL/PNS treatment, composed by 93 trio from Minas Gerais, 74 from Parana, 34 from Bahia and 22 from Paraiba. A predominance of men, leucoderma and affected by cleft lip and palate (CLP) was observed. In relation to systemic alterations, the otorhinolaryngologics were significantly more prevalent in in patients with cleft palate clefts (CP) in comparison with patients affected by cleft lip (CL, p=0.013). More than 80% of the mothers of the NSCL/P patients reported that they did not use vitamin supplements during their first trimester of the pregnancy. Among the 12 polymorphisms, rs3213245 in XRCC1 demonstrated a significant association with the CL (p=0.03) and rs13181 inERCC2 demonstrated atendency of association with CLP (p=0.06). The haplotype formed by XRCC1 polymorphisms rs25487, rs25489, rs321345 and rs1799782 1 was significantly associated with CL (p=0.02). The results of this study show the epidemiologic profile of the patients with NSCL/P assisted in the west of Parana state and demonstrated that polymorphisms in the genes XRCC1 and ERCC2, which encode enzymes of the DNA repair system, may be associated with the susceptibility of the NSCL/P development / Doutorado / Patologia / Doutora em Estomatopatologia

Orientador: Carmen Silvia Bertuzzo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-28T01:52:48Z (GMT). No. of bitstreams: 1 Vencatto_RobyWill_M.pdf: 2327467 bytes, checksum: 906caea2ae7f9182396080360689d4a1 (MD5) Previous issue date: 2015 / Resumo: O câncer de mama é classificado mundialmente, como o segundo tipo de câncer mais frequente e comum entre as mulheres e tem natureza multifatorial. O tratamento sistêmico do câncer é denominado quimioterapia antineoplásica, e o seu efeito terapêutico depende da concentração plasmática e tempo de exposição à droga. A resposta aos medicamentos possui variação entre os pacientes, podendo decorrer de fatores como: outras patologias, farmacocinética e farmacodinâmica diferenciadas, fatores ambientais e genéticos, sendo este último, estudado na farmacogenética e farmacogenômica, onde se verifica a resposta ao tratamento pela variação herdada de cada indivíduo. Em determinadas populações, polimorfismos podem levar a respostas diferenciadas de um medicamento por induzir peculiaridades na genética, farmacocinética ou farmacodinâmica. Em muitos medicamentos em uso, os transportadores de drogas são importantes na questão de absorção, acumulação no tecido e eliminação do organismo. O gene 'ABCB1' codifica a glicoproteína-P que é um transportador de membrana, responsável pelo efluxo celular de uma variedade de drogas, xenobióticos, metabólitos celulares e agentes anticancerígenos, estruturalmente independentes. Assim, o objetivo do presente estudo foi verificar se os polimorfismos C1236T e G2677T/A do gene 'ABCB1', possuem associação com a resposta ao tratamento quimioterápico. Foi realizado estudo com 146 pacientes do sexo feminino, que usaram os quimioterápicos doxorrubicina e ciclofosfamida de forma adjuvante, independente se fizeram quimioterapia neoadjuvante. A presença de recidiva foi utilizada como parâmetro de avaliação do tratamento. A genotipagem dos polimorfismos foi feita por reação da polimerase em cadeia alelo específica. As análises de sobrevivência livre da doença e global foram realizadas pelo software SPSS (Statistical Package for the Social Sciences) vs20.0 e as demais avaliações no SAS (StatisticalAnalysis Software) vs 9.3. ... Observação: O resumo, na íntegra, poderá ser visualizado no texto completo da tese digital / Abstract: Breast cancer is classified worldwide as the second most frequent and common cancer among women and has a multifactorial nature. The systemic treatment of the cancer is called chemotherapy, and its therapeutic effect depends on the plasma concentration and length of exposure to the drug. The drug response varies among patients and may be the result of many things, such as: other diseases, different pharmacokinetics and pharmacodynamics, environmental and genetic factors. Genetic factors are studied in pharmacogenetics and pharmacogenomics, which verify the response to the treatment through the inherited variation of each individual. In some populations, polymorphisms can lead to different responses to a drug by inducing peculiarities on the pharmacokinetics or pharmacodynamics. In many drugs in use, the drug carriers are important in terms of absorption, accumulation in tissue and elimination from the body. The 'ABCB1' gene, which encodes the P-glycoprotein that is a membrane transporter, responsible for the cellular efflux of a variety of drugs, xenobiotics, cellular metabolites and anticancer agents, structurally independent. Thus, the objective of this study was to determine whether the C1236T and G2677T/A of the 'ABCB1' genes could be associated with the response to chemotherapy. The study featured 146 female patients who used doxorubicin and cyclophosphamide adjuvantly, independent of the neoadjuvant chemotherapy. The relapse was used as a parameter of association to the treatment. Genotyping of polymorphisms was performed by polymerase in specific allele chain reaction. The free and global survival analysis, was performed by the SPSS (Statistical Package for Social Sciences) software vs 20.0. Other statistical tests were performed by SAS (Statistical Analysis Software) vs 9.3.... Note: The complete abstract is available with the full electronic digital thesis or dissertation / Mestrado / Ciencias Biomedicas / Mestre em Ciências Médicas

O gênero Phytomonas compreende parasitas cosmopolitas de plantas transmitidos por hemípteros. Não há estudos recentes sobre a taxonomia de Phytomonas e apenas duas espécies foram filogeneticamente validadas: P. serpens e P. françai. Até o momento 6 - 11 grupos (A-K) constituem esse gênero, dependendo do marcador molecular. O principal objetivo deste estudo foi caracterizar uma grande amostragem de isolados de Phytomonas de plantas e hemípteros com inferências filogenéticas de sequências de SSUrDNA, gGAPDH e ITSrDNA. Foi confirmado a subdivisão em 7 clados: Grupo A - 59 isolados de látex e insetos (Coreidae) de diferentes países. Grupo B - 7 isolados de látex e insetos (Scutelleridae e Pentatomidae) do Brasil e Suriname. Grupo C - 4 isolados de látex da Espanha. Grupo D - 11 isolados de Euphorbiaceae e Coreidae de diferentes países. Grupo E - 16 isolados de Solanaceae e Pentatomidae do Brasil. Grupo F - restrito a P. françai do Brasil. Grupo H - isolados de floema da América do Sul. Os resultados permitiram identificar candidatos a novas espécies de Phytomonas. / The Phytomonas genus is comprised by cosmopolitan plant´s parasites and is transmitted by Hemiptera. There are no recent studies based on the taxonomy of Phytomonas and only two species were phylogenetically validated: P. serpens and P. françai. To date this genus is constitute by 6 - 11 groups (A-K), depending on the molecular marker. The aim of this study was to characterize large samples of Phytomonas isolated from plants and Hemiptera throught phylogenetic inferences sequences SSUrDNA, gGAPDH and ITSrDNA. 7 Subclades was confirmed: Group A - 59 isolates of latex and insects (Coreidae) from different countries. Group B - 7 isolates of latex and insects (Scutelleridae and Pentatomidae) from Brazil and Suriname. Group C - 4 isolates of Spain latex. Group D - 11 isolates of Euphorbiaceae and Coreidae from different countries. Group E - 16 isolates of Solanaceae and Pentatomidae from Brazil. Group F - restricted to Brazilian P. françai isolates. Group H - Isolates of phloem from South America. Our results shown possible candidates for new species of Phytomonas.

Implantation requires complex molecular and cellular events involving coagulation, angiogenesis and immunological processes that need to be well regulated for a pregnancy to establish and progress normally.  The overall aim of this thesis was to study different models associated with atypical angiogenesis, impaired implantation and/or placentation, such as recurrent miscarriage (RM), oocyte donation (OD) and preeclampsia. Histidine-rich glycoprotein (HRG), a serum protein with angiogenic potential has been previously shown to have an impact on implantation and fertility.  In two retrospective case-control studies, women suffering from RM (Study I) and gestational hypertensive disorders (GHD) (Study IV) have been compared to healthy control women, regarding carriership of HRG genotypes (HRG A1042G and C633T SNP, respectively).  According to the findings of this thesis, heterozygous carriers of the HRG A1042G SNP suffer from RM more seldom than homozygous carriers (Study I).  Additionally, the presence of the HRG 633T allele was associated with increased odds of GHD (GHD IV).  Studies II and III comprised a national cohort of relatively young women with optimal health status conceiving singletons with donated oocytes versus autologous oocytes (spontaneously or via IVF).  We explored differences in various obstetric (Study II) and neonatal (Study III) outcomes from the Swedish Medical Birth Register.  Women conceiving with donated oocytes had a higher risk of GHD, induction of labor and cesarean section, as well as postpartum hemorrhage and retained placenta, when compared to autologously conceiving women.  OD infants had higher odds of prematurity and lower birthweight and length when born preterm, compared to neonates from autologous oocytes.  With regard to the indication of OD treatment, higher intervention but neverthelss favourable neonatal outcomes were observed in women with diminished ovarian reserve; the risk of GHD did not differ among OD recipients after adjustment. In conclusion, HRG genetic variation appears to contribute to placental dysfunction disorders.  HRG is potential biomarker that may contribute in the prediction of the individual susceptibility for RM and GHD.  Regarding OD in Sweden, the recipients-despite being of optimal age and health status- need careful preconceptional counselling and closer prenatal monitoring, mainly due to increased prevalence of hypertensive disorders and prematurity.