Évoquer ou convoquer l’animal en art met en perspective et accroît l’œuvre humaine jusque dans les excès et les incohérences qu’elle peut manifester, mais rend également possible un accès à ce qui est extérieur au domaine humain. Les artistes s’inspirent de la symbolique à laquelle donne corps l’animal, de sa morphologie aussi bien que des modes d’expressions qu’il invente et déploie. Il leur arrive même de se laisser tenter par le mimétisme. Les créateurs et les poètes ne disposent-ils pas d’autres outils que les penseurs ou les scientifiques pour atteindre cette étrangeté qui semble pourtant souvent si familière ? Qu’il s’agisse d’œuvres plastiques, de performances, de créations musicales ou scéniques, y convier l’animal offre une matrice infinie de figurations et de reconfigurations. L’expression de ce répertoire corporel et comportemental s’articule comme un véritable langage structuré. Au-delà d'une transposition qui permet de donner à entrevoir l’humanité de l'homme, les artistes peuvent aussi tenter la percée vers la réalité d’autres êtres vivants. Entre confrontation, dialogue et transfert, les pratiques artistiques reflètent la diversité, le polymorphisme même du rapport humain aux animaux, et l’histoire d’une attitude foncièrement ambivalente, prise entre empathie et exploitation. L’expérience de l’animal, voire l’expérience animale, donne lieu à un partage de territoire(s) et de conscience(s), ouvrant sur le dévoilement d’un penser en images. / To evoke or to summon the animal in art puts in perspective the work of mankind, increases its excesses and its inconsistencies as well as it makes possible to open up the door to what is outside the human reach. Artists inspire themselves from the symbolic to which the animal is giving flesh, its morphology up to the ways of communication it creates and deploys. Artists may even be tempted by mimicry. Don’t creators and poets dispose of no other means than this one to address that strangeness that seems however so familiar to them ?To invite the animal in art work, could it be plastic arts, performances, musicals works or on stage, is to offer an infinite matrix for figuring or reconfiguring. The expression of its physical and behavioral repertory is articulated as a structured language. Beyond a transposition that allows a glimpse at human kind’s humanity, artists can also try to break through the reality of other living beings. Half way through confrontation, dialogue and transference, artistic practices mirror the diversity of human relationships with animals and even its polymorphism, as well as they reflect on the history of a human attitude fundamentally ambivalent, torn between empathy and exploitation. The animal experience, the animalistic experiment even, gives rise to a territory and consciousness sharing, opening up to the unveiling of a thinking through images.

Single nucleotide polymorphisms (SNPs) are single base variations among groups of individuals. In order to study their properties in fine gene mapping, I considered their occurrence as transitions and transversions. The aim of the study was to classify each polymorphism depending upon whether it was a transition or transversion and to calculate the proportions of transitions and transversions in the SNP data from the public databases. This ratio was found to be 2.35 for data from the Whitehead Institute for Genome Research database, 2.003 from the Genome Database, and 2.086 from the SNP Consortium database. These results indicate that the ratio of the numbers of transitions to transversions was very different than the expected ratio of 0.5. To study the effect of different transition to transversion ratios in fine gene mapping, a simulation study was performed to generate nucleotide sequence data. The study investigated the effect of different transition to transversion ratios on linkage disequilibrium parameter (LD), which is frequently used in association analysis to identify functional mutations. My results showed no considerable effect of different transition to transversion ratios on LD. I also studied the distribution of allele frequencies of biallelic SNPs from the Genome Database. My results showed that the most common SNPs are normally distributed with mean allele frequency of 0.7520 and standard deviation of 0.1272. These results can be useful in future studies for simulating SNP behavior. I also studied the simulated data provided by the Genetic Analysis Workshop 12 to identify functional SNPs in candidate genes by using the genotype-specific linkage disequilibrium method.

Controlling crystal polymorphism, the ability of a molecule to crystallise in different solid forms, is one of the grand, ongoing challenges in materials science. In the pharmaceutical industry particularly, where up to half of the active pharmaceutical active ingredients exhibit polymorphic behaviour, it is of paramount importance to rationalise the impact of experimental conditions, such as the nature of the solvent, on the obtainment of a specific c crystal form. As strategies for the selection of polymorphs is still, by and large, based on a trial-and-error approach, it is necessary to acquire a fundamental understanding of the factors controlling the formation of a speci fic solid-state structure during crystallisation from solution. During this doctoral research project, we have conducted a computer simulation study of the early stages of crystallisation of meta-aminobenzoic acid, an important model system in the investigation of polymorphic phenomena. This molecule can in fact form five different polymorphic forms whose selective crystallisation from solution chiefly depends on the nature of the solvent. Molecular models and computational chemistry methods, based on density functional theory and molecular dynamics, have been developed and applied to quantify the processes surrounding the crystallisation of meta-aminobenzoic acid: solvent-solute separation, solute aggregation and surface reactivity. The aim was to identify what controls, at the molecular level, the polymorphic selection process during crystallisation from solution of this important active pharmaceutical ingredient. The results show that the solvent play a signi cant role during the key stages of meta-aminobenzoic acid crystallisation by controlling both the kinetics and thermodynamics of solute desolvation, formation of prenucleation clusters and surface reactivity. This work represents a paradigm of the role of molecular processes during the early stages of nucleation in affecting polymorph selection during crystallisation from solution.

O objetivo do presente estudo foi avaliar, em humanos, a associação entre polimorfismos genéticos no sistema RANK/RANKL/OPG com as dimensões craniofaciais. Foram incluídos neste estudo um total de 100 indivíduos caucasianos brasileiros não relacionados. O DNA foi extraído da saliva de cada um dos participantes e os polimorfismos rs3826620, rs9594738 e rs2073618 em RANK, RANKL e OPG, respectivamente, foram analisados por PCR em tempo real. Para avaliação das dimensões craniofaciais foram avaliadas três medidas angulares (SNA, SNB e ANB) e quatro medidas lineares (Co-Gn, Go-Pg, Co-Go e PTM-A), obtidas de traçados cefalométricos. Para avaliar a distribuição dos genótipos de acordo com os padrões esqueléticos faciais (Classe I, Classe II e Classe III) foi utilizado o teste do qui-quadrado. Para comparar as médias das dimensões maxilares e mandibulares de acordo com os genótipos utilizou-se o teste de Kruskal-Wallis, com o pós-teste de Dunn para comparações múltiplas, ou ANOVA com pós-teste de Tukey. Foi utilizada a análise de regressão linear multivariada para ajustar a possível influência da idade e do gênero em cada medida. O equilíbrio de Hardy-Weinberg foi avaliado utilizando o teste do qui-quadrado para cada polimorfismo. O nível de significância adotado para todas as análises foi 5%. Os resultados obtidos evidenciaram que não houve associação estatisticamente significante entre a distribuição genotípica de RANK, RANKL e OPG com as médias dos ângulos SNA e SNB, nem com a distribuição fenotípica (padrão esquelético Classe I, II ou III) (p>0,05). Observou-se diferença estatisticamente significante entre a distribuição das medidas do comprimento da base mandibular, de acordo com os genótipos de RANK, onde o genótipo GG apresentou maior medida de Go-Pg (p=0,039). Na análise multivariada, observou-se associação significante para os polimorfismos em RANK e medidas mandibulares (Go-Pg e Co-Gn) (p<0,05). A medida da maxila não foi associada a nenhum polimorfismo. Conclui-se que houve associação entre o polimorfismo genético rs3826620 em RANK com maiores dimensões mandibulares, onde o comprimento da mandíbula (Co-Gn) e o comprimento da base da mandíbula (Go-Pg) estavam aumentados. / The objective of the present study was to evaluate, in humans, the association between genetic polymorphisms in the RANK/ RANKL/OPG system with alterations in craniofacial dimensions. A total of 100 unrelated Brazilian Caucasians were included in this study. DNA was extracted from the saliva of each of the participants and the polymorphisms rs3826620, rs9594738 and rs2073618 in RANK, RANKL and OPG, respectively, were analyzed by real-time PCR. To evaluate the craniofacial dimensions, three angular measurements (SNA, SNB and ANB) and four linear measurements (Co-Gn, Go-Pg, Co-Go and PTM-A) were obtained from cephalometric tracings. To compare the difference between the means of the linear and angular measurements according to the genotype, Kruskal-Wallis followed by Dunn post test or ANOVA followed by the Tukey post test was used. Linear regression analysis was also used to adjust the possible influence of age and gender on each linear maxillary and mandibular measure. The Hardy-Weinberg equilibrium was also evaluated using the chi-square test within each polymorphism. The level of significance was 5%. The results demonstrated that there were no statistically significant association between the genotypic distribution of RANK, RANKL, OPG, and the angules SNA, SNB and according to the phenotypic distribution (Class I, Class II or Class III of skeletal pattern) (p> 0.05). A statistically significant difference was observed between the distribution of mandibular base length measurements according to the RANK genotypes, where the GG genotype showed a higher Go-Pg measurement (p=0.039). In the multivariate analysis, a statistically significant association was found for RANK and mandibular (Go-Pg and Co-Gn) polymorphisms (p<0.05). Measurement of the maxilla was not associated with any polymorphism. It was concluded that there was an association between genetic polymorphism rs3826620 in RANK with a greater mandibular dimension, in which the length of the mandible (Co-Gn) and the length of the base of the mandible (Go-Pg) were increased.

Electrophoretic techniques were utilized for the demonstration of variation in 22 proteins from 24 natural populations of four species ( G. bursarius, G. pinetis, G. arenarius and G. personatus ) of the Geomys complex of pocket gophers. Of the 24 structural loci , 19 were considered to be polymorphic. Five of the six esterases contributed greatested to the polymorphism while non-esterase proteins generally showed low values. In the GeoMys complex of pocket gophers in this study,selection appeared to be the most important influence on genetic structure with some evidence of random drift in two of the four species. Populations of G. arenarius and G. personatus had the highest average interspecific genetic similarities to G. bursarius and . pinetis was the most divergent. Biochemical evidence supports the phylogeny of Geomys based on morphological and fossil data.

Os tumores extra-axias do sistema nervoso são de localização extra-cerebral e na maioria das vezes benignos; meningiomas, schwanomas e metástases fazem parte deste grupo. O aparecimento de um tumor ocorre a partir do acúmulo de alterações genéticas e epigenéticas nas células. Para entender o mecanismo molecular da progressão tumoral e a formação de metástases é indispensável identificar os genes que acumulam essas alterações. Sendo assim, este trabalho teve como objetivo analisar o perfil de metilação dos genes TP16, TP53, DAL-1, GSTP-1, MEN-1, NDRG2 e das DNA metiltransferases 3A, 3B e 3L e sua associação com os tumores extra-axiais e ainda, avaliar, através de um estudo caso-controle, a influência dos SNPs TP53 Pro47Ser e Arg72Pro, EGF + 61, GSTP-1 Ile105Val e WRN Cys1367Arg no desenvolvimento e prognóstico desses tumores. A técnica utilizada para a análise de hipermetilação foi a MSP, e através dela observamos que a atividade das DNMTs não está associada à metilação dos tumores extra-axiais e ainda, os perfis de metilação das DNMTs de novo não estão associados com alterações no padrão de metilação dos genes TP16, TP53, DAL-1, GSTP-1, MEN- 1 e NDRG2. Observamos que a metilação do gene TP53 está associada principalmente aos tumores de maior grau de malignidade, a uma deficiência na resposta a tratamentos e, conseqüentemente, a um maior número de óbitos. A metilação do gene TP16 está envolvida mais freqüentemente na formação de schwanomas e a de NDRG2 na progressão dos meningiomas. A análise de polimorfismos foi realizada através da técnica de PCR-RFLP e observamos diferenças nas distribuições genotípicas entre pacientes e controles nos SNPs TP53 Pro47Ser e Arg72Pro, EGF + 61 e GSTP-1 Ile105Val, onde as variantes Ser47, Pro72, EGF G61 e Val105 foram observadas com maior freqüência entre os portadores de tumores extra-axiais. Dessa forma, estas variantes podem ser fatores de susceptibilidade para o desenvolvimento dos tumores. / The extra-axial brain tumors have extra-brain localization and in most of the time they are benign, meningiomas, schwannomas and metastasis are included in this group. The appearance of a tumor occurs because of the accumulation of genetic and epigenetic alterations in the cells. In order to understand the molecular mechanism of the tumor progression and the metastasis formation it is important to identify the genes that accumulate the alterations. Thereby, the objective of this study was to analyze the methylation profile of the genes TP16, TP53, DAL-1, GSTP-1, MEN-1, NDRG2 and the DNA methyltransferases 3A, 3B and 3L and their association with the extra-axial brain tumors. Another purpose was to determine, in a case-control study, the roles of the TP53 Pro47Ser and Arg72Pro, EGF + 61, GSTP-1 Ile105Val and WRN Cys1367Arg SNPs in the development and prognosis of these tumors. We used the MSP to screen the hypermethylation profile and we observed no association between the DNMTs activity and the hypermethylation of the tumors. We also did not find association between the methylation of the DNMTs de novo and alterations in the methylation profile of the genes TP16, TP53, DAL-1, GSTP-1, MEN-1 and NDRG2. We observed that TP53 hypermethylation was associated with the high grade tumors, a poor response to the treatments and, consequently, the high number of obits. The TP16 methylation was involved with the shwannomas formation and the NDRG2 gene was involved in the meningiomas progression. For the polymorphism analysis, we used the PCR-RFLP technique and we observed differences in the genotype distributions between cases and controls of TP53 Pro47Ser and Arg72Pro, EGF + 61 and GSTP-1 Ile105Val SNPs, where the variants Ser47, Pro72, EGF G61 and Val105 were more frequent in patients than in controls. Thus, these variants can be important factors of susceptibility to the tumor development.

Type 2 diabetes and metabolic syndrome are the metabolic disorders which constitute a major public health problem in both developed and developing countries. Various studies have suggested the genetic susceptibility to the disorders. The main aim of the thesis was to investigate the putative association of single nucleotide polymorphisms with Type 2 diabetes (T2D), metabolic syndrome (MetS) and the major components of metabolic syndrome. This study used meta‐analysis, polymerase chain reaction (PCR) based restriction fragment length polymorphism (RFLP) and Sanger sequencing methods to analyse the results. The single nucleotide polymorphism rs57829442 of peroxisome proliferator‐activated receptor‐γ coactivator‐1 (PPARGC1A) gene and its relation to risk of type 2 diabetes has been studied in the United Kingdom population. A meta‐analysis of genetic variant rs8192678 (Gly482Ser) of peroxisome proliferator‐activated receptor‐γ coactivator‐1 (PPARGC1A) gene and its association with the components of metabolic syndrome has been studied. An association of the genetic variants rs8192678 (Gly482Ser) of the PPARGC1A gene, rs7903146 of Transcription Factor 7 Like 2 (TCF7L2) gene, rs9939609 of Fat mass and obesity‐associated (FTO) gene and rs1801282 (Pro12Ala) of peroxisome proliferator‐activated receptor gamma (PPARG) gene with the metabolic syndrome and its components has been studied in the Nepalese population. The results showed that variant rs57829442 of PPARGC1A is not associated with T2D in the United Kingdom population. Further investigation with increased sample size is warranted. In the meta‐analysis, the variant rs8192678 (Gly482Ser) of PPARGC1A gene was found to be significantly associated with body mass index (BMI) in Asian populations under dominant genetic model, total cholesterol (TC) in non‐Asian population under recessive genetic model and with fasting plasma glucose (FPG) under a recessive model in overall and non‐Asian populations. No significant association of the variants rs8192678 (Gly482Ser), rs7903146, rs9939609 and rs1801282 (Pro12 Ala) was found associated with MetS under dominant, recessive, co‐dominant and additive models in the Nepalese population. However, the genotypes (AG and AA) of rs8192678 (Gly482Ser) had a statistically significant protective effect on systolic blood pressure. The genotypes with the risk allele of rs9930609 of FTO gene was significantly associated with weight, waist circumference and diastolic blood pressure under dominant genetic model and with BMI under both dominant and recessive genetic models in the Nepalese population. To the best of our knowledge, this is the first study to report the findings in the Nepalese population.

O polimorfismo é um fenômeno relativamente comum entre os compostos farmacêuticos, e um dos principais problemas na produção e desenvolvimento de medicamentos. Sua presença pode acarretar prejuízos para a saúde da população e forte impacto econômico para as indústrias farmacêuticas ao influenciar diretamente nas propriedades mecânicas e biofarmacêuticas de insumos e produtos acabados. A investigação de polimorfismo associado aos oxicans, grupo pertencente à classe dos fármacos anti-inflamatórios não esteróides (AINEs), sofreu um incremento nos últimos anos, e para o fármaco tenoxicam relata-se a existência de quatro formas polimórficas na literatura. O objetivo deste trabalho foi caracterizar a presença de diferentes formas polimórficas de tenoxicam em insumos farmacêuticos ativos e formulações farmacêuticas orais e avaliar a influência sobre a dissolução in vitro. Com base na literatura, a análise inicial com a substância química de trabalho (SQT) demonstrou padrão difratométrico forma III. Diferentes condições para formação de formas polimórficas de tenoxicam foram aplicadas conforme Cantera et al. (2002), contudo, os diferentes processos de preparação de polimorfos do tenoxicam não conduziram a modificações na estrutura cristalina original da SQT. Dados complementares foram obtidos através da espectrofotometria no IV e DSC, que indicaram a inexistência de modificações no espectro original e ausência de eventos térmicos atípicos. A caracterização dos insumos farmacêuticos do tenoxicam, provenientes de diferentes fornecedores, por IV, DRX e perfil de dissolução indicou presença da estrutura cristalina (forma III), sem apresentar diferenças significativas entre os perfis de dissolução in vitro. As especialidades farmacêuticas do tenoxicam disponíveis no mercado nacional, ao serem submetidas à análise por DRX, também apresentaram estruturas cristalinas forma III. Apesar de ocorrer perfis de dissolução diferentes entre as formulações, estas não apresentaram comprometimento em sua qualidade. Contudo, muitas vezes as transformações pós-processamento podem induzir a alterações na estrutura cristalina e por consequência, problemas biofarmacêuticos. Tal fato demonstra a importância do estudo do polimorfismo, ao avaliar e correlacionar a presença de estruturas cristalinas com alterações na qualidade e desempenho de insumos e produtos farmacêuticos. / Polymorphism is a relatively common phenomenon among pharmaceutical compounds, and one of the main problems in producing and developing drugs. Its presence may harm people’s health and have a strong economic impact on the pharmaceutical industries by directly influencing the mechanical and biopharmaceutical properties of inputs and finished products. The investigation of polymorphism associated with oxycans, a group belonging to the class of non-steroidal anti-inflammatory drugs (NSAIDs), has grown in recent years, and it is reported that there are four polymorphic forms for tenoxycam the literature. The purpose of this study was to characterize the presence of different polymorphic forms in active pharmaceutical inputs and oral pharmaceutical formulations and evaluate their influence on in vitro dissolution. Based on the literature, the initial analysis with the reference standard showed a form III diphractometric pattern. Polymorphic forms of tenoxycam were prepared according to Cantera et al. (2002), but the different preparation processes of tenoxycam polymorphs did not lead to modifications of the original crystalline structure of the reference standard. Complementary data were obtained by spectrophotometry in IV and DSC, indicating that there were no modifications in the original spectrum and no atypical thermal events. The characterization of the pharmaceutical inputs of tenoxycam, from different suppliers, as IV, DRX and dissolution profile indicated the presence of a crystalline structure (Form III), without presenting significant differences between the in vitro dissolution profiles. The pharmaceutical specialties of tenoxycam available on the national market, on being analyzed by DRX also presented crystalline structures Form III. Although there was different dissolution profiles between the formulations, they showed no impairment in their quality. However, often the post-processing transformations can induce alterations in the crystalline structure and, consequently, biopharmaceutical problems. This shows the importance of the study of polymorphism with the evaluation and correlation of the presence of crystalline structures with alterations in the quality and performance of pharmaceutical inputs and products.

Submitted by Natalia Vieira (natalia.vieira@famerp.br) on 2016-05-19T19:07:09Z No. of bitstreams: 1 sabrinamayaracezario_dissert.pdf: 2283086 bytes, checksum: 130b0d9b52a17f23980c982605a0dad9 (MD5) / Made available in DSpace on 2016-05-19T19:07:10Z (GMT). No. of bitstreams: 1 sabrinamayaracezario_dissert.pdf: 2283086 bytes, checksum: 130b0d9b52a17f23980c982605a0dad9 (MD5) Previous issue date: 2015-05-27 / Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP / Background - Age-related macular degeneration (AMD) is a complex disease. The identification of risk factors may contribute to the prognosis and treatment. Objectives – Evaluate the influence of genetic variants related to lipid metabolism, angiogenesis and inflammation and its relation with clinical and lipid profile and lifestyle beyond their gene expression in patients with AMD. Casuistic and Methods – We studied 333 individuals aged ≥50 years, 108 with AMD in exudative form (G1); 45 with AMD in dry form (G2), and 180 individuals without clinical and angiographic signs of the disease (G3). The polymorphisms of apolipoprotein E (APOE-rs429358/rs7412), triphosphate binding cassette sub -family A transport -member 4 (ABCA4-rs472908), complement factor H (CFH-rs1061170) and vascular endothelial growth factor (VEGF-rs3025039/rs1570360) were analyzed by PCR/RFLP (polymerase chain reaction/restriction fragments length polymorphism), while the respective gene expression in blood by PCR/RT (reverse transcription-PCR) and serum levels of apo E, ABCR, CFH, VEGF by ELISA (enzyme linked immuno sorbent assay). Clinical and lipid profile data in addition to lifestyle were obtained from medical records and questionnaire. Level of significance was accepted for P<0.05. Results – Systemic arterial hypertension (SAH) and smoking prevailed in patients with AMD exudative form (P<0.05). Genetic polymorphisms: APOE- rs429358/rs7412 - APOE*3/3 was noted in all groups, followed by APOE*3/4, as well APOE*4 (P>0.05). ABCA4-rs472908 - Genotype A/G was more frequent in G3 (68%) versus G2 (44%; P<0.0001), while A/A in G2 (36%) versus G1 (19%; P=0.04) and G3 (14%; P=0.003). The mutant genotype (G/G) prevailed in combination (G1+G2) versus G3 (P< 0.0001), and allele G in all groups (P>0.05). CFH-rs1061170 – The wild homozygous (TT) was in evidence in G3 (58%) versus G1 (39%, P=0.003); as well the homozygous mutant (CC) in G1 (27 %) versus G3 (14%; P=0.002) and allele T in G3 (0.72) versus G1 (0.56; P=0.0002). VEGF-rs3025039 – Genotypic and allelic distribution were similar between groups (P>0.05), highlighting the CC genotype and allele C. VEGF-rs1570360 – Mutant homozygote (A/A) prevailed in G2 (21%) versus G1 (5%; P=0.002) and G3 (8%; P=0.015) as well as the wild type allele (G) G1 (0.75) and G3 (0.71) versus G2 (0.57, P=0.004, P=0.020, respectively). Gene expression – Similar values between groups for all analyzed genes (P>0.05). Serum levels (median values in ng/mL) – ApoE– Increased level in G1 (270.6) versus G2 (196.5; P<0.0001) and G3 (242.8; P=0.035), and G3 versus G2 (P=0.0002). ABCR – High levels in G1 (0.30) versus G2 (0.25; P=0.003) and G3 (0.25, P<0.0001). CFH – Increase in G1 (1198.9) versus G2 (859.8; P=0.0069), both higher than in G3 (618.3; P<0.001, P=0.001, respectively). VEGF – Similar values between groups (P>0.05). Lipid profile – G3 showed the highest level of HDLc (median=71mg /dL), compared to G2 (60mg/dL), and G1 (45mg/dL; P=0.003, P=0.029, respectively). Conclusion – Genetic variants of ABCA4, VEGF and CFH, besides SAH, smoking and increased serum levels of apo E, ABCR and CFH are associated with AMD, whereas the expression of the respective genes not differentiate AMD exsudative and dry forms, in contrast CFH (homozygous wild-type) has a protective character, as well as serum levels of HDLc. / Introdução – Degeneração macular relacionada à idade (DMRI) é uma doença complexa. A identificação de fatores de risco poderá contribuir no seu prognóstico e tratamento. Objetivos – Avaliar a influência de variantes genéticas, relacionadas com o metabolismo de lipídios, angiogênese e inflamação e sua relação com perfil clínico e lipídico e hábitos de vida, além das respectivas expressões gênicas em pacientes com DMRI. Casuística e Métodos – Foram estudados 333 indivíduos com idade ≥50 anos, sendo 108 com DMRI na forma exsudativa (G1); 45 com DMRI na forma seca (G2) e 180 indivíduos sem sinais clínicos e angiográficos da doença (G3). Os polimorfismos de apolipoproteína E (APOE-rs429358/rs7412), triphosphate binding cassette transporte sub-family A-member 4 (ABCA4-rs472908), complemento do fator H (CFH-rs1061170) e fator de crescimento endotelial vascular (VEGF-rs3025039/rs1570360) foram analisados por PCR/RFLP (polymerase chain reaction/restriction fragments lengh polymorphism), enquanto as respectivas expressões gênicas no sangue por PCR/RT (reverse transcription-PCR) e níveis séricos de apo E, ABCR, CFH, VEGF por ELISA (enzyme linked immuno sorbent assay). Dados de perfil clínico e lipídico, além de hábitos de vida, foram obtidos em prontuário médico e questionário. Admitiu-se nível de significância para P<0,05. Resultados – Hipertensão arterial sistêmica (HAS) e tabagismo prevaleceram em pacientes com DMRI exsudativa (P<0,05). Polimorfismos genéticos: APOE-rs429358/rs7412–APOE*3/3 destacou-se em todos os grupos, seguido de APOE*3/4, assim como o alelo APOE*4 (P>0,05). ABCA4-rs472908–O genótipo A/G foi mais frequente em G3 (68%) versus G2 (44%; P<0,0001), enquanto A/A em G2 (36%) versus G1 (19%; P=0,043) e G3 (14%; P=0,003). O genótipo mutante (G/G) prevaleceu na combinação (G1+G2) versus G3 (P<0,0001), e o alelo G em todos os grupos (P>0,05). CFH-rs1061170–O homozigoto selvagem (TT) destacou-se em G3 (58%) versus G1 (39%, P=0,003); já o homozigoto mutante (CC) em G1 (27%) versus G3 (14%; P=0,002), e o alelo T em G3 (0,72) versus G1 (0,56; P=0,0002). VEGF-rs3025039–Distribuição genotípica e alélica semelhante entre os grupos (P>0,05), destacando-se o genótipo CC e alelo C. VEGF-rs1570360–Homozigoto mutante (A/A) prevaleceu em G2 (21%) versus G1 (5%; P=0,002) e G3 (8%; P=0,015), assim como o alelo selvagem (G) em G1 (0,75) e G3 (0,71) versus G2 (0,57; P=0,004; P=0,020, respectivamente). Expressão gênica–Valores semelhantes entre os grupos para todos os genes analisados (P>0,05). Níveis séricos (valores de mediana em ng/mL)–ApoE–Aumento em G1 (270,6) versus G2 (196,5; P<0,0001) e G3 (242,8; P=0,035), e G3 versus G2 (P=0,0002). ABCR–Níveis elevados em G1 (0,30) versus G2 (0,25; P=0,003) e G3 (0,25; P<0,0001). CFH–Aumento em G1 (1.198,9) versus G2 (859,8; P=0,0069), ambos com acréscimo em relação a G3 (618,3; P<0,001; P=0,001, respectivamente). VEGF–Valores semelhantes entre os grupos (P>0,05). Perfil lipídico–G3 mostrou valor mais elevado de HDLc (mediana=71mg/dL), comparado a G2 (60mg/dL) e G1 (45mg/dL) (P=0,003; P=0,029, respectivamente). Conclusão – Variantes genéticas de ABCA4, VEGF e CFH, além de HAS, tabagismo e nível sérico elevado de apoE, ABCR e CFH associam-se a DMRI, enquanto a expressão dos referidos genes não diferencia DMRI exsudativa e seca, em contrapartida CFH (homozigoto selvagem) tem caráter protetor, assim como nível sérico de HDLc.

Submitted by Carvalho Dias João Paulo (joao.dias@famerp.br) on 2018-04-05T17:26:15Z No. of bitstreams: 1 gracieledtenani_dissert.pdf: 2576225 bytes, checksum: 0b7b634cafdb9080cbe9b93e41c2e195 (MD5) / Made available in DSpace on 2018-04-05T17:26:15Z (GMT). No. of bitstreams: 1 gracieledtenani_dissert.pdf: 2576225 bytes, checksum: 0b7b634cafdb9080cbe9b93e41c2e195 (MD5) Previous issue date: 2016-12-07 / Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP / Background - Hepatocellular carcinoma (HCC) is highlighted as the most aggressive malignant liver tumor. The identification of candidate genes to become biomarkers may help to clarify the pathophysiology of HCC, as well as the diagnosis of the disease at early stage, leading to new therapeutic interventions. Objectives -To evaluate the association of genetic variants and the gene expression involved in the cell signaling process, apoptosis, and angiogenesis with HCC, to characterize risk subgroups and identify biological markers for early diagnosis, prognosis and treatment of the disease. Casuistics and Methods – We studied 343 subjects, 102 with HCC (SG = study group) and 215 controls (CG = control group) for the analysis of PTEN polymorphisms (rs10490920, rs532678 and rs701848) and VEGF-A (rs3025039 and rs1570360). For gene expression analysis of PTEN and PIK3CA, 24 patients with HCC were selected (SGge = Study Group of gene expression), 16 with cirrhosis (CiGge = Cirrhosis Group of gene expression) and 10 controls who underwent bileo and digestive surgery (CGge = Group control of gene expression). The polymorphisms of related genes were analyzed by PCR/RFLP (polymerase chain reaction/restriction fragments length polymorphism), while the gene expression (fresh liver tissue) by qPCR (quantitative/polymerase chain reaction). Data from the clinical profile, lifestyle and comorbidities were obtained from medical records and questionnaire. Alpha error level was set at 5%. Results - Male gender, advanced age, smoking, alcohol consumption and diabetes mellitus (DM) prevailed in the group with HCC compared to the control (P<0.05). Genetic polymorphisms: PTEN- rs10490920 - The T/T genotype was noted in both groups followed by T/C, and the T allele (P>0.05). PTEN- rs532678- The genotype T/C was the most common in both groups, followed by C/C, and the C allele was predominant in GE compared to control group (P>0.05). PTEN- rs701848- The genotype C/C was highlighted in SG compared to CG, followed by T/T, and the T allele (P>0.05). VEGF-A- rs3025039- The C/C genotype is distinguished in both groups (P>0.05), the same occured for the C allele (P = 0.4226). VEGF-A- rs1570360 – G/G genotype prevailed in both groups (P>0.05), as well as the G allele (P=0.6387). Although similarity between the groups for genotypic and allelic distribution was observed, mutant PTEN and VEGF-A alleles prevailed in patients with HCC and tobacco and alcohol consumption, compared to the control group (P <0.05). PTEN and VEGF haplotype analysis was similar among the groups (P>0.05). Gene expression - PTEN expression levels was decreased in patients with HCC (median= 0,908) compared to cirrhotic patients (median = 5.93, P=0.0347). PIK3CA expression levels (median- HCC= 0,108; cirrhosis= 0,493) were similar between groups (P> 0.05). Conclusion – PTEN and VEGF-A genetic variants, as well as their haplotypes were not associated to HCC. Reduced gene expression of PTEN in tumor tissue can be associated with HCC, while PIK3CA does not differentiate between patients with HCC from those with cirrhosis. It stands out as independent risk factors for HCC, smoking, alcohol consumption, male sex, advanced age and DM. PTEN and VEGF-A mutant alleles, particularly in the presence of smoking and alcohol consumption may enhance the risk for HCC. / Introdução - O carcinoma hepatocelular (CHC) destaca-se como o mais agressivo tumor maligno do fígado. A identificação de genes candidatos a biomarcadores pode contribuir para esclarecer a fisiopatologia do CHC e auxiliar no diagnóstico precoce da doença com novas intervenções terapêuticas. Objetivos - Avaliar a associação de variantes genéticas e expressão gênica envolvidas no processo de sinalização celular, apoptose e angiogênese com CHC, visando caracterizar subgrupos de risco e identificar marcadores biológicos para diagnóstico precoce, prognóstico e tratamento da doença. Casuística e Métodos – Foram estudados 343 indivíduos, sendo 102 com CHC (GE= Grupo de estudo) e 215 controles (GC= Grupo controle) para a análise dos polimorfismos de PTEN (rs10490920, rs532678, rs701848) e VEGF-A (rs3025039 e rs1570360). Para a análise de expressão gênica de PTEN e PIK3CA, foram selecionados 24 pacientes com CHC (GEeg= Grupo de estudo da expressão gênica), 16 com cirrose (GCieg= Grupo cirrose da expressão gênica) e 10 controles submetidos a cirurgias bileo-digestivas (GCeg= Grupo controle da expressão gênica). Os polimorfismos dos referidos genes foram analisados por PCR/RFLP (polymerase chain reaction/restriction fragments lengh polymorphism), enquanto a expressão gênica (tecido hepático fresco) por qPCR (quantitative/polymerase chain reaction). Dados do perfil clínico, hábitos de vida e comorbidades foram obtidos em prontuário médico e questionário. Admitiu-se erro α de 5%. Resultados - O gênero masculino, idade avançada, tabagismo, etilismo e diabetes mellitus (DM) prevaleceram no grupo com CHC, comparado ao controle (P<0,05). Polimorfismos genéticos: PTEN- rs10490920- O genótipo T/T destacou-se em ambos os grupos, seguido de T/C, assim como o alelo T (P>0,05). PTEN- rs532678- O genótipo T/C foi o mais frequente em ambos os grupos, seguido de C/C, assim como o alelo C predominou em GE comparado a GC (P>0,05). PTEN- rs701848- O genótipo T/C destacou-se em GE compado a GC, seguido de T/T, assim como o alelo T (P>0,05). VEGF-A- rs3025039- O genótipo C/C destacou-se em ambos os grupos (P>0,05), o mesmo ocorreu para o alelo C (P=0,4226). VEGF-A- rs1570360 – O genótipo G/G prevaleceu em ambos os grupos (P>0,05), assim como o alelo G (P=0, 6387). Embora detectada semelhança entre os grupos para distribuição genotípica e alélica, alelos mutantes de PTEN e VEGF-A prevaleceram em pacientes com CHC e hábitos tabagista e etilista, comparado ao controle (P<0,05). A análise de haplótipos de PTEN e VEGF-A mostrou semelhança entre os grupos (P>0,05). Expressão gênica- Houve diminuição dos níveis de expressão de PTEN em pacientes com CHC (mediana= 0,908) comparado aos cirróticos (mediana= 5,93; P= 0, 0347). Níveis de expressão de PIK3CA (mediana- CHC=0,108; cirrose= 0,493) foram semelhantes entre os grupos (P>0,05). Conclusão – Variantes genéticas de PTEN e VEGF-A, assim como seus haplótipos, não se associam ao CHC. Expressão gênica reduzida de PTEN no tecido tumoral hepático pode estar associado a CHC, enquanto PIK3CA não diferencia pacientes com CHC daqueles com cirrose. Destacam-se como fatores de risco independentes para CHC tabagismo, etilismo, sexo masculino, idade avançada e DM. Alelos mutantes de PTEN e VEGF-A, particularmente na presença de tabagismo e etilismo podem potencializar o risco para CHC.