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Adenoid ameloblastoma is a very rare benign epithelial odontogenic tumor characterized microscopically by epithelium resembling conventional ameloblastoma, with additional duct-like structures, epithelial whorls, and cribriform architecture. Dentinoid deposits, clusters of clear cells, and ghost-cell keratinization may also be present. These tumors do not harbor BRAF or KRAS mutations and their molecular basis appears distinct from conventional ameloblastoma but remains unknown. We assessed CTNNB1 (beta-catenin) exon 3 mutations in a cohort of 11 samples of adenoid ameloblastomas from 9 patients. Two of the 9 patients were female and 7 male and in 7/9 patients the tumors occurred in the maxilla. Tumors of 4 of these 9 patients harbored CTNNB1 mutations, specifically p.Ser33Cys, p.Gly34Arg, and p.Ser37Phe. Notably, for one patient 3 samples were analyzed including the primary tumour and two consecutive recurrences, and results were positive for the mutation in all three tumors. Therefore, 6/11 samples tested positive for the mutation. In the 6 mutation-positive samples, ghost cells were present in only 2/6, indicating beta-catenin mutations are not always revealed by ghost cell formation. Dentinoid matrix deposition was observed in 5/6 mutation-positive samples and clear cells in all 6 cases. None of the cases harbored either BRAF or KRAS mutations. Beta-catenin immunoexpression was assessed in the samples of 8 patients. Except for one wild-type case, all cases showed focal nuclear expression irrespective of the mutational status. Together with the absence of BRAF mutation, the detection of beta-catenin mutation in adenoid ameloblastomas supports its classification as a separate entity, and not as a subtype of ameloblastoma. The presence of this mutation may help in the diagnosis of challenging cases.

BACKGROUND/AIM Familial multiple endocrine neoplasia type 1 (MEN1) is a rare disorder mostly associated with germline MEN1 mutations. MATERIALS AND METHODS Genotyping of the MEN1, cyclin-dependent kinase inhibitor 1B (CDKN1B) and aryl hydrocarbon receptor-interacting protein (AIP) genes using Sanger sequencing was carried out in a family with MEN1 and the resulting germline variants genotyped in an additional 95 ethnically matched controls. RESULTS A missense variant in AIP (p.Arg16His) gene and a truncating mutation (p.Gly111fs*8) in MEN1 gene were both detected in the proband and his father, showing limited co-segregation with phenotype. p.Arg16His AIP missense variant was detected in one control. CONCLUSION There are conflicting data regarding the functional effects of AIP p.Arg16His and its role in disease development. We demonstrated the co-occurrence of p.Arg16His AIP missense variant in a patient with a bona fide MEN1 mutation. Our finding of p.Arg16His AIP in one of the 95 controls and its co-occurrence with MEN1 in a patient suggests that it is more likely that this variant is a rare polymorphism, unrelated to MEN1 pathogenesis.

Introduction: Congenital megaureter constitutes the second most frequent cause of hydronephrosis in children. There is still much debate on what extent environmental or genetic factors are involved in the pathogenesis of congenital megaureter. Objectives: This study aimed at investigating a pair of monozygotic twins discordant for the expression of bilateral congenital megaureter using the whole exome sequencing technique. Methods: Peripheral blood DNA was extracted and then sequenced using the whole exome technique from a pair of twins discordant for the presence of bilateral congenital refluxing unobstructed megaureter, his parents and a set of 11 non-related individuals with confirmed diagnosis of congenital megaureter. The DNA of the set of 11 non-related individuals was pooled in three groups. The monozygotic twins and their parents had DNA samples sequenced separately. Sanger validation was performed after data was filtered. Results: In the proband were identified 256 candidate genes, including TBX3, GATA6, DHH, LDB3, and HNF1, which are expressed in the urinary tract during the embryonic period. After Sanger validation, the SNVs found in the genes TBX3, GATA6, DHH and LDB3 were not confirmed in the proband. The SNV chr17:36104650 in the HNF1b gene was confirmed in the proband, his twin brother and the mother, however was not found in the pool of 11 non-related individuals with congenital megaureter. Conclusion: Due to the possible complex causative network of genetic variations and the challenges regarding the use of the whole exome sequencing technique we could not unequivocally associate the genetic variations identified in this study with the development of the congenital megaureter.

Background/Aim: Ethnicity has an effect on survival in patients with pancreatic adenocarcinoma (PDAC), which may be reflected in the rate of somatic driver mutations. The Brazilian population represents au extensive interethnic admixture and little is known about the spectrum and rates of somatic driver mutations in Brazilian PDAC cases. Materials and Methods: Direct sequencing of six genes in 23 PDAC cases was performed and the ancestry of patients was determined using a validated panel of ancestry-informative insertion/ deletion DNA polymorphisms. Results: KRAS proto-oncogene (KRAS) was the most commonly mutated gene (60%). A novel putatively pathogenic mutation in phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) (c.2948T>A; p.M983K) was identified. Mutations in epidermal growth factor receptor (EGFR) (4%), PIK3CA (4%), cyclin-dependent kinase inhibitor 2A (CDKN2A) (4%) and TP53 (8%) were noted, in rates that are less frequent than those reported for other populations. Mutations of B-Raf proto-oncogene, serine/threonine kinase (BRAF) were not present. All individuals with high African ancestral component (allelic frequency, >0.45) exhibited KRAS mutations. Conclusion: Our results highlight the importance of the effect of ethnicity on somatic mutations in Brazilian patients with PDAC.

Familial multiple endocrine neoplasia type 1 (MEN1) is a rare disorder mostly associated with germline MEN1 mutations.Genotyping of the MEN1, cyclin-dependent kinase inhibitor 1B (CDKN1B) and aryl hydrocarbon receptor-interacting protein (AIP) genes using Sanger sequencing was carried out in a family with MEN1 and the resulting germline variants genotyped in an additional 95 ethnically matched controls.A missense variant in AIP (p.Arg16His) gene and a truncating mutation (p.Gly111fs*8) in MEN1 gene were both detected in the proband and his father, showing limited co-segregation with phenotype. p.Arg16His AIP missense variant was detected in one control.There are conflicting data regarding the functional effects of AIP p.Arg16His and its role in disease development. We demonstrated the co-occurrence of p.Arg16His AIP missense variant in a patient with a bona fide MEN1 mutation. Our finding of p.Arg16His AIP in one of the 95 controls and its co-occurrence with MEN1 in a patient suggests that it is more likely that this variant is a rare polymorphism, unrelated to MEN1 pathogenesis.

Lung cancer (LC) is a leading cause of cancer-related mortality. Although smoking is the major risk factor, ~15% of all cases occur in never-smokers, suggesting that genetic factors play a role in LC predisposition. Indeed, germline mutations in the TP53 gene predispose to multiple cancer types, including LC. To date, few studies compared the somatic and germline mutational profiles of LC cases by smoking status, and none was reported in Brazilians. Whole-exome sequencing (WES) was performed on two pools (seven smokers and six non-smokers) of tumor-derived DNA using the Illumina HiSeq2000 platform. Files from pools were analyzed separately using Ingenuity®Variant AnalysisTM and Mendel,MD. Validation of all candidate variants was performed by Sanger sequencing. Subsequently, validated mutations were analyzed in germline DNA from the same patients and in ethnically matched controls. In addition, a single recurring Brazilian TP53 germline mutation (R337H) was genotyped in 45 non-small-cell lung cancer patients.Four novel germline variants in the ATAD2, AURKA, PTPRD and THBS1 genes were identified exclusively in smoker patients, and four germline missense variants in PLCD1, RAD52, CP and CDC6 genes were identified solely in non-smokers. There were 4/45 (8.9%) germline carriers of the R337H TP53 mutation. In conclusion, the recurring Brazilian TP53 mutation should be genotyped in all non-small-cell lung cancer in Brazil, regardless of smoking status. Distinct pathogenic mutations and novel sequence variants are detected in Brazilian non-small-cell lung cancer patients, by smoking status. The contribution of these sequence variants to LC pathogenesis remains to be further explored.

Circadian rhythms comprise of daily oscillations in a variety of biological processes and are regulated by an endogenous clock. Disruption of these rhythms has been associated with cancer progression, and understanding natural oscillations in cellular growth control, tumor suppression and cancer treatment, may reveal how clock and clock‑controlled genes are regulated in normal physiological functioning. To investigate the association between clock genes and non‑small cell lung cancer (NSCLC), we genotyped three tag SNPs (rs938836, rs17050680, rs3805213) in the Nocturnin gene (CCRN4L), five SNPs (rs228727, rs228644, rs228729, rs707467, rs104620202) in the period 3 (PER3) gene and one SNP (rs6855837) in the CLOCK gene, in 78 Brazilian patients with NSCLC. One tag SNP in CCRN4L (rs3805213) and another tag SNP from PER3 (rs228729) demonstrated a significant correlation with genotype and allele frequency in lung cancer (P=4.4x10‑3 and P=5.7x10‑2; P=0.004 and P=0.02, respectively). The results of our study suggest these polymorphisms in the CCRN4L and PER3 genes may represent a risk factor in the occurrence and development of NSCLC in Brazilian patients.

O presente trabalho teve como objetivo mostrar a utilidade da biologia molecular para o diagnóstico da síndrome de Bartter (SB) por meio do relato de caso de duas irmãs e propor um algoritmo para abordagem molecular dessa síndrome. Os dois casos relatados apresentaram prematuridade, gestação complicada com poli-hidrâmnio e baixo peso ao nascer. Durante o primeiro ano de vida, as crianças apresentaram poliúria, polidipsia e atraso no crescimento, o que levou à investigação de doenças tubulares renais e erros inatos do metabolismo. Os exames laboratoriais sugeriram SB, mas a confirmação diagnóstica só foi obtida pela detecção de mutação em homozigose no exon 5 do gene KCNJ1, resultando em substituição do aminoácido alanina por valina no códon 214 (A214V) nas duas fitas de DNA nas duas irmãs e de mutação em heterozigose em seus pais. O diagnóstico de certeza da SB muitas vezes é difícil de ser obtido. Dessa forma, por meio dos casos relatados, mostrou-se a utilidade de métodos moleculares para o diagnóstico de certeza da SB, e foi proposto um algoritmo para a utilização racional dessas técnicas.

We review studies from our laboratories using different molecular tools to characterize the ancestry of Brazilians in reference to their Amerindian, European and African roots. Initially we used uniparental DNA markers to investigate the contribution of distinct Y chromosome and mitochondrial DNA lineages to present-day populations. High levels of genetic admixture and strong directional mating between European males and Amerindian and African females were unraveled. We next analyzed different types of biparental autosomal polymorphisms. Especially useful was a set of 40 insertion-deletion polymorphisms (indels) that when studied worldwide proved exquisitely sensitive in discriminating between Amerindians, Europeans and Sub-Saharan Africans. When applied to the study of Brazilians these markers confirmed extensive genomic admixture, but also demonstrated a strong imprint of the massive European immigration wave in the 19th and 20th centuries. The high individual ancestral variability observed suggests that each Brazilian has a singular proportion of Amerindian, European and African ancestries in his mosaic genome. In Brazil, one cannot predict the color of persons from their genomic ancestry nor the opposite. Brazilians should be assessed on a personal basis, as 190 million human beings, and not as members of color groups.

SummaryBackground: Paraganglioma syndrome type 1 (PGL1) is a rare autosomal dominant syndrome associated with multiple, overwhelmingly benign, pheochromocytomas and paragangliomas, attributed to SDHD gene mutations. Objective: Clinically and molecularly characterize a family with uncommon malignant phenotype of paragangliomas attributed to two seemingly pathogenic SDHD germline mutations. Materials & methods: The proband presented with large bilateral carotid body tumours and family history of cervical masses in his five siblings. All family members underwent clinical examination, imaging studies (18F-FDG PET/CT) and genotyping of relevant genes. The proband was diagnosed with locally advanced paraganglioma; his hypertensive, otherwise asymptomatic father, had locally advanced pheochromocytoma and his three siblings showed multiple head and neck masses, confirmed to be paragangliomas with local metastasis. All affected patients carried two germline mutations in the SDHD gene; a previously reported nonsense mutation in exon 1 (p.Trp5X) and a novel missense mutation in exon 2 (p.Pro53Leu), highly deleterious by in silico analysis. Allelic loss at the SDHD locus was not shown for any of the analysed tumours. Conclusions: This is a rare case of malignant PGL1 with seemingly double pathogenic mutations in the SDHD gene, highlighting the possibility that the presence of both mutations is associated with the more aggressive phenotype.

The Y chromosome carries several spermatogenesis genes distributed in three regions: AZFa, AZFb and AZFc. Microdeletions in these regions have been seen in 10% of sterile males with azoospermia or oligozoospermia, the most frequent of them being characterized by a complete deletion of AZFc region. A partial AZFc deletion named gr/gr has been singled out as a risk factor for spermatogenic failure. However, other authors have diagnosed it as a polymorphic deletion with no clinical relevance. We decided to investigate the association of gr/gr deletion and infertility in Brazilian males. We analysed 350 individuals (110 azoospermic, 122 fertile and 118 presumably fertile) and observed 12 g/gr deletions: five in infertile men (4.5%), three among fertile males (2.5%) and four in probably fertile individuals (3.4%). These differences were not statistically significant. Then, we decided to ascertain whether the clinical impact of the gr/gr deletion was associated with the type of Y chromosome. We have identified Y-chromosome haplogroups using 22 unique event polymorphisms (UEPs). Among the individuals with the gr/gr deletion, we found haplogroups R, K*, F*, E1, E3b2 and E3b*, all of which are common in white Brazilian males, and none revealed a particular association with infertility. Taken together, these results show no evidence of association between the occurrence of gr/gr deletion and male infertility.

Background Obesity has become a common human disorder associated with significant morbidity and mortality and adverse effects on quality of life. Sequence variants in two candidate genes, FTO and UCP-1, have been reported to be overrepresented in obese Caucasian population. The association of these genes polymorphisms with the obesity phenotype in a multiethnic group such as the Brazilian population has not been previously reported. Methods To assess the putative contribution of both FTO and UCP-1 to body mass index (BMI) and cardiovascular risk we genotyped SNPs rs9939609 (FTO) and rs6536991, rs22705565 and rs12502572 (UCP-1) from 126 morbidly obese subjects (BMI 42.9 ± 5.6 kg/m2, mean ± SE) and 113 normal-weight ethnically matched controls (BMI 22.6 ± 3.5 kg/m2, mean ± SE). Waist circumference, blood pressure, glucose and serum lipids were also measured. Each sample was also genotyped for 40 biallelic short insertion/deletion polymorphism (indels) for ethnic assignment and to estimate the proportion of European, African and Amerindian biogeographical ancestry in the Brazilian population. Results Cases did not differ from controls in the proportions of genomic ancestry. The FTO SNP rs9939609 and UCP-1 SNP rs6536991 were significantly associated with BMI (p= 0.04 and pFTO and UCP-1 SNPs with obesity were noted. There was not an association between rs9939609 (FTO) and rs6536991 (UCP-1) in with maximum weight loss after 1 year in 94 obese patients who underwent bariatric surgery. Conclusion Our data are consistent with FTO rs9939609 and UCP-1 rs6536991 common variants as contributors to obesity in the Brazilian population.

This paper aims to show the utility of molecular biology for diagnose Bartter syndrome (BS) by the case report of two sisters and to propose a diagram for the molecular approach of this syndrome. The two reported cases presented prematurity, pregnancy complicated with polyhydramnio and low birth weight. During the first year of life, children exhibited polyuria, polydipsia and failure to thrive, leading to the investigation of renal tubular diseases and innate errors of metabolism. The laboratorial exams suggested BS, but the definitive diagnostic was only obtained by the detection of homozygous mutation on the exon 5 of the gene KCNJ1, resulting in a substitution of the aminoacid alanin for valin on codon 214 (A214V) in both DNA stripes in the two sisters and a heterozygous mutation in their parents. The definitive diagnostic of BS is frequently very difficult to be obtained. Consequently, considering the reported cases, we showed the utility of molecular techniques for the definitive diagnostic of BS and we proposed a diagram for the rational use of these techniques.

In the contemporary world, "race" narratives are so multifaceted that at times, different views of the concept appear mutually incompatible. In recent decades biologists, especially geneticists, have repeatedly stated that the notion of race does not apply to the human species. On the other hand, social scientists claim that race is highly significant in cultural, historical, and socioeconomic terms because it molds everyday social relations and because it is a powerful motivator for social and political movements based on race differences. In this paper we present the results of an interdisciplinary research project incorporating approaches from genetics and anthropology. Our objective is to explore the interface between information about biology/genetics and perceptions about color/ race in Rio de Janeiro, Brazil. We argue that the data and interpretation of our research resonate far beyond the local level, stimulating discussion about methodological, theoretical, and political issues of wider national and international relevance. Topics addressed include the complex terminology of color/race classification in Brazil, perceptions about ancestry in the context of ideologies of Brazilian national identity, and the relationship between genetic information about the Brazilian population and a sociopolitical agenda that turns on questions of race and racism.

The methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism is associated with the expression of a thermolabile enzyme with decreased activity that influences the pool of methyl-donor molecules. Several studies have reported an association between C677T polymorphism and susceptibility to colorectal cancer (CRC). Considering that methylation abnormalities appear to be important for the pathogenesis of CRC, we examined the correlation between the genotype of the MTHFR C677T polymorphism, hypermethylation of the promoter region of five relevant genes (DAPK, MGMT, hMLH1, p16(INK4a), and p14(ARF)), and microsatellite instability, in 106 patients with primary CRCs in Brazil. We did not find significant differences in the genotypic frequencies of the MTHFR C677T polymorphism when one or more loci were hypermethylated. However, we did find a significant excess of 677TT individuals among patients with CRC who had microsatellite instability. This strong association was independent of the methylation status of hMLH1 and of the biogeographical genomic ancestry of the patients. Although the mechanism responsible for the link between the C677T polymorphism and microsatellite instability was not apparent, this finding may provide a clue towards a better understanding of the pathogenesis of microsatellite instability in human colorectal cancer.

Acquisition of detailed knowledge of the structure and evolution of Trypanosoma cruzi populations is essential for control of Chagas disease. We profiled 75 strains of the parasite with five nuclear microsatellite loci, 24Sα RNA genes, and sequence polymorphisms in the mitochondrial cytochrome oxidase subunit II gene. We also used sequences available in GenBank for the mitochondrial genes cytochrome B and NADH dehydrogenase subunit 1. A multidimensional scaling plot (MDS) based in microsatellite data divided the parasites into four clusters corresponding to T. cruzi I (MDS-cluster A), T. cruzi II (MDS-cluster C), a third group of T. cruzi strains (MDS-cluster B), and hybrid strains (MDS-cluster BH). The first two clusters matched respectively mitochondrial clades A and C, while the other two belonged to mitochondrial clade B. The 24Sα rDNA and microsatellite profiling data were combined into multilocus genotypes that were analyzed by the haplotype reconstruction program PHASE. We identified 141 haplotypes that were clearly distributed into three haplogroups (X, Y, and Z). All strains belonging to T. cruzi I (MDS-cluster A) were Z/Z, the T. cruzi II strains (MDS-cluster C) were Y/Y, and those belonging to MDS-cluster B (unclassified T. cruzi) had X/X haplogroup genotypes. The strains grouped in the MDS-cluster BH were X/Y, confirming their hybrid character. Based on these results we propose the following minimal scenario for T. cruzi evolution. In a distant past there were at a minimum three ancestral lineages that we may call, respectively, T. cruzi I, T. cruzi II, and T. cruzi III. At least two hybridization events involving T. cruzi II and T. cruzi III produced evolutionarily viable progeny. In both events, the mitochondrial recipient (as identified by the mitochondrial clade of the hybrid strains) was T. cruzi II and the mitochondrial donor was T. cruzi III., Synopsis The parasite protozoan Trypanosoma cruzi causes Chagas disease, a malady that afflicts almost 20 million people in South America and Central America. Although the genome sequencing of T. cruzi has been recently completed, little is known about its population structure and evolution. Since 1999, two major evolutionary lineages presenting distinct epidemiological characteristics have been recognized in the parasite: T. cruzi I and T. cruzi II, the latter being much more associated with severe chronic cases of the disease. We describe new and important aspects of the population structure of the parasite, especially the characterization of a third ancestral lineage that we propose to call T. cruzi III. Through careful dissection of the genetic constitution of blocks of genes that are stably transmitted from generation to generation of the parasite we deduced at least two occurrences of the formation of hybrid strains from the parental lineages T. cruzi II and T. cruzi III, including the strain CLBrener, whose genome was sequenced. We did not find any hybrids originating from T. cruzi I. A fascinating finding was that both hybrids studied had the same mitochondrial DNA type as the T. cruzi III ancestral lineage, which was quite different from T.cruzi II.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior A síndrome da apneia obstrutiva do sono (SAOS) é caracterizada pela obstrução completa ou parcial das vias aéreas superiores, resultando em períodos de apneia associada ao sono. A SAOS aumenta o risco de morbidade e mortalidade por doenças cardiovasculares e cerebrovasculares. Embora a herdabilidade da SAOS seja estimada em ~40%, os genes subjacentes precisos permanecem indefinidos. O objetivo do presente estudo é definir a base genética da SAOS aparentemente monogênica em duas famílias brasileiras. Essas famílias seguem um padrão sugestivo de herança autossômica dominante. O sequenciamento completo do exoma do DNA da linhagem germinativa e os dados gerados foram analisados usando o software Mendel,MD. As variantes selecionadas foram analisadas usando Varstation®, com análises subsequentes que incluíram validação por sequenciamento de Sanger, avaliação de pontuação patogênica pelos critérios ACMG, análises de cosegregação (quando possível), frequência alélica, padrões de expressão tecidual, análises de vias, efeito na modelagem de dobramento de proteínas usando SwissModelo e RaptorX. Duas famílias (seis casos de SAOS afetados e três controles não afetados) foram analisadas. Uma análise abrangente de várias etapas apontou variantes em COX20 (rs946982087) (família A), PTPDC1 (rs61743388) e TMOD4 (rs141507115) (família B) que parecem ser genes fortes candidatos a serem associados à OSAS nessas famílias. Em conclusão, as variantes encontradas em COX20, PTPDC1 e TMOD4 aparentemente estão correlacionadas ao fenótipo da SAOS nessas famílias. Mais estudos em famílias etnicamente diversas e casos de SAOS não familiares são necessários para melhor definir o papel dessas variantes como contribuintes para o fenótipo da SAOS. Obstructive sleep apnea syndrome (OSAS) is characterized by complete or partial obstruction of the upper airways, resulting in periods of sleep associated apnea. OSAS increases morbidity and mortality risk from cardiovascular and cerebrovascular diseases. While heritability of OSAS is estimated at ~40%, the precise underlying genes remain elusive. The aim of present study is define the genetic basis of seemingly monogenic OSAS in two Brazilian families. Brazilian families with OSAS that follows as seemingly autosomal dominant inheritance pattern were recruited. Whole exome sequencing of germline DNA and the generated data were analyzed using Mendel,MD software. Variants selected were analyzed using Varstation® with subsequent analyses that included validation by Sanger sequencing, pathogenic score assessment by ACMG criteria, cosegregation analyses (when possible) allele frequency, tissue expression patterns, pathway analyses, effect on protein folding modeling using SwissModel and RaptorX. Two families (six affected OSAS cases and three unaffected controls) were analyzed. A comprehensive multistep analyses yielded variants in COX20 (rs946982087) (family A), PTPDC1 (rs61743388) and TMOD4 (rs141507115) (family B) that seemed to be strong candidate genes for being OSAS associated genes in these families. In conclusion, sequence variants in COX20, PTPDC1 and TMOD4 seemingly are associated with OSAS phenotype in these families. Further studies in more, ethnically diverse families and nonfamilial OSAS cases are needed to better define the role of these variants as contributors to OSAS phenotype.

INTRODUÇÃO: A cirurgia de derivação gástrica em Y de Roux (DGYR) é considerada padrão para o tratamento da obesidade grave. Apesar de ser um tratamento eficaz na maioria dos casos, uma parcela dos pacientes não obtém uma perda de peso bem sucedida nos dois primeiros anos. Vários fatores têm sido demonstrados como preditores da resposta de perda de peso, porém mais estudos ainda são necessários. OBJETIVO: Avaliar os preditores da resposta de perda de peso em pacientes submetidos à DGRY. MÉTODOS: Trata-se de um estudo longitudinal retrospectivo em 63 pacientes submetidos à DGYR entre 2004 e 2014 e acompanhados no pós-operatório por pelo menos 24 meses. Dados do pré- e pós-operatório de 12 e 24 meses foram coletados em prontuários, incluindo presença de comorbidades antes da cirurgia, histórico familiar de obesidade, medidas antropométricas, ingestão de energia e macronutrientes e prática de exercícios físicos. Polimorfismos nos genes BDNF (Val66Met - rs6265) e LYPLAL1 (rs4846567) foram genotipados utilizando a metodologia TaqMan™ SNP Genotyping assays. A perda de excesso de peso (PEP) >50% foi adotada como critério de perda de peso bem sucedida e os participantes foram distribuídos em grupo perda de peso bem sucedida (PPBS) e perda de peso insuficiente (PPI). Modelos de regressão logística foram utilizados para verificar variáveis preditoras do sucesso da cirurgia. O nível de significância adotado foi p

Introdução: A síndrome da apneia obstrutiva do sono (SAOS) é um distúrbio comum associado ao aumento do risco de doenças cardiovasculares, metabólicas e mortalidade. Fatores de risco para SAOS têm uma base genética conhecida ou suspeita e a descoberta dos marcadores genéticos é de extrema importância para sua prevenção e tratamento personalizado. O objetivo desse estudo foi avaliar a correlação entre alterações genéticas, fenótipos e dados clínicos que possam contribuir para o desenvolvimento da SAOS e propor um modelo preditivo para a síndrome. Métodos: Avaliamos dados polissonográficos, clínicos e genotípicos de duzentos e quarenta e dois pacientes, cento e oitenta e um casos e oitenta e um controles. As variantes genéticas: dos genes PER3 (rs228697, rs228727, rs228729 e rs10462020), BDNF (rs6265), NRXN3 (rs10146997), APOE (rs7412 e rs429358), HCRTR2 (rs2653349), MC4R (rs17782313) foram genotipadas e analisadas no programa estatístico R (versão 3.6.3). Resultados: A média de idade dos participantes foi 51,48 ± 14,29, com predomínio do sexo masculino (57,02%) e a média de IMC foi 26,63 ± 4,36. A análise de associação entre os genótipos e a SAOS não evidenciou associação entre os polimorfismos PER3 (rs228697, rs228727 e rs10462020), NRXN3 (rs10146997), APOE (rs7412 e rs429358), HCRTR2 (rs2653349) e MC4R (rs17782313) com SAOS. Entretanto, os polimorfismos do BDNF (rs6265) e do PER3 (rs228729) mostraram associação significante com SAOS (p = 0,01317 e p = 0,000094, respectivamente). No entanto, após ajuste para idade e IMC, BDNF (rs6265) e do PER3 (rs228729) não continuaram mostrando associação significante (p =0,06201 e p= 0,0659, respectivamente). Um algoritmo proposto mostrou capacidade adequada para predizer SAOS apresentando boa sensibilidade e especificidade. Para valores de pontos de corte crescentes de 0 a 1, obteve-se a curva ROC cuja área baixa foi de 79,4% (IC 95% 72,2% - 86,6%). O melhor valor de corte foi de 0,763 com sensibilidade de 71,6% e especificidade de 76,5%. Conclusão: O algoritmo gerado neste trabalho, o qual inclui dados de características sociodemográficas, clínicas e genéticas dos pacientes poderá ser aplicado à prática clínica e permitirá a estratificação de risco para a SAOS. A aplicabilidade desse algoritmo deve ser comprovada em estudos com maior número de pacientes e em diferentes etnias. Introduction: Obstructive sleep apnea syndrome is a common disorder associated with an increased risk of cardiovascular, metabolic diseases and mortality. Risk factors for OSAS have a known or suspected genetic basis and the discovery of genetic markers is extremely important for their prevention and personalized treatment. The aim of this study was to assess the correlation between genetic changes, phenotypes and clinical data that can contribute to the development of OSAS and propose a predictive model for OSAS. Methods: case-control study with two hundred and forty-two patients, one hundred and eighty-one cases and eighty-one controls included. Polysomnographic, clinical, and genotypic data for the PER3 genes (rs228697, rs228727, rs228729 and rs10462020), BDNF (rs6265), NRXN3 (rs10146997), APOE (rs7412 and rs429358), HCRTR2 (rs2653349), MC4R (rs17782313) were analyzed (rs7) statistic R (version 3.6.3). Results: Two hundred and forty-two patients, 161 cases and 81 controls were included in the research. The average age of the participants was 51.48 ± 14.29, with a male predominance (57.02%) and the average BMI was 26.63 ± 4.36. The analysis of association between genotypes and OSAS did not show any association between the PER3 (rs228697, rs228727 and rs10462020) polymorphisms, NRXN3 (rs10146997), APOE (rs7412 and rs429358), HCRTR2 (rs2653349) and MC4R (rs17782313) with SA However, the BDNF (rs6265) and PER3 (rs228729) polymorphisms showed a significant association with OSAS (p = 0.01317 and p = 0.000094, respectively). However, after adjusting for age and BMI, BDNF (rs6265) and PER3 (rs228729) did not continue to show a significant association (p = 0.06201 and p = 0.0659, respectively). A proposed algorithm showed an adequate ability to predict OSAS with good sensitivity and specificity. For values of increasing cutoff points from 0 to 1, we obtained ROC curve whose low area was 79.4% (CI 95% 72.2% - 86.6%). The best cutoff value was 0.763 with sensitivity of 71.6% and specificity of 76.5%. Conclusion: The algorithm generated in this study, which includes data on sociodemographic, clinical, and genetic characteristics of patients, can be applied to clinical practice and will allow risk stratification for OSAS. The applicability of this algorithm must be proven in studies with a larger number of patients and in different ethnicities.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico O câncer de tireoide é o tumor maligno mais comum do sistema endócrino e representa 2,1% dos diagnósticos de câncer em todo o mundo. O carcinoma papilífero da tireoide (PTC) corresponde a mais de 80% dos casos desse tumor e é predominantemente esporádico. Casos de PTC hereditários consistem em 5%, tendem a ter início mais precoce, além de maiores riscos de metástase e de recorrência. O PTC familiar é uma doença geneticamente heterogênea, cuja patogênese não está associada às variantes observadas no carcinoma papilífero de tireoide esporádico. Assim, ainda permanecem desconhecidas as alterações moleculares associadas à carcinogênese da tireoide na maioria das famílias acometidas pelo PTC. Nesse contexto, o objetivo deste trabalho foi determinar a base genética de PTC em três famílias brasileiras (Famílias A, B e C) compostas por dois ou mais parentes de primeiro grau com o fenótipo do tumor de tireoide. Mutações germinativas foram pesquisadas em pelo menos um integrante de cada família utilizando-se o sequenciamento completo do exoma, enquanto o sequenciamento de Sanger foi utilizado para validação das mutações encontradas nos exomas e para genotipagem dos genes candidatos nos outros indivíduos de cada família. Foram identificadas sete variantes que podem estar associadas à suscetibilidade para PTC familiar. Dentre as variantes candidatas, foram observadas duas na Família A (p.D283N no gene ANXA3 e p.Y157S no gene NTN4), uma na Família B (p.G172W no gene SERPINA1), quatro na Família C (p.G188S no gene FKBP10, p.R937C no gene PLEKHG5, p.L32Q no gene P2RX5, e p.Q76* no gene SAPCD1). Essas mutações foram validadas pelo Sequenciamento de Sanger e também estavam presentes nos demais membros das respectivas famílias. Não foram validadas variantes candidatas comuns a todas as famílias. Mais estudos são necessários para a compreensão do papel exato dessas variantes, bem como a avaliação da presença destas mutações em outras famílias com PTC.

OBJETIVOS: Investigar a ação do uso de Bevacizumabe, Mitomicina C (MMC) e Acetato de Triancinolona (AT) (isolados e em associação) na cicatrização pós-operatória após trabeculectomia (TREC) em coelhos. MÉTODOS: Quarenta e cinco coelhos albinos da raça Nova Zelândia foram submetidos à TREC em ambos os olhos e receberam, no transoperatório, tratamentos diferentes: solução salina; MMC; AT; AT+MMC; Bevacizumabe; Bevacizumabe+MMC. A MMC e a solução salina foram utilizadas abaixo do retalho conjuntival; AT e Bevacizumabe foram utilizados imediatamente após a cirurgia em injeção subconjuntival. Os coelhos foram avaliados em dias diferentes durante o pós-operatório; mediu-se a pressão intraocular (Po) e fez-se a análise da bolha baseada no sistema de classificação de bolha de Moorfields. Os animais foram mortos nos dias 3, 14 e 30 de pós-operatório (DPO). Histologia e imunohistoquímica foram realizadas para avaliar a cicatrização. RESULTADOS: O grupo Bevacizumabe+MMC apresentou menor Po ao final do estudo (7,7 ± 0,8 mmHg), seguido pelos grupos MMC (8,8 ± 1,85 mmHg) e AT+MMC (8,9 ± 0,9 mmHg) (p=0,001). O grupo Bevacizumabe+MMC apresentou melhores resultados clínicos em relação à altura máxima da bolha, à área central da bolha e à área máxima da bolha (p

Adenomas hipofisários são tumores intracranianos comuns que ocorrem de forma esporádica. Em alguns casos raros, esta doença é identificada em grupos familiares e não tem envolvimento com outros tumores endócrinos, uma condição identificada como adenomas de hipófise familiais isolados (FIPA). O desenvolvimento de FIPA tem sido associado a anormalidades genéticas, especialmente no gene AIP, no qual mutações germinativas foram relatadas em aproximadamente 30% dos casos. Mutações no gene MEN1 foram descritas em famílias com adenoma de hipófise, mas com fenótipo específico para neoplasia endócrina múltipla tipo 1. Mutações no receptor de prolactina (PRLR) também têm sido associadas ao desenvolvimento de adenoma hipofisário em modelos animais. Assim, na maioria dos casos de FIPA o defeito genético exato que leva ao desenvolvimento da doença permanece desconhecido. Portanto, o objetivo deste trabalho é determinar a base genética de FIPA em uma família brasileira. A família estudada é composta por três irmãos diagnosticados com adenoma hipofisário secretor de prolactina através de testes clínicos, bioquímicos e de imagem. O sequenciamento Sanger foi utilizado para genotipagem dos genes candidatos AIP, MEN1 e PRLR. Além disso, uma pesquisa por novas mutações foi realizada utilizando-se a técnica de sequenciamento de todo exoma. Uma análise in silico e algoritmos de previsão foram aplicados para priorizar variantes provavelmente patogênicas. Nenhuma mutação foi identificada na região codificadora e reguladora de MEN1, AIP e PRLR. O exoma revelou variantes novas e provavelmente patogênicas em três genes diferentes: RXRG, REXO4 e TH. As variantes em RXRG e TH podem estar associadas ao prolactinoma isolado na família estudada e a possível contribuição desses genes para outras famílias FIPA devem ser exploradas. Pituitary adenomas are common intracranial tumors that occur sporadically. In some rare cases this condition is identified in familial clusters and has no involvement with other endocrine tumors, a disorder identified as Familial Isolated Pituitary Adenoma (FIPA). FIPA development has been associated with genetic abnormalities, especially in AIP gene, where germline mutations have been reported in approximately 20% of cases. Mutations in the MEN1 gene have been described in a subset of pituitary adenoma families, but with bona fide multiple endocrine neoplasia type 1 feature. Mutations in prolactin receptor (PRLR) have also been associated to pituitary adenoma in animal models. Thus, in most FIPA cases the exact genetic defect that lead to disease development remains unknown. Therefore, the aim of this work is to determine the genetic basis of FIPA in a Brazilian family. The studied family is composed of three siblings presented with isolated prolactinsecreting pituitary adenoma diagnosed through clinical, biochemical and imaging testing. Sanger sequencing was used to genotype candidate genes AIP, MEN1 and PRLR. Further mutation screening was performed using whole-exome sequencing. In silico analysis and additional predictive algorithms were applied to prioritize likely pathogenic variants. No mutations in the coding and flanking intronic regions in the MEN1, AIP and PRLR genes were detected. Whole-exome sequencing revealed novel, predicted damaging, heterozygous variants in three different genes: RXRG, REXO4 and TH. The RXRG and TH possibly pathogenic variants may be associated with isolated prolactinoma in the studied family and the possible contribution of these genes to additional FIPA families should be explored.

A origem das anomalias congênitas do rim e do trato urinário (CAKUT) é complexa e diversificada. O megaureter é uma entidade clínica caracterizada pela dilatação do ureter associada ou não às alterações do trato urinário. Constitui a segunda causa mais comum de hidronefrose no recém-nascido, apesar disso, pouco se sabe a respeito dos mecanismos fisiopatológicos do megaureter. Historicamente, estudos em animais ou envolvendo gêmeos humanos têm sido usados como modelo para determinar a participação de fatores ambientais e/ou genéticos no desenvolvimento de doenças congênitas. Neste contexto, este estudo pretendeu, através da técnica WES, ampliar os conhecimentos a respeito dos possíveis fatores genéticos associados ao desenvolvimento do megaureter. Entre junho de 2012 e junho de 2013, todos os portadores de megaureter em acompanhamento no ambulatório Bias Fortes do Hospital das Clínicas da UFMG foram recrutados mediante o fluxo constante de atendimento em nefrologia. O diagnóstico do megaureter foi assegurado durante os períodos pré-natal e neonatal por nefrologistas do Serviço de Nefrologia Pediátrica do Hospital das Clínicas da UFMG. Propôs-se estudar os variantes gênicos em uma família de gêmeos monozigóticos discordantes quanto à expressão do megaureter e em um pool de 11 pacientes não aparentados portadores de megaureter. Este estudo seguiu as diretrizes da Declaração de Helsinki e foi aprovado pelo COEPE UFMG. Após aplicados os filtros para priorização de genes candidatos identificamos 81 SNVs exclusivas do probando em relação aos seus familiares e 5 SNVs exclusivas do probando em relação aos seus familiares e comuns ao pool de portadores de megaureter. Utilizando a ferramenta STRING identificamos como sendo de particular interesse as variações encontradas nos genes TBX3, GATA6, GATA1, DHH, ACVRL1 e SOX2. As SNVs identificadas nos genes TBX3, GATA6 e DHH não foram confirmadas pelo método Sanger, sugerindo se tratarem de falsos positivos. A validação das SNVs identificadas nos genes GATA1,ACVRL1 e SOX2 encontra-se em andamento. Ainda existe um longo caminho a ser percorrido antes que os mecanismos envolvidos no desenvolvimento do megaureter sejam totalmente esclarecidos. O presente estudo representa um passo inicial no sentido de identificar possíveis genes envolvidos na gênese desta condição. Congenital anomalies of the kidney and urinary tract (CAKUT) represent a broad range of disorders that result from abnormalities of the urinary collecting system, abnormal embryonic migration of the kidneys or abnormal renal parenchyma development. Megaureter constitutes one of the phenotypes of CAKUT and represents a condition whereby the diameter of the ureter is abnormally dilated. Currently, much is beingstudied regarding the pathogenesis of CAKUT. While many CAKUT cases are apparently sporadic, familial clustering is common, suggesting that CAKUT pathogenesis is influenced by genetic factors. Some authors have hypothesized that variations in specific regions of the genome could imbalance the mechanisms involved in the expressions of cytokines needed to modulate the ureter morphogenesis and thusresult in the development of megaureter. Despite this hypothesis, its still unknown to what extent environmental or genetic factors are implicated. Therefore, this study aimed to investigate a pair of monozygotic twins discordant for the expression of bilateral congenital megaureter for the presence of genetic variations using the whole exome technique. This study also included a set of 11 non-related individuals with a confirmeddiagnosis of congenital megaureter. All patients were recruited from June 2012 to June 2013 at the Pediatric Nephrology Unit, Hospital Bias Fortes, at UFMG. This study followed the Declaration of Helsinki and was approved the local ethics committee. After applying filters for SNVs prioritization we identified 81 SNVs present exclusively in the proband compared to his family and 5 SNVs present exclusively in the probandand in common to the pool of non-related megaureter patients. After using the STRING tool we prioritize the SNVs found in the genes TBX3, GATA6, GATA1, DHH, ACVRL1 and SOX2 for validation using the Sanger technic. The SNVs found at the genes TBX3, GATA6 and DHH were not confirmed in the proband. Validation of SNVs identified at the genes GATA1, ACVRL1 and SOX2 are in progress. There is still a long way to go before we fully uncover the mechanisms involved in the development the megaureter. This study has taken one step closer in identifying candidate genes involved in the genesis of megaureter.

Acidose tubular renal (ATR) distal ou ATR tipo 1 compreende um grupo heterogêneo de afecções resultantes da disfunção dos túbulos distais, que pode levar a um déficit de crescimento, nefrocalcinose, raquitismo e, raramente, doença renal crônica. Essa tubulopatia pode ser transmitida tanto na forma autossômica dominante quanto na autossômica recessiva. A forma dominante da doença ocorre tipicamente na adolescência ou na idade adulta, enquanto a variante recessiva desenvolve-se predominantemente na infância. O objetivo deste estudo foi detectar e caracterizar asalterações genéticas presentes em duas famílias, não relacionadas, de pacientes portadores de ATR distal atravésda metodologia de whole-exome sequencing (WES). A familia 1 écomposta por uma menina e seu irmão que apresentavam ATR distal e audição normal, enquanto na família 2 havia duas irmãs gêmeas com ATR distal associada a surdez neurosensorial. A técnica de WES foi realizada em duas amostras agrupadas e, para confirmar os resultados, foi utilizado o método de sequenciamento de Sanger. Duas mutações foram identificadas nos genes ATP6VOA4 e ATP6V1B1: uma mutação nova no exon 13 do gene ATP6V0A4 (c.1232G>T) e uma mutação no exon 12 do gene ATP6.V1B1 que já foi previamente descrita na literatura (c.1149_1152insC). Nosso estudo indica que os resultados obtidos com o whole-exome sequencing podem ser úteis para o diagnóstico e a abordagem clinica de pacientes ATR distal, especialmente porque, além de ser uma doença Mendeliana, é rara e com herança complexa. Nossos resultados confirmaram o valor do whole-exome sequencing para o estudo de doenças raras e complexas, permitindo a identificação de mutações novas e recorrentes. Além disso, nosso estudo mostra a aplicabilidade deste método molecular no estudo de doenças renais tubulares. renal tubular acidosis (RTA) or distal ATR type 1 comprises a heterogeneous group of disorders arising from dysfunction of the distal tubules, which can lead to a growth deficit, nephrocalcinosis, rickets and rarely chronic kidney disease. This tubulopathy may be transmitted either in an autosomal dominant manner and in autosomal recessive. The dominant form of the disease typically occurs in adolescence or adulthood, while the recessive variant develops predominantly in childhood. The aim of this study was to detect and characterize genetic asalterações present in two families, unrelated to patients with distal ATR atravésda methodology of whole-exome sequencing (WES). The family is 1 composed of a girl and her brother who had distal ATR and normal hearing, while the family had two twin sisters with distal ATR associated with sensorineural deafness. WES technique was performed on two pooled samples and to confirm the results was performed using the Sanger sequencing method. Two mutations were identified in ATP6VOA4 and ATP6V1B1 genes: a novel mutation in exon 13 of the gene ATP6V0A4 (c.1232G> T) and a mutation in exon 12 of the ATP6.V1B1 gene has been previously described in the literature (c.1149_1152insC). Our study indicates that the results obtained with whole-exome sequencing may be useful for the diagnosis and clinical management of patients distal ATR, especially because in addition to being a Mendelian disease is rare and complex inheritance. Our results confirm the value of whole-exome sequencing for the study of rare and complex diseases, allowing the identification of new and recurrent mutations. Furthermore, our study shows the applicability of this method to study the molecular Tubular renal disease. .

O câncer de vesícula é a neoplasia mais comum do trato biliar e a quinta mais frequente no trato gastrointestinal. Esse câncer é considerado muito raro na maioria dos países ocidentais, mas apresenta alta incidência na Ásia e em alguns países da América do Sul. Seu principal fator de risco é a colelitíase, mas também está associado à da idade avançada, sexo feminino, alguns grupos étnicos, inflamação crônica na vesícula e junção pancreatobiliar anômala (JPBA). O carcinoma vesicular apresenta sinais e sintomas inespecíficos e por isso o diagnóstico na maioria dos casos é tardio, quando o tumor já se encontra irressecável. O prognóstico é, em geral, ruim com sobrevida de cinco anos em apenas 5 a 10% dos pacientes. A carcinogênese vesicular ainda não é bem compreendida, principalmente no que tange aos mecanismos genéticos. Com o objetivo de melhor elucidar esse mecanismo o presente trabalho avaliou o papel de oncogenes e genes supressores de tumor na tumorigênese vesicular. Sendo assim, a presença e frequência de mutações nos oncogenes KRAS, PIK3CA e BRAF e nos supressores tumorais P53 e CDKN2A foi avaliada em 12 pacientes com diagnóstico de carcinoma de vesícula. Os dados clínicos foram avaliados e a ancestralidade genômica desses indivíduos investigada. Os pacientes apresentaram média de idade de 65 anos, 75% do sexo feminino e a grande maioria (92%) apresentava colelitíase. A maior parte deles (75%) recebeu o diagnóstico com o tumor em estágio avançado. A análise de ancestralidade genômica sugere relação do componente ancestral africano com a ocorrência da doença. As análises genéticas mostraram que 25% dos pacientes apresentaram mutação no oncogene KRAS, sendo que um deles apresentou duas alterações. Uma dessas alterações (I55T) encontradas é uma mutação ainda não descrita na literatura. CDKN2A foi o gene que apresentou maior frequência de mutação (41,7%). Deleções em TP53 foram encontradas em 16,7% das amostras. Não foram encontradas mutações em BRAF e PIK3CA. De acordo com os resultados desse trabalho pode-se indicar uma importante associação do gene CDKN2A no desenvolvimento de tumores de vesícula. Os genes TP53 e KRAS, apesar de apresentarem uma frequência menor de mutações, também estão associados à carcinogênese desses tumores. Apesar de os resultados não terem demonstrado alterações nos genes BRAF e PIK3CA mais estudos são necessários para avaliar sua participação na tumorigênese vesicular. Palavras chave: Câncer de vesícula, CDKN2A, P53 Gallbladder cancer is the most common malignancy of the biliary tract and the fifth most common tumor in the gastrointestinal tract. This tumor is rare in most Western countries but has high incidence in Asia and in some South American countries. Its main risk factor is cholelithiasis but it is also associated with advanced age, female sex, some ethnic groups, chronic inflammation in gallbladder and anomalous pancreaticobiliary junction (APBJ). Gallbladder carcinoma presents nonspecific signs and symptoms; in most cases the tumor is not easily diagnosed and not probe to resection. The prognosis is generally poor with a five-year survival in only 5-10 % of patients. The gallbladder carcinogenesis is not well understood, especially regarding genetic mechanisms. Aiming to further elucidate this mechanism the present study evaluated the role of oncogenes and tumor suppressor genes in vesicular tumorigenesis. Thus, the presence and frequency of mutations in the oncogenes KRAS, BRAF, and PIK3CA, and the tumor suppressors p53 and CDKN2A were determined in 12 patients with gallbladder carcinoma. Clinical data were assessed and genomic ancestry of these individuals performed. Patients had a mean age of 65 years old, 75% were female and the vast majority (92%) had cholelithiasis. Most of them (75%) had been diagnosed with advanced-stage tumor. The analysis of genomic ancestry suggests a relationship of African component ancestry with the occurrence of the disease. Genetic analysis showed that 25% of patients had mutations in KRAS oncogene, one of which presented two changes. One of these changes (I55T) is a novel mutation. The CDKN2A gene was the one with higher mutation frequency (41.7%). P53 deletions were found in 16.7% of samples. There were no mutations in BRAF and PIK3CA. According to the results of this study a significant association of the CDKN2A gene in the development of tumors of the gallbladder may be suggested. KRAS and p53 genes, despite having a lower mutation frequency, are also associated with carcinogenesis of these tumors. Although the results have not shown alterations in BRAF and PIK3CA more studies are needed to establish their involvement in gallbladder tumorigenesis.

O Transtorno Afetivo Bipolar (TAB) e um transtorno psiquiatrico grave, cronico e com progressivo aumento da gravidade dos episodios. O prognostico para os pacientes com TAB e pobre, com altas taxas de recaida, de sintomas residuais, disfuncoes cognitivas e de diminuicao da qualidade de vida. Um crescente corpo de estudos suporta a hipotese de que o TAB surge de anormalidades em cascatas de plasticidade celular, levando a processamento deficiente de informacao nas sinapses e circuitos de mediacao de funcoes afetivas e cognitivas. Varias linhas de evidencias sugerem que a via Wnt pode, tambem, estar envolvida na etiologia do TAB, pois esta via esta relacionada a processos de desenvolvimento essenciais tais como crescimento celular, migracao e diferenciacao celular. Por esse motivo, o objetivo do nosso estudo foi avaliar possiveis associacoes entre polimorfismos de alguns genes da via Wnt e a susceptibilidade ao TAB e comorbidades. Nossa amostra consistiu de 546 individuos, sendo 306 pacientes com TAB e 240 controles saudaveis. Utilizamos os genes WNT2B, WNT3A, WNT5A, WNT7A, APC1 e FZD3. Nas analises de associacao entre caso e controle, nao encontramos correlacao entre os tagSNPs estudados e o TAB. Encontramos associacao positiva entre o gene APC1, rs7419262, e tentativa de suicidio violenta e entre o gene WNT2B, rs3790606 e rs351372 e o transtorno de ansiedade generalizada como comorbidade em pacientes com TAB. Concluimos que a via Wnt pode estar associada a susceptibilidade ao TAB e suicidio. Bipolar Disorder (BD) is a severe and chronic psychiatric illness with progressive increase in the severity of episodes. The prognosis for patients with BD is poor, with high rates of relapse in residual symptoms, cognitive impairment and decreased quality of life. A growing body of research supports the assumption that BD arises from abnormalities in neuronal plasticity. Several lines of evidence suggests that the Wnt family is involved in this pathway since it is related to key developmental processes such as cell growth, migration and cell differentiation. Therefore, the aim of our study was to evaluate whether genes of the Wnt pathway are associated to BD and its comorbidities. Our sample consisted of 546 individuals: 306 patients with BD and 240 healthy controls. We used the genes WNT2B, WNT3A, WNT5A, WNT7A, APC1 and FZD3. We found no correlation between the studied tagSNPs and BD. However, we found a positive association between ACP1 tagSNP, rs7419262, and violent suicide attempt, and between the WNT2B tagSNPs, rs3790606 and rs351372, and generalized anxiety disorder as a comorbidity in patients with BD. We conclude that the Wnt pathway may be associated with susceptibility to BD and suicide.

Introdução: O adenocarcinoma é o tipo histológico mais prevalente entre os tumores de pâncreas (90%) e possui alta taxa de mortalidade devido ao seu mau prognostico. A importância de se compreender a carcinogênese pancreática é determinar novos alvos terapêuticos e biomarcadores que auxiliem no diagnostico precoce. A progressão do adenocarcinoma ductal pancreático (ADP) é relacionada ao acúmulo de mutações em lesões pré cancerígenas como a Neoplasia Intraepitelial Pancreática (PanIN). Alterações em oncogenes como: K-ras, EGFR, BRAF, PIK3CA e perda de função em supressores tumorais como CDKN2A e p53 estão envolvidas no desenvolvimento do tumor. Nosso objetivo foi avaliar a frequência de mutações nos genes envolvidos com a progressão do câncer e correlacioná-las com os achados clínicos e a ancestralidade genômica Material e Método: A análise das mutações foi feita por sequenciamento e a ancestralidade através de 40 indels bialélicos em 23 pacientes brasileiros com adenocarcinoma pancreático. Resultados: Dentre todos os genes estudados, as mutações no códon 12 de K-ras foram as mais freqüentes (60%) em nossa amostragem. Em PIK3CA, identificamos uma mutação não descrita (M983K) em um paciente. Mutações nos genes EGFR, BRAF, PIK3CA, CDKN2A e p53 não foram frequentes em nosso estudo. A ancestralidade genômica dos nossos pacientes não diferiu do grupo controle, contudo, observamos que os pacientes com maior componente ancestral africano apresentaram mutações em K-ras. Conclusão: Observamos em nossos pacientes uma menor frequência de mutações em K-ras em relação à maioria dos resultados já descritos, porém mais aproximada à da população coreana, além disto, nossas amostras apresentaram menor freqüência de mutações em CDKN2A e p53 em relação a outras populações já estudadas, reforçando a importância de se levar em consideração diferenças étnicas nos estudos envolvendo alterações genéticas em pacientes com adenocarcinoma pancreático. Devido à alta miscigenação da população brasileira, é interessante que outros estudos como este sejam realizados em outras regiões do país, para caracterizar o perfil genético da população brasileira com adenocarcinoma pancreático. Introduction: Adenocarcinoma is the most prevalent histological type among tumors of the pancreas ( 90 % ) and has high mortality. The importance of understanding the pancreatic carcinogenesis is to determine new therapeutic targets and biomarkers to assist in early diagnosis. The progression of pancreatic ductal adenocarcinoma involved the accumulation of mutations in pre- cancerous lesions, for example, the pancreatic intraepithelial neoplasia ( PanIN ) . Molecular alterations in oncogenes such as K-ras, EGFR , BRAF , PIK3CA and loss function of tumor suppressor, such as CDKN2A and p53 are involved in tumor development. Our objective was to evaluate the frequency of mutations in genes involved in cancer progression and correlate them with the clinical and genomic ancestry Material and Methods: Mutation analysis was made by direct sequencing in 23 samples with pancreatic adenocarcinoma and ancestry was analyzed using 40 biallelic indels. Results: Among all the genes, mutations in codon 12 of K - ras were the most frequent (60 %). In PIK3CA, was identified a new mutation in one patient, do not described in the literature ( M983K ). Mutations in EGFR , BRAF , PIK3CA , CDKN2A and p53 were infrequent in our study . The genomic ancestry of our patients did not differ from the control group, however, we observed that all patients with higher African ancestral component showed mutations in K - ras . Conclusion: We observed in our patients a lower frequency of mutations in K - ras in relation to most of the results already described in other countries, but closer to the Korean population , and our samples showed a lower frequency of mutations in CDKN2A and p53 in relation to other populations, reinforcing the importance of th ethnic differences in studies involving genetic alterations in patients with pancreatic adenocarcinoma . Due to the high miscegenation of the Brazilian population, it is interesting that other studies like this are done in other regions of the country, with the objective of to characterize the genetic profile of Brazilian population with pancreatic adenocarcinoma .

A obesidade mórbida (OBM) é um distúrbio metabólico comum na população mundial. A doença representa um alto risco para a saúde já que, frequentemente, encontra-se associada a diversas comorbidades, como doenças cardiovasculares e diabetes tipo 2. Além de fatores ambientais, diversos estudos também têm sugerido que determinados polimorfismos genéticos predispõem ao desenvolvimento da OBM. Os genes FOXO3a (fator de transcrição forkhead box O 3a), AMPK (proteína quinase ativada por AMP) e POMC (proopiomelanocortina) codificam proteínas reguladoras do metabolismo energético e comportamento alimentar e, desta forma, é possível que suas variantes genéticas possam modular a susceptibilidade à OBM. Por esse motivo, os polimorfismos dos genes FOXO3a (rs1536057, rs2802292, rs3813498, rs1935952), AMPK (rs1442760, rs1036851, rs1348316, rs11584787) e POMC (rs934778, rs6545975) foram investigados a fim de conferir o risco de desenvolver OBM. Para tanto, foram recrutados 242 pacientes obesos mórb idos (IMC 40 Kg/m²) e 283 indivíduos saudáveis (IMC 24,99 Kg/m²). Em seguida, os polimorfismos de interesse foram avaliados através do método TaqMan Genotypinga partir de DNA isolado do sangue. As análises genéticas do estudo demonstraram que as frequências alélicas e genotípicas de quatro dos polimorfismos estudados, a saber, rs1536057 (FOXO3a), rs103685 (AMPK), rs934778 e rs6545975 (POMC), foram distribuídas diferentemente entre os grupos de obesos mórbidos e indivíduos saudáveis. Nenhuma associação significativa foi encontrada quando as análises de interação gene-gene foram empregadas. Os resultados deste estudo fornecem dados inéditos sobre a frequência genética dos marcadores estudados na população brasileira e sustentam o princípio de que variações genéticas específicas podem conferir o risco de desenvolver a OBM. Morbid obesity (MO) is a metabolic disorder that affects people worldwide. Excessive fat accumulation in MO patients is frequently found to increase the risk for several co-morbidities, including cardiovascular disease and type 2 diabetes. Although environmental factors are well-known to play a role in MO risk, accumulate evidence has also highlighted that inherited factors may predispose subjects to MO. Because the genes FOXO3a (forkhead box O 3a transcription factor), AMPK (activated protein kinase) and POMC (proopiomelanocortin) encode regulatory proteins that act on both energy metabolism and feeding beha vior, here we have addressed whether polymorphisms in these genes are risk factors to MO. Therefore, the genetic frequencies of rs1536057, rs2802292, rs3813498, rs1935952 (FOXO3a), rs1442760, rs1036851, rs1348316, rs11584787 (AMPK ) and rs934778, rs6545975 (POMC) were evaluated in 242 morbidly obese patients (BMI 40 kg/m²) and 283 healthy subjects (BMI 24.99 kg/m²). DNA samples were isolated from blood and genotyping was performed by TaqMan-based assays. Our findings revealed that out of 10 polymorphisms analyzed, only allele and genotype frequencies of rs1536057 (FOXO3a), rs103685 (AMPK), rs934778 and rs6545975 (POMC) were heterogeneously distributed between morbid obesity and healthy subjects groups. Furthermore, epistasis analysis did not show any significant model of genetic interaction influencing the susceptibility to morbid obesity. Results of this study for the first time provide data on distribution of FOXO3a, AMPK and POMC polymorphisms in the Brazilian population and suggest that selected geneti c variants may confer risk of morbid obesity in humans.

O câncer de pulmão é a neoplasia que apresenta maior taxa de mortalidade mundial, sendo o prognóstico dos pacientes acometidos muito ruim. O desenvolvimento de novas estratégias terapêuticas e a melhor compreensão da tumorigênese relacionada a este tipo de câncer são necessários para aumentar a sobrevida dos doentes. Neste sentido, EGFR, KRAS, BRAF e PTEN são genes que atuam em vias de crescimento, proliferação, migração, sobrevivência celular e inibição de apoptose enquanto NTH1 é uma glicosilase que atua na via de reparo de DNA. Alterações somáticas nesses genes podem estar associadas ao desenvolvimento do câncer de pulmão . Para avaliar essas possíveis alterações, analisamos as regiões-alvo de NTH1 (exons 1-6), PTEN (exons 1-9), KRAS (exons 2 e 3), BRAF (exon 15) e EGFR (exon 18-21) em oitenta e dois pacientes brasileiros portadores de carcinoma de não-pequenas células de pulmão (NSCLC). A expressão da proteína PTEN foi investigada em vinte e uma amostras pelo método de imunohistoquímica. Analisamos a ancestralidade genômica dos nossos pacientes através de 40 indels bialélicos. Não foram detectadas mutações ou polimorfismos em PTEN e BRAF. A expressão de PTEN nuclear foi maior em células neoplásicas quando comparado com células benignas. Na análise de KRAS detectamos a presença da mutação G228A no códon 12 em um paciente (1.2%) e dois pacientes (2,4%) apresentaram polimorfismo no códon 61 (rs17851045). Em EGFR encontramos a presença da deleção 746_750 (domínio LREA) no exon 19 em 3.6% pacientes e não detectamos alterações nos exons 18 e 21. No exon 20, dois polimorfismos foram identificados: a substituição silenciosa (CAG> CAA) na posição c.2538 (rs1050171) detectada em 54,9% dos casos e a substituição 2284-60T>C a montante do exon 20 (rs10241451) encontrada em 13,4% dos casos. A substituição 2284-60T>C apresentou associação significativa com a doença tanto para freqüência alélica quanto genotípica (p = 0,03 e p = 0,02, respectivamente). Na análise de ancestralidade encontramos um maior componente africano em nossa amostra. Em NTH1 encontramos os polimorfismos G99T (rs2302102), 140-17C>T (rs2233518) e 131 (C> A) em apenas dois pacientes (4%) que podem estar relacionados ao desenvolvimento da doença. Como demonstrado, nossos dados apresentam uma baixa frequência de mutações em EGFR, KRAS, BRAF e PTEN, o que pode ser explicado pelo alto componente africano encontrado em nossos pacientes. Nossos dados destacam que a heterogeneidade genética da via de EGFR em NSCLC pode diferir entre as populações, ressaltando assim a necessidade de reavaliar a utilização de agentes terapêuticos destinados a inibição desta via. É importante destacar que esses resultados devem ser expandidos e validados em estudos de escopo mais largo, com foco em pacientes brasileiros em geral. Lung cancer is the leading global cause of cancer -related mortality and the patients have poor prognosis . To improve the survival rate of lung cancer patients, a better understanding of tumor biology is required as well as the subsequent development of new therapeutic strategies. EGFR, KRAS, BRAF and PTEN genes play important role in the regulation of cell growth, proliferation, migration, survival and inhibition of apoptosis, while NTH1 function as a DNA glycosylase in DNA repair pathway. Somatic alterations in these genes may be associated with the development of lung cancer. To investigate these molecular alterations, we analyzed the targeted regions of NTH1 (exons 1-6), PTEN (exons 1-9), KRAS (exons 2 and 3), BRAF (exon 15) and EGFR (exons 18-21) in eighty-two Brazilian patients with non-small cell lung cancer (NSCLC). PTEN protein expression was evaluated in twenty -one samples by immunohistochemistry. We analyzed the genomic ancestry of our patients through 40 biallelic indels. No mutations or polymorphisms were detected in PTEN and BRAF. The nuclear PTEN expression was higher in neoplastic cells compared to benign cells. The analysis detected the presence of KRAS mutation G228A in codon 12 in one patient (1.2%) and two patients (2.4%) showed polymorphism at codon 61 (rs17851045). In EGFR sequences we found 746_750 deletion (domain LREA) in exon 19 in 3.6% patients and no alterations in exons 18 and 21. In exon 20, two polymorphisms were identified: a silent substitution ( CAG> CAA) at position c.2538 (rs1050171) was detected in 54.9% of cases and a substitution 2284-60T> C upstream of exon 20 (rs10241451) was found in 13.4% of cases. The substitution 2284 -60T> C was significantly associated with the disease for both, allele and genotype frequency (p = 0.03 and p = 0.02, respectively). Ancestry analysis showed a greater African component in our sample. Analysis of NTH1, showed the G99T polymorphism (rs2302102), 140-17C> T (rs2233518) and 131 (C> A) in only two patients (4%), wich may be related to the disease. As shown, our data show a low frequency of mutations in EGFR, KRAS, BRAF and PTEN, which can be explained by the high African component of our patients. Our data highlight that the genetic heterogeneity of the EGFR pathw ay in NSCLC may differ between populations. Therefore, it underscores the need to reassess the use of therapeutic agents for the inhibition of this pathway in different populations. Futhermore, these results should be expanded and validated in studies of larger scope, focusing on Brazilian patients in general.

A depressão é uma das doenças neuropsiquiátricas com uma das maiores prevalências na população em geral, atingindo cerca de 20% dos indivíduos ao longo da vida. A sua alta prevalência associada com mudanças signigicativas no comportamento acarreta em um importante prejuízo socio-econômico e individual. Quando o primeiro episódio atinge indivíduos com mais de 60 anos é considerada depressão de início tardio (DIT). A população acima desta faixa etária tem aumentado significativamente no Brasil e estima-se que alcance, no ano de 2025, cerca de 32 milhões de idosos. Várias linhas de pesquisas sugerem que a depressão está associada com alterações do sistema serotoninérgico. Nesta via, os genes envolvidos na síntese, transporte e degradação de serotonina têm recebido especial atenção nos estudos que tentam desvendar as bases genéticas da depressão. A enzima triptofano hidroxilase-2 (Tph2) está envolvida na biosíntese de serotonina no cérebro e polimorfismos no gene TPH2 podem promover alterações na síntese deste neurotransmissor. Apesar de algumas variantes genéticas do TPH2 já estarem associadas à diversas doenças psiquiátricas, em especial à depressão maior e suicídio, poucos estudos investigaram a associação de polimorfismos deste gene e a susceptibilidade à depressão de início tardio. Neste contexto, o objetivo deste estudo foi avaliar se existe associação entre um ou mais polimorfismos do TPH2 e a DIT em uma população idosa do município de Belo Horizonte. Foram convocados 84 pacientes idosos com DIT e 79 idosos não portadores de nenhum distúrbio psiquiátrico atendidos no Centro de Referência do Idoso Professor Caio Benjamin Dias do Estado de Minas Gerais. Para a genotipagem foram selecionados previamente oito tagSNPs do TPH2, os quais foram identificados através do método de PCR em tempo real TaqMan seguindo o protocolo do fabricante. Nossos resultados mostraram uma associação genotípica entre dois tagSNPs do gene TPH2, rs4565946 e rs11179000, e a depressão de início tardio. Para o tagSNP rs11179000 foi observado maior risco para o desenvolvimento de depressão de início tardio para indivíduos portando o alelo ancestral. Nossos resultados reforçam a hipótese neurobiológica do envolvimento do sistema serotoninérgico na etiopatogênese da depressão de início tardio. Major depression is one of the most common psychiatric disorders, and in elderly its prevalence impact in terms of high rates of suicide and worst prognosis in comorbidities. Major depression that happens after 60 years is considered late-onset depression. The population above this age has increased significantly in Brazil, and it is estimated that in 2025 will reach approximately 32 million elderly. Many studies suggest that early-onset depression and late-onset depression have many different clinical and also genetic aspects. Some tryptophan hydroxylase type II gene (TPH2) variants have been associated with many psychiatric disorders in special with the major depression. TPH2 is the rate-limiting enzyme in the biosynthesis of serotonin. Polymorphisms in TPH2 might promote changes in brain serotonin synthesis. Thus, we aimed to investigate the frequencies of TPH2 polymorphisms in a group of elderly patients with late-onset depression in a brazilian population and whether these frequencies are associated with depressive events. Here, we genotyped eight tagSNPs from the gene TPH2 in 84 samples with late-onset depression and 79 controls. Genotyping was performed using TaqMan Real Time PCR. Unphased 3.0.13 program was used for genetic association. Overall, we identified an association between tagSNPs rs4565946 (C/T) and rs11179000(A/A) and late-onset depression. For allelic association of tagSNP rs11179000 we observed a strong association of allele A and risk of late-onset depression. Our findings suggest the involvement of genetic variants in the TPH2 gene in the etiology of late onset depression in brazilian population.