Vivian Nogueira Silbiger

Genetic and non‐genetic factors that increase the risk of non‐syndromic cleft lip and/or palate development

OBJECTIVESnWe investigated the relationship between non-syndromic cleft lip/palate (NSCLP) and polymorphisms in methylenetetrahydrofolate reductase (MTHFR), methionine synthase (MTR), methionine synthase reductase (MTRR), and RFC1, as well as the corresponding interactions with environmental factors.nnnSUBJECTS AND METHODSnOne hundred and forty NSCLP patients and their mothers, as well as 175 control individuals and their mothers, were recruited. Information regarding smoking and alcohol consumption was recorded. Blood samples were obtained in order to measure serum folate and cobalamin, as well as, plasma total homocysteine concentrations and to extract DNA. Polymorphisms in MTHFR(677C>T and 1298A>C), MTR(2756A>G), MTR(66A>G), and RFC1(80A>G) were analyzed by PCR-restriction fragment length polymorphism.nnnRESULTSnAmong the patients, 59.5% had cleft lip and palate, 22.0% had cleft palate, and 18.5% had cleft lip only. Maternal alcohol consumption and reduced folic acid concentrations in both children and mothers (Pxa0<xa00.001 and Pxa0=xa00.003, respectively) were risk factors for NSCLP. Patients and their mothers carrying the MTHFR 667T allele showed lower serum folate than CC (Pxa0=xa00.011 and Pxa0=xa00.030, respectively). Mothers who carried the MTHFR 1298C allele exhibited increased risk of having a child with NSCLP, after adjusting for alcohol consumption (OR: 1.75, 95% CI: 1.03-2.99, Pxa0=xa00.038).nnnCONCLUSIONSnReduced folic acid levels, alcohol consumption, and the MTHFR 677T and 1298C alleles may have contributed to NSCLP development in this sample population from Rio Grande do Norte.

Increased clopidogrel response is associated with ABCC3 expression: a pilot study.

BACKGROUNDnThe aim of this study was investigate the relationship between ABCB1 and ABCC3 gene expressions in peripheral blood cells (PBC) and the response to clopidogrel in patients with coronary arterial disease (CAD).nnnMETHODSnTwenty-six male CAD patients (50-70 years) under treatment with clopidogrel (75 mg/day) for at least 5 days were selected. Blood samples were obtained to evaluate platelet reactivity and ABCB1 and ABCC3 mRNA expression. Platelet reactivity was measured in P2Y12 Reaction Units (PRU) using VerifyNow. RNA was extracted from PBC and mRNA levels were measured by qPCR, using GAPD as a reference gene.nnnRESULTSnPlatelet response to clopidogrel was categorized in to PRU quartiles. Individuals with PRU values within the first quartile (Q1, <151 units) were considered good responders, while those who had PRU within the fourth quartile (Q4, PRU>260) were considered non-responders. ABCC3 was 1.7 times more expressed in Q4 than in Q1 PRU group (p=0.048). Moreover, CAD patients with low ABCC3 expression (Qe1, <2.5×10(-3)) had higher probability to have a good response to clopidogrel (OR: 18.00, 95%CI: 1.90-169.99, p=0.001). Univariate linear regression analysis demonstrated that low ABCC3 mRNA expression contributed with a reduction of 73 PRU in relation to the patients with expression value higher than 2.5×10(-3) (p=0.027). Neither ABCB1 mRNA levels nor clinical variables studied influenced PRU values.nnnCONCLUSIONSnLow ABCC3 mRNA expression in peripheral blood cells is associated with increased clopidogrel response, but further studies are needed to describe the functional relationship of clopidogrel with the ABCC3.

Exploring iris colour prediction and ancestry inference in admixed populations of South America

New DNA-based predictive tests for physical characteristics and inference of ancestry are highly informative tools that are being increasingly used in forensic genetic analysis. Two eye colour prediction models: a Bayesian classifier - Snipper and a multinomial logistic regression (MLR) system for the Irisplex assay, have been described for the analysis of unadmixed European populations. Since multiple SNPs in combination contribute in varying degrees to eye colour predictability in Europeans, it is likely that these predictive tests will perform in different ways amongst admixed populations that have European co-ancestry, compared to unadmixed Europeans. In this study we examined 99 individuals from two admixed South American populations comparing eye colour versus ancestry in order to reveal a direct correlation of light eye colour phenotypes with European co-ancestry in admixed individuals. Additionally, eye colour prediction following six prediction models, using varying numbers of SNPs and based on Snipper and MLR, were applied to the study populations. Furthermore, patterns of eye colour prediction have been inferred for a set of publicly available admixed and globally distributed populations from the HGDP-CEPH panel and 1000 Genomes databases with a special emphasis on admixed American populations similar to those of the study samples.

Integrated analysis of miRNA and mRNA gene expression microarrays: Influence on platelet reactivity, clopidogrel response and drug-induced toxicity.

Genetic and epigenetic variability may influence the efficacy and safety of antiplatelet therapies, including clopidogrel. Therefore, the miRNA-mRNA interactions and drug toxicity were investigated in silico using available microarray data. Expressions profiles of platelet miRNA (GSE59488) from acute coronary syndrome and mRNA in peripheral blood cells (GSE32226) from coronary artery disease patients were used to miRNA-target mRNA integrated analysis by Ingenuity Pathways Analysis 6 software (IPA). Results showed that ST13 mRNA is regulated by hsa-miR-107 (miR-103-3p); BTNL3 and CFD mRNAs are regulated by hsa-miR-4701-3p (miR-1262); SLC7A8 is regulated by hsa-miR-145-5p (miR-145-5p); and SENP5 mRNA is regulated by hsa-miR-15b-5p (miR-16-5p) and hsa-miR-26a-5p (miR-26a-5p). Drug toxicity IPA tool showed that these miRNAs/mRNAs are associated with clopidogrel-related liver and renal injury. In conclusion, these results demonstrate that differential expression of miRNAs in platelets and interactions with their target mRNAs are associated with variability in platelet reactivity, clopidogrel response and drug-induced toxicity.

Association of insertion-deletions polymorphisms with colorectal cancer risk and clinical features

AIM To investigate the association between 16 insertion-deletions (INDEL) polymorphisms, colorectal cancer (CRC) risk and clinical features in an admixed population. METHODS One hundred and forty patients with CRC and 140 cancer-free subjects were examined. Genomic DNA was extracted from peripheral blood samples. Polymorphisms and genomic ancestry distribution were assayed by Multiplex-PCR reaction, separated by capillary electrophoresis on the ABI 3130 Genetic Analyzer instrument and analyzed on GeneMapper ID v3.2. Clinicopathological data were obtained by consulting the patients’ clinical charts, intra-operative documentation, and pathology scoring. RESULTS Logistic regression analysis showed that polymorphism variations in IL4 gene was associated with increased CRC risk, while TYMS and UCP2 genes were associated with decreased risk. Reference to anatomical localization of tumor Del allele of NFKB1 and CASP8 were associated with more colon related incidents than rectosigmoid. In relation to the INDEL association with tumor node metastasis (TNM) stage risk, the Ins alleles of ACE, HLAG and TP53 (6 bp INDEL) were associated with higher TNM stage. Furthermore, regarding INDEL association with relapse risk, the Ins alleles of ACE, HLAG, and UGT1A1 were associated with early relapse risk, as well as the Del allele of TYMS. Regarding INDEL association with death risk before 10 years, the Ins allele of SGSM3 and UGT1A1 were associated with death risk. CONCLUSION The INDEL variations in ACE, UCP2, TYMS, IL4, NFKB1, CASP8, TP53, HLAG, UGT1A1, and SGSM3 were associated with CRC risk and clinical features in an admixed population. These data suggest that this cancer panel might be useful as a complementary tool for better clinical management, and more studies need to be conducted to confirm these findings.

Circulating MicroRNAs as Potential Biomarkers of Atrial Fibrillation

Atrial fibrillation (AF) is the most common supraventricular arrhythmia in the population. MicroRNAs (small endogenous noncoding RNAs) are attractive candidates as biomarkers for AF, especially considering that miRNAs are stable and are detected within easily accessible biofluids such as blood. In this review, we selected twelve studies (2012 to 2016) that were classified according to the sample type. We aimed to provide an overview of the role of circulating miRNAs in AF and to discuss the variability of the results, seeking to improve the perspective of the use of miRNAs as potential noninvasive biomarkers for this heart disease.

The relationship of the oleic acid level and ECHDC3 mRNA expression with the extent of coronary lesion

BackgroundThe fatty acid profile is associated with the risk and progression of several diseases, probably via mechanisms including its influence on gene expression. We previously reported a correlation between ECHDC3 upregulation and the severity of acute coronary syndrome. Here, we assessed the relationship of serum fatty acid profile and ECHDC3 expression with the extent of coronary lesion.MethodsFifty-nine individuals aged 30 to 74xa0years and undergoing elective cinecoronariography for the first time were enrolled in the present study. The extent of coronary lesion was assessed by the Friesinger index and patients were classified as without lesion (nu2009=u200918), low lesion (nu2009=u200917), intermediate lesion (nu2009=u200917) and major lesion (nu2009=u20097). Serum biochemistry, fatty acid concentration, and ECHDC3 mRNA expression in blood were evaluated.ResultsElevated serum levels of oleic acid and total monounsaturated fatty acids were observed in patients with low and intermediate lesion, when compared to patients without lesion (pu2009<u20090.05). ECHDC3 mRNA expression was 1.2 fold higher in patients with low lesion than in patients without lesion (pu2009=u20090.020), and 1.8 fold lower in patients with major lesion patients than in patients with low lesion (pu2009=u20090.023).ConclusionIncreased levels of monounsaturated fatty acids, especially oleic acid, and ECHDC3 upregulation in patients with coronary artery lesion suggests that these are independent factors associated with the initial progression of cardiovascular disease.

Glucose metabolism in discordant monozygotic twins for cardiorespiratory fitness

OBJECTIVEnTo determine if glucose and insulin concentrations are regulated by cardiorespiratory fitness (VO2max) regardless of their genetic effects.nnnMETHODSnThis cross-sectional study enrolled 38 pairs of young monozygotic twins (11 to 18 years-old). All subjects underwent a progressive maximal exercise test on a treadmill to determine the VO2max with gas exchange analysis (MedGraphics VO2000® - Medical Graphics Corp., St. Paul, MN). Blood samples were drawn after fasting to determine glucose and insulin levels. Monozygosity was confirmed by genotyping 15 informative genetic markers. Nine pairs had at least 10mL.kg-1.min-1 difference in VO2max and were divided into the more and less active group, according to their VO2max. Mean differences between more and less active groups were evaluated by Wilcoxons test for paired data.nnnRESULTSnOn average, twins from the more active group presented a 17% (13.5±3.7mL.kg-1.min-1) higher VO2max compared to their less active siblings. No significant differences were observed between the groups for fasting insulin (36.5±34.6 versus 25.3±13.7mg/dL; p<0.813). However, the more active twins had lower fasting glucose than the less active ones (82.9±7.3 versus 86.7±7.6mg/dL; p<0.010).nnnCONCLUSIONSnIn this case-control study (discordant monozygotic twins), the less active co-twins were characterized by higher fasting plasma glucose levels. This implies that poor cardiorespiratory fitness can be associated with defective glucose metabolism regardless of genetic factors.OBJETIVO: Verificar si las concentraciones de glucosa e insulina en ayuno son reguladas por la aptitud cardiorrespiratoria (VO2max), independiente de los efectos geneticos. METODOS: Datos de 38 pares de gemelos monocigoticos (11 a 18 anos) fueron analizados transversalmente. Los participantes fueron sometidos a una prueba de esfuerzo maximo con ergoespirometria abierta (MedGraphics VO2000® Medical Graphics Corp., St. Paul, MN) y a la recoleccion de sangre para estimar la concentracion de glucosa e insulina en ayuno. La cigosidad fue determinada por medio de la investigacion de concordancia de los gemelos respecto a 15 marcadores geneticos polimorficos. Nueve pares demostraron diferencia mediana intrapares para el consumo maximo de oxigeno ≥10mL.kg-1.min-1 y fueron divididos en dos grupos, de alta y baja aptitud. Los grupos fueron comparados a partir de la prueba pareada de Wilcoxon, teniendo en vista la asimetria de los datos. RESULTADOS: En promedio, los gemelos del grupo de alta aptitud presentaron consumo maximo de oxigeno el 17% superior (13,5±3,7mL.kg-1.min-1) a sus hermanos menos aptos. No hubo diferencia entre los grupos para las concentraciones de insulina (36,5±34,6 versus 25,3±13,7mg/dL; p<0,813), pero los gemelos mas aptos demostraron menor concentracion de glucosa que sus contrapares menos aptos (82,9±7,3 versus 86,7±7,6mg/dL; p<0,010, respectivamente). CONCLUSIONES: En este estudio, caracterizado como caso-control (gemelos monocigoticos discordantes), el hermano con menor aptitud cardiorrespiratoria presento mayor concentracion de glucosa en ayuno, sugiriendo que la baja aptitud cardiorrespiratoria esta asociada a disturbios en el metabolismo de glucosa.

Metabolismo de glicose em gêmeos monozigóticos discordantes para aptidão cardiorrespiratória

OBJECTIVEnTo determine if glucose and insulin concentrations are regulated by cardiorespiratory fitness (VO2max) regardless of their genetic effects.nnnMETHODSnThis cross-sectional study enrolled 38 pairs of young monozygotic twins (11 to 18 years-old). All subjects underwent a progressive maximal exercise test on a treadmill to determine the VO2max with gas exchange analysis (MedGraphics VO2000® - Medical Graphics Corp., St. Paul, MN). Blood samples were drawn after fasting to determine glucose and insulin levels. Monozygosity was confirmed by genotyping 15 informative genetic markers. Nine pairs had at least 10mL.kg-1.min-1 difference in VO2max and were divided into the more and less active group, according to their VO2max. Mean differences between more and less active groups were evaluated by Wilcoxons test for paired data.nnnRESULTSnOn average, twins from the more active group presented a 17% (13.5±3.7mL.kg-1.min-1) higher VO2max compared to their less active siblings. No significant differences were observed between the groups for fasting insulin (36.5±34.6 versus 25.3±13.7mg/dL; p<0.813). However, the more active twins had lower fasting glucose than the less active ones (82.9±7.3 versus 86.7±7.6mg/dL; p<0.010).nnnCONCLUSIONSnIn this case-control study (discordant monozygotic twins), the less active co-twins were characterized by higher fasting plasma glucose levels. This implies that poor cardiorespiratory fitness can be associated with defective glucose metabolism regardless of genetic factors.OBJETIVO: Verificar si las concentraciones de glucosa e insulina en ayuno son reguladas por la aptitud cardiorrespiratoria (VO2max), independiente de los efectos geneticos. METODOS: Datos de 38 pares de gemelos monocigoticos (11 a 18 anos) fueron analizados transversalmente. Los participantes fueron sometidos a una prueba de esfuerzo maximo con ergoespirometria abierta (MedGraphics VO2000® Medical Graphics Corp., St. Paul, MN) y a la recoleccion de sangre para estimar la concentracion de glucosa e insulina en ayuno. La cigosidad fue determinada por medio de la investigacion de concordancia de los gemelos respecto a 15 marcadores geneticos polimorficos. Nueve pares demostraron diferencia mediana intrapares para el consumo maximo de oxigeno ≥10mL.kg-1.min-1 y fueron divididos en dos grupos, de alta y baja aptitud. Los grupos fueron comparados a partir de la prueba pareada de Wilcoxon, teniendo en vista la asimetria de los datos. RESULTADOS: En promedio, los gemelos del grupo de alta aptitud presentaron consumo maximo de oxigeno el 17% superior (13,5±3,7mL.kg-1.min-1) a sus hermanos menos aptos. No hubo diferencia entre los grupos para las concentraciones de insulina (36,5±34,6 versus 25,3±13,7mg/dL; p<0,813), pero los gemelos mas aptos demostraron menor concentracion de glucosa que sus contrapares menos aptos (82,9±7,3 versus 86,7±7,6mg/dL; p<0,010, respectivamente). CONCLUSIONES: En este estudio, caracterizado como caso-control (gemelos monocigoticos discordantes), el hermano con menor aptitud cardiorrespiratoria presento mayor concentracion de glucosa en ayuno, sugiriendo que la baja aptitud cardiorrespiratoria esta asociada a disturbios en el metabolismo de glucosa.

Application of high-resolution array platform for genome-wide copy number variation analysis in patients with nonsyndromic cleft lip and palate

Although more than 14 loci may be involved in the development of nonsyndromic cleft lip and palate (NSCLP), the etiology has not been fully elucidated due to genetic and environmental risk factor interactions. Despite advances in identifying genes associated with the NSCLP development using traditional genetic mapping strategies of candidate genes, genome‐wide studies, and epidemiologic and linkage analysis, microarray techniques have become important complementary tools in the search for potential causative oral clefts genes in genetic studies. Microarray hybridization enables scanning of the whole genome and detecting copy number variants (CNVs). Although common benign CNVs are often smaller, with sizes smaller than 20 kb, here we reveal small exonic CNVs based on the importance of the encompassed genes in cleft lip and palate phenotype.