Resumo Introdução: As fendas orofaciais não sindrômicas possuem uma etiologia complexa devido à contribuição de fatores de risco genéticos e ambientais, assim como a interação entre eles. Dentre os mais de 15loci de susceptibilidade para as fendas orofaciais não sindrômicas com considerável suporte estatístico e biológico, o IRF6 é o gene mais validado pela maioria dos estudos. Apesar disso, em populações geneticamente heterogêneas como a brasileira, a confirmação da associação entre as fendas orofaciais não sindrômicas e as variantes mais comuns do IRF6 ainda não é um fato consolidado e outras variantes não tão conhecidas IRF6 são pouco investigadas. Objetivo: O objetivo deste estudo foi investigar a associação de variados polimorfismos do IRF6 com o desenvolvimento das fendas orofaciais não sindrômicas em uma população do nordeste do Brasil. Método: Amostras de sangue de 186 pacientes com fendas orofaciais não sindrômicas e 182 controles do estado do Rio Grande do Norte, Brasil, foram obtidas para analisar os polimorfismos do IRF6 (rs2235371, rs642961, rs2236907, rs861019 e rs1044516) por reação em cadeia da polimerase em tempo real. Os pacientes com fendas orofaciais não sindrômicas foram classificados em fenda labiopalatina, fenda palatina isolada e fenda labial isolada. Resultados: As frequências genotípica e alélica do polimorfismo de único nucleotídeo rs2235371 no IRF6 mostraram-se significativamente diferentes em pacientes com fenda palatina isolada quando comparadas às dos controles, enquanto que nenhuma associação foi encontrada entre rs642961, rs2236907, rs861019 e rs1044516 e risco para o desenvolvimento das fendas orofaciais não sindrômicas. Conclusão: A associação encontrada entre rs2235371 e fenda palatina isolada deve ser interpretada com cautela devido ao baixo número de indivíduos investigados, sendo necessários mais estudos com um tamanho amostral maior para confirmar essa associação. Além disso, não foram encontradas associações significativas entre os demais polimorfismos do IRF6 rs642961, rs2236907, rs861019 e rs1044516 e a susceptibilidade às fendas orofaciais não sindrômicas.

Abstract Introduction: Non-syndromic orofacial clefts have a complex etiology due to the contribution from both genetic and environmental risk factors, as well as the interaction between them. Among the more than 15 susceptibility loci for non-syndromic orofacial clefts with considerable statistical and biological support, the IRF6 is the most validated gene by the majority of studies. Nonetheless, in genetically heterogeneous populations such as Brazilian, the confirmation of association between non-syndromic orofacial clefts and IRF6 common variants is not a consolidated fact and unrecognized IRF6 variants are poorly investigated. Objective: The aim of this study was to investigate the association of IRF6 polymorphisms with non-syndromic orofacial clefts development in a population from northeast Brazil. Methods: Blood samples of 186 non-syndromic orofacial clefts patients and 182 controls from Rio Grande do Norte, Brazil, were obtained to analyze IRF6 polymorphisms (rs2235371, rs642961, rs2236907, rs861019, and rs1044516) by real-time polymerase chain reaction. Non-syndromic orofacial clefts patients were classified in cleft lip and palate, cleft palate only and cleft lip only groups. Results: The genotype and allele frequencies of single nucleotide polymorphism rs2235371 in IRF6 showed significant differences in patients with cleft palate when compared to the controls, whereas no association was shown between rs642961, rs2236907, rs861019, and rs1044516 and non-syndromic orofacial clefts. Conclusion: The association found between rs2235371 and isolated cleft palate should be interpreted with caution due to the low number of individuals investigated, and more studies with larger sample size are needed to confirm these association. In addition, there is a lack of association of the rs642961, rs2236907 and rs861019 polymorphisms with non-syndromic orofacial clefts susceptibility.

ABSTRACT Objective: The present study has investigated the association between low-density lipoprotein receptor-related protein 5 (LRP5) 4037C>T polymorphism and type 1 diabetes mellitus (T1DM) susceptibility in a Brazilian population. Subjects and methods: A total number of 134 T1DM patients and 180 normoglycemic individuals (NG) aged 6-20 years were studied. Glycated hemoglobin and glucose levels were determined. Genotyping of LRP5 4037C>T (rs3736228) was performed. Results: T1DM patients showed poor glycemic control. Genotypes in the codominant (CT: OR = 2.99 [CI 95%: 1.71-5.24], p < 0.001; TT: OR = 5.34 [CI 95%: 1.05-2702], p < 0.001), dominant (CT + TT: OR = 3.16 [CI 95%: 1.84-5.43], p < 0.001) and log-additive (OR = 2.78 [CI 95%: 1.70-4.52], p < 0.001) models, and LRP5 4037T allele (OR = 2.88, [CI 95%: 1.78-4.77], p < 0.001) were associated with an increased risk of developing T1DM. LRP5 4037CT and CT+TT carriers in T1DM group showed higher concentrations of serum glucose and glycated hemoglobin when compared with CC carriers. Conclusion: The LRP5 4037C>T may represent a candidate for T1DM susceptibility, as well as poor glycemic control.

Abstract: Considering that environmental risk factors substantially contribute to the etiology of orofacial clefts and that knowledge about the characteristics and comorbidities associated with oral clefts is fundamental to promoting better quality of life, this study aimed to describe the risk factors, main characteristics, and comorbidities of a group of patients with cleft lip and/or cleft palate (CL/P) from Rio Grande do Norte (RN), Brazil. Data were obtained from 173 patients with CL/P using a form from the Brazilian database on Orofacial Clefts. Most patients were male with cleft lip and palate and had a normal size and weight at birth; presented few neonatal intercurrent events; and had anemia and respiratory and cardiovascular diseases as main associated comorbidities. They also required timely surgical rehabilitation and multidisciplinary care to stimulate their neuropsychomotor development. In addition, a high frequency of familial recurrence and of parental consanguinity was evidenced in the studied population, especially for the cleft lip and cleft palate type. Other relevant findings were the considerable maternal exposure to alcohol, infections, smoking, and hypertension, as well as low supplementation with vitamins and minerals and deliberate consumption of analgesics, antibiotics, and antihypertensives during pregnancy. Characterization of the CL/P patient profile is essential for the planning of health services and integration among the health professionals involved in the diagnosis and treatment of these malformations. Our results reinforce the need for additional research to confirm the association between environmental factors and the development of orofacial clefts.

OBJECTIVES: Inflammatory molecules play a role in the development of atherosclerosis, which is the primary origin of cardiovascular disorders. However, to the best of our knowledge, no study has attempted to investigate the relationship between these circulating molecules and the prediction of cardiovascular risk. The present study aimed to investigate the relationships of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, E-selectin and matrix metalloproteinase 9 serum concentrations with the extent of coronary lesions. METHODS: Seventy-four individuals who were undergoing coronary angiography for the first time for diagnostic purposes were enrolled in this study. The extent of the coronary lesion was assessed using the Friesinger Index, and subjects were classified into four groups: no lesions, minor lesions, intermediate lesions and major lesions. Serum biochemical parameters and serum concentrations of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, E-selectin and matrix metalloproteinase 9 were analyzed. RESULTS: The vascular cell adhesion molecule-1 concentration was higher than 876 ng/mL in individuals with intermediate and major lesions (p<0.001 and p=0.020, respectively). Moreover, logistic regression analysis showed that these patients had an increased risk of having an intermediate lesion (p=0.007). Interestingly, all individuals with major lesions had vascular cell adhesion molecule-1 concentrations higher than 876 ng/mL. No association was found between the concentrations of the other proteins and the Friesinger Index. CONCLUSIONS: Serum vascular cell adhesion molecule-1 may be associated with the extent of coronary lesions. Moreover, it may represent an alternative to improve the cardiovascular risk classification in patients without acute coronary syndrome.

OBJETIVO: Verificar se as concentrações de glicose e insulina em jejum são reguladas pela aptidão cardiorrespiratória (VO2máx), independentemente dos efeitos genéticos. MÉTODOS: Dados de 38 pares de gêmeos monozigóticos (11 a 18 anos) foram analisados transversalmente. Os participantes foram submetidos a um teste de esforço máximo com ergoespirometria aberta (MedGraphics VO2000® - Medical Graphics Corp., St. Paul, MN) e à coleta de sangue para estimar a concentração de glicose e insulina em jejum. A zigosidade foi determinada por intermédio da investigação de concordância dos gêmeos em relação a 15 marcadores genéticos polimórficos. Nove pares demonstraram diferença média intrapar para o consumo máximo de oxigênio ≥10mL.kg-1.min-1 e foram divididos em dois grupos, de alta e baixa aptidão. Os grupos foram comparados a partir do teste pareado de Wilcoxon, tendo em vista a assimetria dos dados. RESULTADOS: Em média, os gêmeos do grupo de alta aptidão apresentaram consumo máximo de oxigênio 17% superior (13,5±3,7mL.kg-1.min-1) a seus irmãos menos aptos. Não houve diferença entre os grupos para as concentrações de insulina (36,5±34,6 versus 25,3±13,7mg/dL; p<0,813), porém, os gêmeos mais aptos demonstraram menor concentração de glicose do que seus contrapares menos aptos (82,9±7,3 versus 86,7±7,6mg/dL; p<0,010). CONCLUSÕES: Neste estudo, caracterizado como caso-controle (gêmeos monozigóticos discordantes), o irmão com menor aptidão cardiorrespiratória apresentou maior concentração de glicose em jejum, sugerindo que a baixa aptidão cardiorrespiratória está associada a distúrbios no metabolismo de glicose.

OBJECTIVE: To determine if glucose and insulin concentrations are regulated by cardiorespiratory fitness (VO2max) regardless of their genetic effects. METHODS: This 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 Wilcoxon's test for paired data. RESULTS: On 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). CONCLUSIONS: In 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 (VO2máx), independiente de los efectos genéticos. MÉTODOS: Datos de 38 pares de gemelos monocigóticos (11 a 18 años) fueron analizados transversalmente. Los participantes fueron sometidos a una prueba de esfuerzo máximo con ergoespirometría abierta (MedGraphics VO2000® Medical Graphics Corp., St. Paul, MN) y a la recolección de sangre para estimar la concentración de glucosa e insulina en ayuno. La cigosidad fue determinada por medio de la investigación de concordancia de los gemelos respecto a 15 marcadores genéticos polimórficos. Nueve pares demostraron diferencia mediana intrapares para el consumo máximo de oxígeno ≥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 asimetría de los datos. RESULTADOS: En promedio, los gemelos del grupo de alta aptitud presentaron consumo máximo de oxígeno 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 más aptos demostraron menor concentración 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 monocigóticos discordantes), el hermano con menor aptitud cardiorrespiratoria presentó mayor concentración de glucosa en ayuno, sugiriendo que la baja aptitud cardiorrespiratoria está asociada a disturbios en el metabolismo de glucosa.

Quantitative polymerase chain reaction-high-resolution melting (qPCR-HRM) analysis was used to screen for mutations related to drug resistance in Mycobacterium tuberculosis. We detected the C526T and C531T mutations in the rifampicin resistance-determining region (RRDR) of the rpoB gene with qPCR-HRM using plasmid-based controls. A segment of the RRDR region from M. tuberculosis H37Rv and from strains carrying C531T or C526T mutations in the rpoB were cloned into pGEM-T vector and these vectors were used as controls in the qPCR-HRM analysis of 54 M. tuberculosis strains. The results were confirmed by DNA sequencing and showed that recombinant plasmids can replace genomic DNA as controls in the qPCR-HRM assay. Plasmids can be handled outside of biosafety level 3 facilities, reducing the risk of contamination and the cost of the assay. Plasmids have a high stability, are normally maintained in Escherichia coli and can be extracted in large amounts.

A Organização Mundial de Saúde (OMS) tem por objetivo o controle político de acesso, de qualidade/segurança e de uso racional, no que se refere a medicamentos. Este último item salienta a relevante questão do uso incorreto de medicamentos induzido por promoções que visam exclusivamente a lucros. O presente trabalho apresenta os resultados de um projeto cujo objetivo foi acompanhar e analisar, do ponto de vista legal e farmacológico, as informações veiculadas por diferentes meios de comunicação através da propaganda, auxiliando a ANVISA na monitoração de propaganda e publicidade de medicamentos de venda livre ou com receita médica, bem como, dando suporte às medidas corretivas pertinentes, visando a garantir a segurança sanitária de produtos, serviços e, principalmente, da população. O projeto abrangeu cinco regiões brasileiras, sob a responsabilidade de quatorze universidades. Para atingir o objetivo proposto, realizou-se, durante 12 meses, a coleta do material publicitário: 55,1% retirado de (dois pontos) consultórios médicos, hospitais públicos e congressos, 15,8% de jornais e revistas, 8,9% de emissoras de rádio e 20,2% de televisão. A análise do material foi realizada tendo por base a RDC nº 102, publicada em 01/12/2000 (em vigor desde o dia 01/06/2001), e as demais legislações sanitárias vigentes, bem como a bibliografia científica aplicável. Os resultados obtidos para produto de venda livre revelaram que 3,6% dos produtos veiculados nos anúncios analisados não eram registrados na ANVISA (Artigo 4º, inciso I) e que 17,5% das campanhas publicitárias não apresentaram a principal contra-indicação do medicamento (Artigo 3º, inciso I). Observou-se, ainda, que 15,5% do material analisado estimulavam e/ou induziam ao uso indiscriminado de medicamentos, cujo consumo exige prescrição médica e que 38,5% das campanhas de vendas com receita médica não apresentavam o número do registro no Ministério da Saúde. Tais resultados serão importantes para o aperfeiçoamento das medidas regulatórias e a implementação de políticas de educação e conscientização referentes à publicidade de medicamentos. Os dados obtidos igualmente evidenciam a importância do monitoramento desse tipo diferenciado de publicidade a fim de garantir informações reais aos consumidores, cuja saúde pode ser posta em risco em conseqüência de propaganda enganosa.

World Health Organization (WHO) has the goal of politically controlling the access, quality/safety and rational use of medicines. The later is concerned with the important matter of the incorrect use of medicines due to advertising campaigns whose single objective is profit. This work shows the results of a project carried out with the intention of following and critically analyzing, under the legal and pharmacological points of view, the information issued by different media, thus helping ANVISA monitoring either under strict medical control or over the counter medicine advertisings. It also gives support to the restrictive measures related to the sanitary safety of products, services and, most of all, of the population. The project included five Brazilian regions, under the responsibility of fifteen universities. The advertising material was collected for 12 months: 5.1% were from medical offices, public hospitals and congresses, 15.8% from newspapers and magazines, 8.9% from radio stations, and 20.2% from television. The analysis was carried out according to RDC n. 102, published in December 1st, 2000, and to other sanitary regulation and those based on scientific bibliography as well. This study showed that from over the counter medicine advertised 3.6% have not been registered by ANVISA (4th Article, item I), and also that 17.5% of the advertisings have not informed about their main side effects (Article 3, item I). It has also been stated that the analyzed material stimulated and/or induced the indiscriminate use of medicines whose consumption demands a prescription and that 38.5% of sale campaigns of medical prescription products did not have the registry number from the Health Ministry. Such results are important as they point out to the need of better control and of an awareness and education policy relatively to medicine advertising. The data obtained also show the importance of monitoring this kind of publicity so that consumers are given real information and are therefore protected from deceitful propaganda.