RESUMO
This study aimed to assess whether genetic polymorphisms in MTR and MTRR are potential biomarkers of oral health-related quality of life (OHRQoL) in children with caries. A cross-sectional study was designed wherein pairs of parents/caregivers and children (aged two-five years) were selected. Clinical examination was used to detect dental caries, which were classified as low-severity and high-severity caries. The Early Childhood Oral Health Impact Scale (ECOHIS) questionnaire was used to assess OHRQoL. Genomic DNA extracted from the saliva was used to analyze two missense genetic polymorphisms: MTR (rs1805087) and MTRR (rs1801394). Mann-Whitney non-parametric test was used to analyze candidate genes with OHRQoL scale and domain, with a significance level of p≤0.05. MTR (rs1805087) was found associated (p = 0.05) with children's OHRQoL subscale scores in the dominant model (GG + AG). Genetic polymorphisms in MTR may increase the risk of poor OHRQoL in children with caries. Further studies are needed to investigate genetics, molecular factors, and OHRQoL.
Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase , Cárie Dentária , Ferredoxina-NADP Redutase , Criança , Pré-Escolar , Humanos , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , Estudos Transversais , Cárie Dentária/genética , Saúde Bucal , Qualidade de Vida , Inquéritos e Questionários , Ferredoxina-NADP Redutase/genéticaRESUMO
Genetic polymorphisms could explain the inter-individual differences in the oral health-related quality of life (OHRQoL) of children with anterior open bite (AOB). OBJECTIVE: To assess the impact of AOB on OHRQoL in children and to evaluate whether MTR (rs1805087), MTRR (rs1801394), TGFß1 (rs1800469) and TNF-α (rs1799964, rs1799724 and rs1800629) genes are potential biomarkers for OHRQoL in children with AOB. STUDY DESIGN: A cross-sectional study was performed with 173 children aged between 2-6 years. The Brazilian version of Early Childhood Oral Health Impact Scale (ECOHIS) was applied. Genetic polymorphisms were analyzed using real-time PCR. Mann-Whitney U-test and Chi-square were used. RESULTS: The overall mean ECOHIS scores were 5.49 (SD= 5.72) and 3.45 (SD = 4.49) (p < 0.01) in the AOB and control groups, respectively. Children with the CC genotype of TNF-α (rs1799724) had a significantly higher psychological QoL level. The MTRR AA genotype group showed a lower QoL level in the child subscale (p = 0.006), function (p = 0.017), and psychological (p = 0.006) domains. There was no significant difference between OHRQoL and the genetic polymorphisms in MTR and TGFß1. CONCLUSIONS: Genetic polymorphisms in TNF-α and MTRR are associated with the impact on the OHRQoL in children with AOB.
Assuntos
Ferredoxina-NADP Redutase , Mordida Aberta , Saúde Bucal , Fator de Necrose Tumoral alfa , Criança , Pré-Escolar , Estudos Transversais , Ferredoxina-NADP Redutase/genética , Humanos , Mordida Aberta/genética , Polimorfismo Genético , Qualidade de Vida , Inquéritos e Questionários , Fator de Necrose Tumoral alfa/genéticaRESUMO
Background: Homocysteine levels can be impacted by enzymes variations. Aim: To correlate MTHFR, MTR and MTRR variants with homocysteine levels in the blood and follicular fluid and assisted reproduction results. Material & methods:MTHFR (rs2274976, rs1801131, rs1801133), MTR (rs1805087) and MTRR (rs1801394) genotyping was performed by TaqMan assays and compared with homocysteine levels, measured by ELISA, to oocytes retrieved and to the pregnancy status of women with endometriosis and controls. Results: The MTR G allele and GG genotype were more common in patients with endometriosis. They also showed lower levels of homocysteine and more clinical gestations. Epistasis analysis showed a model associated with gestational results, composed of MTHFR+MTR variants (CC+AG). Conclusion: The summation effect of variants in genes participating in folate metabolism was associated with pregnancy status in Brazilian women. MTR variants were more observed in endometriosis patients, as well as lower follicular Hcy levels and increased clinical pregnancy results.
What was the aim of the study? To correlate genetic variants to homocysteine levels in the blood and oocyte surrounding fluid, and the results of assisted reproduction techniques. How was the study done? A total of 152 women with endometriosis and controls with male infertility were evaluated. DNA was extracted from blood for genetic analysis, and homocysteine levels were measured from the blood and oocyte surrounding fluid. Genetic results were correlated to homocysteine levels, oocyte quality and pregnancy status. What were the results? A specific genetic marker occurred more in endometriosis patients. They also showed lower levels of homocysteine and a tendency to more clinical gestations than controls. What do the results of the study mean? Endometriosis patients showed specific genetic markers and different levels of homocysteine compared with controls. These results can be helpful to predict gestational results.
Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase , Endometriose , Ferredoxina-NADP Redutase , Homocisteína , Metilenotetra-Hidrofolato Redutase (NADPH2) , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , Alelos , Endometriose/complicações , Endometriose/genética , Feminino , Ferredoxina-NADP Redutase/genética , Ácido Fólico/metabolismo , Genótipo , Homocisteína/sangue , Humanos , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Polimorfismo de Nucleotídeo Único , GravidezRESUMO
OBJECTIVE: The study aim to investigate MTHFR C677T, MTHFR A1298C, RFC1 A80G, MTR A2756G, CBS 844ins68, MTRR A66G polymorphisms in Down syndrome (DS) parents. METHODS: Polymorphisms were evaluated in 35 mothers and 24 fathers of individuals with free trisomy of chromosome 21 confirmed by karyotype. The control group included 26 mothers and 26 fathers who had no children with DS. The molecular analysis was performed by polymerase chain reaction and restriction fragment length polymorphism (reaction chain polymerase restriction fragment length polymorphism) or polymerase chain reaction. The χ2 test (chi-square) was used to compare allele's differences among the study and the control group. Hardy-Weinberg equilibrium model was performed by χ2 testing. Multiple logistic regression models and binary logistic regression used to determine the association between polymorphisms and parental DS risk. RESULTS: This study did not reveal any significant difference in frequencies of polymorphisms. The haplotype analysis did not reveal linkage disequilibrium. The logistic regression analysis did not demonstrate differences between the groups. However, the binary logistic regression showed a higher frequency of the polymorphic homozygote genotype in DS parent group to codominant and dominant model in the RFC1 A80G. CONCLUSION: In conclusion, although the screening results were significant only to the RFC1 A80G polymorphism, the other determinations of the genetic factors associated with abnormal chromosome segregation could be helpful in future studies, including other polymorphisms involved in folate metabolism.
Assuntos
Síndrome de Down , Ferredoxina-NADP Redutase , Ácido Fólico , Síndrome de Down/genética , Feminino , Ferredoxina-NADP Redutase/genética , Ácido Fólico/metabolismo , Humanos , Masculino , Pais , Polimorfismo de Nucleotídeo ÚnicoRESUMO
BACKGROUND: The S-adenosyl-methionine (SAM) availability is crucial for DNA methylation, an epigenetic mechanism involved in nonsyndromic cleft lip with or without cleft palate (NSCL/P) expression. The aim of this study was to assess the association between single-nucleotide polymorphisms (SNPs) of genes involved in SAM synthesis and NSCL/P in a Chilean population. METHODS: In 234 cases and 309 controls, 18 SNPs in AHCY, MTR, MTRR, and MAT2A were genotyped, and the association between them and the phenotype was evaluated based on additive (allele), dominant, recessive and haplotype models, by odds ratio (OR) computing. RESULTS: Three deep intronic SNPs of MTR showed a protective effect on NSCL/P expression: rs10925239 (OR 0.68; p = 0.0032; q = 0.0192), rs10925254 (OR 0.66; p = 0.0018; q = 0.0162), and rs3768142 (OR 0.66; p = 0.0015; q = 0.0162). Annotations in expression database demonstrate that the protective allele of the three SNPs is associated with a reduction of MTR expression summed to the prediction by bioinformatic tools of its potentiality to modify splicing sites. CONCLUSIONS: The protective effect against NSCL/P of these intronic MTR SNPs seems to be related to a decrease in MTR enzyme expression, modulating the SAM availability for proper substrate methylation. However, functional analyses are necessary to confirm our findings. IMPACT: SAM synthesis pathway genetic variants are factors associated to NSCL/P. This article adds new evidence for folate related genes in NSCL/P in Chile. Its impact is to contribute with potential new markers for genetic counseling.
Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , Adenosil-Homocisteinase/genética , Fenda Labial/genética , Fissura Palatina/genética , Ferredoxina-NADP Redutase/genética , Metionina Adenosiltransferase/genética , Polimorfismo de Nucleotídeo Único , S-Adenosilmetionina/metabolismo , Alelos , Chile/epidemiologia , Fenda Labial/fisiopatologia , Fissura Palatina/fisiopatologia , Feminino , Frequência do Gene , Genes Dominantes , Genes Recessivos , Predisposição Genética para Doença , Genótipo , Haplótipos , Humanos , Masculino , Metionina/genética , Razão de ChancesRESUMO
BACKGROUD: Ferredoxin NADP(H) oxidoreductases (EC 1.18.1.2) (FNR) are flavoenzymes present in photosynthetic organisms; they are relevant for the production of reduced donors to redox reactions, i.e. in photosynthesis, the reduction of NADP+ to NADPH using the electrons provided by Ferredoxin (Fd), a small FeS soluble protein acceptor of electrons from PSI in chloroplasts. In rhodophyta no information about this system has been reported, this work is a contribution to the molecular and functional characterization of FNR from Gracilaria chilensis, also providing a structural analysis of the complex FNR/Fd. METHODS: The biochemical and kinetic characterization of FNR was performed from the enzyme purified from phycobilisomes enriched fractions. The sequence of the gene that codifies for the enzyme, was obtained using primers designed by comparison with sequences of Synechocystis and EST from Gracilaria. 5'RACE was used to confirm the absence of a CpcD domain in FNRPBS of Gracilaria chilensis. A three dimensional model for FNR and Fd, was built by comparative modeling and a model for the complex FNR: Fd by docking. RESULTS: The kinetic analysis shows KMNADPH of 12.5 M and a k cat of 86 s-1, data consistent with the parameters determined for the enzyme purified from a soluble extract. The sequence for FNR was obtained and translated to a protein of 33646 Da. A FAD and a NADP+ binding domain were clearly identified by sequence analysis as well as a chloroplast signal sequence. Phycobilisome binding domain, present in some cyanobacteria was absent. Transcriptome analysis of Gch revealed the presence of two Fd; FdL and FdS , sharing the motif CX5CX2CX29X. The analysis indicated that the most probable partner for FNR is FdS. CONCLUSION: The interaction model produced, was consistent with functional properties reported for FNR in plants leaves, and opens the possibilities for research in other rhodophyta of commercial interest.
Assuntos
Ferredoxina-NADP Redutase/química , Ferredoxinas/metabolismo , Gracilaria/enzimologia , Sequência de Aminoácidos , Eletroforese em Gel de Poliacrilamida , Ferredoxina-NADP Redutase/genética , Ferredoxina-NADP Redutase/farmacocinética , Gracilaria/química , Oxirredução , Fotossíntese/fisiologiaRESUMO
AIM: To investigate the relationships of polymorphisms in genes whose protein products are related in the metabolic pathway of folic acid, particularly MTRR A66G, RFC1 G80A, and MTHFR C677T and A1298C, and disease activity in Mexican patients with rheumatoid arthritis (RA) treated with methotrexate (MTX). MATERIALS AND METHODS: Sixty-eight patients with RA were included in the study who were being treated with MTX, either with or without other drugs. In addition to general data, disease activity was measured by the disease activity score 28 (DAS28). Single nucleotide polymorphisms (SNPs) genotyping was performed by allelic discrimination using real-time polymerase chain reaction. RESULTS: Differences in genotype (homozygotic or heterozygotic for each allele), allele distributions, and phenotype were not statistically different between the RA group and control populations. We did not find any association between the studied polymorphisms and disease activity nor with the intragroup variables (e.g., clinical activity, body mass index, and single- or combined-drug treatment) or between genetic markers; we also did not find any association within the RA group or between the RA group and control populations. CONCLUSION: Additional studies of more polymorphisms related to this or other metabolic pathways are required to determine the influence of genetics on disease activity in RA.
Assuntos
Artrite Reumatoide/genética , Ferredoxina-NADP Redutase/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Proteína de Replicação C/genética , Adulto , Idoso , Alelos , Etnicidade/genética , Feminino , Ferredoxina-NADP Redutase/metabolismo , Ácido Fólico/genética , Ácido Fólico/metabolismo , Frequência do Gene/genética , Genótipo , Humanos , Masculino , Metotrexato , Metilenotetra-Hidrofolato Redutase (NADPH2)/metabolismo , México , Pessoa de Meia-Idade , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , Proteína de Replicação C/metabolismoRESUMO
Folate and other B vitamins are essential co-factors of one-carbon metabolism, and genetic variants, such as polymorphisms, can alter the metabolism. Furthermore, the adoption of food fortification with folic acid showed a decrease of homocysteine concentration. The aim of this study was to investigate the frequencies of the polymorphisms of enzymes and carrier proteins involved in one-carbon metabolism, and to evaluate homocysteine concentrations in the presence of these genetic variants in a population exposed to mandatory food fortification with folic acid. Using data from a population-based cross-sectional study in São Paulo, Brazil, the study population comprised 750 participants above 12 years of age of both genders. A linear regression model was used to evaluate the homocysteine concentrations according to genetic variants and folate level. The results showed that the minor allelic frequencies were 0.33 for MTHFR (rs1801133), 0.24 for MTHFR (rs1801131), 0.19 for MTR (rs1805087), 0.42 for MTRR (rs1801394), 0.46 for RFC1 (rs1051266), and 0.47 for DHFR (19-bp deletion). The genetic variants of MTHFR 677C>T, MTRR 66A>G and RFC-1 80G>A were different according to race. The homocysteine concentrations increased in the CT and TT compared to CC genotypes of polymorphism MTHFR 677C>T in all populations, and differences between the homocysteine concentrations according to the genotypes of MTHFR 677C>T were observed regardless of folate level.
Assuntos
Ferredoxina-NADP Redutase/metabolismo , Ácido Fólico/farmacologia , Homocisteína/sangue , Metilenotetra-Hidrofolato Redutase (NADPH2)/metabolismo , Polimorfismo Genético , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/metabolismo , Estudos Transversais , Inquéritos sobre Dietas , Feminino , Ferredoxina-NADP Redutase/genética , Ácido Fólico/administração & dosagem , Alimentos Fortificados , Genótipo , Humanos , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Pessoa de Meia-Idade , Proteína de Replicação C/genética , Proteína de Replicação C/metabolismoRESUMO
Polymorphisms in genes encoding the enzymes involved in the metabolism of homocysteine, such as methionine synthase (MTR) and methionine synthase reductase (MTRR), play an important function in the metabolism of folic acid and vitamin B12. The present study aimed to evaluate the association of polymorphisms in genes MTR (rs1805087) and MTRR (rs1801394) with susceptibility of early childhood caries (ECC) and with body mass index alterations. A cross-sectional study was performed in 488 children aged from 2 to 6 years from 25 public day care centers in Rio de Janeiro, Brazil. Demographic data and oral health habits were obtained through a questionnaire. Anthropometric measurements and caries experience data were collected by 2 examiners (κ = 0.80). Genotyping of the selected polymorphisms was carried out by TaqMan real-time PCR using genomic DNA extracted from buccal cells. Allele and genotype frequencies were compared between groups with and without disease. The t test, χ2 test, odds ratio, Pearson correlation tests, and logistic regression analysis were used (p ≤ 0.05). The mean white spot lesion score was 1.18 (±2.57) in normal weight children and 2.50 (±3.87) in underweight children (p = 0.05). For MTRR polymorphisms, significant differences were observed for allele and genotype frequency distributions between caries-free and caries-affected children (p = 0.03 and 0.04 for allele and genotype frequencies, respectively) and in the genotype frequencies between normal weight and underweight children (p = 0.04). Our results suggest an association between underweight and ECC; in addition it is suggested that MTRR is a common genetic risk factor for ECC and underweight.
Assuntos
Cárie Dentária/genética , Ferredoxina-NADP Redutase/genética , Polimorfismo de Nucleotídeo Único , Magreza/genética , Pré-Escolar , Estudos Transversais , Feminino , Humanos , MasculinoRESUMO
BACKGROUND: Ferredoxin NADP(H) oxidoreductases (EC 1.18.1.2) (FNR) are flavoenzymes present in photosynthetic organisms; they are relevant for the production of reduced donors to redox reactions, i.e. in photosynthesis, the reduction of NADP+ to NADPH using the electrons provided by Ferredoxin (Fd), a small FeS soluble protein acceptor of electrons from PSI in chloroplasts. In rhodophyta no information about this system has been reported, this work is a contribution to the molecular and functional characterization of FNR from Gracilaria chilensis, also providing a structural analysis of the complex FNR/Fd. METHODS: The biochemical and kinetic characterization of FNR was performed from the enzyme purified from phycobilisomes enriched fractions. The sequence of the gene that codifies for the enzyme, was obtained using primers designed by comparison with sequences of Synechocystis and EST from Gracilaria. 5'RACE was used to confirm the absence of a CpcD domain in FNRPBS of Gracilaria chilensis. A three dimensional model for FNR and Fd, was built by comparative modeling and a model for the complex FNR: Fd by docking. RESULTS: The kinetic analysis shows KMNADPH of 12.5 M and a kcat of 86 s-1, data consistent with the parameters determined for the enzyme purified from a soluble extract. The sequence for FNR was obtained and translated to a protein of 33646 Da. A FAD and a NADP+ binding domain were clearly identified by sequence analysis as well as a chloroplast signal sequence. Phycobilisome binding domain, present in some cyanobacteria was absent. Transcriptome analysis of Gch revealed the presence of two Fd; FdL and FdS, sharing the motif CX5CX2CX29X. The analysis indicated that the most probable partner for FNR is FdS. CONCLUSION: The interaction model produced, was consistent with functional properties reported for FNR in plants leaves, and opens the possibilities for research in other rhodophyta of commercial interest.