CHDH-PNPLA3 Gene-Gene Interactions Predict Insulin Resistance in Children with Obesity.

INTRODUCTION: Insulin resistance plays a major role in metabolic syndrome and is recognized as the most common risk factor for non-alcoholic fatty liver disease (NAFLD). Identifying predictors for insulin resistance could optimize screening and prevention. PURPOSE: To evaluate the contribution of multiple single nucleotide polymorphisms across genes related to NAFLD and choline metabolism, in predicting insulin resistance in children with obesity. METHODS: One hundred fifty-three children with obesity (73 girls), aged 7-18 years, were evaluated within the NutriGen Study (ClinicalTrials.gov-NCT02837367). Insulin resistance was defined by Homeostatic Model Assessment for insulin-resistance cut-offs that accommodated pubertal and gender differences. Anthropometric, metabolic, intake-related variables, and 55 single nucleotide polymorphisms related to NAFLD and choline metabolism were evaluated. Gene-gene interaction effects were assessed using Multiple Data Reduction Software. RESULTS: Sixty percent (93/153) of participants showed insulin resistance (58.7% of boys, 63% of girls). Children with insulin resistance presented significantly higher values for standardized body mass index, triglycerides, transaminases and plasma choline when compared to those without insulin resistance. Out of 52 single nucleotide polymorphisms analysed, the interaction between genotypes CHDH(rs12676) and PNPLA3(rs738409) predicted insulin resistance. The model presented a 6/10 cross-validation consistency and 0.58 testing accuracy. Plasma choline levels and alanine aminotransferase modulated the gene interaction effect, significantly improving the model. CONCLUSION: The interaction between genotypes in CHDH and PNPLA3 genes, modulated by choline and alanine aminotransferase levels, predicted insulin-resistance status in children with obesity. If replicated in larger cohorts, these findings could help identify metabolic risk in children with obesity.

Reliability and validity of a General Nutrition Knowledge Questionnaire for adults in a Romanian population.

BACKGROUND: Nutritional knowledge assessment is an important component in nutrition research, and a prerequisite for the implementation of many policies and programs aimed at improving eating behavior. In order to generate objective results, validated tools for a given population must be employed. The aim of this study was to determine the validity and reliability of a nutrition knowledge questionnaire for Romanian adults. METHODS: Kleimann's version of a General Nutrition Knowledge Questionnaire, was translated and adapted to Romanian language, culture, and cuisine. The final format was developed in several steps and used four components: internal and external reliability were assessed in a general population sample (n1 = 412), respectively in a subgroup (n2 = 46) from Component 1; Component 3 assessed construct validity (n3 = 96) using the "known-groups" method; Component 4 (convergent validity, n4 = 508) tested the association between socio-demographic characteristics and nutrition knowledge. RESULTS: The overall internal reliability was 0.878 and the external reliability was >0.880 in all sections, and overall. Specialists had higher scores than nonspecialists, with a very large effect size. In the general population, females scored higher than males, and middle-aged and older adults scored higher than young adults. Higher scores were associated with higher levels of education. The characteristics of individuals prone to giving wrong answers were: males (beta = 0.170), high school or less (beta = 0.167), and no training in nutrition (beta = 0.154). CONCLUSIONS: The Romanian version of the General Nutrition Knowledge Questionnaire is a reliable and valid tool for measuring nutrition knowledge in adults.

Single Nucleotide Polymorphisms in PEMT and MTHFR Genes are Associated with Omega 3 and 6 Fatty Acid Levels in the Red Blood Cells of Children with Obesity.

Polyunsaturated fatty acids (PUFAs) play important roles in health and disease. PUFA levels are influenced by nutrition and genetic factors. The relationship between PUFA composition in red blood cells (RBCs) and genetic variations involved in PUFA metabolism has not been investigated in children with obesity. This study evaluated the association between several genetic variations and PUFA levels in RBCs in children with obesity. One hundred ninety-six children with obesity (101 females, 95 males) were evaluated using anthropometric measurements, dietary intakes, plasma and RBC PUFA quantification, blood biochemistry, and 55 single nucleotide polymorphisms within 14 genes. phosphatidylethanolamine N-methyltransferase (PEMT) rs1109859 and methylenetetrahydrofolate reductase gene (MTHFR) rs4846052 genotypes were associated with PUFA levels in RBCs. PUFA intake did not influence the RBC eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels. Higher RBC DHA and EPA levels were observed for PEMT rs1109859 GG and GA genotypes versus the AA genotype. Higher levels of RBC DHA, EPA, arachidonic acid (ARA), and linoleic acid (LA) and were observed for MTHFR rs4846052 TT genotype versus TC and CC genotypes. Genetic variations in PEMT rs1109859 and MTHFR rs4846052 were associated with different PUFA levels in RBC membranes and are estimators for PUFA species in RBCs. Further research is needed to establish whether these genotype-specific alterations are specific to overweight children.

Assessment of Nutritional Intakes in Individuals with Obesity under Medical Supervision. A Cross-Sectional Study.

People with obesity in Romania are often under medical supervision, which is aimed to decrease body weight and treat accompanying metabolic disorders and cardiovascular implications. However, there is limited information regarding the implementation of dietary recommendations in adults with obesity. We aimed to evaluate the prevalence of reaching the recommended intakes of macro- and micro-nutrients in adults with obesity under medical supervision. Individuals with obesity, recruited in the context of a study with a larger scope (NutriGen ClinicalTrials.gov NCT02837367), who were under medical supervision underwent four 24 h recalls in order to assess daily food intakes. Macro- and micro-nutrient intakes were computed, and the prevalence of reaching recommended dietary allowances (RDAs) for each nutrient was calculated. The majority of subjects did not meet the recommended intakes for most nutrients. Energy from fat exceeded the threshold of 35% recommended intake, even in the lowest quartile of energy intake. The micronutrients with less than 5% of individuals reaching the RDAs were vitamin D, vitamin E, fluoride, and omega-3 fatty acids for both males and females, and choline, magnesium, and potassium in females. The burden of inadequate nutrition in individuals with obesity should be acknowledged and properly addressed within efforts to reduce obesity rates and associated disorders.

Development and Validation of a LC⁻MS/MS-Based Assay for Quantification of Free and Total Omega 3 and 6 Fatty Acids from Human Plasma.

Few high-performance liquid chromatography⁻tandem mass spectrometry (LC-MS/MS) methods have been developed for the full quantitation of fatty acids from human plasma without derivatization. Therefore, we propose a method that requires fewer sample preparation steps, which can be used for the quantitation of several polyunsaturated fatty acids in human plasma. The method offers rapid, accurate, sensitive, and simultaneous quantification of omega 3 (α-linolenic, eicosapentaenoic, and docosahexaenoic acids) and omega 6 fatty acids (arachidonic and linoleic acids) using high-performance LC-MS/MS. The selected fatty acids were analysed in lipid extracts from both free and total forms. Chromatographic separation was achieved using a reversed phase C18 column with isocratic flow using ammonium acetate for improving negative electrospray ionization (ESI) response. Mass detection was performed in multiple reaction monitoring (MRM) mode, and deuterated internal standards were used for each target compound. The limits of quantification were situated in the low nanomolar range, excepting linoleic acid, for which the limit was in the high nanomolar range. The method was validated according to the U.S. Department of Health and Human Services guidelines, and offers a fast, sensitive, and reliable quantification of selected omega 3 and 6 fatty acids in human plasma.