Highlights of three metabolites HDL and reduction in blood pressure values after dietary fiber supplementation in overweight and obese normotensive women: a metabolomic study.

INTRODUCTION: The prevalence of hypertension and obesity are a worldwide concern. OBJETIVES: Assess the metabolites profile after intervention with mixed dietary fiber in overweight and obese normotensive women. METHODS: This is a randomized double blind placebo-controlled study. Through a simple randomization process, two groups were allocated, with eleven women (group 1) receiving 12 g of mixed dietary fiber and thirteen women (group 2) receiving 12 g of placebo (corn starch) for eight weeks. Anthropometric and biochemical tests and lifestyle were analyzed. As for evaluation metabolomics, used a 1H NMR. The data matrix generated 96 samples and 225 variables, which was exported in the ASCII format for the "The Unscrumbler" statistics software (version 9.7, CAMO Process). RESULTS: After the intervention with mixed dietary fiber, significant differences were observed between the main types of metabolites, referring to the increase in the relative peak areas of in three HDL metabolites 4.94 ppm (0.0086*), HDL 1.28 ppm (0 .0337*), HDL 0.88 ppm (0.0224*) and an α-glucose metabolite 4.90 ppm (0.0106) and the reduction in systolic blood pressure (SBP) (0.0292*) of 7 mmHg in the reference range and in the placebo group there was a reduction in SBP (0.0118*) of 4 mmHg and of a choline metabolite 3.65 ppm (0.0266*), which does not call into question the validity of these results in the literature. CONCLUSION: The synergism of the functions of these statistically highlighted metabolites contributed to prevention the increase in SBP after fiber intervention in overweight and obese normotensive women.

Intoxication by 3-MeO-PCP and O-desmethyltramadol: an unusual NPS mix.

Over the past few years, the new psychoactive substances' phenomenon has been continuously studied. Its dynamic context is characterized by a broad diversity of substances, including several groups, such as synthetic cathinones, synthetic opiates, and synthetic cannabinoids. However, and due both to this diversity and to the low number of detected cases, information on intoxication reports is always important, in order to understand their biological mechanisms. In this case, a male individual was found unresponsive, with some different powders and paraphernalia near him. After toxicological analysis to the powders, paraphernalia, and whole blood samples, five different compounds were identified. From these, two of them (3-MeO-PCP and o-desmethyltramadol) were identified and quantitated in the whole blood sample. The obtained results suggested that death was due to the presence and action of these two substances, in what may be considered an unusual mix of NPS. This case highlights the value of evaluating all the traces found in the scene investigation and the need of sending all the paraphernalia found for toxicological examination, together with all the possible information obtained on the scene, namely by relatives or witnesses. On the other hand, this case shows the significance of broad-spectrum analytical methods, in order to detect and identify, as specifically as possible, eventual substances present and used by victims.

Willingness to pay for a hypothetical malaria vaccine in Brazil: a cross-sectional study and the implications.

Aim: Malaria is an infection caused by protozoa of genus Plasmodium, considered the one associated with increasingly large outbreaks. Methods: A cross-sectional study was conducted with residents in the northern region of Brazil on the willingness to pay (WTP) for a hypothetical vaccine against malaria (effective protection of 80%). Results: Of 616 people interviewed, most interviewees were female (61%) and were employed (97%). The median individual maximum WTP for a hypothetical malaria vaccine was US$11.90 (BRL 50). Conclusion: The northern region of Brazil is one of the largest markets for a malaria vaccine due to its epidemiological relevance. Consequently, economic studies will be important to assist in the assessment of the potential price and value of new vaccines.

Evaluation of anthropometry as an alternative to DXA as predictor of low bone mineral density in children and adolescents with cystic fibrosis.

BACKGROUND & AIMS: Low bone mineral density (BMD) for age in people with Cystic Fibrosis (CF) is associated with worse nutritional status. The aim of this study is to assess body composition by anthropometry as a predictor of BMD in people with CF. METHODS: Multicenter cross-sectional study with 39 people aged 5 and 20 years with CF. BMD was assessed by dual energy x-ray emission (DXA) in the incidence of the total body less head (TBLH) and the TBLH Z-score (Z-TBLH) was calculated, adjusted by sex, age, height and ethnicity. Anthropometry was assessed by weight, height, mid-upper arm circumference (MUAC) and triceps skinfold (TSF). Arm muscle area (AMA) and Body Mass Index (BMI) were calculated. Lean mass (LM), fat mass (FM) and free-fat mass (FFM) were identified by DXA. The molecular analysis method by sequencing was used to identify and classify the participants regarding the presence of the F508del pathogenic variant of the CFTR gene. Statistical models of simple and multiple linear regression were created to establish the predictive power of Z-TBLH in the variables. RESULTS: Average age of the participants was 13.31 ± 3.86 years, 59% of whom were male. They showed more LM (30.97 Kg ± 11.29) than females (23 Kg ± 6.73). 20 of 30 participants (66.7%) had at least copy of F508del. Among the multiple models, adjusted by height, age and sex, it found BMI (R2 = 0.367), Weight (R2 = 0.220), AMA (R2 = 0.338) as significant predictors of Z-TBLH. The final model composed of AMA, TSF and Age (p = 0.001; R2 = 0.381) had AMA and Age as significant predictors. AMA was associated with an increase in the BMD Z-score in the participants studied. 66.7% of genetically tested participants had the F508del pathogenic variant. The presence of the F508del variant was associated with worse nutritional status. CONCLUSION: A statistical model composed of the values of AMA, TSF and Age can predict Z-TBLH, as well as anthropometric variables Weight, or BMI, or AMA associated with height, age and sex, in children and adolescents aged 5-20 years old, of both sexes. Anthropometric markers, as they are easy and relatively inexpensive to obtain, it is a promising alternative to the use of DXA in predicting BMD in these people with CF.

Relationship between BsmI polymorphism and VDR gene methylation profile, gender, metabolic profile, oxidative stress, and inflammation in adolescents / Relación entre polimorfismo BsmI y perfil de metilación del gen VDR, género, perfil metabólico, estrés oxidativo e inflamación en adolescentes

Background: the biological activity of vitamin D depends on the activity of its receptor or VDR. On the other hand, the activity of this receptor is influenced by its state of methylation. The objective of this study was to verify if the BsmI polymorphism of the VDR gene influences its methylation profile in adolescents. Secondly, it was to verify if the status of some metabolic factors (oxidative stress, inflammation, lipid profile, and glycemia) in the serum, and gender-adjusted vitamin D levels are independent factors with an influence on the VDR methylation profile. Methods and results: the study included 198 adolescents of both sexes, aged 15-19 years, who underwent testing for VDR gene methylation polymorphisms, serum vitamin D levels, and metabolic, oxidative stress, and systemic inflammation markers. It was observed that the BB genotype was less methylated than the other groups (26.1 % versus 30.3 %, and 29.3 % for Bb and bb, respectively), although without statistical differences between them. The odds ratio indicated a protection of 13 % (partially methylated) for vitamin D status, while alpha glycols increased the risk ratio (of being partially methylated) by 3 %. MDA was protective at a 28 % chance of risk that adolescents with higher levels of lipid peroxidation would be hypomethylated. Conclusion: we conclude that the methylation profile of the VDR gene is not influenced by the different BsmI polymorphism genotypes, and that serum vitamin D and serum markers of oxidative stress and inflammation can modulate this profile. (AU)

Antecedentes: la actividad biológica de la vitamina D depende de la actividad de su receptor, el VDR. Por otro lado, la actividad de este receptor está influenciada por su estado de metilación. El objetivo de este estudio es verificar si el polimorfismo BsmI del gen VDR influye en el perfil de metilación del mismo en los adolescentes. En segundo lugar, verificar si los factores metabólicos (estrés oxidativo, inflamación, perfil lipídico y glucemia) del suero y la vitamina D ajustada por sexo actúan independientemente de los polimorfismos sobre el perfil de metilación del VDR. Métodos y resultados: el estudio incluyó a 198 adolescentes de ambos sexos, de 15 a 19 años de edad, que se sometieron a análisis de polimorfismos de metilación del gen VDR, niveles de vitamina D, marcadores metabólicos, estrés oxidativo e inflamación sistémica. Se observó que el genotipo BB estaba menos metilado que los otros grupos (26,1 % contra 30,3 % y 29,3 % para Bb y bb respectivamente), aunque sin diferencias estadísticas entre ellos. El odds ratio indicó una protección del 13 % (parcialmente metilado) para el estado de la vitamina D, mientras que los alfa glicoles aumentaron el índice de riesgo (de estar parcialmente metilado) en un 3 %. La MDA fue protectora con un 28 % de probabilidad de riesgo de que los adolescentes con niveles más altos de peroxidación lipídica fueran hipometilados. Conclusión: concluimos que el perfil de metilación del gen VDR no está influenciado por los diferentes genotipos del polimorfismo BsmI y que la vitamina D y los marcadores de estrés oxidativo e inflamación en el suero pueden modular este perfil. (AU)

Relationship between BsmI polymorphism and VDR gene methylation profile, gender, metabolic profile, oxidative stress, and inflammation in adolescents.

INTRODUCTION: Background: the biological activity of vitamin D depends on the activity of its receptor or VDR. On the other hand, the activity of this receptor is influenced by its state of methylation. The objective of this study was to verify if the BsmI polymorphism of the VDR gene influences its methylation profile in adolescents. Secondly, it was to verify if the status of some metabolic factors (oxidative stress, inflammation, lipid profile, and glycemia) in the serum, and gender-adjusted vitamin D levels are independent factors with an influence on the VDR methylation profile. Methods and results: the study included 198 adolescents of both sexes, aged 15-19 years, who underwent testing for VDR gene methylation polymorphisms, serum vitamin D levels, and metabolic, oxidative stress, and systemic inflammation markers. It was observed that the BB genotype was less methylated than the other groups (26.1 % versus 30.3 %, and 29.3 % for Bb and bb, respectively), although without statistical differences between them. The odds ratio indicated a protection of 13 % (partially methylated) for vitamin D status, while alpha glycols increased the risk ratio (of being partially methylated) by 3 %. MDA was protective at a 28 % chance of risk that adolescents with higher levels of lipid peroxidation would be hypomethylated. Conclusion: we conclude that the methylation profile of the VDR gene is not influenced by the different BsmI polymorphism genotypes, and that serum vitamin D and serum markers of oxidative stress and inflammation can modulate this profile.

INTRODUCCIÓN: Antecedentes: la actividad biológica de la vitamina D depende de la actividad de su receptor, el VDR. Por otro lado, la actividad de este receptor está influenciada por su estado de metilación. El objetivo de este estudio es verificar si el polimorfismo BsmI del gen VDR influye en el perfil de metilación del mismo en los adolescentes. En segundo lugar, verificar si los factores metabólicos (estrés oxidativo, inflamación, perfil lipídico y glucemia) del suero y la vitamina D ajustada por sexo actúan independientemente de los polimorfismos sobre el perfil de metilación del VDR. Métodos y resultados: el estudio incluyó a 198 adolescentes de ambos sexos, de 15 a 19 años de edad, que se sometieron a análisis de polimorfismos de metilación del gen VDR, niveles de vitamina D, marcadores metabólicos, estrés oxidativo e inflamación sistémica. Se observó que el genotipo BB estaba menos metilado que los otros grupos (26,1 % contra 30,3 % y 29,3 % para Bb y bb respectivamente), aunque sin diferencias estadísticas entre ellos. El odds ratio indicó una protección del 13 % (parcialmente metilado) para el estado de la vitamina D, mientras que los alfa glicoles aumentaron el índice de riesgo (de estar parcialmente metilado) en un 3 %. La MDA fue protectora con un 28 % de probabilidad de riesgo de que los adolescentes con niveles más altos de peroxidación lipídica fueran hipometilados. Conclusión: concluimos que el perfil de metilación del gen VDR no está influenciado por los diferentes genotipos del polimorfismo BsmI y que la vitamina D y los marcadores de estrés oxidativo e inflamación en el suero pueden modular este perfil.

Consumer willingness to pay for a hypothetical Chagas disease vaccine in Brazil: a cross-sectional study and the implications.

Aim: Chagas disease is a serious public health problem, endemic in 21 countries in Latin America. A future vaccine can contribute to decreasing the number of cases and its complications. Methods: A cross-sectional study was conducted with residents of the northern region of Brazil, on the willingness to pay for a hypothetical vaccine against Chagas disease (effective protection of 80%). Results: We interviewed 619 individuals and seven were excluded from the analysis and the value of willingness to pay was US$23.77 (100.00 BRL). Conclusion: The Northern region of Brazil is one of the largest markets for this vaccine, due to its epidemiological relevance, so economic studies with this vaccine will be important to assist in the assessment of technologies.

Choline Metabolites, Hydroxybutyrate and HDL after Dietary Fiber Supplementation in Overweight/Obese Hypertensive Women: A Metabolomic Study.

Metabolomics has been increasingly used to evaluate metabolic changes associated with morbidities. The objective of this study is to assess the metabolic profile before and after intervention with mixed dietary fiber in overweight and obese hypertensive women. This is an intervention study, and the sample consists of 14 women aged 28 to 58 years. An intervention with 12 g of mixed soluble and insoluble fiber is performed for a period of eight weeks. Serum metabolites are identified using a Bruker 1H NMR spectrometer at 400 MHz. Multivariate data analysis, including principal component analysis (PCA), is used to differentiate the two groups. After supplementation with dietary fiber, there is a significant increase in the peak intensity values of the metabolites HDL-C (0.0010*), choline (0.0012*) and hydroxybutyrate (0.0010*) as well as a decrease in systolic (0.0013*) and diastolic (0.0026*) blood pressure. The analysis of the metabolomic profile allows the identification of metabolites that have been associated in the literature with hypertension and excess weight (choline, hydroxybutyrate and amino acids) and with fiber intake (choline, hydroxybutyrate and amino acids) in addition to an increase in HDL-C. The increase in the detection of the described metabolites possibly occurs due to the presence of pathologies and the use of fiber in the intervention, which also contributes to elevated HDL-c and reduced blood pressure.

The direct correlation between oxidative stress and LDL-C levels in adults is maintained by the Friedewald and Martin equations, but the methylation levels in the MTHFR and ADRB3 genes differ.

Low-density lipoprotein (LDL-C) concentrations are a standard of care in the prevention of cardiovascular disease and are influenced by different factors. This study compared the LDL-C concentrations estimated by two different equations and determined their associations with inflammatory status, oxidative stress, anthropometric variables, food intake and DNA methylation levels in the LPL, ADRB3 and MTHFR genes. A cross-sectional population-based study was conducted with 236 adults (median age 37.5 years) of both sexes from the municipality of João Pessoa, Paraíba, Brazil. The LDL-C concentrations were estimated according to the Friedewald and Martin equations. LPL, ADRB3 and MTHFR gene methylation levels; malondialdehyde levels; total antioxidant capacity; ultra-sensitive C-reactive protein, alpha-1-acid glycoprotein, homocysteine, cobalamin, and folic acid levels; usual dietary intake; and epidemiological variables were also determined. For each unit increase in malondialdehyde concentration there was an increase in the LDL-C concentration from 6.25 to 10.29 mg/dL (p <0.000). Based on the Martin equation (≥70 mg/dL), there was a decrease in the DNA methylation levels in the ADRB3 gene and an increase in the DNA methylation levels in the MTHFR gene (p <0.05). There was a positive relation of homocysteine and cholesterol intake on LDL-C concentrations estimated according to the Friedewald equation and of waist circumference and age based on the two estimates. It is concluded the LDL-C concentrations estimated by the Friedewald and Martin equations were different, and the Friedewald equation values were significantly lower than those obtained by the Martin equation. MDA was the variable that was most positively associated with the estimated LDL-C levels in all multivariate models. Significant relationships were observed based on the two estimates and occurred for most variables. The methylation levels of the ADRB3 and MTHFR genes were different according to the Martin equation at low LDL-C concentrations (70 mg/dL).

Food Intervention with Folate Reduces TNF-α and Interleukin Levels in Overweight and Obese Women with the MTHFR C677T Polymorphism: A Randomized Trial.

Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism associated with body fat accumulation could possibly trigger an inflammatory process by elevating homocysteine levels and increasing cytokine production, causing several diseases. This study aimed to evaluate the effects of food intervention, and not folate supplements, on the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) in overweight and obese women with the MTHFR C677T polymorphism. A randomized, double-blind eight-week clinical trial of 48 overweight and obese women was conducted. Participants were randomly assigned into two groups. They received 300 g of vegetables daily for eight weeks containing different doses of folate: 95 µg/day for Group 1 and 191 µg/day for Group 2. MTHFR C677T polymorphism genotyping was assessed by digestion with HinfI enzyme and on 12% polyacrylamide gels. Anthropometric measurements, 24-h dietary recall, and biochemical analysis (blood folic acid, vitamin B12, homocysteine (Hcy), TNF-α, IL-1ß, and IL-6) were determined at the beginning and end of the study. Group 2 had a significant increase in folate intake (p < 0.001) and plasma folic acid (p < 0.05) for individuals with the cytosine-cytosine (CC), cytosine-thymine (CT), and thymine-thymine (TT) genotypes. However, only individuals with the TT genotype presented reduced levels of Hcy, TNF-α, IL-6, and IL-1ß (p < 0.001). Group 1 showed significant differences in folate consumption (p < 0.001) and folic acid levels (p < 0.05) for individuals with the CT and TT genotypes. Food intervention with folate from vegetables increased folic acid levels and reduced interleukins, TNF-α, and Hcy levels, mainly for individuals with the TT genotype.

Variants rs1544410 and rs2228570 of the vitamin D receptor gene and glycemic levels in adolescents from Northeast Brazil / Variantes rs1544410 y rs2228570 del gen receptor de la vitamina D y niveles glicémicos en adolescentes del noreste Brasil

Objective: to verify the association of serum concentrations of 25-hydroxyvitamin D and glycemic levels with the genetic variants rs1544410 and rs2228570 of the VDR gene in adolescents from the Northeast region of Brazil. Materials and methods: a cross-sectional epidemiological study with 208 adolescents from public schools in the city of João Pessoa (Paraíba, Brazil) between 15 and 19 years of age. Blood samples were collected for DNA extraction and analysis of polymorphisms rs1544410 and rs2228570, as well as biochemical analyses (25-hydroxyvitamin D, parathyroid hormone, calcium and glycemia). Results: the mean age was 17.7 (± 1.14) years. Half of adolescents had sufficient serum levels of 25-hydroxyvitamin D and the other half had insufficient/deficient vitamin. The most frequent genotypic distribution was bb and Ff and of lesser frequency BB and ff. There was a significant relationship between the genotypes of rs1544410 and glycemia values (p = 0.049) in the relationships between the genotypes BBxbb (p = 0.012) and Bbxbb (p = 0.037); (p = 0.036, OR = 2.15, 95 % CI = 1.05-4.41), and in the BB+Bb group analysis when compared to the bb (p = 0.025, OR = 1.89, 95 % CI = 1.08-3.29) presented higher risk of glycemia above the median. On the other hand, when Bb+bb was analyzed in relation to BB, adolescents had a greater chance of blood glucose below the median (p = 0.025, OR = 0.66, CI = 0.47-0.95). Conclusion: this study showed a significant relation of glycemia with the distribution of rs1544410 polymorphism genotypes

Objetivo: verificar la asociación de las concentraciones séricas de 25-hidroxivitamina D y los niveles de glucemia con las variantes genéticas rs1544410 y rs2228570 del gen VDR en adolescentes de la región noreste de Brasil. Materiales y métodos: se realizó un estudio epidemiológico transversal con 208 adolescentes de escuelas públicas en la ciudad de João Pessoa (Paraíba, Brasil) de entre 15 y 19 años de edad. Se recogieron muestras de sangre para la extracción de ADN y el análisis de los polimorfismos rs1544410 y rs2228570, así como para análisis bioquímicos (25-hidroxivitamina D, hormona paratiroidea, calcio y glucemia). Resultados: la edad media fue de 17,7 (± 1,14) años. La mitad de los adolescentes tenía niveles séricos suficientes de 25-hidroxivitamina D y la otra mitad, vitamina insuficiente/deficiente. La distribución genotípica más frecuente fue bb y Ff y la de menor frecuencia, BB y ff. Hubo una relación significativa entre los genotipos de rs1544410 y los valores de glucemia (p = 0,049) en las relaciones entre los genotipos BBxbb (p = 0,012) y Bbxbb (p = 0,037); (p = 0,036, OR = 2,15, IC 95 % = 1,05-4.41), y el análisis del grupo BB + Bb en comparación con el bb (p = 0,025, OR = 1,89, IC 95 % = 1,08-3,29) mostró un mayor el riesgo de glucemia, por encima de la mediana. Por otro lado, cuando se analizó Bb+bb en relación con la BB, los adolescentes tuvieron una mayor probabilidad de que la glucosa en sangre estuviera por debajo de la mediana (p = 0,025, OR = 0,66, IC = 0,47-0,95). Conclusión: este estudio mostró una relación significativa entre la glucemia y la distribución de genotipos de polimorfismo rs1544410

Impact of sitagliptin on endometrial mesenchymal stem-like progenitor cells: A randomised, double-blind placebo-controlled feasibility trial.

BACKGROUND: Recurrent pregnancy loss (RPL) is associated with the loss of endometrial mesenchymal stem-like progenitor cells (eMSC). DPP4 inhibitors may increase homing and engraftment of bone marrow-derived cells to sites of tissue injury. Here, we evaluated the effect of the DPP4 inhibitor sitagliptin on eMSC in women with RPL, determined the impact on endometrial decidualization, and assessed the feasibility of a full-scale clinical trial. METHODS: A double-blind, randomised, placebo-controlled feasibility trial on women aged 18 to 42 years with a history of 3 or more miscarriages, regular menstrual cycles, and no contraindications to sitagliptin. Thirty-eight subjects were randomised to either 100 mg sitagliptin daily for 3 consecutive cycles or identical placebo capsules. Computer generated, permuted block randomisation was used to allocate treatment packs. Colony forming unit (CFU) assays were used to quantify eMSC in midluteal endometrial biopsies. The primary outcome measure was CFU counts. Secondary outcome measures were endometrial thickness, study acceptability, and first pregnancy outcome within 12 months following the study. Tissue samples were subjected to explorative investigations. FINDINGS: CFU counts following sitagliptin were higher compared to placebo only when adjusted for baseline CFU counts and age (RR: 1.52, 95% CI: 1.32-1.75, P<0.01). The change in CFU count was 1.68 in the sitagliptin group and 1.08 in the placebo group. Trial recruitment, acceptability, and drug compliance were high. There were no serious adverse events. Explorative investigations showed that sitagliptin inhibits the expression of DIO2, a marker gene of senescent decidual cells. INTERPRETATION: Sitagliptin increases eMSCs and decreases decidual senescence. A large-scale clinical trial evaluating the impact of preconception sitagliptin treatment on pregnancy outcome in RPL is feasible and warranted. FUNDING: Tommy's Baby Charity. CLINICAL TRIAL REGISTRATION: EU Clinical Trials Register no. 2016-001120-54.

Variants rs1544410 and rs2228570 of the vitamin D receptor gene and glycemic levels in adolescents from Northeast Brazil.

INTRODUCTION: Objective: to verify the association of serum concentrations of 25-hydroxyvitamin D and glycemic levels with the genetic variants rs1544410 and rs2228570 of the VDR gene in adolescents from the Northeast region of Brazil. Materials and methods: a cross-sectional epidemiological study with 208 adolescents from public schools in the city of João Pessoa (Paraíba, Brazil) between 15 and 19 years of age. Blood samples were collected for DNA extraction and analysis of polymorphisms rs1544410 and rs2228570, as well as biochemical analyses (25-hydroxyvitamin D, parathyroid hormone, calcium and glycemia). Results: the mean age was 17.7 (± 1.14) years. Half of adolescents had sufficient serum levels of 25-hydroxyvitamin D and the other half had insufficient/deficient vitamin. The most frequent genotypic distribution was bb and Ff and of lesser frequency BB and ff. There was a significant relationship between the genotypes of rs1544410 and glycemia values (p = 0.049) in the relationships between the genotypes BBxbb (p = 0.012) and Bbxbb (p = 0.037); (p = 0.036, OR = 2.15, 95% CI = 1.05-4.41), and in the BB+Bb group analysis when compared to the bb (p = 0.025, OR = 1.89, 95% CI = 1.08-3.29) presented higher risk of glycemia above the median. On the other hand, when Bb+bb was analyzed in relation to BB, adolescents had a greater chance of blood glucose below the median (p = 0.025, OR = 0.66, CI = 0.47-0.95). Conclusion: this study showed a significant relation of glycemia with the distribution of rs1544410 polymorphism genotypes.

INTRODUCCIÓN: Objetivo: verificar la asociación de las concentraciones séricas de 25-hidroxivitamina D y los niveles de glucemia con las variantes genéticas rs1544410 y rs2228570 del gen VDR en adolescentes de la región noreste de Brasil. Materiales y métodos: se realizó un estudio epidemiológico transversal con 208 adolescentes de escuelas públicas en la ciudad de João Pessoa (Paraíba, Brasil) de entre 15 y 19 años de edad. Se recogieron muestras de sangre para la extracción de ADN y el análisis de los polimorfismos rs1544410 y rs2228570, así como para análisis bioquímicos (25-hidroxivitamina D, hormona paratiroidea, calcio y glucemia). Resultados: la edad media fue de 17,7 (± 1,14) años. La mitad de los adolescentes tenía niveles séricos suficientes de 25-hidroxivitamina D y la otra mitad, vitamina insuficiente/deficiente. La distribución genotípica más frecuente fue bb y Ff y la de menor frecuencia, BB y ff. Hubo una relación significativa entre los genotipos de rs1544410 y los valores de glucemia (p = 0,049) en las relaciones entre los genotipos BBxbb (p = 0,012) y Bbxbb (p = 0,037); (p = 0,036, OR = 2,15, IC 95% = 1,05-4.41), y el análisis del grupo BB + Bb en comparación con el bb (p = 0,025, OR = 1,89, IC 95% = 1,08-3,29) mostró un mayor el riesgo de glucemia, por encima de la mediana. Por otro lado, cuando se analizó Bb+bb en relación con la BB, los adolescentes tuvieron una mayor probabilidad de que la glucosa en sangre estuviera por debajo de la mediana (p = 0,025, OR = 0,66, IC = 0,47-0,95). Conclusión: este estudio mostró una relación significativa entre la glucemia y la distribución de genotipos de polimorfismo rs1544410.

Methylation profile of the ADRB3 gene and its association with lipid profile and nutritional status in adults.

BACKGROUND: Defects in DNA methylation have been shown to be associated with metabolic diseases such as obesity, dyslipidemia, and hypercholesterolemia. To analyze the methylation profile of the ADRB3 gene and correlate it with lipid profile, lipid intake, and oxidative stress based on malondialdehyde (MDA) and total antioxidant capacity (TAC), homocysteine and folic acid levels, nutritional status, lifestyle, and socioeconomic variables in an adult population. A cross-sectional epidemiological study representative of the East and West regions of the municipality of João Pessoa, Paraíba state, Brazil, enrolled 265 adults of both genders. Demographic, lifestyle, and socioeconomic questionnaires and a 24-h recall questionnaire were applied by trained interviewers' home. Nutritional and biochemical evaluation (DNA methylation, lipid profile, MDA, TAC, homocysteine and folic acid levels) was performed. RESULTS: DNA hypermethylation of the ADRB3 gene, analyzed in leukocytes, was present in 50% of subjects and was associated with a higher risk of being overweight (OR 3.28; p = 0.008) or obese (OR 3.06; p = 0.017), a higher waist-hip ratio in males (OR 1.17; p = 0.000), greater intake of trans fats (OR 1.94; p = 0.032), higher LDL (OR 2.64; p = 0.003) and triglycerides (OR 1.81; p = 0.031), and higher folic acid levels (OR 1.85; p = 0.022). CONCLUSIONS: These results suggest that epigenetic changes in the ADRB3 gene locus may explain the development of obesity and non-communicable diseases associated with trans-fat intake, altered lipid profile, and elevated folic acid. Because of its persistence, DNA methylation may have an impact in adults, in association with the development of non-communicable diseases. This study is the first population-based study of the ADRB3 gene, and the data further support evaluation of ADRB3 DNA methylation as an effective biomarker.

Optimal design, anti-tumour efficacy and tolerability of anti-CXCR4 antibody drug conjugates.

Antibody-drug conjugates (ADCs) are promising therapies for haematological cancers. Historically, their therapeutic benefit is due to ADC targeting of lineage-restricted antigens. The C-X-C motif chemokine receptor 4 (CXCR4) is attractive for targeted therapy of haematological cancers, given its expression in multiple tumour types and role in cancer "homing" to bone marrow. However, CXCR4 is also expressed in haematopoietic cells and other normal tissues, raising safety challenges to the development of anti-CXCR4 ADCs for cancer treatment. Here, we designed the first anti-CXCR4 ADC with favourable therapeutic index, effective in xenografts of haematopoietic cancers resistant to standard of care and anti-CXCR4 antibodies. We screened multiple ADC configurations, by varying type of linker-payload, drug-to-antibody ratio (DAR), affinity and Fc format. The optimal ADC bears a non-cleavable linker, auristatin as payload at DAR = 4 and a low affinity antibody with effector-reduced Fc. Contrary to other drugs targeting CXCR4, anti-CXCR4 ADCs effectively eliminated cancer cells as monotherapy, while minimizing leucocytosis. The optimal ADC selectively eliminated CXCR4+ cancer cells in solid tumours, but showed limited toxicity to normal CXCR4+ tissues, sparing haematopoietic stem cells and progenitors. Our work provides proof-of-concept that through empirical ADC design, it is possible to target proteins with broad normal tissue expression.

The MTHFR promoter hypermethylation pattern associated with the A1298C polymorphism influences lipid parameters and glycemic control in diabetic patients.

BACKGROUND: Polymorphisms in the gene encoding methylenetetrahydrofolate reductase (MTHFR) have been investigated as risk factors for microvascular complications of diabetes; however, simultaneous analysis of these polymorphisms and the methylation pattern of the gene has never been conducted. The objective of the present study was to evaluate the simultaneous relationship between MTHFR methylation and MTHFR C6TT7 and A1298C polymorphisms with metabolic, inflammatory and oxidative stress parameters related to microvascular complications, diabetic retinopathy (DR) and diabetic nephropathy (DN) in diabetic patients. METHODS: A total of 107 patients who were diagnosed in the previous 5 to 10 years were recruited and divided into groups with complications (DR and/or DN) or without complications. Methylation analysis of the gene promoter was conducted using the MSP technique, and analysis of the A1298C and C677T polymorphisms was conducted using the restriction fragment length polymorphism (RFLP) assay. Microalbuminuria was determined using urine samples, and other analytes of interest were determined in blood samples using commercial kits. The Mann-Whitney and Chi square statistical tests were used with significance considered at p < 0.05. RESULTS: Subjects with a hypermethylated profile and the 1298AA genotype showed the highest levels of blood glucose (p = 0.03), total cholesterol (p = 0.0001) and LDL cholesterol (p = 0.0006). The same profile was associated with higher levels of HbA1c (p = 0.025), glycemia (p = 0.04) and total cholesterol (0.004) in the control group and total cholesterol (p = 0.005) and LDL cholesterol (p = 0.002) in the complications group. Serum creatinine was higher in subjects in the hypermethylated group with the genotype 677CC only in the control group (p = 0.0020). The methylated profile in presence of 677CC + 1298AA and the 677CT/TT +1298AA haplotypes showed higher levels of total cholesterol (p = 0.0024; 0.0031) and LDL cholesterol (p = 0.0060; 0.0125) than 1298AC/CC carriers. The fasting glycemia was higher in hypermethylated profile in the presence of 677CC/1298AA haplotype (p = 0.0077). CONCLUSION: The hypermethylated methylation profile associated with the 1298AA genotype appeared to be connected to higher values of glycemia, total cholesterol and LDL cholesterol.

Bacteriophage ϕIBB-PF7A loaded on sodium alginate-based films to prevent microbial meat spoilage.

Despite the recent advances achieved in food industries to fulfil the growing consumer demand for high quality and food safety, microbial contamination remains a serious issue. This study aimed to incorporate ϕIBB-PF7A bacteriophage (phage) onto sodium alginate-based films crosslinked with calcium chloride, to prevent poultry spoilage caused by Pseudomonas fluorescens. Films were prepared by casting and characterized in terms of phage loading, distribution, stability, release profile and antimicrobial performance. Results showed that phages were successfully incorporated as evidenced by their viability and homogeneous distribution within the films as assessed by microscopy. A decrease in phage viability was only detected after 8 weeks when stored under refrigerated conditions. Antimicrobial activity demonstrated that incorporated phages significantly impaired P. fluorescens growth. Films' antimicrobial efficacy was further demonstrated on chicken breast fillets artificially inoculated, decreasing 2Log P. fluorescens viable cell counts in the first two days and reductions were maintained up to 5 days of exposure (1 Log). These results highlight that phage incorporation onto sodium-alginate-based films constitutes a simple approach of preserving the antimicrobial activity of phages in a dried and insoluble format, that can further be applied in food industry for the prevention of microbial spoilage.

Methylation profile of the ADRB3 gene and its association with lipid profile and nutritional status in adults

BACKGROUND: Defects in DNA methylation have been shown to be associated with metabolic diseases such as obesity, dyslipidemia, and hypercholesterolemia. To analyze the methylation profile of the ADRB3 gene and correlate it with lipid profile, lipid intake, and oxidative stress based on malondialdehyde (MDA) and total antioxidant capacity (TAC), homocysteine and folic acid levels, nutritional status, lifestyle, and socioeconomic variables in an adult population. A cross-sectional epidemiological study representative of the East and West regions of the municipality of João Pessoa, Paraíba state, Brazil, enrolled 265 adults of both genders. Demographic, lifestyle, and socioeconomic questionnaires and a 24-h recall questionnaire were applied by trained interviewers' home. Nutritional and biochemical evaluation (DNA methylation, lipid profile, MDA, TAC, homocysteine and folic acid levels) was performed. RESULTS: DNA hypermethylation of the ADRB3 gene, analyzed in leukocytes, was present in 50% of subjects and was associated with a higher risk of being overweight (OR 3.28; p = 0.008) or obese (OR 3.06; p = 0.017), a higher waist-hip ratio in males (OR 1.17; p = 0.000), greater intake of trans fats (OR 1.94; p = 0.032), higher LDL (OR 2.64; p = 0.003) and triglycerides (OR 1.81; p = 0.031), and higher folic acid levels (OR 1.85; p = 0.022). CONCLUSIONS: These results suggest that epigenetic changes in the ADRB3 gene locus may explain the development of obesity and non-communicable diseases associated with trans-fat intake, altered lipid profile, and elevated folic acid. Because of its persistence, DNA methylation may have an impact in adults, in association with the development of non-communicable diseases. This study is the first population-based study of the ADRB3 gene, and the data further support evaluation of ADRB3 DNA methylation as an effective biomarker.

α-Tocopherol influences glycaemic control and miR-9-3 DNA methylation in overweight and obese women under an energy-restricted diet: a randomized, double-blind, exploratory, controlled clinical trial.

BACKGROUND: Excess weight is a strong risk factor for the development of dysglycaemia. It has been suggested that changes in the metabolism microRNAs, small non-coding RNAs that regulate gene expression, could precede late glycaemic changes. Vitamin E in turn may exert important functions in methylation and gene expression processes. This study aimed to determine the effect of α-tocopherol on glycaemic variables and miR-9-1 and miR-9-3 promoter DNA methylation in overweight women. METHODS: A randomized, double-blind, exploratory, placebo-controlled study was conducted in overweight and obese adult women (n = 44) who ingested synthetic vitamin E (all-rac-α-tocopherol), natural source vitamin E (RRR-rac-α-tocopherol) or placebo capsules and were followed up for a period of 8 weeks. Supplemented groups also received dietary guidance for an energy-restricted diet. An additional group that received no supplementation and did not follow an energy-restricted diet was also followed up. The intervention effect was evaluated by DNA methylation levels (quantitative real-time PCR assay) and anthropometric and biochemical variables (fasting plasma glucose, haemoglobin A1C, insulin, and vitamin E). RESULTS: Increased methylation levels of the miR-9-3 promoter region (P < 0.001) and reduced haemoglobin A1C (P < 0.05) were observed in the natural source vitamin E group after intervention. Increased fasting plasma glucose was observed in the synthetic vitamin E group, despite the significant reduction of anthropometric variables compared to the other groups. CONCLUSIONS: α-Tocopherol from natural sources increased methylation levels of the miR-9-3 promoter region and reduced haemoglobin A1C in overweight women following an energy-restricted diet. These results provide novel information about the influence of vitamin E on DNA methylation. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02922491. Registered 4 October, 2016.