Lipidomic analysis identified potential predictive biomarkers of statin response in subjects with Familial hypercholesterolemia

Familial hypercholesterolemia (FH) is a disorder of lipid metabolism that causes elevated low-density lipoprotein cholesterol (LDL-c) and increased premature atherosclerosis risk. Statins inhibit endogenous cholesterol biosynthesis, which reduces LDL-c plasma levels and prevent from cardiovascular events. This study aimed to explore the effects of statin treatment on serum lipidomic profile and to identify biomarkers of response in subjects with FH. Seventeen adult FH patients underwent a 6-week washout followed by 4-week treatment with atorvastatin (80 mg/day) or rosuvastatin (40 mg/day). LDL-c response was considered good (40­70 % reduction, n = 9) or poor (3­33 % reduction, n = 8). Serum lipidomic profile was analyzed by ultra-high-performance liquid chromatography combined with electrospray ionization tandem time-of-flight mass spectrometry, and data were analyzed using MetaboAnalyst v5.0. Lipidomic analysis identified 353 lipids grouped into 16 classes. Statin treatment reduced drastically 8 of 13 lipid classes, generating a characteristic lipidomic profile with a significant contribution of phosphatidylinositols (PI) 16:0/18:2, 18:0/18:1 and 18:0/18:2; and triacylglycerols (TAG) 18:2x2/18:3, 18:1/18:2/18:3, 16:1/18:2x2, 16:1/18:2/18:3 and 16:1/18:2/Arachidonic acid (p-adjusted <0.05). Biomarker analysis implemented in MetaboAnalyst subsequently identified PI 16:1/18:0, 16:0/18:2 and 18:0/18:2 as predictors of statin response with and receiver operating characteristic (ROC) areas under the curve of 0.98, 0.94 and 0.91, respectively. In conclusion, statins extensively modulate the overall serum lipid composition of FH individuals and these findings suggest that phosphatidyl-inositol molecules are potential predictive biomarkers of statin response.

Influence of polymorphisms in IRS1, IRS2, MC3R, and MC4R on metabolic and inflammatory status and food intake in Brazilian adults: An exploratory pilot study

Polymorphisms in genes of leptin-melanocortin and insulin pathways have been associated with obesity and type 2 diabetes. We hypothesized that polymorphisms in IRS1, IRS2, MC3R, and MC4R influence metabolic and inflammatory markers and food intake composition in Brazilian subjects. This exploratory pilot study included 358 adult subjects. Clinical, anthropometric, and laboratory data were obtained through interview and access to medical records. The variants IRS1 rs2943634 A˃C, IRS2 rs1865434 C>T, MC3R rs3746619 C>A, and MC4R rs17782313 T>C were analyzed by real-time polymerase chain reaction. Food intake composition was assessed in a group of subjects with obesity (n = 84) before and after a short-term nutritional counseling program (9 weeks). MC4R rs17782313 was associated with increased risk of obesity (P = .034). Multivariate linear regression analysis adjusted by covariates indicated associations of IRS2 rs1865434 with reduced low-density lipoprotein cholesterol and resistin, MC3R rs3746619 with high glycated hemoglobin, and IRS1 rs2943634 and MC4R rs17782313 with increased high-sensitivity C-reactive protein (P < .05). Energy intake and carbohydrate and total fat intakes were reduced after the diet-oriented program (P < .05). Multivariate linear regression analysis showed associations of IRS2 rs1865434 with high basal fiber intake, IRS1 rs2943634 with low postprogram carbohydrate intake, and MC4R rs17782313 with low postprogram total fat and saturated fatty acid intakes (P < .05). Although significant associations did not survive correction for multiple comparisons using the Benjamini-Hochberg method in this exploratory study, polymorphisms in IRS1, IRS2, MC3R, and MC4R influence metabolic and inflammatory status in Brazilian adults. IRS1 and MC4R variants may influence carbohydrate, total fat, and saturated fatty acid intakes in response to a diet-oriented program in subjects with obesity.

Identification of pathogenic variants in the brazilian cohort with familial hypercholesterolemia using exon-targeted gene sequencing

Familial hypercholesterolemia (FH) is a monogenic disease characterized by high plasma low-density lipoprotein cholesterol (LDL-c) levels and increased risk of premature atherosclerotic cardiovascular disease. Mutations in FH-related genes account for 40% of FH cases worldwide. In this study, we aimed to assess the pathogenic variants in FH-related genes in the Brazilian FH cohort FHBGEP using exon-targeted gene sequencing (ETGS) strategy. FH patients (n = 210) were enrolled at five clinical sites and peripheral blood samples were obtained for laboratory testing and genomic DNA extraction. ETGS was performed using MiSeq platform (Illumina). To identify deleterious variants in LDLR, APOB, PCSK9, and LDLRAP1, the long-reads were subjected to Burrows-Wheeler Aligner (BWA) for alignment and mapping, followed by variant calling using Genome Analysis Toolkit (GATK) and ANNOVAR for variant annotation. The variants were further filtered using in-house custom scripts and classified according to the American College Medical Genetics and Genomics (ACMG) guidelines. A total of 174 variants were identified including 85 missense, 3 stop-gain, 9 splice-site, 6 InDel, and 71 in regulatory regions (3'UTR and 5'UTR). Fifty-two patients (24.7%) had 30 known pathogenic or likely pathogenic variants in FH-related genes according to the American College Medical and Genetics and Genomics guidelines. Fifty-three known variants were classified as benign, or likely benign and 87 known variants have shown uncertain significance. Four novel variants were discovered and classified as such due to their absence in existing databases. In conclusion, ETGS and in silico prediction studies are useful tools for screening deleterious variants and identification of novel variants in FH-related genes, they also contribute to the molecular diagnosis in the FHBGEP cohort.

Influencia de la seropositividad contra adenovirus-36 sobre riesgo de obesidad y resistencia a la insulina en población infanto-juvenil de la Región de la Araucanía / Influence of adenovirus 36 seropositivity on the risk of obesity and insulin resistance in a population of children and adolescents from the Región de la Araucanía

La infección previa por el adenovirus-36 (Ad-36) se ha asociado con el proceso adipogénico y el control glicémico en modelos experimentales de cultivos celulares y animales. En humanos, la presencia de anticuerpos contra Ad-36 ha mostrado aumentar el riesgo de obesidad y, paradójicamente, mejorar el control glicémico en diferentes poblaciones. Se evaluó la influencia de la seropositividad contra Ad-36 sobre riesgo de obesidad, el perfil lipídico y glicémico en una población de niños en edad escolar. Métodos: Doscientos ocho individuos de entre 9 y 13 años se agruparon según estado nutricional como normopeso (IMC z-score de -1 a +1), con sobrepeso (IMC z-score de +1 a +2) y con obesidad (IMC z-score > +3). Se evaluaron medidas antropométricas, desarrollo puberal según Tanner y parámetros bioquímicos (perfil lipídico, glucemia e insulina) y la seropositividad contra Ad-36. Se determinó la resistencia a la insulina (RI) según criterio para la población infantil chilena. La seropositividad contra Ad-36 se determinó mediante ELISA. Resultados: Hubo una alta prevalencia de sobrepeso/obesidad en la población de estudio. La seropositividad contra Ad-36 fue del 5,4% en el grupo total, pero no se observó una asociación con el estado nutricional. No se encontró correlación entre la seropositividad contra Ad-36 y los parámetros del perfil lipídico. La insulina y la HOMA-RI fueron significativamente más bajas en el grupo Ad-36 (+) (p<0,001), no habiendo sido reportados casos de RI en el grupo Ad-36 (+) en nuestra población. Conclusiones: Nuestros resultados sugieren que la infección previa por el adenovirus-36 afecta la secreción de insulina y la resistencia a la insulina, como se ha descrito anteriormente, sin embargo, no se observa correlación con el desarrollo de la obesidad infantil en la población pediátrica del sur de Chile.

Previous infection with Adenovirus-36 (Ad-36) has been associated with adipogenic process and glycemic control in experimental models of cell culture and animals. In humans, the presence of antibodies against Ad-36 has been shown to increase the risk of obesity and, paradoxically, improve glycemic control in different populations. The influence of Ad-36 seropositivity on obesity risk, lipid and glycemic profile was evaluated in a population of school-age children. Methods: Two hundred eight individuals aged 9 to 13 years were grouped according to their nutritional status as normal weight (BMI z-score from -1 to +1), overweight (BMI z-score from +1 to +2) or obese (BMI z-score from -1 to +1). z-score > +3). Anthropometric measurements, pubertal development according to Tanner stage, biochemical parameters (lipid profile, glycemia and insulin) and seropositivity against Ad-36 were evaluated. Insulin resistance (IR) was determined according to criteria for the Chilean child population. Seropositivity against Ad-36 was determined by ELISA. Results: There was a high prevalence of overweight/obesity in the study population. Seropositivity against Ad-36 was 5.4% in the total group, but no association with nutritional status was observed. No correlation was found between Ad-36 seropositivity and lipid profile parameters. Insulin and HOMA-RI were significantly lower in the Ad-36 (+) group (p<0.001), and no cases of RI were reported in the Ad-36 (+) group in our population. Conclusions: Our results suggest that previous adenovirus-36 infection affects insulin secretion and insulin resistance, as previously described, however, no correlation is observed with the development of childhood obesity in the pediatric population. from southern Chile.

¿Existen discrepancias entre la planificación preoperatoria y la evaluación intraoperatoria realizada por el cirujano en la artroplastia total de rodilla con asistencia robótica? / Are there discrepancies between preoperative planning and intraoperative evaluation by the surgeon in robotically assisted total knee arthroplasty?

Introducción: Existe poca evidencia respecto de la concordancia entre el plan preoperatorio mediante artroplastia total de rodilla asistida por robot y el plan posterior al balance protésico realizado por el cirujano. El objetivo de este trabajo es evaluar el grado de concordancia entre la planificación preoperatoria de la artroplastia total de rodilla con asistencia robótica semiactiva (Mako) y la planificación efectuada por el traumatólogo durante la cirugía. Materiales y métodos: estudio retrospectivo y descriptivo de prótesis primarias instaladas entre octubre de 2018 y junio de 2019 con planificación preoperatoria realizada por el software MAKOplasty®. Se excluyeron las prótesis no colocadas por el sistema robótico o con información clínica incompleta. Esto se comparó con la planificación intraoperatoria del traumatólogo. Variables analizadas: alineación coronal y sagital, rotación y tamaño de los componentes e inserto. Los datos se analizaron con el softwareSTATA v.16.0. Se realizó un análisis descriptivo univariante cualitativo, con un intervalo de confianza del 95%. Resultados: se incluyeron cincuenta y una rodillas operadas de cuarenta y nueve pacientes, el 69% fueron mujeres. El nivel de concordancia para el componente femoral fue: axial 86.3% (IC = 73.7 - 94.2), coronal 88.2% (IC = 76.1 - 95.5), sagital 88.2% (IC = 76.1 - 95.5). Componente tibial: axial 98% (IC = 89.5 ­ 99.9), coronal 96.1% (IC = 86.5 ­ 99.5), sagital 96.1% (IC = 86.5 ­ 99.5). Tamaño del componente: fémur 94.1% (IC = 83.7 ­ 98.7), tibia 84.3% (IC = 71.4 ­ 92.9), inserto 27.4% (IC = 15.8 ­ 41.7). Conclusión: la planificación preoperatoria mediante el uso de la asistencia robótica semiactiva de Mako presenta un buen nivel de concordancia con la planificada intraoperatoriamente, a excepción del tamaño del inserto. El traumatólogo es determinante en la modificación del plan preoperatorio. Nivel de Evidencia: III

Introduction: There is little evidence regarding the concordance between the preoperative plan using robotic-assisted total knee arthroplasty and that after the prosthetic balance by the surgeon. Our aim is to evaluate the level of agreement between the preoperative planning of total knee arthroplasty with semiactive robotic assistance (Mako) and the planning made by the orthopedic surgeon during the surgery. Materials and methods: descriptive study of prostheses installed between October 2018 and June 2019 with preoperative planning performed by the MAKOplasty® software. This was compared with intraoperative planning by the Orthopedic Surgeon. Variables analyzed: coronal and sagittal alignment, rotation and size of the components and insert. The data was analyzed with the STATA v.16.0 software. A qualitative univariate descriptive analysis was performed, with a 95% confidence interval. Results: fifty-one operated knees from forty-nine patients were included, 69% were women. The level of agreement was: Femoral component: axial 86.3% [CI = 73.7 - 94.2], coronal 88.2% [CI = 76.1 - 95.5], sagittal 88.2% [CI = 76.1 - 95.5]. Tibial component: axial 98% [CI = 89.5 - 99.9], coronal 96.1% [CI = 86.5 - 99.5], sagittal 96.1% [CI = 86.5 - 99.5]. Component size: femur 94.1% [CI = 83.7 - 98.7], tibia 84.3% [CI = 71.4 - 92.9], insert 27.4% [CI = 15.8 - 41.7]. Conclusion: preoperative planning through the use of Mako semiactive robotic assistance presents a good level of agreement with that planned intraoperatively, with the exception of the insert size. The orthopedic surgeon is decisive in modifying the preoperative plan. Level of Evidence: III

Genetic variant ABCC1 rs45511401 is associated with increased response to statins in patients with familial hypercholesterolemia

Statins are the first-line treatment for familial hypercholesterolemia (FH), but response is highly variable due to genetic and nongenetic factors. Here, we explored the association between response and genetic variability in 114 Brazilian adult FH patients. Specifically, a panel of 84 genes was analyzed by exon-targeted gene sequencing (ETGS), and the functional impact of variants in pharmacokinetic (PK) genes was assessed using an array of functionality prediction methods. Low-density lipoprotein cholesterol (LDL-c) response to statins (reduction ≥ 50%) and statin-related adverse event (SRAE) risk were assessed in carriers of deleterious variants in PK-related genes using multivariate linear regression analyses. Fifty-eight (50.8%) FH patients responded to statins, and 24 (21.0%) had SRAE. Results of the multivariate regression analysis revealed that ABCC1 rs45511401 significantly increased LDL-c reduction after statin treatment (p < 0.05). In silico analysis of the amino-acid change using molecular docking showed that ABCC1 rs45511401 possibly impairs statin efflux. Deleterious variants in PK genes were not associated with an increased risk of SRAE. In conclusion, the deleterious variant ABCC1 rs45511401 enhanced LDL-c response in Brazilian FH patients. As such, this variant might be a promising candidate for the individualization of statin therapy.

Pharmacogenomics of mycophenolic acid in kidney transplantation: Contribution of immune response-related genes

Abstract Mycophenolic acid (MPA) inhibits IMPDH, involved in the guanosine nucleotides synthesis, and prevents DNA replication in immune cells. The repression of cell and humoral immunity by MPA induces allograft tolerance preventing acute rejection in solid organ transplantation. MPA is an effective and safe drug, but genetic and non-genetic factors have been implicated in the interindividual variability of drug response. Several studies have shown the impact of variants of pharmacokinetics or pharmacodynamics-related genes on MPA response in kidney transplantation. This review explored further the influence of genes involved in the immune response on clinical outcomes of kidney recipients on short- or long-term MPA treatment. Variants in genes related to T cell activation (CD28, CTL4, ICOS, PDPC1), pro-inflammatory cytokines (IL2, IL6, IL12A, IL12B, TNF, IFNG), immunomodulatory cytokines (IL4, IL10, TGFB1), and innate immune response (CD14, TLR2, TLR4) were shown to be associated with increased risk of acute rejection, graft function or survival, chronic graft nephropathy, viral infections or MPA-induced myelotoxicity. Some of the significant pharmacogenetic associations were confirmed by meta-analyses of kidney transplantation. These findings are suggestive that variants in immune response-related genes contribute to the variability of MPA response, and have potential application as biomarkers of acute rejection in kidney transplantation.

Genomics, epigenomics and pharmacogenomics of Familial Hypercholesterolemia (FHBGEP): A study protocol

BACKGROUND: Familial hypercholesterolemia (FH) is a genetic disease that affects millions of people worldwide. OBJECTIVES: The study protocol FHBGEP was design to investigate the main genomic, epigenomic, and pharmacogenomic factors associated with FH and polygenic hypercholesterolemia (PH). METHODS: FH patients will be enrolled at six research centers in Brazil. An exon-targeted gene strategy will be used to sequence a panel of 84 genes related to FH, PH, pharmacogenomics and coronary artery disease. Variants in coding and regulatory regions will be identified using a proposed variant discovery pipeline and classified according to the American College Medical Genetics guidelines. Functional effects of variants in FH-related genes will be investigated by in vitro studies using lymphocytes and cell lines (HepG2, HUVEC and HEK293FT), CRISPR/Cas9 mutagenesis, luciferase reporter assay and other technologies. Functional studies in silico, such as molecular docking, molecular dynamics, and conformational analysis, will be used to explore the impact of novel variants on protein structure and function. DNA methylation profile and differential expression of circulating non-coding RNAs (miRNAs and lncRNAs) will be analyzed in FH patients and normolipidemic subjects (control group). The influence of genomic and epigenomic factors on metabolic and inflammatory status will be analyzed in FH patients. Pharmacogenomic studies will be conducted to investigate the influence of genomic and epigenomic factors on response to statins in FH patients. SUMMARY: The FHBGEP protocol has the potential to elucidate the genetic basis and molecular mechanisms involved in the pathophysiology of FH and PH, particularly in the Brazilian population. This pioneering approach includes genomic, epigenomic and functional studies, which results will contribute to the improvement of the diagnosis, prognosis and personalized therapy of FH patients.

High serum miR-421 is associated with metabolic dysregulation and inflammation in patients with metabolic syndrome

ABSTRACT: To explore the association of circulating miRNAs with adiposity, metabolic status and inflammatory biomarkers in patients with metabolic syndrome (MetS). METHODS: Serum levels of 372 miRNAs were measured in patients with (n = 6) and without MetS (n = 6) by quantitative PCR array, and dysregulated miRNAs were validated in a larger cohort (MetS, n = 89; non-MetS, n = 144). RESULTS: In the screening study, seven miRNAs were dysregulated in patients with MetS, and miR-421 remained increased in the validation study. miR-421 was associated with a high risk of MetS and insulin resistance and hypertension and correlated with glycated hemoglobin, triacylglycerols, high-sensitivity CRP, IL-6, resistin and adiponectin (p < 0.05). CONCLUSION: Circulating miR-421 is a potential biomarker for insulin resistance, metabolic dysregulation and inflammatory status in patients with MetS.

Characterization of the adipogenic protein E4orf1 from denovirus 36 through an in silico approach

Abstract: Adenovirus 36 (Ad-36) is related to human obesity due to its adipogenic activity mediated by the early 4 open reading frame 1 (E4orf1) protein. Mechanisms underlying the adipogenic effect of E4orf1 are not completely understood; however, the proliferation and differentiation of fat cells are increased through the activation of the phosphatidyl inositol 3 kinase pathways by binding proteins containing PDZ domain. This study characterized E4orf1 tridimensional structure and analyzed its interactions with PDZ domain containing proteins in order to provide new information about the behavior of this viral protein and its targets, which could provide an interesting druggable target for obesity-related cardiometabolic alterations. In silico strategies such as homology modeling, docking, and molecular dynamics (MD) were used to study the interaction of E4orf1 with five PDZ domains of disk large homolog 1 (PDZ-1 and PDZ-2), membrane-associated guanylate kinase 1 (PDZ-3), and multi-PDZ domain protein 1 (PDZ-7 and PDZ-10). Mutagenesis analysis of selected residues was performed to evaluate their effects on the stabilization of E4orf1: PDZ complexes. MD simulations showed that the E4orf1: PDZ10 complex was more stable than the others ones. The highly hydrophobic residues at the C-terminal region (114­125) of the E4orf1 are essential in the initial phase stabilization of the complexes. Moreover, the residues 80­85 in the core region contribute to longer stabilization of the E4orf1: PDZ10 complex, a result that was confirmed by in silico mutagenesis. In conclusion, E4orf1 forms a stable complex with PDZ10 domain, and the residues 80­85 are of particular importance. The characterization of E4orf1 interactions with PDZ domains provides an initial approach to discover druggable targets for Ad-36-induced obesity.

Caracterização da proteína adipogênica E4orf1 do Adenovirus 36 através de técnicas de modelagem molecular / Characterization of Adenovirus 36 adipogenic protein E4orf1 by molecular modeling techniques

INTRODUÇÃO: O termo "Infectobesidade" vem tornando-se mais relevância em estudos sobre obesidade, devido à descoberta de alguns vírus responsáveis por produzir um aumento na proliferação e diferenciação de células adiposas. O adenovírus 36 (AdV-36) ganhou mais atenção devido à sua associação com a obesidade humana, onde uma maior prevalência de anticorpos anti-AdV-36 foi descrita em indivíduos obesos em comparação com não obesos. Sugere-se que os efeitos de E4orf1 são produzidos pela ativação da via Fosfatidilinositol 3 quiinase (PI3K), translocando GLUT4, em um processo regulado pela ativação de Ras. A formação de um complexo proteico entre E4orf1 e proteínas pertencentes ao citoplasma celular que contêm um domínio PDZ, como DLG-1, MAGI-1, MUPP1, PATJ e ZO-2, é necessária para a ativação de PI3K. MÉTODOS: Estratégias in silico como modelagem por homologia, dinâmica molecular (DM) e docking molecular foram utilizadas para entender a complementariedade molecular entre E4orf1 e 5 diferentes domínios da PDZ: DLG1 (PDZ1 e PDZ2), MAGI-1 (PDZ3), MUPP1 (PDZ7 e PDZ10). Assim, foram construídos os modelos 3D (código PDB de Adenovírus (E4orf1): 3C3I; 3C2T e 5F9K) baseado grau de identidade (28-30%), a construção dos modelos 3D foi realizada no software MODELLER v9.21. O gráfico de Ramachamdran do modelo de menor energia foi utilizado como principal validação. Para os cálculos de DM, as simulações foram realizadas no programa Gromacs utilizando o campo de força moleculares (amber99 + ILDN), caixa água cúbica 0,5, íons Cl e Na foram adicionados, a concentração utilizada foi o pH fisiológico. RESULTADOS: Estudos de docking molecular realizados pelo servidor web clusPRO para buscar a melhor conformação para cada complexo. Assim, realizou-se uma simulação por DM de 300 ns e análise desvio médio quadrático (RMSD); Flutuação média quadrada da raiz (RMSF) e contatos de frequência durante a DM. No nível molecular, foi possível observar a interação com frequência > 75% do tempo de simulação, interação entre Phe79 (E4orf1) e His289 (PDZ1 e PDZ10) do tipo hidrofóbico e 4 Å da distância de interação, sugerindo que a A estabilidade do complexo é obtida pela interação entre os resíduos nesta região hidrofóbica em E4orf1 (Val77 e Ile78) e PDZ1-PDZ10 (Pro245, His246 e Val287). CONCLUSÃO: Os resultados das interações entre E4orf1 e os diferentes domínios testados sugerem que os resíduos são importantes na interação e estabilização, trazendo inovação à um alvo pouco explorado e despertando a caracterização do mesmo como um druggable target para de novos fármacos contra a obesidade. (AU)

Alta prevalencia de dislipidemias y riesgo aterogénico en una población infanto-juvenil / High prevalence of dyslipidemia and high atherogenic index of plasma in children and adolescents

Background: Dyslipidemias in childhood increase the risk of cardiovascular events in adult life. Aim: To evaluate the prevalence of dyslipidemia and risk of atherogenicity based in the atherogenic index of plasma (AIP) in a sample of school children and adolescents. Material and Methods: Cross-sectional study of 208 children aged 10.4 ± 1.0 years (107 women). Demographic data were obtained, and a clinical evaluation was conducted, including pubertal development according to Tanner and anthropometric parameters. A fasting blood sample was obtained to measure total cholesterol (CT), HDL cholesterol (cHDL) and triglycerides (TG), glucose and insulin. LDL cholesterol (cLDL), Non-HDL cholesterol and the indices CT/cHDL, cLDL/cHDL and AIP (log[TG/cHDL]) were calculated. Risk categories according to AIP for the pediatric population were also determined (low: AIP < 0.11, intermediate: AIP 0.11-0.21, high: AIP > 0.21). Results: Thirty eight percent of participants had dyslipidemia, without differences by gender and pubertal development. The frequency of dyslipidemia was significantly higher in children with obesity (54%, p < 0.01) and a waist circumference over percentile 90 (61%; p < 0.01). The later conditions had also higher CT/cHDL, cLDL/cHDL and AIP. According to AIP, 54% of children had a high atherogenicity risk along with alterations in anthropometric parameters and insulin resistance. All anthropometric and insulin resistance parameters were significantly correlated with the AIP. Conclusions: There is a high prevalence of dyslipidemia in the studied population, which is associated with an increased cardiometabolic risk. The indices of atherogenicity and particularly AIP are correlated with nutritional status, abdominal obesity and parameters of insulin resistance.

Alta prevalencia de trastornos nutricionales por exceso, resistencia insulínica y síndrome metabólico en escolares de la comuna de Carahue, Región de la Araucanía / High prevalence of overweight, obesity, insulin resistance and metabolic syndrome in rural children and adolescents

Background: Childhood and adolescent obesity is a major public health problem in Chile. Aim: To characterize cardiometabolic risk factors in a population of schoolchildren from Carahue, Chile. Material and Methods: Cross-sectional assessment of 208 children aged 10.4 ± 1.0 years (106 women). A clinical evaluation was carried out including pubertal development according to Tanner and anthropometric parameters. A fasting blood sample was obtained to measure glucose, insulin and lipid profile. HOMA-IR and Quicki indices were calculated. Insulin resistance (IR) was established according to Burrows criteria and Barja criteria, previously proposed for the Chilean pediatric population. The metabolic syndrome (MetS) was established using the modified Cook criteria. Results: Thirty eight percent of children had overweight and 33.1% obesity. MetS was only observed in obese subjects and the frequency in this subgroup was 38%. The prevalence of IR was 51% according to the Burrows criteria and 19% according to Barja criteria. It was more common in participants who were overweight, obese or had abdominal obesity. Children with insulin resistance according to Barja criteria, had worse anthropometric measures than their counterparts without resistance. When Burrows criteria was used, no differences in anthropometric measures were observed between participants with or without resistance. The frequency of MetS was 26 and 18% in children with insulin resistance according to Barja and Burrows criteria, respectively. Insulin levels and insulin sensitivity indexes were positively correlated with anthropometric parameters. Conclusions: There was a high prevalence of overweight, obesity and MetS in these participants. Our results suggest that the IR criteria according to Barja allows to identify cases with higher metabolic risk.

Polymorphisms in Genes Involved in the Leptin-Melanocortin Pathway are Associated with Obesity-Related Cardiometabolic Alterations in a Southern Chilean Population

BACKGROUND:Polymorphisms in genes encoding proteins of the leptin-melanocortin pathway have been associated with obesity. The involvement of these polymorphisms with changes in body mass index (BMI) and anthropometric measures could also imply a contribution to the risk of metabolic syndrome (MetS) and metabolic alterations. We evaluated the relationship of leptin-melanocortin system polymorphisms with obesity, MetS, and other metabolic alterations in Southern Chilean individuals.METHODS:Two-hundred individuals were grouped as normoweight (BMI 18.0-24.9 kg/m2), overweight (BMI 25.0-29.9 kg/m2), and obese (BMI ≥ 30 kg/m2) or according to MetS status. Anthropometric measures (BMI, abdominal circumference, waist-to-hip ratio [WHR]) and biochemical parameters (glycemia and lipid profile) were evaluated. Polymorphisms LEP rs7799039, LEPR rs1137101, MC3R rs3746619 and rs3827103, and MC4R rs17782313 were evaluated by real-time PCR using allelic discrimination assays.

Pharmacogenetic implications in the management of metabolic diseases in Brazilian populations

Dyslipidemia, diabetes, obesity and hypertension are common metabolic diseases. In the last decades, unhealthy lifestyle and aging have leads to an increased incidence of these diseases, increasing morbidity and mortality by cardiovascular causes. The treatment of metabolic diseases includes life-style interventions as healthy diet and physical exercise, as well as pharmacological interventions. Several drugs are available for the management of metabolic diseases including among others lipid-lowering antidiabetics and antihypertensive drugs. Variability in response to these drugs is influenced by both genetic and non-genetic factors. Polymorphisms in genes related to drug pharmacokinetics and pharmacodynamics have been shown to influence drug efficacy and safety. This review is focused on pharmacogenetic studies related to the management of metabolic diseases in samples of the Brazilian population. Associations of variants in drug metabolizing enzymes and transporters, drug target and metabolism-related genes with the efficacy and safety of lipid-lowering, antidiabetic and antihypertensive drugs are described. Most pharmacogenetic studies in Brazil have focused in pharmacological response to a small group of drugs, as statins and some antihypertensives, while there are almost no studies on antidiabetic and antiobesity drugs. Some studies reported significant associations of gene polymorphisms with drug response confirming previous data from other populations, whereas other works did not replicate, which may relay on the genetic admixture of our population. In conclusion, further studies are necessary considering larger sample sizes, new unexplored drugs and more genetic variants to obtain stronger conclusions to explore clinical applications of pharmacogenetic studies in our population.

Transport of cowpea bean derived peptides and their modulator effects on mRNA expression of cholesterol-related genes in Caco-2 and HepG2 cells

This work studied the cell transport of peptidase-generated peptides from cowpea bean proteins and their effects on mRNA expression of cholesterol-related genes in intestinal and liver cells. The <= 3 kDa hydrolysate was obtained and incubated with Caco-2 intestinal cells using Transwell center dot plates. HepG2 liver cells were incubated with synthetic analogues of peptides (MELNAVSVVHS and MELNAVSVVSH) identified by "de novo" peptide sequencing in the Caco-2 monolayer permeates. The mRNA expression of NPC1L1, ABCA1 and ABCG1 was measured in Caco-2 cells, in the presence or absence of <= 3 kDa hydrolysate and the expression of HMGCR, SREBP2, LXRa, AMPK1, was determined in the HepG2 cells in the presence or absence of synthetic peptides. Exposure of Caco-2 cells to cowpea <= 3 kDa hydrolysate (2.5 and 5 mg/mL) increased ABCG1 expression at 6 h and 12 h. SREBP2, HMGCR and LDLR mRNA levels were reduced in HepG2 cells after 24 h of treatment with MELNAVSVVHS peptide (50 mu M and 100 mu M). These results suggest that MELNAVSVVHS peptide is able to cross intestinal barrier and to modulate genes involved in cholesterol homeostasis.

Transport of cowpea bean derived peptides and their modulator effects on mRNA expression of cholesterol-related genes in Caco-2 and HepG2 cells

This work studied the cell transport of peptidase-generated peptides from cowpea bean proteins and their effects on mRNA expression of cholesterol-related genes in intestinal and liver cells. The <= 3 kDa hydrolysate was obtained and incubated with Caco-2 intestinal cells using Transwell center dot plates. HepG2 liver cells were incubated with synthetic analogues of peptides (MELNAVSVVHS and MELNAVSVVSH) identified by "de novo" peptide sequencing in the Caco-2 monolayer permeates. The mRNA expression of NPC1L1, ABCA1 and ABCG1 was measured in Caco-2 cells, in the presence or absence of <= 3 kDa hydrolysate and the expression of HMGCR, SREBP2, LXRa, AMPK1, was determined in the HepG2 cells in the presence or absence of synthetic peptides. Exposure of Caco-2 cells to cowpea <= 3 kDa hydrolysate (2.5 and 5 mg/mL) increased ABCG1 expression at 6 h and 12 h. SREBP2, HMGCR and LDLR mRNA levels were reduced in HepG2 cells after 24 h of treatment with MELNAVSVVHS peptide (50 mu M and 100 mu M). These results suggest that MELNAVSVVHS peptide is able to cross intestinal barrier and to modulate genes involved in cholesterol homeostasis.

Effects of clopidogrel on inflammatory cytokines and adhesion molecules in human endothelial cells: Role of nitric oxide mediating pleiotropic effects

NTRODUCTION: Clopidogrel is commonly used in prevention and treatment of atherothrombosis. Some previous studies have suggested a pleiotropic effect of clopidogrel; however, when this drug causes platelet-independent effects on endothelial function remains unclear. AIMS: To evaluate the influence of clopidogrel on inflammatory biomarkers and adhesion molecules in human endothelial cells and the role of nitric oxide (NO) in this process. METHODS: TNF-α-induced human umbilical vein endothelial cells (HUVEC) were exposed to clopidogrel. Gene expression and protein expression of ICAM-1, P-selectin, IL-8, IL-6, and MCP-1 were evaluated by qPCR, flux cytometry, or milliplex technology. Expression of endothelial nitric oxide synthase (NOS3) and NO release were also evaluated. Influence of clopidogrel was further evaluated in NOS3 downregulated HUVEC by RNAi. RESULTS: Clopidogrel at 20 μmol/L induced NO release in HUVEC after 24-hours treatment. Gene expressions of inflammatory markers IL-8 and MCP1 were reduced after clopidogrel treatment (P<.05); however, only MCP-1 remained reduced at protein level. IL-6 was not modified by clopidogrel treatment. Gene expression and protein expression of ICAM-1 were diminished by 24-hours clopidogrel exposure, whereas P-selectin was not modified. NOS3 downregulated HUVEC model revealed that ICAM-1 modification by clopidogrel is dependent of this via, whereas MCP-1 is modulated in an NO-independent form...

Effects of short-term add-on ezetimibe to statin treatment on expression of adipokines and inflammatory markers in diabetic and dyslipidemic patients

AIM: The influence of short-term add-on ezetimibe to simvastatin treatment on expression of adipokines and inflammatory markers was investigated in diabetic and nondiabetic patients with hypercholesterolemia. METHOD: Hypercholesterolemic nondiabetic (HC, n = 37) and diabetic (DM, n = 47) patients were treated with simvastatin (SV, 10 or 20 mg/d/8-wk) and then SV plus ezetimibe (SV + EZ, 10 mg each/d/4 wk). Serum lipids, glycemic profile, and inflammatory markers (hsCRP, adiponectin, resistin, VCAM-1, and ICAM-1) were evaluated before and after the add-on ezetimibe therapy. mRNA expression of ADIPOR1, ADIPOR2, RETN, VCAM1, and ICAM1 was measured by real-time PCR in peripheral blood mononuclear cells (PBMC). RESULTS: Serum concentrations of LDL and HDL cholesterol, and adiponectin were higher in HC than DM patients (P < .05). The add-on ezetimibe therapy reduced total and LDL cholesterol, apoB and adiponectin serum levels in HC and DM groups, and resistin in HC subjects (P < .05). DM patients showed higher expression of ADIPOR1, ADIPOR2, RETN, and VCAM1 in PBMC than subjects in HC group, before and after add-on ezetimibe therapy (P < .05). PBMC RETN mRNA expression was reduced by add-on ezetimibe therapy in HC individuals (P < .05), but not in DM subjects...