Profiling of cell-free DNA methylation and histone signatures in pediatric NAFLD: A pilot study.

Nonalcoholic fatty liver disease (NAFLD) has become the most common chronic liver disease in children and adolescents, increasing the risk of its progression toward nonalcoholic steatohepatitis (NASH), cirrhosis, and cancer. There is an urgent need for noninvasive early diagnostic and prognostic tools such as epigenetic marks (epimarks), which would replace liver biopsy in the future. We used plasma samples from 67 children with biopsy-proven NAFLD, and as controls we used samples from 20 children negative for steatosis by ultrasound. All patients were genotyped for patatin-like phospholipase domain containing 3 (PNPLA3), transmembrane 6 superfamily member 2 (TM6SF2), membrane bound O-acyltransferase domain containing 7 (MBOAT7), and klotho-ß (KLB) gene variants, and data on anthropometric and biochemical parameters were collected. Furthermore, plasma cell-free DNA (cfDNA) methylation was quantified using a commercially available kit, and ImageStream(X) was used for the detection of free circulating histone complexes and variants. We found a significant enrichment of the levels of histone macroH2A1.2 in the plasma of children with NAFLD compared to controls, and a strong correlation between cfDNA methylation levels and NASH. Receiver operating characteristic curve analysis demonstrated that combination of cfDNA methylation, PNPLA3 rs738409 variant, coupled with either high-density lipoprotein cholesterol or alanine aminotransferase levels can strongly predict the progression of pediatric NAFLD to NASH with area under the curve >0.87. Conclusion: Our pilot study combined epimarks and genetic and metabolic markers for a robust risk assessment of NAFLD development and progression in children, offering a promising noninvasive tool for the consistent diagnosis and prognosis of pediatric NAFLD. Further studies are necessary to identify their pathogenic origin and function.

Galectin-3 in patients with atrial fibrillation and restored sinus rhythm.

INTRODUCTION: Cardiac fibrosis is the hallmark of atrial remodeling in atrial fibrillation. Galectin-3 (Gal-3) is a biomarker of fibrosis. It is well studied in heart failure, but the data about its role in atrial fibrillation are sparse. AIM: The aim of the study was to evaluate the levels of Gal-3 in patients with atrial fibrillation after sinus rhythm restoration, to examine the association between this biomarker and other factors for developing atrial fibrillation and to assess its prognostic role. MATERIALS AND METHODS: We included 67 patients (35 male) at the mean age of 67.36±7.25 years, with Gal-3 test after sinus rhythm restoration, a subgroup of participants in placebo-controlled randomized clinical trial of treatment with spironolactone. They were followed up for atrial fibrillation recurrence and hospitalizations. The effect of demographic parameters and other factors on Gal-3 levels were evaluated before and one year after treatment. RESULTS: Mean Gal-3 at baseline was 16.9±6.8 ng/ml. Higher levels of Gal-3 were associated with female gender (р=0.008), increasing age (р=0.005), renal dysfunction (p<0.0001) and gout (р=0.002). Higher thromboembolic risk as assessed by CHA2DS2-VASc score was significantly related to Gal-3. The levels of biomarker did not affect the number of atrial fibrillation recurrences (p=0.9) and hospitalizations. No correlation was found with treatment with spironolactone, antiarrhythmic and antihypertensive drugs. CONCLUSIONS: Higher Gal-3 in atrial fibrillation was associated with female sex, renal dysfunction, and history of gout. The levels of Gal-3 were not related to rhythm control. Treatment with spironolactone did not affect the biomarker of fibrosis Gal-3 in AF patients. Higher Gal-3 was related to high embolic risk.

Risk factors for recurrence of atrial fibrillation.

OBJECTIVE: Atrial fibrillation (AF) is a progressive disease, associated with increased risk of mortality, stroke, heart failure, and worsens quality of life. There is a high incidence of AF recurrence despite the treatment. The aim of the study was to assess the time to recurrence of AF after sinus rhythm restoration with electrical or pharmacological cardioversion and to identify the risk factors. METHODS: This study included 101 patients with AF (56% females) at a mean age of 68.02±7 years, after sinus rhythm restoration in a clinical observation of 1-year placebo-controlled treatment with spironolactone (1: 1). The patients were analyzed on the basis of AF recurrence, hospitalization, demographic parameters, comorbidities, embolic risk, and value of biomarker galectin-3 (Gal-3). RESULTS: The average number of AF recurrences was1.62 per patient per year. The median time of occurrence of at least one new episode was 48 days, 95% confidence interval (CI) 14.24-81.76. Female patients experienced significantly more recurrences than male-53.3% vs. 28.6% hazard ration (HR) =1.76, 95% CI 1.02-3.03, p=0.036. The recurrences were more common with increased age, although not significantly. Patients with arterial hypertension had a threefold risk of recurrences than those without hypertension (p=0.025), independently of the treatment. CHA2DS2-VASc score was significantly associated with AF recurrent episodes. Patients with gout had a twofold increased risk, without statistical significance (p=0.15). There was no difference in the AF episodes according to treatment with spironolactone. The levels of Gal-3 did not affect the number of AF recurrences (p=0.9). CONCLUSION: AF is associated with frequent recurrences after restoration of sinus rhythm in the majority of the patients. Most of them occurred within the first 3 months. Female sex, arterial hypertension, and CHA2DS2-VASc score were significant predictors of AF recurrence. Spironolactone did not reduce AF recurrences.