Assessment of DNA extraction methods for human gut mycobiome analysis.

The gut mycobiome plays an important role in the health and disease of the human gut, but its exact function is still under investigation. While there is a wealth of information available on the bacterial community of the human gut microbiome, research on the fungal community is still relatively limited. In particular, technical methodologies for mycobiome analysis, especially the DNA extraction method for human faecal samples, varied in different studies. In the current study, two commercial kits commonly used in DNA extraction, the QIAamp® Fast DNA Stool Mini Kit and DNeasy PowerSoil Pro Kit, and one manual method, the International Human Microbiome Standards Protocol Q, were compared. Furthermore, the effectiveness of two different bead-beating machines, the Mini-Beadbeater-16 and FastPrep-24TM 5G, was compared in parallel. A mock fungal community with a known composition of fungal strains was also generated and included to compare different DNA extraction methods. Our results suggested that the method using the DNeasy PowerSoil Pro Kit and Mini-Beadbeater-16 provides the best results to extract DNA from human faecal samples. Based on our data, we propose a standard operating procedure for DNA extraction from human faecal samples for mycobiome analysis.

Echocardiographic Assessment of Patients with Glycogen Storage Disease in a Single Center.

Glycogen storage disease (GSD) is a hereditary metabolic disorder caused by enzyme deficiency resulting in glycogen accumulation in the liver, muscle, heart, or kidney. GSD types II, III, IV, and IX are associated with cardiac involvement. However, cardiac manifestation in other GSD types is unclear. This study aimed to describe whether energy deprivation and the toxic effects of accumulated glycogen affect the heart of patients with GSD. We evaluated the left ventricle (LV) wall mass, LV systolic and diastolic function and myocardial strain with conventional echocardiography and two-dimensional speckle-tracking echocardiography (2D STE) in 62 patients with GSD type I, III, VI and IX who visited the Wonju Severance Hospital in 2021. Among the GSD patients, the echocardiographic parameters of 55 pediatrics were converted into z-scores and analyzed. Of the patients, 43 (62.3%), 7 (11.3%) and 12 (19.4%) patients were diagnosed with GSD type I, type III, and type IX, respectively. The median age was 9 years (range, 1-36 years), with 55 children under 18 years old and seven adults over 18 years. For the 55 pediatric patients, the echocardiographic parameters were converted into a z-score and analyzed. Multiple linear regression analysis showed that the BMI z-score (p = 0.022) and CK (p = 0.020) predicted increased LV mass z-score, regardless of GSD type. There was no difference in the diastolic and systolic functions according to myocardial thickness; however, 2D STE showed a negative correlation with the LV mass (r = -0.28, p = 0.041). Given that patients with GSD tend to be overweight, serial evaluation with echocardiography might be required for all types of GSD.

Handgrip Strength Cutoff Value Among Korean Adolescents with Metabolic Syndrome Components: Korean National Health and Nutrition Examination Survey Data 2014-2017.

Abstract Objectives: To propose the cutoff value of handgrip strength (HGS) for each metabolic syndrome component in Korean adolescents. Methods: Total of 2303 adolescents (1226 boys and 1077 girls; age 10-18 years) who participated in the Korea National Health and Nutrition Examination Survey from 2014 to 2017. We used the International Diabetes Federation metabolic syndrome guideline for children to define metabolic syndrome. The highest HGS for both hands were recorded. The optimal HGS cutoff for predicting metabolic syndrome components was determined by receiver operating characteristic curve analysis. Results: Adolescents with metabolic syndrome components defined by each criterion had higher HGS, systolic blood pressure, waist circumference, body mass index, fasting glucose, cholesterol, triglyceride (TG), alanine aminotransferase, and aspartate aminotransferase compared with average participants' data. The cutoff value of HGS defining waist circumference, TG level, high-density lipoprotein, and blood pressure was 24, 21.5, 30.9, and 30.2, respectively. The cutoff value of HGS defining metabolic syndrome was 28.9. For HGS to body weight ratio, the cutoff value defining metabolic syndrome was 0.38. Conclusions: This study showed that cutoff values of HGS have relation with metabolic syndrome in adolescents. Although cutoff has been suggested, it may not be sufficient for clinical use. Additional data are need to be accumulated in actual clinical trials for more accurate cutoff HGS value.

Quantitative MRI Assessment of Pancreatic Steatosis Using Proton Density Fat Fraction in Pediatric Obesity.

OBJECTIVE: To assess the feasibility of quantitatively assessing pancreatic steatosis using magnetic resonance imaging (MRI) and its correlation with obesity and metabolic risk factors in pediatric patients. MATERIALS AND METHODS: Pediatric patients (≤ 18 years) who underwent liver fat quantification MRI between January 2016 and June 2019 were retrospectively included and divided into the obesity and control groups. Pancreatic proton density fat fraction (P-PDFF) was measured as the average value for three circular regions of interest (ROIs) drawn in the pancreatic head, body, and tail. Age, weight, laboratory results, and mean liver MRI values including liver PDFF (L-PDFF), stiffness on MR elastography, and T2* values were assessed for their correlation with P-PDFF using linear regression analysis. The associations between P-PDFF and metabolic risk factors, including obesity, hypertension, diabetes mellitus (DM), and dyslipidemia, were assessed using logistic regression analysis. RESULTS: A total of 172 patients (male:female = 125:47; mean ± standard deviation [SD], 13.2 ± 3.1 years) were included. The mean P-PDFF was significantly higher in the obesity group than in the control group (mean ± SD, 4.2 ± 2.5% vs. 3.4 ± 2.4%; p = 0.037). L-PDFF and liver stiffness values showed no significant correlation with P-PDFF (p = 0.235 and p = 0.567, respectively). P-PDFF was significantly associated with obesity (odds ratio 1.146, 95% confidence interval 1.006-1.307, p = 0.041), but there was no significant association with hypertension, DM, and dyslipidemia. CONCLUSION: MRI can be used to quantitatively measure pancreatic steatosis in children. P-PDFF is significantly associated with obesity in pediatric patients.

Quick assessment with controlled attenuation parameter for hepatic steatosis in children based on MRI-PDFF as the gold standard.

BACKGROUND: Controlled attenuation parameter (CAP) is a recently introduced, non-invasive and quantitative method to evaluate hepatic steatosis demonstrated in adults, but limited in obesity and not well evaluated in children. The aim of this study was to investigate the diagnostic performance for assessing hepatic steatosis grades using CAP in children based on MR proton density fat fraction (PDFF). METHODS: Children evaluated for non-alcoholic fatty liver disease (NAFLD) who were assessed for PDFF and CAP were enrolled retrospectively. Hepatic steatosis grades 0-3 were classified according to PDFF using cutoff values of 6, 17.5, and 23.3%. Subgroup analyses were performed in non-obese and obese groups using the 95th percentile body mass index (BMI) as a cutoff and BMI30 group when BMI > 30 kg/m2. Pearson's correlations between variables were also analyzed. RESULTS: In a total of 86 children, there were 53 in the obese group including 17 of the BMI30 group. CAP demonstrated 98.7% sensitivity and 80% specificity for diagnosing grades 1-3 vs. grade 0 using a cutoff value of 241 dB/m (area under the curve = 0.941, p < 0.001). The diagnostic performance for higher steatosis grades was suboptimal. CAP correlated with abdominal wall thickness in both obese (r = 0.549, p = 0.001) and non-obese (r = 0.386, p = 0.004) groups and did not correlate with PDFF in BMI30 group. CONCLUSION: In children with NAFLD, CAP showed excellent diagnostic performance for differentiating presence and absence of hepatic steatosis using a cutoff value of 241 dB/m. However, CAP was limited in evaluating grades of steatosis, especially in children with BMI > 30 kg/m2.

Liver intravoxel incoherent motion diffusion-weighted imaging for the assessment of hepatic steatosis and fibrosis in children.

AIM: To evaluate the correlation between intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) parameters and the degree of hepatic steatosis and fibrosis in children. METHODS: This retrospective study was approved by the institutional review board. The children (≤ 18 years) who underwent liver IVIM DWI with 8 b-values under the suspicion of hepatic steatosis or fibrosis from February 2013 to November 2016 were included. Subjects were divided into normal, fatty liver (FAT), and fibrotic liver (FIB) groups. The slow diffusion coefficient (D), fast diffusion coefficient (D*), perfusion fraction (f), and apparent diffusion coefficient (ADC) were measured. MR proton density fat fraction (PDFF), MR elastography (MRE), and IVIM values were compared. RESULTS: A total of 123 children (median age of 12 years old, range: 6-18 years) were included, with 8 in the normal group, 93 in the FAT group, and 22 in the FIB group. The D* values were lower in the FIB group compared with those of the normal (P = 0.015) and FAT (P = 0.003) groups. The f values were lower in the FIB group compared with the FAT group (P = 0.001). In multivariate analyses, PDFF value was positively correlated with f value (ß = 3.194, P < 0.001), and MRE value was negatively correlated with D* value (ß = -7.031, P = 0.032). The D and ADC values were not influenced by PDFF or MRE value. CONCLUSION: In liver IVIM DWI with multiple b-values in children, there was a positive correlation between hepatic fat and blood volume, and a negative correlation between hepatic stiffness and endovascular blood flow velocity, while diffusion-related parameters were not affected.

MRI-based decision tree model for diagnosis of biliary atresia.

OBJECTIVES: To evaluate MRI findings and to generate a decision tree model for diagnosis of biliary atresia (BA) in infants with jaundice. METHODS: We retrospectively reviewed features of MRI and ultrasonography (US) performed in infants with jaundice between January 2009 and June 2016 under approval of the institutional review board, including the maximum diameter of periportal signal change on MRI (MR triangular cord thickness, MR-TCT) or US (US-TCT), visibility of common bile duct (CBD) and abnormality of gallbladder (GB). Hepatic subcapsular flow was reviewed on Doppler US. We performed conditional inference tree analysis using MRI findings to generate a decision tree model. RESULTS: A total of 208 infants were included, 112 in the BA group and 96 in the non-BA group. Mean age at the time of MRI was 58.7 ± 36.6 days. Visibility of CBD, abnormality of GB and MR-TCT were good discriminators for the diagnosis of BA and the MRI-based decision tree using these findings with MR-TCT cut-off 5.1 mm showed 97.3 % sensitivity, 94.8 % specificity and 96.2 % accuracy. CONCLUSIONS: MRI-based decision tree model reliably differentiates BA in infants with jaundice. MRI can be an objective imaging modality for the diagnosis of BA. KEY POINTS: • MRI-based decision tree model reliably differentiates biliary atresia in neonatal cholestasis. • Common bile duct, gallbladder and periportal signal changes are the discriminators. • MRI has comparable performance to ultrasonography for diagnosis of biliary atresia.

Assessment of liver fibrosis and cirrhosis by aspartate aminotransferase-to-platelet ratio index in children with biliary atresia.

BACKGROUND: In patients with biliary atresia (BA), liver fibrosis and cirrhosis commonly occur even after Kasai hepatoportoenterostomy. Although liver biopsy is the gold standard to evaluate liver fibrosis, it is invasive and may result in life-threatening complications. The aspartate aminotransferase-to-platelet ratio index (APRI) is a safe and simple method to assess liver fibrosis in patients with chronic liver diseases. To use APRI as a postoperative follow-up tool, we validated the diagnostic power of APRI for the degree of liver fibrosis in postoperative patients with BA. PATIENTS AND METHODS: Patients with newly diagnosed BA who underwent the Kasai procedure between March 2006 and May 2009 were analyzed. Several laboratory tests including APRI were performed. Liver wedge biopsy specimens were obtained during the surgical procedure, and histopathologic analyses were performed using the Metavir classification. RESULTS: Thirty-five patients (12 boys, median age of 1.9 months) were enrolled. Metavir scores were F1 in 0, F2 in 11, F3 in 11, and F4 in 13 patients. The areas under the receiver operating characteristics curves for F > or = 3 and F = 4 were 0.92 and 0.91, respectively. Distinct optimal cutoff values of APRI for F > or = 3 and F = 4 were obtained (1.01 and 1.41, respectively). Clinical outcomes of patients were significantly different between 2 groups (noncirrhosis vs cirrhosis) based on APRI before and 3 months after the Kasai procedure. CONCLUSION: APRI may be used as a simple and readily available tool for assessing liver fibrosis without additional risks in patients with BA during postoperative follow-up care.