Hepatitis B virus (HBV) infection is a major risk factor for hepatocellular carcinoma (HCC). Although nucleoside/nucleotide analogue (NA) therapy significantly reduces this risk, a subset of patients still develop HCC. This study aimed to identify high-risk groups by evaluating the comprehensive utility of multidimensional factor combinations. We analyzed 565 patients with chronic HBV infection who underwent liver stiffness (LS) measurement between 2009 and 2024. Independent risk factors for HCC were identified using multivariate logistic regression. In 84 patients who underwent testing before and after NA therapy, the annual rate of change in parameters was evaluated to assess dynamic risk prediction. Multivariate analysis identified male sex, LS, total bilirubin, gamma-glutamyl transpeptidase, and fibrosis-4 index as independent predictors, with LS being the most significant (cut-off value: 1.54 m/s; area under the curve: 0.811). Diagnostic accuracy (69.4

Scalp high-frequency oscillations (HFOs) are promising noninvasive biomarkers of epileptogenicity, but their phenotypic diversity and clinical relevance in absence epilepsy (AE) remain unclear. This study aimed to classify scalp HFOs in AE using k-means clustering based on multiple morphological characteristics, and to evaluate their distribution across electroencephalogram (EEG) epochs and seizure control statuses. We analyzed scalp EEG recordings from 14 children and adolescents with AE. After excluding outliers, 163 scalp HFOs were characterized by average frequency, duration, amplitude, and number of cycles. Amplitude and cycle count were log-transformed prior to clustering, and k-means clustering was applied to identify distinct HFO phenotypes. Three clusters were identified: Cluster 1 (short duration, low amplitude), Cluster 2 (low frequency), and Cluster 3 (long duration, high cycle count). Cluster 2 and Cluster 3 were significant predictors of ictal HFOs in active AE, with odds ratios (ORs) of 0.33 (95% confidence interval [CI]: 0.14-0.74) and 5.00 (CI: 2.02-17.73), respectively. Cluster 2 also predicted interictal HFOs in active AE (OR [95% CI] = 2.71 [1.23-5.67]). These findings support the utility of scalp HFO phenotypes as EEG-based biomarkers for seizure detection and disease monitoring, potentially guiding treatment strategies in pediatric AE.

Infantile epileptic spasms syndrome (IESS) is characterized by epileptic spasms (ES) in infants and hypsarrhythmia on EEG. High-frequency oscillations (HFOs), defined as oscillatory events at >80 Hz, reflect rapid spatiotemporal dynamics of cortical activity and might serve as novel biomarkers for epilepsy. In this study, we investigated the association among hypsarrhythmia, ES and scalp HFOs in IESS, focusing on their spatiotemporal and morphological characteristics. We analysed scalp EEG recordings from 22 paediatric patients with IESS (12 males; age range, 0-12 years), classified into four groups: hypsarrhythmia with ES; hypsarrhythmia without ES; multifocal interictal epileptiform discharges (IED); and non-multifocal IED. The proportions of EEG recordings with at least one detected scalp HFO differed significantly among the groups, being 100.0% in hypsarrhythmia with ES, 90.9% in hypsarrhythmia without ES, 36.0% in multifocal IED and 29.4% in non-multifocal IED (P < 0.001). Age-adjusted odds ratios [95% confidence intervals] for high HFO detection rates were significantly greater in the hypsarrhythmia groups than in the IED groups (with ES, 24.66 [4.86-125.10], P < 0.001; and without ES, 8.38 [1.94-36.20], P = 0.004). Receiver operating characteristic analysis showed that scalp HFO detection rates discriminated ES activity with an area under the curve of 0.92, 100% sensitivity and 77.4% specificity. Compared with the IED groups, the hypsarrhythmia groups exhibited lower-frequency HFOs, with the lowest HFO frequencies being observed in the hypsarrhythmia with ES group. These findings suggest that the spatiotemporal and morphological characteristics of scalp HFOs might provide complementary multidimensional biomarkers for hypsarrhythmia and assessing ictal states in IESS.

Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) is a rare mitochondrial disease with a prevalence of 16-18 per 100,000 persons. Most patients with MELAS develop epilepsy and require effective control of recurrent attacks. High-frequency oscillations (HFOs) on scalp EEG are transient bursts of EEG activity with frequencies beyond 80 Hz and are a promising biomarker for seizure control in epilepsy. However, reports on scalp HFOs are limited to some epilepsy syndromes. Herein, we report on the appearance of scalp HFOs in a pediatric patient with MELAS. The patient, a 13-year-old boy, presented with a stroke-like episode at age 9 years and was diagnosed with MELAS. The main symptom was visual disturbances, but epilepsia partialis continua (EPC) was also observed. Scalp EEG recordings were made six times: 8 days before the stroke-like episode (day -8 EEG), the day of appearance (day 1 EEG), and 4, 8, 10, and 100 days after the episode. Analysis of scalp HFOs showed that no scalp HFOs were detected in the day -8 EEG, whereas 1.20 scalp HFOs per minute were detected in the day 1 EEG at the appearance of the stroke-like episode with EPC. The scalp HFO detection rate decreased with the loss of EPC, and no scalp HFOs appeared on EEG, although visual disturbances continued to be observed. By contrast, epileptic discharges remained on EEG after EPC disappearance. Scalp HFOs have the potential to be a useful biomarker for reflecting epileptic seizure in patients with MELAS.

Shear wave velocity (Vs) measurements can be unstable in individuals with obesity or with a skin-capsule distance (SCD) of ≥ 20 mm. However, instability of Vs has also been observed in cases where SCD is not markedly increased; therefore, the present study aimed to investigate the factors contributing to reduced VsN (net amount of effective shear wave velocity), an index that reflects the reliability of individual Vs measurements. In total, 317 individuals who underwent community-based health screening were included in this study. We investigated the relationship between VsN and physical findings, laboratory data, SCD, and structure of the SCD (single vs. multilayered) to identify factors associated with decreased VsN. Multiple regression analysis revealed that body fat percentage (β = − 0.135, p = 0.011), SCD (β = − 0.347, p < 0.0001), and the structure of the SC region (β = − 0.295, p < 0.0001) were independently associated with decreased VsN. In cases where the SCD exceeded 20 mm, the VsN within the SCD was significantly lower in the multilayer structural type [52 (28–75)] than in the single-layer structural type [95 (77–99)] (p = 0.004). Furthermore, even when the SCD was 20 mm or less, the VsN within the SCD was significantly lower in the multilayer structure type [80 (68–89)] than in the single-layer structure type [99 (92–100)] (p < 0.0001). The results show that, in addition to SCD thickness, its internal structure reduces the VsN and contributes to Vs measurement instability.

OBJECTIVE:The accuracy of self-reported seizure-freedom duration are essentially limited. Scalp high-frequency oscillations (HFOs) are more tightly linked to seizures than spikes alone and are a promising new biomarker. The purpose of this study is to determine the relationship between scalp HFO and time since the last reported seizure. METHODS:The study population consisted of 169 pediatric epilepsy patients (91 males; age range, 0-20 years). A holdout method was used to develop and validate a predictive model (multivariate HFO model) to estimate the time since the last reported seizure. RESULTS:The multivariate HFO model was created with four variables: scalp HFO detection rate, developmental delay, epilepsy duration, and the use of antiepileptic drugs. The area under the curve (AUC) of the multivariate HFO model was higher than that for the HFO and spike models in all four discriminations for time since the last reported seizure (≥ 2 years: AUC = 0.95, ≥ 1 year: 0.91, ≥ 2 months: 0.82, and ≥ 2 weeks: 0.76). CONCLUSIONS:The multivariate HFO model showed higher performance in patients with a longer time since the last reported seizure (≥ 1 year). SIGNIFICANCE:This model may help establish a new measure of epilepsy remission.

BACKGROUND:Mitochondria, which harbor their own genome (mtDNA), have attracted attention due to the potential of mtDNA copy number (mtDNA-CN) as an indicator of mitochondrial dysfunction. Although mtDNA-CN has been proposed as a simple and accessible biomarker for metabolic disorders such as metabolic dysfunction-associated steatotic liver disease, the underlying mechanisms and the causal relationship remain insufficiently elucidated. In this investigation, we combined longitudinal epidemiological data, animal studies, and in vitro assays to elucidate the potential causal relationship between reduced mtDNA-CN and the development of steatotic liver disease (SLD). METHODS:We conducted a longitudinal study using data from a health examination cohort initiated in 1981 in Yakumo, Hokkaido, Japan. Data from examinations performed in 2015 and 2022 were analyzed, focusing on 76 subjects without SLD at baseline (2015) to assess the association between baseline mtDNA-CN and subsequent risk of SLD development. In addition, 28-day-old SD rats were fed ad libitum on a 45% high-fat diet and dissected at 2 and 8 weeks of age. Blood and liver mtDNA-CN were measured and compared at each feeding period. Additionally, in vitro experiments were performed using HepG2 cells treated with mitochondrial function inhibitors to induce mtDNA-CN depletion and to examine its impact on intracellular lipid accumulation. RESULTS:Epidemiological analysis showed that the subjects with low mtDNA-CN had a significantly higher odds ratio for developing SLD compared to high (odds ratio [95% confidence interval]: 4.93 [1.08-22.50]). Analysis of the animal model showed that 8 weeks of high-fat diet led to the development of fatty liver and a significant decrease in mtDNA-CN. A further 2 weeks of high-fat diet consumption resulted in a significant decrease in hepatic mtDNA-CN, despite the absence of fatty liver development, and a similar trend was observed for blood. Complementary in vitro experiments revealed that pharmacologically induced mitochondrial dysfunction led to a significant reduction in mtDNA-CN and was associated with increases in intracellular lipid accumulation in HepG2 cells. CONCLUSIONS:Our findings suggest that reduced mtDNA-CN may contribute causally to SLD development and could serve as a convenient, noninvasive biomarker for early detection and risk assessment.

Purpose: High-frequency oscillation (HFO) in scalp electroencephalography is a promising new noninvasive prognostic epilepsy biomarker, but further data are needed to ascertain the utility of this parameter. The present work investigated the association between epileptic activity and scalp HFO in pediatric patients with various types of epilepsy, using multivariable regression models to correct for possible confounding factors. Methods: The authors analyzed 97 subjects who were divided into groups with active epilepsy (within 1 year of seizure), seizure-free epilepsy (>1 year without seizure), and nonepilepsy. Regarding the frequency of seizure occurrence as an indicator of epileptic activity, we categorized subjects into four groups (Daily/Weekly, Monthly, Yearly, and Rarely). Results: Multiple linear regression analysis showed that the scalp HFO detection rate was significantly higher in patients with active epilepsy than in those with nonepilepsy (beta [95% confidence interval] = 2.77 [1.79-4.29]; P < 0.001). The association between scalp HFO detection rate and frequency of seizure occurrence was highest in the Daily/Weekly group (beta [95% confidence interval] = 3.38 [1.57-7.27]; P = 0.002), followed by Monthly and Yearly groups (beta [95% confidence interval] = 2.42 [1.02-5.73]; P = 0.046 and 0.36 [0.16-0.83]; P = 0.017). In addition, HFO duration, number of peaks, and number of channels detected were significantly higher in patients with active epilepsy. Conclusions: Pediatric patients with active epilepsy and high frequency of seizure occurrence exhibited a higher scalp HFO detection rate. These results may help to establish HFO detectable by noninvasive scalp electroencephalography as a biomarker of active epilepsy in pediatric patients.

Tools for measuring the likelihood of relapse in infantile epileptic spasms syndrome (IESS) treatment could aid clinicians in making critical management decisions. High-frequency oscillations (HFOs), transient bursts of electroencephalography (EEG) activity with frequencies beyond 80 Hz, are a new and promising noninvasive biomarker. The present study aimed to investigate the association between the Burden of Amplitudes and Epileptiform Discharges (BASED) scores, an interictal EEG grading scale for IESS, and scalp HFOs in patients with IESS. The study enrolled 50 patients, 25 with a clinical diagnosis of IESS and 25 without epilepsy. The percentage of patients with at least one scalp HFO detected, stratified by BASED scores, differed significantly: for BASED scores ≤ 2, 7.7%; for 3, 16.7%; for 4, 87.5%; and for 5, 100% (p < 0.001). Compared with BASED scores ≤ 2, the median scalp HFO detection rate was significantly highest for BASED scores of 5 (median [IQR]: 6.24 [2.25–8.32], p < 0.001), followed by BASED scores of 4. The scalp HFO detection rates showed a better performance in estimating patients with BASED scores of 4 and 5. It is hoped that scalp HFOs can be used as an objective indicator to validate the results of BASED scores.

AbstractObjectiveEpilepsy treatment with anti‐seizure medications (ASMs) is based on careful assessment of the balance between the likelihood of further seizures and the risk of side effects of treatment. However, there is currently no established biomarker to ascertain seizure control status with ASMs. High‐frequency oscillations (HFOs), transient bursts of EEG activity with frequencies beyond 80 Hz, are a new and promising noninvasive epilepsy biomarker. We compared the risk of scalp HFO appearance between pediatric patients with good and poor seizure control by treatment with ASMs.MethodsA total of 72 epilepsy patients (aged 0–18 years, 39 males) with good and poor seizure control with ASMs participated in this study. We applied a validated automated detector to determine HFO and spike. We calculated the odds ratios (ORs) for scalp HFO and spike appearance according to seizure control status by multiple logistic regression analysis.ResultsScalp HFO was seen more commonly and with a significantly higher detection rate in patients with poor seizure control as compared with patients with good seizure control for both ripple and fast ripple. These significant associations were found for both focal and generalized epilepsy. The ORs for scalp HFO appearance adjusted for confounding factors were significantly higher in patients with poor seizure control compared to those with good seizure control (ripple: OR [95% CI] = 11.91 [2.21–64.30], p = 0.004; fast ripple: 4.98 [1.03–24.09], p = 0.046). There were no significant associations between spike appearance and seizure control status.SignificanceWe found an increased risk of scalp HFO appearance in patients with poor seizure control. The results of this study support that scalp HFO is associated with patients having frequent seizures after treatment in both ripple and fast ripple.Plain Language SummaryThis study analyzed scalp high‐frequency oscillations and spikes in pediatric patients with various types of epilepsy who were being treated using ASMs. The results showed that an increased risk of scalp HFO appearance was observed in patients with poor seizure control compared to those with good seizure control. These findings were observed in both the ripple (80–250 Hz) and fast ripple (250–500 Hz) bands. The scalp HFO is associated with patients having frequent seizures after treatment in both ripple and fast ripple.

Background: The increasing prevalence of non-alcoholic fatty liver disease (NAFLD) has become a global health problem. NAFLD has few initial symptoms and may be difficult to detect early, so there is need for a minimally invasive early detection marker. We hypothesized that miR-122 and miR-20a levels combined, as the miR-122/miR-20a ratio might detect NAFLD more sensitively. Methods: This study involved 167 participants with low alcohol intake. Those who had an increase in echogenicity of the liver parenchyma and hepato-renal contrast on ultrasonography were classified as the NAFLD group (n = 44), which was further classified into mild (n = 26) and severe (n = 18) groups based on echogenic intensity and hepatic vessel and diaphragm visualization. Participants without fatty liver were included in the normal group, except for those with an abnormal body mass index, glycated hemoglobin, and systolic blood pressure (n = 123) values. Serum miR-122 and miR-20a expression levels in participants were measured by real-time polymerase chain reaction, and the miR-122/miR-20a was calculated. Results: In the NAFLD group, miR-122 expression was significantly higher and the miR-20a was significantly lower than in the normal group, in agreement with previous studies. miR-122/miR-20a was also significantly higher in the NAFLD group. Receiver operating characteristic curve analysis was performed with miR-122/miR-20a as an NAFLD detection marker, and the area under the curve of miR-122/miR-20a was significantly larger than that of miR-122 or miR-20a alone. Conclusions: The miR-122/miR-20a ratio, combined with miR-122 and miR-20a levels, is a useful biomarker to detect NAFLD with high sensitivity.

Background There is limited evidence regarding the relationship between Diabetes mellitus (DM) in middle age and mild cognitive impairment after a follow-up. Therefore, we investigated the relationship between fasting blood glucose (FBG) levels in middle age and cognitive function assessed using the Japanese version of the Montreal Cognitive Assessment (MoCA-J) in later life, following over 15 years of follow-up in the Aichi Workers’ Cohort Study in Japan. Methods Participants were 253 former local government employees aged 60–79 years in 2018 who participated in a baseline survey conducted in 2002. Using baseline FBG levels and self-reported history, participants were classified into the normal, impaired fasting glucose (IFG) and, and DM groups. Total MoCA-J score ranges from 0 to 30, and cognitive impairment was defined as MoCA-J score ≤25 in this study. A general linear model was used to estimate the mean MoCA-J scores in the FBG groups, adjusted for age, sex, educational year, smoking status, alcohol consumption, physical activity, body mass index, systolic blood pressure, total cholesterol, and estimated glomerular filtration rate. Results The mean MoCA-J score in the total population was 25.0, and the prevalence of MoCA-J score ≤25 was 49.0%. Multivariable-adjusted total MoCA-J scores were 25.2, 24.8, and 23.4 in the normal, IFG, and DM groups, respectively. The odds ratio of MoCA-J score ≤25 in the DM group was 3.29. Conclusion FBG level in middle age was negatively associated with total MoCA-J scores assessed later in life, independent of confounding variables.

INTRODUCTION:DNA methylation in the CpG sites of intron 1 of HIF3A is associated with body mass index (BMI). This cross-sectional study investigated correlations between DNA methylation of HIF3A and BMI or adiposity parameters in the Japanese population.METHOD:DNA methylation of HIF3A was quantified via pyrosequencing.RESULT:DNA methylation of HIF3A differed only in women; DNA methylation level at cg27146050 was associated with visceral adipose tissue thickness and correlated with BMI and percent (%) body fat after excluding smokers.CONCLUSION:Peripheral blood DNA methylation at the CpG site (cg27146050) of HIF3A correlated with VAT thickness in Japanese women.

The increasing prevalence of nonalcoholic fatty liver disease (NAFLD) is a global health problem. In recent years, the inhibitory effect of brain-derived neurotrophic factor (BDNF) on diabetes mellitus and fatty liver has been clarified. The purpose of this study was to analyze the relationship between serum BDNF and NAFLD which caused by abnormal metabolism of glucose and lipids. This cross-sectional study involved 429 participants (mean age, 63.5 years: men, 38.5%) with low alcohol intake. Of the participants, those who had an increase in echogenicity of the liver parenchyma and hepato-renal contrast on ultrasonography were classified as the NAFLD group (n = 88), and the others were classified as the normal (n = 341) group. The NAFLD group was further classified into a mild group (n = 60) and a severe group (n = 28) based on the intensity of echogenicity and visualization of the hepatic vessels and diaphragm. Median BDNF levels were higher in the NAFLD group than the normal group (35.5 vs. 42.3 ng/mL, p < 0.01). Furthermore, BDNF levels tended to be associated with the severity of NAFLD (p < 0.01). In addition to the univariate analysis, in the sex- and age-adjusted model, there was a significant association between the BDNF levels and NAFLD severity (p < 0.01). The fully adjusted regression analysis also showed a positive association between the serum BDNF level and NAFLD (p < 0.01). These results suggest that NAFLD patients have a compensatory increase in circulating BDNF levels.

Objectives: Development of the autonomic nervous system may play a role in myocardial repolarization lability in infants, but its relationship to repolarization abnormalities remains unclear. Thus, the aim of the present study was to evaluate the relationship between gestational age and ventricular repolarization lability using the variability ratio (VR). Methods: Infants who underwent electrocardiography at a 1-month check-up were included (n=209; 125 males). Gestational age and the following four VR parameters at 1 month of age were compared: VR-I, SDQT/SDRR; VR-II, SDQT/rMSSD; VR-III, SDQTc/SDRR; and VR-IV, SDQTc/rMSSD; where SD, QTc, and rMSSD are standard deviation, QT interval corrected using Fridericia’s formula, and root mean square difference of successive RR intervals, respectively. Twenty-eight preterm infants born at <37 weeks of gestation and 181 full-term infants were included. Results: Significant correlations were observed between gestational age and VR-I, -III, and -IV (all p<0.05). All VR values were significantly higher in preterm infants compared with full-term infants (I: 0.54 vs 0.48, II: 1.15 vs 0.96, III: 0.88 vs 0.68, IV: 1.59 vs 1.39; median, all p<0.05). Conclusion: VR assessed at 1 month after birth was impaired in preterm infants, suggesting immaturity of their cardiac autonomic nervous system and ventricular myocardial repolarization.

Obesity is a major public health problem worldwide owing to the substantial increase in risk of metabolic diseases. Hypoxia-inducible factors (HIFs) regulate transcriptional responses to hypoxic stress. DNA methylation in the CpG sites of intron 1 of HIF3A is associated with body mass index in the whole blood and adipose tissue. This study investigates the correlation between DNA methylation of HIF3A and parameters of obesity, including thickness of visceral (VAT) and subcutaneous adipose tissues, in the general Japanese population. Participants (220 men and 253 women) who underwent medical examination were enrolled in this cross-sectional study. We used pyrosequencing to quantify DNA methylation (CpG sites of cg16672562, cg22891070, and cg27146050) in HIF3A . DNA methylation of HIF3A was only different in women. Multiple regression analysis showed that DNA methylation level at cg27146050 was associated with thickness of VAT in women. DNA methylation level at cg27146050 also correlated with body mass index and percentage of body fat in women after excluding smokers and non-smokers who quit smoking with the last 5 years. DNA methylation in the CpG site (cg27146050) of HIF3A correlated with parameters of obesity in Japanese women.

Nonalcohol fatty liver disease (NAFLD) is one of the considerable health problems in the world. NAFLD is a representative phenotype in the liver reflecting individual's lifestyle. Previous studies suggested that a single nucleotide polymorphism (SNP) in the CD36 gene was associated with dietary fat intake and fat metabolism. Therefore, we hypothesized that this SNP may be associated with fatty liver (FL) and NAFLD. We conducted this cross-sectional study to examine the association between two CD36 polymorphisms (rs1761667 and rs1527483) and FL and NAFLD in a Japanese population. A total of 525 local residents participated in the health check-up in Yakumo town, Hokkaido, Japan. This cross-sectional study was conducted based on clinical information collected in this health check-up. Among all participants, 57 people were excluded due to the inadequate informed consent, clinical history of liver disease. Genotyping of two polymorphisms in the CD36 gpgbrkkene was performed by using the PCR-CTPP method. Ultrasound examination was performed by four qualified ultrasonographers. In this present, NAFLD was defined as 1) presence of FL and 2) a daily alcohol consumption <10 g in women and <20 g in men. Multivariable regression analysis was performed after adjusting for age, sex, waist circumference, medicine for dyslipidemia. The mean age of participants was 63.4 (10.0). The minor allele frequency of rs1761667 and rs1527483 was 0.26 and 0.21, respectively. The presence of FL and NAFLD was 90 (19.2%) and 67 (14.3%), respectively. Multiple regression analyses showed a significant association between rs1761667 and presence of FL (P = 0.04), and FL grade (P = 0.03). However, rs1761667 was not significantly associated with NAFLD (P = 0.11). No significant associations were found between rs1527483 and FL and NAFLD. A polymorphism (rs1761667) in the CD36 gene was associated with the presence of FL, but we did not find a significant association between this SNP and NAFLD. This study was supported by the Mishima Kaiun Memorial Foundation.

Non-alcoholic fatty liver disease (NAFLD) is closely associated with obesity, metabolic syndrome, and type II diabetes mellitus. Recently, circulating microRNAs (miRNAs) have been proposed as useful disease biomarkers. We examined whether circulating miRNAs, such as miR-20a, miR-27a, and miR-126, were useful biomarkers for NAFLD. We conducted a cross-sectional analysis of 527 subjects aged 39 years or older who had undergone a health examination in the Yakumo Study. Of the residents, 92 were diagnosed with NAFLD using a registered medical sonographer. Serum miR-20a, miR-27a and miR-126 levels were measured by quantitative real-time PCR. We then calculated the odds ratios for serum miRNA level changes according to the severity of NAFLD using normal liver status as the reference group. Serum levels of miR-20a and 27a, but not miR-126, were significantly lower in NAFLD subjects than normal subjects. Serum miR-20a and miR-27a levels were significantly lower in both male and female severe NAFLD subjects. Logistic regression analysis showed a significant relationship between low circulating miR-20a and 27a levels and severe NAFLD. Down-regulated circulating miR-20a and 27a levels were significantly associated with severe NAFLD in the general population. Circulating miR-20a and miR-27a may be useful biomarkers for severe NAFLD.