Molecular Simulation Analysis of the Structure Complex of C2 Domains of DKK Family Members and β-propeller Domains of LRP5/6:Explaining Why DKK3 Does Not Bind to LRP5/6

Dickkopf (DKK) proteins interact with low-density lipoprotein receptor-related protein 5/6 (LRP5/6) to modulate WNT signaling. The interaction is mediated by a cysteine-rich domain (C2) in the DKK protein and β-propeller domains (PD) of LRP5/6. However, the third member of the DKK family (DKK3) does not bind to LRP5/6. To determine why DKK3 does not bind to the receptor domains, we performed a molecular modeling simulation study including homology modeling, protein-protein docking and molecular dynamics (MD). The computed affinities (ΔGbinding) between the C2 and PD models were consistent with the previously reported experimental results. The C2 model of DKK3 showed the lowest affinity for PD models. Multiple sequence alignment of C2 domains revealed that the DKK3 genes have a unique 7-amino-acid insertion (L249-E255 in human DKK3) and P258 in a finger loop 1 (FL1). Interestingly, the insertion sequence is evolutionally conserved. MD simulations of high-affinity complex models of C2 and PD showed that FL1 directly interacts with the PD models and stabilizes the complex models. We also built a 7-amino-acid-deletion/P258G mutant model of DKK3C2 and estimated its affinities for the PD models. The affinity for human LRP5PD2 was increased by the substitution (ΔGbinding＝－48.9kcal/mol) and the affinity was compatible with that of high-affinity ligands. The results suggested that the lack of affinity between human DKK3 and human LRP5/6 results from: i) insertion of the 7 amino acids, and ii) P258 in human DKK3. The sequence differences thus suggest an explanation for this unique property of DKK3

General Consumer Awareness of Warnings Regarding the Consumption of Alcoholic Beverages

Over the past two decades, the liquor industry in Japan has strived to address alcohol-related problems through initiatives such as warnings in the various media. In this study, we conducted an Internet-based questionnaire survey to examine general consumer awareness of such warnings, and the media by which they are conveyed, on the consumption of alcoholic beverages. A total of 985 subjects (males:487, females:498) in age groups ranging from 20s to 70s responded (response rate:22.4%). The awareness rates for warnings regarding underage drinking, drunk driving, and drinking during pregnancy, and those for messages encouraging moderation in drinking, were 96.4%, 83.7%, 59.6%, and 45.5%, respectively. Logistic regression analysis adjusted for habitual alcohol consumption demonstrated significant gender- and/or age-based differences in the rates of awareness of warnings and the media publicizing them. For example, the odds ratio of awareness among women of warnings against underage drinking was significantly higher than that of awareness among men. Issues that must be addressed in the future include:(1) increasing public awareness about messages regarding drinking during pregnancy and drinking in moderation;(2) reviewing the wording of warnings to make them more effective;and (3) devising and employing, on a regular basis, more effective means of transmitting messages in consideration of gender and age

Sleep Duration and Overweight among Elementary Schoolchildren:A Population-based Study in Japan

Although a number of studies have investigated the relationship of sleep duration to overweight and obesity, studies conducted among population-based elementary schoolchildren have been limited in Japan. The aim of the present study was to investigate the relationship between sleep duration and overweight among elementary schoolchildren in Japan. The study subjects were all fourth-grade schoolchildren (9 or 10 years of age) in Ina-town, Saitama Prefecture, Japan from 1999 to 2008. Information concerning each subjectʼs sex, age, and lifestyle was obtained using a self-administered questionnaire, while measurements of his or her height and weight were carried out. Childhood overweight was determined according to the definition established by the International Obesity Task Force. Data from 3,433 children were analyzed. In logistic regression analysis, a statistically significant dose-response relationship was observed between sleep duration and overweight among boys (p for trend＝0.014) but not among girls (p for trend＝0.149). Short sleep duration was associated with childhood overweight, and the sex difference in the association was observed. These findings suggested that it is important to consider sleep duration as part of any program to prevent overweight among elementary schoolchildren, especially among boys

Relationship between Eating Quickly and Overweight : A Cohort Study of Schoolchildren in Japan

Several cross-sectional studies have shown that eating quickly is associated with overweight in children and adolescents. However, few cohort studies have examined this relationship. Here we investigated the relationship between eating quickly and overweight in a cohort study of fourth-grade schoolchildren (aged 9 or 10 years) who attended elementary schools in Ina-town, Japan. The children were followed for 3 years from 2001-2004 (at baseline) to 2004-2007. A questionnaire survey including information about eating speed (fast, medium, or slow) was administered, and height and weight measurements were obtained at baseline and follow-up. Overweight was defined according to the body mass index cutoff points proposed by the International Obesity Task Force. We used a Poisson regression model to examine the association between eating quickly at baseline and being overweight 3 years later. Data from 934 non-overweight children (465 boys, 469 girls) were analyzed. Eating quickly was not significantly associated with being overweight in the boys, whereas it was significantly associated with being overweight in the girls. Our analysis indicates that among girls, eating quickly leads to overweight, and that the modification of eating quickly could help prevent overweight in adolescents

B-Cell-Activating Factor Affects the Occurrence of Thyroid Autoimmunity in Chronic Hepatitis C Patients Treated with Interferon Alpha

Chronic hepatitis C (CHC) patients frequently suffer from thyroid disorders during interferon therapy. However, the mechanism remains unclear. In this study, we investigated the association between serum B-cell-activating factor belonging to the TNF family (BAFF) levels and the presence of antithyroid peroxidase antibody (anti-TPO) in CHC patients treated with pegylated interferon alpha and ribavirin combination therapy. Six months after the therapy, anti-TPO antibody was detected in 10 (males, 1; females, 9) of 50 patients. The mean age of these patients was higher than that of the anti-TPO-negative patients (61 yr versus 55 yr). Before treatment, the serum BAFF levels of the anti-TPO-positive patients were higher than those of the anti-TPO-negative patients. After starting therapy, the serum BAFF levels of both the anti-TPO-positive and -negative patient groups were elevated. Our findings suggest that the serum BAFF concentration before therapy can predict the risk of thyroid autoimmunity in elderly female patients with CHC

Physiological and Pathological Mitochondrial Clearance Is Related to Pectoralis Major Muscle Pathogenesis in Broilers With Wooden Breast Syndrome

Wooden breast syndrome (WB) constitutes an emerging myopathy in the pectoralis major muscle (PM) of broiler chickens, characterized by myofiber hypertrophy and degeneration along with severe fibrosis. WB pathogenesis has been considered to involve hypoxia induced by rapid growth of the PM. In this study, we focused on mitochondrial morphology and dynamics in the myofibers, as these organelles are sensitive to damage by hypoxia, and examined the effects on WB pathogenesis. Specifically, the PMs of a flock of 35 broilers at 50 days of age were evaluated. First, the severity of disease in each bird was determined by measuring histopathological indices including the fibrotic area (FA) in the muscle and circularity of myofibers (CM). These values were 29.4 ± 9.6% and 0.70 ± 0.042, respectively, showing variety among the flock. Myofiber vacuolization was observed in all birds including numerous small- or large-rimmed vacuoles, with the former consisting of ultrastructurally autophagosome-like vacuoles engulfing degenerated mitochondria. The large-rimmed vacuoles frequently occurred in the PMs with more severe FA and CM, indicating a relationship between altered autophagy/mitophagy and WB severity. Next, the expression levels of hypoxia-adaptive and mitochondrial dynamics-related genes were analyzed, and their correlations with the histopathological indices were examined. The histopathological indices were negatively correlated with the expression of vascular endothelial growth factor A (VEGFA), indicating that less angiogenesis owing to weakened hypoxia-inducible factor signaling induces more severe WB pathology. In addition, the observed negative correlation with mitochondrial dynamics-related genes implied that WB pathology deteriorates concomitant with reduced mitochondrial dynamics. Furthermore, the expression of mitochondrial dynamics-related genes showed strong positive correlation with that of VEGFA and autophagy-/mitophagy-related genes. These results revealed that the PMs of broilers possess the mechanism of physiological clearance of mitochondria damaged by the hypoxia resulting from the continuous mitochondrial dynamics and autophagy/mitophagy accompanying rapid PM growth. In turn, the altered mitochondrial clearance induced by chronic hypoxia and the accumulation of damaged mitochondria likely underly the severe pathological features of WB.ArticleFrontiers in Pharmacology.11:579(2020)journal articl