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Human exposure to persistent organic pollutants (POPs) is reflected by POP concentrations in breast milk. Many studies of POPs in breast milk have been performed in Japan, but insufficient information is available about some legacy POPs (e.g., mirex and toxaphenes, included in the Stockholm Convention in 2001) and novel POPs (e.g., dicofol and endosulfans, included in the Stockholm Convention in 2019 and 2011, respectively). In this study, dicofol, endosulfan, mirex, and toxaphene concentrations in breast milk from 10 prefectures in Japan were determined. The samples were collected between 2005 and 2010, before Stockholm Convention restrictions on endosulfans and mirex were implemented. Common POPs (e.g., polychlorinated biphenyls) were also analyzed to allow the contamination statuses to be compared. The α-endosulfan and β-endosulfan concentrations were 0.26-13 and 0.012-0.82 ng/g lipid, respectively. The toxaphene #26 and #50 concentrations were0.08-5.6 and 0.1-8.5 ng/g lipid, respectively. The dicofol concentrations were0.01-4.8 ng/g lipid. The mirex concentrations were0.2-3.5 ng/g lipid. The α- and β-endosulfan concentrations on a lipid weight basis negatively correlated with the lipid contents of the milk samples (ρ = -0.65, p 0.01 for α-endosulfan; ρ = -0.58, p 0.01 for β-endosulfan). The toxaphene concentrations positively correlated with the lipid contents. The mirex concentrations positively correlated with the maternal age but negatively correlated with the maternal body mass index. No correlations between the dicofol concentrations and the factors were found. Principal component analysis divided the data into four groups, (1) chlordanes, dichlorodiphenyltrichloroethanes and related compounds, hexachlorobenzene, hexachlorocyclohexanes, hexachloroethane, and polychlorinated biphenyls, (2) endosulfans, (3) dicofol, dieldrin, and toxaphenes, and (4) bromodiphenyl ether 47. This indicated that bromodiphenyl ether 47, dicofol, endosulfans, and toxaphenes have different exposure routes or different kinetics to the other legacy POPs.

Objectives After the Fukushima Daiichi nuclear power plant disaster in 2011, residents of Kawauchi village who experienced evacuation had a high risk of suffering from diabetes and metabolic syndrome compared with non-evacuees. In addition to evacuation, lifestyle characteristics can be important factors influencing the development and prognosis of diabetes or glucose tolerance. The current study aimed to evaluate the effects of evacuation (i.e., lifestyle changes) on the incidence of diabetes among the non-diabetic residents of Kawauchi village. Methods Design is retrospective cohort study. Annual health examination data of residents of Kawauchi village and control area (Ono town) in Fukushima prefecture from 2008 to 2017, as available from the Japanese National Health Insurance system. Participants were classified into three groups: “Diabetes (DM)” (FBG ≥ 126 mg/dL or HbA1c ≥ 6.5% or hospital visit for DM or usage of diabetic medication), “Borderline DM” (126 mg/dL > FBG ≥ 110 mg/dL or 6.5% > HbA1c ≥ 6.0%, and without hospital visit, and without diabetic medication), and “Normoglycemic” (FBG < 110 mg/dL and HbA1c < 6.0%, and without hospital visit, and without diabetic medication). New onset of diabetes was evaluated and the events or missing data were occurred at health checkup. For this survival analysis, 339 residents in Kawauchi and 598 residents in Ono were included. Average follow-up periods after 2010 were 3.9 years in Kawauchi village and 3.6 years in Ono town. Results Compared with the normoglycemic group, incidence of DM was much greater in the borderline DM group, where DM occurred among 38.2% of the group in 2012 and increased to over 60% cumulatively through 2017 in Kawauchi village. DM had a prevalence of 16.3% in 2012, and below 30% in 2017 in borderline DM group of Ono town. Cox proportional hazard regression analysis was applied to non-DM groups at both study sites separately to evaluate the effects of lifestyle changes at each site. While BMI, BMI change, and the lack of regular exercise (HR = 1.29, 1.72, and 5.04, respectively) showed significant associations with the onset of diabetes in Ono town, only BMI and late-night dinner (HR = 1.21 and 4.86, respectively) showed significant associations with diabetes onset in Kawauchi village. Conclusions The current results confirmed that diabetes incidence was increased 6 years after the Daiichi nuclear power plant disaster in Kawauchi. We also found changes in lifestyle habits, suggesting that diabetes prevention with promotion of healthy lifestyle behaviors is an urgent priority.

[Objectives] It is sometimes difficult to differentiate middle cerebral artery disease from moyamoya disease because the two can present similarly yet have different treatment strategies. We investigated whether the presence of a narrow carotid canal and the RNF213 mutation can help differentiate between the two phenotypes. [Population and Methods] We analyzed 78 patients with moyamoya disease, 27 patients with middle cerebral artery disease, and 79 controls from 2 facilities. The carotid canal diameter was measured using computed tomography. The p.R4810K mutation was genotyped by TaqMan assay. A receiver operating characteristics analysis was performed to assess the significance of the carotid canal diameter for the accurate diagnosis of moyamoya disease. [Results] The carotid canal diameter was significantly narrower in patients with moyamoya disease than in controls. The optimal cutoff values were 5.0 mm for adult males and 4.5 mm for adult females and children (sensitivity: 0.82; specificity: 0.92). Among the patients with middle cerebral artery disease, 18.5% and 25.0% of the affected hemispheres had the p.R4810K mutation and narrow canal (i.e., below the cutoff), respectively, whereas only 3.1% of those had both. Contrastingly, 68.8% of the affected hemispheres in patients with moyamoya disease had both these characteristics. Among the patients with moyamoya disease, those with the p.R4810K mutation tended to have narrower carotid canals. [Conclusions] Although the presence of a narrow carotid canal or the p.R4810K mutation alone could not be used to distinguish those with moyamoya disease from those with middle cerebral artery disease, the combination of these factors could better characterize the two phenotypes.

Fluoride is an element that is widely distributed in the environment. The involvement of fluoride in pathogenesis of Chronic Kidney Disease of uncertain aetiology (CKDu) in Sri Lanka is a much-debated topic. This study aimed to investigate the fluoride concentration in drinking water in CKDu affected areas in Sri Lanka and to evaluate the possible effect of renal impairment on serum fluoride levels in CKDu patients. Drinking water (n = 60) from the common water sources from two CKDu prevalent areas and serum samples of CKDu patients (n = 311) and healthy controls (n = 276) were collected. Both environmental and biological samples were analysed for the concentration of fluoride. The fluoride concentration in over 95% of drinking water samples was below the WHO guideline of 1.5 mg/L. Serum fluoride concentrations in majority of the unaffected and early-stage CKDu patients (stages 1 and 2, eGFR >60 ml/min/1.73m2) were below the normal upper concentration of 50 μg/l and significantly higher levels were observed in patients in late stages of CKDu compared to the healthy controls. The available guidelines for drinking water are solely based on healthy populations with normal renal function. But, it is evident that once the kidney function is impaired, patients enter a vicious cycle as fluoride gradually accumulates in the body, further damaging the kidney tissue. Thus, close monitoring of serum fluoride levels in CKDu patients and establishing health-based target guidelines for fluoride in drinking water for the CKDu patients are recommended to impede the progression to end stage renal disease.

Levels and profiles of naturally produced halogenated bipyrroles (Br4Cl2-DBP and Cl7-MBP), methoxylated tetrabromodiphenyl ethers (6-MeO-BDE47), anthropogenic perfluoroalkyl substances (PFASs) and legacy persistent organic pollutants (POPs) were investigated in the livers of 14 cetaceans from the Sea of Japan and the North Pacific Ocean. The concentrations of Br4Cl2-DBP (4 to 4900 ng/g-wet), Cl7-MBP (16 to 3960 ng/g-wet) and 6-MeO-BDE47 (7 to 190 ng/g-wet) were higher in the order of killer whales > toothed whales > baleen whales. Profiles of PFASs were dominated by perfluoroundecanoic and perfluorotridecanoic acids (10 to 540 ng/g-wet), sum of which accounted for 70% of total measured PFASs. Regional difference was observed for Cl7-MBP and PFASs, which were higher in the Sea of Japan, whereas Br4Cl2-DBP was in the North Pacific Ocean. Specific accumulation pattern of these natural contaminants in cetaceans around northern Japan could help compare the exposure profile of PFASs and POPs among other geographic regions.

This study investigated contamination trends and factors affecting the levels of brominated flame retardants (BFRs), including hexabromocyclododecane (HBCD) diastereomers, tetrabromobisphenol A (TBBP-A), and 2,4,6-tribromophenol (2,4,6-TBP), in breast milk in Japan. Breast milk samples (n = 64) were collected from mothers living in six prefectures in Japan. The mean concentrations were 2.2, 0.19, 0.29, 3.0, and 0.59 ng/g lipid weight for α-HBCD, β-HBCD, γ-HBCD, TBBP-A, and 2,4,6-TBP, respectively. Based on the previous studies, the levels of ΣHBCD in Japanese women's milk appear to be increasing, and the levels of TBBP-A are higher than those in other Asian countries. Although ΣHBCD were not correlated to phenolic BFRs, the concentration of β-HBCD was significantly correlated to the concentrations of TBBP-A (r = 0.440, p 0.01) and 2,4,6-TBP (r = 0.320, p 0.01). The concentration of γ-HBCD increased significantly with maternal age (r = 0.378, p 0.01), but the concentrations of the other analytes were not dependent on age. The concentration of α-HBCD was higher in primiparae than in multiparae (p 0.05), while TBBP-A was higher in multiparae. No significant correlation was found between the phenolic BFR levels in milk and mothers' age, working place, and drinking/smoking habits. These results suggest that exposure to α- and γ-HBCD diastereomers could be affected by maternal age and parity, respectively, because of their different kinetics and sources. Therefore, these factors should be considered when conducting infant risk assessments.

Background This study aimed to determine the effectiveness of genetic testing for the p.R4810K variant (rs112735431) of the Mysterin/RNF213 gene, which is associated with moyamoya disease and other intracranial vascular diseases, in the family members of patients with moyamoya disease. Methods We performed genotyping of the RNF213 p.R4810K polymorphism and magnetic resonance angiography on 59 relatives of 18 index patients with moyamoya disease. Nineteen individuals had follow-up magnetic resonance angiography with a mean follow-up period of 7.2 years. Results Six of the 34 individuals with the GA genotype (heterozygotes for p.R4810K) showed intracranial steno-occlusive lesions in the magnetic resonance angiography, whereas none of the 25 individuals with the GG genotype (wild type) showed any abnormalities. Follow-up magnetic resonance angiography revealed de novo lesions in 2 and disease progression in 1 of the 11 individuals with the GA genotype, despite none of the 8 individuals with the GG genotype showing any changes. Accordingly, 8 individuals had steno-occlusive lesions at the last follow-up, and all had the p.R4810K risk variant. The prevalence of steno-occlusive intracranial arterial diseases in family members with the p.R4810K variant was 23.5% (95% confidence interval: 9.27%-37.78%), which was significantly higher than in those without the variant (0%, P = .0160). Conclusions Genotyping of the p.R4810K missense variant is useful for identifying individuals with an elevated risk for steno-occlusive intracranial arterial diseases in the family members of patients with moyamoya disease.

Background and Purpose— The ring finger protein 213 gene ( RNF213 ) is a susceptibility gene for moyamoya disease and large-artery ischemic stroke in East Asia. We examined the prevalence and correlates of the RNF213 p.R4810K variant in patients with early-onset ischemic stroke in a Japanese single-center cohort. Methods— We analyzed 70 early-onset stroke patients with intracranial arterial stenosis who developed a noncardioembolic stroke or transient ischemic attack from 20 to 60 years of age. Patients with moyamoya disease were excluded. Results— The RNF213 p.R4810K variant was found in 17 patients (24%), and more often in women than men (38% versus 16%, odds ratio 3.3; 95% CI, 1.1–10.2, P =0.04). The variant was identified in 35% of patients with stenosis in the M1 segment of the middle cerebral artery or the A1 segment of the anterior cerebral artery (odds ratio, 25.0; 95% CI, 1.4–438; P Conclusions— The RNF213 p.R4810K variant is common in early-onset ischemic stroke with anterior circulation stenosis in Japan. Further investigation of the RNF213 gene will provide new insights into pathogenetic mechanisms of early-onset stroke.

Background: Bradyarrhythmia is a common clinical manifestation. Although the majority of cases are acquired, genetic analysis of families with bradyarrhythmia has identified a growing number of causative gene mutations. Because the only ultimate treatment for symptomatic bradyarrhythmia has been invasive surgical implantation of a pacemaker, the discovery of novel therapeutic molecular targets is necessary to improve prognosis and quality of life. Methods: We investigated a family containing 7 individuals with autosomal dominant bradyarrhythmias of sinus node dysfunction, atrial fibrillation with slow ventricular response, and atrioventricular block. To identify the causative mutation, we conducted the family-based whole exome sequencing and genome-wide linkage analysis. We characterized the mutation-related mechanisms based on the pathophysiology in vitro. After generating a transgenic animal model to confirm the human phenotypes of bradyarrhythmia, we also evaluated the efficacy of a newly identified molecular-targeted compound to upregulate heart rate in bradyarrhythmias by using the animal model. Results: We identified one heterozygous mutation, KCNJ3 c.247A>C, p.N83H, as a novel cause of hereditary bradyarrhythmias in this family. KCNJ3 encodes the inwardly rectifying potassium channel Kir3.1, which combines with Kir3.4 (encoded by KCNJ5 ) to form the acetylcholine-activated potassium channel ( I KACh channel) with specific expression in the atrium. An additional study using a genome cohort of 2185 patients with sporadic atrial fibrillation revealed another 5 rare mutations in KCNJ3 and KCNJ5 , suggesting the relevance of both genes to these arrhythmias. Cellular electrophysiological studies revealed that the KCNJ3 p.N83H mutation caused a gain of I KACh channel function by increasing the basal current, even in the absence of m 2 muscarinic receptor stimulation. We generated transgenic zebrafish expressing mutant human KCNJ3 in the atrium specifically. It is interesting to note that the selective I KACh channel blocker NIP-151 repressed the increased current and improved bradyarrhythmia phenotypes in the mutant zebrafish. Conclusions: The I KACh channel is associated with the pathophysiology of bradyarrhythmia and atrial fibrillation, and the mutant I KACh channel ( KCNJ3 p.N83H) can be effectively inhibited by NIP-151, a selective I KACh channel blocker. Thus, the I KACh channel might be considered to be a suitable pharmacological target for patients who have bradyarrhythmia with a gain-of-function mutation in the I KACh channel.

Human exposure to polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) may result in retention of specific congeners of hydroxylated PCBs (OH-PCBs) and hydroxylated/methoxylated PBDEs (OH-/MeO-PBDEs) in serum. However, dietary sources and biotransformation of OH-/MeO-PBDEs in humans are poorly understood. Therefore, this study was conducted to investigate the levels, profiles, and exposure sources of OH-/MeO-PBDEs along with OH-PCBs present in human serum. Twenty serum samples pooled from women of four age groups (30s/40s/50s/60s) living in four districts of Japan were analyzed for OH-/MeO-PBDEs, and their profiles were then compared with those of seafood (seaweed and fish). The major component of OH-PCBs in the phenolic fraction of serum was 4-OH-CB187 (mean: 85 pg g−1 wet weight (ww)). Total OH-PCBs accounted for about 1/20 of the total PCBs (mean; 1800 pg g−1 ww). In contrast, the predominant component of OH-PBDEs in serum was 6-OH-BDE47 (mean: 183 pg g−1 ww), which was about 20-fold higher than BDE-47 (mean; 8.7 pg g−1 ww). In the neutral fraction, 2′-MeO-BDE68 was primarily found at a similar concentration (mean 5.6 pg g−1 ww) to BDE-47. Both 4-OH-PCB187 and 2′-MeO-BDE68 were significantly correlated with woman's age (p

Background Quasi-moyamoya disease is an angiographical moyamoya disease equivalent accompanied by known underlying diseases. Mysterin/RNF213 is a major susceptibility gene for moyamoya disease, of which the p.R4810K variant is a founder polymorphism. The genetics of quasi-moyamoya disease is poorly understood, therefore, this study investigated a potential association between the p.R4810K polymorphism and quasi-moyamoya disease. Methods Genotyping of the p.R4810K variant was performed on 18 quasi-moyamoya disease cases and 91 controls, who visited Kyoto University Hospital or Kobe City Medical Center, Japan, between 2006 and 2015. Results The p.R4810K variant was found in 12 of 18 quasi-moyamoya disease patients. The frequency of p.R4810K carriers was significantly higher in quasi-moyamoya disease cases than in controls (66.7% versus 2.2%, odds ratio 89.0, 95% confidence interval: 19.2-669.4). Conclusions Our data showed that the RNF213 p.R4810K polymorphism was significantly associated with quasi-moyamoya disease.

Objectives Neonicotinoid insecticides have been widely used around the world since the 1990s. Reports have been made since the 1990s of rice paddy farmers in the North Central Region (NCR) of Sri Lanka suffering from chronic kidney disease with unknown etiology (CKDu). A preliminary evaluation of the exposure of local farmers in the NCR of Sri Lanka to neonicotinoids was performed. Methods We analyzed neonicotinoid and neonicotinoid metabolite concentrations in spot urine samples. We selected 40 samples, 10 from farmers with CKDu and 10 from controls from each of two areas, Medawachchiya and Girandurukotte. Results Imidacloprid and desmethyl-acetamiprid were found at significantly higher concentrations in the control samples (with medians of 51 ng/l and 340 ng/l, respectively) than in the CKDu samples (medians of 15 ng/l and 150 ng/l, respectively) when the results were not adjusted for the creatinine contents. None of the six compounds that were measured in the urine samples were found at significantly higher concentrations in the CKDu samples than in the control samples. None of the neonicotinoid concentrations in the samples analyzed in this study exceeded the concentrations that have been found in samples from the general population of Japan. Conclusions Farmers (both with and without CKDu) living in CKDu-endemic areas in the NCR of Sri Lanka are exposed to lower neonicotinoid concentrations than non-occupationally exposed residents of Japan.

Background:Helicobacter pylori eradication rates have decreased worldwide. Gastric acid inhibition during treatment is important to eradicate these bacteria successfully. A new potassium-competitive acid blocker, vonoprazan (VPZ), has been shown to achieve high eradication rates in a previous randomized controlled trial. Objective: To determine the efficacy of VPZ for H. pylori eradication. Methods: A total of 874 patients were enrolled; 431 received esomeprazole (EPZ) and 443 received VPZ. First-line regimens contained clarithromycin (CAM) 200 mg b.i.d., amoxicillin 750 mg b.i.d., and either EPZ 20 mg b.i.d. or VPZ 20 mg b.i.d. for 7 days. Metronidazole 250 mg b.i.d. replaced CAM in the second-line regimens. The eradication of H. pylori was assessed by 13C-urea breath tests 4-8 weeks after each therapy. Results: The overall first-line eradication rate was 79.9% (341/427) with EPZ vs. 86.3% (377/439) with VPZ (p = 0.019). The second-line eradication rate was 83.3% (45/51) with EPZ vs. 91.1% (41/45) with VPZ (p = 0.900). Conclusion: VPZ was significantly more effective than EPZ for first-line treatment. However, for second-line treatment, there was no significant difference between EPZ and VPZ.

In 1952, the Japanese Society for Hygiene had once passed a resolution at its 22nd symposium on population control, recommending the suppression of population growth based on the idea of cultivating a healthier population in the area of eugenics. Over half a century has now passed since this recommendation; Japan is witnessing an aging of the population (it is estimated that over 65-year-olds made up 27.7% of the population in 2017) and a decline in the birth rate (total fertility rate 1.43 births per woman in 2017) at a rate that is unparalleled in the world; Japan is faced with a “super-aging” society with low birth rate. In 2017, the Society passed a resolution to encourage all scientists to engage in academic researches to address the issue of the declining birth rate that Japan is currently facing. In this commentary, the Society hereby declares that the entire text of the 1952 proposal is revoked and the ideas relating to eugenics is rejected. Since the Society has set up a working group on the issue in 2016, there have been three symposiums, and working group committee members began publishing a series of articles in the Society’s Japanese language journal. This commentary primarily provides an overview of the findings from the published articles, which will form the scientific basis for the Society’s declaration. The areas we covered here included the following: (1) improving the social and work environment to balance between the personal and professional life; (2) proactive education on reproductive health; (3) children’s health begins with nutritional management in women of reproductive age; (4) workplace environment and occupational health; (5) workplace measures to counter the declining birth rate; (6) research into the effect of environmental chemicals on sexual maturity, reproductive function, and the children of next generation; and (7) comprehensive research into the relationship among contemporary society, parental stress, and healthy child-rearing. Based on the seven topics, we will set out a declaration to address Japan’s aging society with low birth rate.

Over the last two decades, usage of neonicotinoid (NEO) insecticides has increased due to their high selectivity for insects versus mammals and their effectiveness for extermination of insects resistant to conventional pesticides such as pyrethroids and organophosphates (OPs). However, historical change of the NEO exposure level in humans is poorly understood. The aim of this study is to reveal changes in the levels of NEO and OP exposure in the human body over the last two decades using biomonitoring technique. We quantified urinary concentrations of 7 NEOs (acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam) and 4 metabolites of OPs (dimethylphosphate, dimethylthiophosphate, diethylphosphate, and diethylthiophosphate) in 95 adult females aged 45-75 in 1994, 2000, 2003, 2009, and 2011 (n = 17-20 different individuals in each year). The results show that the detection rates of urinary NEOs in Japanese women increased significantly between 1994 and 2011, suggesting that intakes of NEOs into the human body rose during that period. In contrast, exposure to OPs having O,O-dimethyl moieties decreased steadily according to a finding that geometric means of urinary dimethylphosphate concentrations kept diminishing considerably. These changes may reflect the amounts of NEOs and OPs used as insecticides in Japan.

ObjectiveTo investigate the efficacy of recombinant human soluble thrombomodulin (rTM) in preventing the development of walled-off necrosis (WON) in severe acute pancreatitis (SAP) patients.MethodsWe retrospectively analyzed 54 SAP patients divided into two groups: SAP patients treated by rTM (rTM group, 24 patients) and not treated by rTM (control group, 30 patients). rTM was administered to patients with disseminated intravascular coagulation (DIC). Initially, on the admission day, we recorded patient severity and pancreatic necrosis/ischemia positive or negative. Then we investigated development of WON using 4 weeks later CT/MRI. Finally we compared the proportions of patients developing WON in the rTM group and the control group.ResultsOn the admission day, the condition of patients treated by rTM was significantly worse than patients in the control group; rTM group vs. control: 71.8 ± 13.9 vs. 59.8 ± 15.3 years for age, 10.7 ± 3.5 vs. 8.0 ± 4.4 for Acute Physiology and Chronic Health Evaluation II (APACHE II) score, and 3.3 ± 1.8 vs. 2.2 ± 1.8 for sequential organ failure assessment (SOFA) score (p

Background and Purpose— Rupture of intracranial aneurysms (IAs) causes subarachnoid hemorrhage, leading to immediate death or severe disability. Identification of the genetic factors involved is critical for disease prevention and treatment. We aimed to identify the susceptibility genes for IAs. Methods— Exome sequencing was performed in 12 families with histories of multiple cases of IA (number of cases per family ≥3), with a total of 42 cases. Various filtering strategies were used to select the candidate variants. Replicate association studies of several candidate variants were performed in probands of 24 additional IA families and 426 sporadic IA cases. Functional analysis for the mutations was conducted. Results— After sequencing and filtering, 78 variants were selected for the following reasons: allele frequencies of variants in 42 patients was significantly ( P GPR63 , ADAMST15 , MLL2 , IL10RA , PAFAH2 , THBD , IL11RA , FILIP1L , and ZNF222 ) to form 78 candidate variants by considering commonness in families, known disease genes, or ontology association with angiogenesis. Replicate association studies revealed that only p.E133Q in ADAMTS15 was aggregated in the familial IA cases (odds ratio, 5.96; 95% confidence interval, 2.40–14.82; P =0.0001; significant after the Bonferroni correction [ P =0.05/78=0.0006]). Silencing ADAMTS15 and overexpression of ADAMTS15 p.E133Q accelerated endothelial cell migration, suggesting that ADAMTS15 may have antiangiogenic activity. Conclusions— ADAMTS15 is a candidate gene for IAs.

1Department of Health and Environmental Sciences, Kyoto University Graduate School of Medicine, Yoshida Konoecho, Kyoto 606-8501, Japan 2Faculty of Medical Sciences, Department of Health Science, University of Fukui, Fukui 910-1193, Japan 3Department of Genetics, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya 663-8501, Japan 4Ishikawa Prefectural Central Hospital, Kuratsukihigashi, Kanazawa, Ishikawa 920-8530, Japan 5Department of Pediatrics (NICU), Kyoto University Hospital, 54 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan

Background/Aims: Currently, there is no study evaluating the effect of acotiamide on transient lower esophageal sphincter relaxations (TLESRs). The aim of this study was to evaluate the effect of acotiamide on TLESRs using simultaneous high-resolution manometry (HRM) and impedance-pH monitoring. Methods: Ten healthy subjects were enrolled. On day 1, subjects underwent HRM and impedance-pH recordings as a baseline. Subjects ate a 750-kcal liquid meal; recording was continued for 2 h while the subjects were in a sitting position. After the administration of acotiamide 100 mg three times a day for 1 week, subjects underwent HRM and impedance-pH recording under the same protocol. Results: A total of 208 TLESRs were identified at baseline. Acotiamide decreased the total number of TLESRs from 208 to 143 (p < 0.05). The rate of reflux events during TLESRs after acotiamide administration was similar to that at baseline (57% after acotiamide vs. 58% at baseline). Bolus clearance time was significantly reduced by acotiamide. Conclusions: Acotiamide was believed to have the potential for reducing TLESRs and for enhancing esophageal bolus clearance in healthy volunteers. Future research is needed to determine whether the effects of acotiamide that reduce TLESRs and enhance esophageal motility could improve symptoms in patients with refractory gastroesophageal reflux disease.

Aims The aim of this study was to clarify the genetic background of a family with multiple cases of diabetes accompanied by absolute insulin deficiency using whole-exome sequencing (WES). Methods In a Japanese family, WES was performed in four affected members with absolute insulin deficiency and two unaffected members. We focused on variants that were predicted to be disease-causing by bioinformatics and were shared by all of the four affected members but were not present in the two unaffected members. We assumed that the familial clustering of diabetes was caused by rare variants excluding those with allele frequency of more than 0.01 in the 1000 Genomes Project, the Human Genetic Variation Database, or a cohort of 105 normoglycemic controls in Japan. The rare variants were then genotyped in 2102 Japanese without diabetes and 119 Japanese with diabetes. Results Among the variants detected by WES and predicted to be disease-causing, 16 variants shared by all of the four of the affected members and not present in the two unaffected members were confirmed to be rare. Genotyping of the 16 rare variants revealed that only A137T in ADAMTSL3 (rs181914721) was observed more frequently in the 119 subjects with diabetes than in the 105 normoglycemic controls, and the allele frequency of the variant was significantly higher in the 119 subjects with diabetes than in another cohort of 2102 Japanese without diabetes. Conclusions We propose that A137T in ADAMTSL3 is a candidate mutation for susceptibility to diabetes in this family and in the Japanese population.

This study investigated human exposure to neutral, phenolic, and methoxylated organohalogen contaminants (OHCs) in a duplicate diet study to evaluate their concentrations in breast milk and serum of Okinawan people from Japan during 2004–2009. Dietary intakes of phenolic OHCs were predominantly 2,4,6-tribromophenol (TriBP), followed by tetrabromobisphenol A (TBBPA), and 6-hydroxy-2,2′,4,4′-tetrabromodiphenyl ether (6-OH-BDE47). After exposure, TriBP and TBBPA were transferred to breast milk, whereas 6-OH-BDE47 was selectively retained in serum. Despite a lower dietary exposure to pentachlorophenol and 4-hydroxy-CB187, both were retained in serum. For the methoxylated OHCs, 2,4,6-tribromoanisole (TriBA) and 6-methoxy-BDE47 were the predominant dietary contaminants, of which TriBA was present in both breast milk and serum, whereas 6-methoxy-BDE47 was selectively transferred to breast milk. These findings suggest that dietary exposure to phenolic and methoxylated OHCs may result in differential partitioning between breast milk and serum with different pharmacokinetic or exposure routes. Keywords: Dietary exposure, Breast milk, Serum, 6-Hydroxy-BDE47, 6-Methoxy-BDE47

We investigated the incidence of human T-cell leukemia virus type I (HTLV-1) infection in a total of 17 regions in four countries, including 13 regions in Japan, as well as Korea (Seoul and Busan), China, and Vietnam, by testing breast milk using a particle agglutination assay (PA) and line immunoassay (LIA). Among 266 samples from Japan, 24 (9.0%) were positive on PA and 3 (1.1%) were positive on LIA. Among 50 samples from Seoul, 2 were positive on PA and 1 was positive on LIA. In contrast, all 50 samples from Busan were negative on both tests, suggesting the maldistribution of HTLV-1 infectants in South Korea. The numbers of positive samples were 2/91 on PA and 1/91 on LIA for China and 1/88 on both PA and LIA for Vietnam. In China, one sample with a high probability of HTLV-2 infection was identified by LIA and synthetic peptide enzyme-linked immunosorbent assay (ELISA). We examined HTLV-1 antibody in breast milk samples using commercially available test kits, suggesting the existence of HTLV-1 carriers in endemic areas in Southeast Asia and an HTLV-2 infectant in China. As a part of human ethno-epidemiological research, these results constitute valuable epidemiological data. Further studies on the sensitivity, specificity, and reliability of assays using antibodies to HTLV-1 and 2 in breast milk will be necessary for large-scale epidemiological surveys of HTLV infection.

Familial goiter is a genetic disease showing heterogeneous expression. To identify causative genes, we investigated three multigenerational goiter families with an autosomal dominant inheritance pattern. We performed genome-wide linkage analysis on all the families, combined with whole-exome sequencing in two affected individuals from each family. For linkage analysis, we considered loci with logarithm of odds (LOD) scores >1.5 as candidate regions for identification of rare variants. In one of the families, we found two rare heterozygous missense variants, p.V56M in RGS12 and p.G37D in GRPEL1, which segregate with goiter and are both located within the same haplotype on 4p16. This haplotype was not observed in 150 controls. In the other two families, we identified two additional rare missense variants segregating with goiter, p.A551T in CLIC6 on 21q22.12 and p.V412A in WFS1 on 4p16. In controls, the minor allele frequency (MAF) of p.V412A in WFS1 was 0.017 while p.A551T in CLIC6 was not detected. All identified genes (RGS12, GRPEL1, CLIC6 and WFS1) show expression in the human thyroid gland, suggesting that they may play a role in thyroid gland function. Moreover, these four genes are novel with regard to their involvement in familial goiter, supporting genetic heterogeneity of this disease.

Since the nuclear power plant accident in Fukushima in March 2011, the Japanese government has conducted screening and removal of contaminated foods from the market that exceed provisional regulation limits for radionuclides. This study aimed to provide an urgent estimate of the dietary exposure of adult residents recruited from three areas in Japan to cesium 134 ((134)Cs), cesium 137 ((137)Cs), and, for comparison, natural potassium 40 ((40)K) on December 4, 2011. Fifty-three sets of 24-h food-duplicate samples were collected in Fukushima Prefecture and neighboring regions. The (134)Cs, (137)Cs, and (40)K levels in the samples were measured using a germanium detector. Items in the food-duplicate samples were recorded and analyzed for radiocesium intake. Radiocesium was detected in 25 of 26 samples from Fukushima. The median dietary intake of radiocesium was 4.0 Bq/day (range0.26-17 Bq/day). The estimated annual dose from radiocesium was calculated assuming that the daily intake of radiocesium was constant throughout the year. The median estimated dose level was 23 μSv/year (range2.6-99 μSv/year). The estimated dose level of radiocesium was significantly higher in Fukushima than in the Kanto region and western Japan. Stepwise multiple linear regression analyses demonstrated that the intake of fruits and mushrooms produced in Fukushima were significant factors for the dietary intake of (137)Cs in the 26 participants from Fukushima. The average radioactivity (±SD) of locally produced persimmons and apples (n = 16) were 23 ± 28 and 30 ± 35 Bq/kg for (134)Cs and (137)Cs, respectively. The preliminary estimated dietary dose levels among Fukushima residents were much lower than the maximum permissible dose 1 mSv/year, based on new Japanese standard limits for radiocesium in foods (100 Bq/kg for general foods). In future studies, the exposure estimates should be refined by probability sampling to eliminate biases.

Tumours exist in a hypoxic microenvironment and must limit excessive oxygen consumption. Hypoxia-inducible factor (HIF) controls mitochondrial oxygen consumption, but how/if tumours regulate non-mitochondrial oxygen consumption (NMOC) is unknown. Protein-tyrosine phosphatase-1B (PTP1B) is required for Her2/Neu-driven breast cancer (BC) in mice, although the underlying mechanism and human relevance remain unclear. We found that PTP1B-deficient HER2(+) xenografts have increased hypoxia, necrosis and impaired growth. In vitro, PTP1B deficiency sensitizes HER2(+) BC lines to hypoxia by increasing NMOC by α-KG-dependent dioxygenases (α-KGDDs). The moyamoya disease gene product RNF213, an E3 ligase, is negatively regulated by PTP1B in HER2(+) BC cells. RNF213 knockdown reverses the effects of PTP1B deficiency on α-KGDDs, NMOC and hypoxia-induced death of HER2(+) BC cells, and partially restores tumorigenicity. We conclude that PTP1B acts via RNF213 to suppress α-KGDD activity and NMOC. This PTP1B/RNF213/α-KGDD pathway is critical for survival of HER2(+) BC, and possibly other malignancies, in the hypoxic tumour microenvironment.

We investigated temporal changes of perfluoroalkyl carboxylic acids (PFCAs) with 8-14 carbon atoms (C8 to C14) in duplicate diet and serum samples in Japan. The sum dietary intakes of PFCAs (C8 to C13) in the Kansai and Tohoku region were highest in the 2010s (mean; 177 ng/day for Kansai, 107 ng/day for Tohoku) followed by the 2000s (77 ng/day for Kansai, 34 ng/day for Tohoku) and the 1990s (53 ng/day for Kansai, 58 ng/day for Tohoku), then the 1980s (19 ng/day for Kansai, 23 ng/day for Tohoku). The sum of the serum concentartions (C8 to C13) was also highest in the 2010s (mean; 17 ng/mL for Kansai, 7.4 ng/mL for Tohoku), followed by the 2000s (12 ng/mL for Kansai, 6.3 ng/mL for Tohoku), then the 1990s (6.8 ng/mL for Kansai, 5.5 mg/mL for Tohoku) and the 1980s (3.8 ng/mL for Kansai, 0.4 ng/mL for Tohoku). A positive correlation was observed between dietary intakes and serum concentration for C8 to C11 (r = 0.94, p 0.05 for C8; r = 0.80, p 0.05 for C9; r = 0.98, p 0.05 for C10; and r = 0.84, p 0.05 for C11). The levels of C8, C9 and C10 in serum and dietary intake in the 2010s were much higher in Kansai than those in Tohoku, although those of C11 did not show such differences. Kansai has a fluoropolymer manufacture known as a specific source of PFOA (C8), and is more urbanized than Tohoku, which may be attributed to the higher levels of PFCAs (C8 to C10). On the other hand, C11 is common to residents in Kansai and Tohoku.

To investigate short-chain chlorinated paraffin (SCCP) levels in human breast milk, pooled breast milk samples (BMSs) collected between 2007 and 2010 from Chinese (Beijing, n = 17), Korean (Seoul, Busan, n = 16), and Japanese (Kyoto, Sendai, n = 44) women were analyzed. SCCPs found in air samples in Beijing ( n = 4, in 2008) were also analyzed and compared with BMSs to estimate the possible source of contamination in Beijing. The electron-capture negative ionization method demonstrated the different sensitivities for SCCPs, and pentachlorinated alkanes had the highest method detection limit (MDL) among congeners. In Beijing, SCCPs were detected in 8 of 17 pooled BMSs at concentrations more than the highest MDL of each homolog. The total SCCP concentration ranged from below the MDL to 54 ng g −1 lipid weight. Among the SCCP homologs, polychlorinated tridecanes were most frequently detected in Beijing. In Korea and Japan, no samples contained detectable total SCCP concentrations at more than the highest MDL. In Seoul, only two samples showed trace levels of polychlorinated undecanes. In Kyoto and Sendai, congeners of polychlorinated dodecanes were most frequently detected. C 10 components were the major contributors to the SCCPs in the atmosphere of Beijing. Congener profiles in breast milk in Beijing provided a clear contrast to the profiles found in food and air. The unique congener profiles necessitate the monitoring of breast milk for exposure of infants to SCCPs. The calculated mean exposure of SCCPs in 1-year-olds in China was 337 ng (kg body weight) −1 d −1 . These results demonstrate the body burden of SCCPs in the study areas and potential lactational exposure to SCCPs in Asian countries.

Objectives: Kawauchi Village lies 20 km west of the Fukushima Daiichi nuclear power plant. On 16 March 2011, evacuation was ordered due to the threat of radiological exposure, and was lifted in April 2012. In this study, we aimed to evaluate the predisaster and postdisaster health status of the Kawauchi Villagers, measured by routine yearly physical examinations. Methods: We analysed the annual health examination data of residents of Kawauchi Village from 2008 to 2013, as available from the Japanese National Health Insurance system. Data from 2011 were not available due to the disaster. Since the health data included the same participants repeatedly from year to year, the sample was non-independent and generalised estimated equation modelling was used. A predisaster time period (2008–2010) was categorised for comparison with postdisaster 2012 and 2013. The outcome examined was the prevalence of metabolic disease, and was adjusted for confounding factors. Results: Data for 20.6%–25.9% of the total residents were available in this period. In 2013, the prevalence of metabolic syndrome (from 17.0% to 25.2%, p

Cardiovascular complications are the leading cause of death in autosomal dominant polycystic kidney disease (ADPKD), and intracranial aneurysm (ICA) causing subarachnoid hemorrhage is among the most serious complications. The diagnostic and therapeutic strategies for ICAs in ADPKD have not been fully established. We here generated induced pluripotent stem cells (iPSCs) from seven ADPKD patients, including four with ICAs. The vascular cells differentiated from ADPKD-iPSCs showed altered Ca2+ entry and gene expression profiles compared with those of iPSCs from non-ADPKD subjects. We found that the expression level of a metalloenzyme gene, matrix metalloproteinase (MMP) 1, was specifically elevated in iPSC-derived endothelia from ADPKD patients with ICAs. Furthermore, we confirmed the correlation between the serum MMP1 levels and the development of ICAs in 354 ADPKD patients, indicating that high serum MMP1 levels may be a novel risk factor. These results suggest that cellular disease models with ADPKD-specific iPSCs can be used to study the disease mechanisms and to identify novel disease-related molecules or risk factors.

RNF213 p.R4810K is a founder polymorphism that confers genetic susceptibility to moyamoya disease in East Asia. Only a few studies have investigated the symptoms and disease histories of RNF213 p.R4810K carriers in Japan. This study investigated the frequency of RNF213 p.R4810K in the general Japanese population and the health characteristics of the carriers. Through a health-promotion campaign in the city of Uji, Japan, 519 subjects (120 males and 399 females) of the general Japanese population were genotyped for RNF213 p.R4810K and interviewed to determine health characteristics. Nine RNF213 p.R4810K heterozygous carriers (GA genotype) and no RNF213 p.R4810K homozygous carriers (AA genotype) were found among the 519 individuals. The estimates of the genotypes and allele frequencies for RNF213 p.R4810K were 1.73 and 0.87 %, respectively. There were no obvious differences in age, gender ratio, body mass index, hypertension, dyslipidemia, diabetes, kidney disease, liver disease, heart disease, or drinking or smoking habits between carriers and non-carriers. Interestingly, one patient with moyamoya disease was found among the nine RNF213 p.R4810K carriers. This study showed the genotypes and allele frequencies of RNF213 p.R4810K in the general Japanese population to be similar to results of previous reports.

Intracranial aneurysms (IAs) are the result of focal weakness in the artery wall and have a complex genetic makeup. To date, genome-wide association and sequencing studies have had limited success in identifying IA risk factors. Distinct populations, such as the French-Canadian (FC) population, have increased IA prevalence. In our study, we used exome sequencing to prioritize risk variants in a discovery cohort of six FC families affected by IA, and the analysis revealed an increased variation burden for ring finger protein 213 (RNF213). We resequenced RNF213 in a larger FC validation cohort, and association tests on further identified variants supported our findings (SKAT-O, p = 0.006). RNF213 belongs to the AAA+ protein family, and two variants (p.Arg2438Cys and p.Ala2826Thr) unique to affected FC individuals were found to have increased ATPase activity, which could lead to increased risk of IA by elevating angiogenic activities. Common SNPs in RNF213 were also extracted from the NeuroX SNP-chip genotype data, comprising 257 FC IA-affected and 1,988 control individuals. We discovered that the non-ancestral allele of rs6565666 was significantly associated with the affected individuals (p = 0.03), and it appeared as though the frequency of the risk allele had changed through genetic drift. Although RNF213 is a risk factor for moyamoya disease in East Asians, we demonstrated that it might also be a risk factor for IA in the FC population. It therefore appears that the function of RNF213 can be differently altered to predispose distinct populations to dissimilar neurovascular conditions, highlighting the importance of a population’s background in genetic studies of heterogeneous disease.

Painful peripheral neuropathy has been correlated with various voltage-gated sodium channel mutations in sensory neurons. Recently Nav1.9, a voltage-gated sodium channel subtype, has been established as a genetic influence for certain peripheral pain syndromes. In this study, we performed a genetic study in six unrelated multigenerational Japanese families with episodic pain syndrome. Affected participants (n = 23) were characterized by infantile recurrent pain episodes with spontaneous mitigation around adolescence. This unique phenotype was inherited in an autosomal-dominant mode. Linkage analysis was performed for two families with 12 affected and nine unaffected members, and a single locus was identified on 3p22 (LOD score 4.32). Exome analysis (n = 14) was performed for affected and unaffected members in these two families and an additional family. Two missense variants were identified: R222H and R222S in SCN11A. Next, we generated a knock-in mouse model harboring one of the mutations (R222S). Behavioral tests (Hargreaves test and cold plate test) using R222S and wild-type C57BL/6 (WT) mice, young (8-9 weeks old; n = 10-12 for each group) and mature (36-38 weeks old; n = 5-6 for each group), showed that R222S mice were significantly (p < 0.05) more hypersensitive to hot and cold stimuli than WT mice. Electrophysiological studies using dorsal root ganglion neurons from 8-9-week-old mice showed no significant difference in resting membrane potential, but input impedance and firing frequency of evoked action potentials were significantly increased in R222S mice compared with WT mice. However, there was no significant difference among Nav1.9 (WT, R222S, and R222H)-overexpressing ND7/23 cell lines. These results suggest that our novel mutation is a gain-of-function mutation that causes infantile familial episodic pain. The mouse model developed here will be useful for drug screening for familial episodic pain syndrome associated with SCN11A mutations., 乳幼児期に特異的な手足の痛み発作を起こす病気を見つけ原因を解明 －この病気を小児四肢疼痛発作症と命名－. 京都大学プレスリリース. 2016-05-27.

Background Neonicotinoids, which are novel pesticides, have entered into usage around the world because they are selectively toxic to arthropods and relatively non-toxic to vertebrates. It has been suggested that several neonicotinoids cause neurodevelopmental toxicity in mammals. The aim was to establish the relationship between oral intake and urinary excretion of neonicotinoids by humans to facilitate biological monitoring, and to estimate dietary neonicotinoid intakes by Japanese adults. Methodology/Principal Findings Deuterium-labeled neonicotinoid (acetamiprid, clothianidin, dinotefuran, and imidacloprid) microdoses were orally ingested by nine healthy adults, and 24 h pooled urine samples were collected for 4 consecutive days after dosing. The excretion kinetics were modeled using one- and two-compartment models, then validated in a non-deuterium-labeled neonicotinoid microdose study involving 12 healthy adults. Increased urinary concentrations of labeled neonicotinoids were observed after dosing. Clothianidin was recovered unchanged within 3 days, and most dinotefuran was recovered unchanged within 1 day. Around 10% of the imidacloprid dose was excreted unchanged. Most of the acetamiprid was metabolized to desmethyl-acetamiprid. Spot urine samples from 373 Japanese adults were analyzed for neonicotinoids, and daily intakes were estimated. The estimated average daily intake of these neonicotinoids was 0.53–3.66 μg/day. The highest intake of any of the neonicotinoids in the study population was 64.5 μg/day for dinotefuran, and this was

RNF213/Mysterin has been identified as a susceptibility gene for moyamoya disease, a cerebrovascular disease characterized by occlusive lesions in the circle of Willis. The p. R4810K (rs112735431) variant is a founder polymorphism that is strongly associated with moyamoya disease in East Asia. Many non-p.R4810K rare variants of RNF213 have been identified in white moyamoya disease patients, although the ethnic mutations have not been investigated in this population. In the present study, we screened for RNF213 variants in 19 Slovakian and Czech moyamoya disease patients. A total of 69 RNF213 coding exons were directly sequenced in 18 probands and one relative who suffered from moyamoya disease in Slovakia and the Czech Republic. We previously reported one proband harboring RNF213 p.D4013N. Results from the present study identified four rare variants other than p.D4013N (p.R4019C, p.E4042K, p.V4146A, and p.W4677L) in four of the patients. P.V4146A was determined to be a novel de novo mutation, and p.R4019C and p. E4042K were identified as double mutations inherited on the same allele. P.W4677L, found in two moyamoya disease patients and an unaffected subject in the same pedigree, was a rare single nucleotide polymorphism. Functional analysis showed that RNF213 p.D4013N, p.R4019C and p.V4146A-transfected human umbilical vein endothelial cells displayed significant lowered migration, and RNF213 p.V4146A significantly reduced tube formation, indicating that these are disease-causing mutations. Results from the present study identified RNF213 rare variants in 22.2% (4/18 probands) of Slovakian and Czech moyamoya disease patients, confirming that RNF213 may also be a major causative gene in a relative large population of white patients.

Exposure of mothers to organochlorine pesticides (OCPs) was assessed by measuring the levels of 20 OCPs in 70 human breast milk samples pooled from 210 individuals from China, Korea and Japan. The OCPs were analyzed using gas chromatography/mass spectrometry (GC/MS) in electron capture negative ionization (ECNI) monitoring. The results showed that β-hexachlorocyclohexane and hexachlorobenzene were one order of magnitude higher in China than in the other nations, whereas chlordanes and polychlorinated biphenyl levels were highest in Japan. Heptachlor epoxide, dieldrin, endrin, toxaphenes and mirex were detected in most samples, and levels of these chemicals were significantly higher in Japan (0.8-4.5 ng g(-1) lipid), followed by Korea (0.2-4.7 ng g(-1) lipid), and lowest in China (less than 1.0 ng g(-1) lipid). α- and β-endosulfans were detected at a range of 0.9-1.5 ng g(-1) lipid levels in all samples analyzed, and their levels were higher in Korean than in Chinese samples.

Moyamoya disease-an idiopathic vascular disorder of intracranial arteries-is often accompanied by hypertension. RNF213 has been identified as a susceptibility gene for moyamoya disease. In the present study, the association of p.R4810K (GA) with blood pressure (BP) was investigated in a Japanese population.Three independent study populations, the Nyukawa (n = 984), Noshiro (n = 2,443) and Field (n = 881) studies, joined this study. BP, body weight and height were measured. Past and present symptoms and disease and medication histories were assessed by interview. Associations of p.R4810K (rs112735431, ss179362673) of RNF213 with BP were investigated. Two linkage disequilibrium blocks were constructed for moyamoya patients with p.R4810K (n = 140) and the general population (n = 384) using 39 single nucleotide polymorphisms (SNPs) spanning 390 kb around RNF213. A total of 60 carriers (3 for AA genotype and 57 for GA genotype) were found in these samples, and the minor allele frequencies were 1.4 % in the Nyukawa and Field studies and 0.2 % in the Noshiro study. Regression analyses adjusted for age, sex and body mass index based on an additive model demonstrated significant associations with systolic BP (mmHg/allele): β (standard error) was 8.2 (2.9) in the Nyukawa study (P = 4.7 × 10(-3)), 18.7 (5.4) in the Noshiro study (P = 4.6 × 10(-4)) and 8.9 (2.0) (P = 1.0 × 10(-5)) in the three populations. In contrast, diastolic BP showed significant associations only in the Noshiro study. Linkage disequilibrium blocks contained none of the BP-associated proxy SNPs reported by previous studies.Our study suggests that p.R4810K of RNF213 is associated strongly with systolic BP.

In this study, 90 human breast milk samples collected from Japan, Korea, and China were analyzed for perfluorooctanoic acid (PFOA) (C8), perfluorononanoic acid (PFNA) (C9), perfluorodecanoic acid (PFDA) (C10), perfluoroundecanoic acid (PFUnDA) (C11), perfluorododecanoic acid (PFDoDA) (C12), and perfluorotridecanoic acid (PFTrDA) (C13). In addition, infant formulas (n = 9) obtained from retail stores in China and Japan were analyzed. PFOA was the predominant compound and was detected in more than 60% of samples in all three countries. The PFOA, PFNA, PFDA, and PFUnDA levels in Japan were significantly higher than those in Korea and China (p0.05). The PFTrDA level was highest in Korea (p0.05). The median PFOA concentrations were 89 pg mL(-1) (48% of total perfluorinated carboxylic acids (PFCAs) (C8-C13)) in Japan, 62 pg mL(-1) (54%) in Korea, and 51 pg mL(-1) (61%) in China. The remaining ∑PFCAs (C9-C13) were 95 pg mL(-1) in Japan, 52 pg mL(-1) in Korea, and 33 pg mL(-1) in China. Among the long-chain PFCAs, odd-numbered PFCAs were more frequently detected than even-numbered PFCAs, except for PFDA in Japan. There were no evident correlations between the mother's demographic factors and the PFCA concentrations. PFOA, PFNA, and PFDA were frequently detected in both Japan and China, but there were no significant differences between the two countries. The total PFCA concentrations in the infant formulas were lower than those in the breast milk samples in Japan (p0.05), but not in China (p0.05). In conclusion, various PFCAs were detected in human breast milk samples from East Asian countries.

Japanese breast milk samples were tested for antibodies to human T-cell leukemia virus type I (HTLV-1) by particle agglutination (PA) and a line immunoassay (LIA). In the PA method, the agglutination reaction between the HTLV-1 antibody and sensitized particles occurred at a 1 : 128 dilution of some breast milk samples. The average antibody titer was one order of magnitude lower than that in the serum positive control. A total of 243 human breast milk specimens were assayed by PA, of which 21 samples from Okinawa, Hyogo, Miyagi and Hokkaido were positive or deferred. The results of the 21 positive samples were subsequently assayed by LIA (INNO-LIA™ HTLV I/II) for confirmation; and one sample was positive, and two were indeterminate. We attempted to use polymerase chain reaction (PCR) to detect HTLV-1 provirus DNA, but we did not detect PCR products for the pX1 region of the HTLV-1 genome in the LIA-positive samples. These negative PCR results are most likely due to the lower sensitivity of the PCR for amplification from milk than from HTLV-1-positive monocytes. In conclusion, the PA method to breast milk samples appears to be a suitable tool to screen for antibodies to HTLV-1 in the breast milk of carrier mothers in cases in which it would be difficult to use serum for the test. Although LIA may be able to confirm HTLV-1 infection, the presence of HTLV-1 provirus should be confirmed in the breast milk.

Perfluorooctanoic acid (PFOA) has recently attracted attention as a potential health risk following environmental contamination. However, information detailing exposure to perfluorinated carboxylic acids (PFCAs) other than PFOA is limited. We measured the concentrations of PFCAs (from perfluorohexanoic acid to perfluorotetradecanoic acid) in serum samples obtained from patients in Japan (Sendai, Takayama, Kyoto and Osaka) between 2002 and 2009, Korea (Busan and Seoul) between 1994 and 2008 and Vietnam (Hanoi) in 2007/2008. Total PFCA levels (geometric mean) were increased from 8.9 ng mL−1 to 10.3 ng mL−1 in Japan; from 7.0 ng mL−1 to 9.2 ng mL−1 in Korea; and were estimated at 4.7 ng mL−1 in Vietnam. PFCAs of greater length than PFOA were significantly increased in Sendai, Takayama and Kyoto, Japan, and levels of long-chain PFCAs exceeded PFOA levels in serum. Among these PFCAs, perfluoroundecanoic acid (PFUnDA) was the predominant component (28.5%), followed by perfluorononanoic acid (PFNA 17.5%), perfluorodecanoic acid (PFDA 7.9%), perfluorotridecanoic acid (PFTrDA 6.1%) and perfluorododecanoic acid (PFDoDA 1.8%). Odd-numbered PFCAs (PFNA, PFUnDA and PFTrDA) were also observed in Korea and Vietnam and their presence increased significantly in Korea between 1994 and 2007/2008. The proportion of long-chain PFCAs in serum was relatively high compared to reports in Western countries. Further investigations into the sources and exposure routes are needed to predict the future trajectory of these serum PFCA levels. Keywords: Perfluorocarboxylate, Perfluorooctanoic acid, Serum, Temporal trend, East Asia

The alarming rise in the prevalence of chronic kidney disease of uncertain etiology (CKDu) among the low socioeconomic farming community in the North Central Province of Sri Lanka has been recognized as an emerging public health issue in the country.This study sought to determine the possible factors associated with the progression and mortality of CKDu. The study utilized a single-center cohort registered in 2003 and followed up until 2009 in a regional clinic in the endemic region, and used a Cox proportional hazards model.We repeatedly found an association between disease progression and hypertension. Men were at higher risk of CKDu than women. A significant proportion of the patients in this cohort were underweight, which emphasized the need for future studies on the nutritional status of these patients.Compared with findings in western countries and other regions of Asia, we identified hypertension as a major risk factor for progression of CKDu in this cohort.

This was a screening study that aimed to determine the presence of nephrotoxic mycotoxins in urine samples from patients with chronic kidney disease of uncertain etiology in the North Central Province of Sri Lanka. The percentage detection of aflatoxins, ochratoxins and fumonisins in 31 patients were 61.29%, 93.5% and 19.4%, respectively. Geometric means of urinary aflatoxins and ochratoxins were 30.93 creatinine and 34.62 ng/g creatinine in chronic kidney disease of uncertain etiology stage 1–2 patients and 84.12 ng/g creatinine and 63.52 ng/g creatinine in unaffected relatives of patients. In chronic kidney disease of uncertain etiology stage 3–5 patients, geometric means of urinary aflatoxins and ochratoxins were 10.40 and 17.08 ng/g creatinine, respectively. Non-affected relatives of patients (n = 6) had comparable levels of these mycotoxins, but healthy Japanese individuals (n = 4) had lower levels than in Sri Lanka. The higher detection rate of urinary ochratoxins in Sri Lankans indicates that exposure is common in the region.

Recently, the Stockholm Convention prohibited the use of toxaphene and has been reviewing endosulfan. The historical use of these pesticides may contaminate food and tend to accumulate in the food chain. In this study, to evaluate the spatial and temporal trends of food contamination, the endosulfan and toxaphene levels were measured in pooled 24-h food composite samples from Chinese (n=10), Korean (n=10) and Japanese (n=40) adults in the 1990s and 2007-2009. Endosulfan was detected in 32 of 40 samples from Japan, but its levels (sum of α- and β-isomers) were low in both the 1990s and 2009 (range as geometric mean (geometric standard deviation) [GM (GSD)]: 0.96 (1.6)-1.42 (1.4) ng kg(-1)d(-1)). The dietary intakes of endosulfan in Seoul as GM (GSD) were 38.68 (1.3) ng kg bw(-1)d(-1) in 1994 and 92.17 (4.4) ng kg bw(-1)d(-1) in 2007, and significantly higher than those in Japan (p

Objective The aim was to investigate the genetic background of familial clustering of type 2 diabetes. Subjects and methods We recruited Japanese families with a 3-generation history of diabetes. Genome-wide linkage analysis was performed assuming an autosomal dominant model. Genes in the linkage region were computationally prioritized using Endeavour. We sequenced the candidate genes, and the frequencies of detected nucleotide changes were then examined in normoglycemic controls. Results To exclude known genetic factors, we sequenced 6 maturity onset diabetes of the young (MODY) genes in 10 familial cases. Because we detected a MODY3 mutation HNF1A R583G in one case, we excluded this case from further investigation. Linkage analysis revealed a significant linkage region on 2p25-22 (LOD score = 3.47) for 4 families. The 23.6-Mb linkage region contained 106 genes. Those genes were scored by computational prioritization. Eleven genes, i.e., top 10% of 106 genes, were selected and considered primary candidates. Considering their functions, we eliminated 3 well characterized genes and finally sequenced 8 genes. GCKR ranked highly in the computational prioritization. Mutations (minor allele frequency less than 1%) in exons and the promoter of GCKR were found in index cases of the families (3 of 18 alleles) more frequently than in controls (0 of 36 alleles, P = 0.033). In one pedigree with 9 affected members, the mutation GCKR g.6859C>G was concordant with affection status. No mutation in other 7 genes that ranked highly in the prioritization was concordant with affection status in families. Conclusions We propose that GCKR is a susceptibility gene in Japanese families with clustered diabetes. The family based approach seems to be complementary with a large population study.

Environmental ecological modeling (EEM), which unifies models simulating transport of chemicals and exposure of humans to chemicals, was used to simulate long-term trends of female adult human blood lead levels (BLLs) and historical exposure to the atmospheric lead in four East Asian countries: Japan, Korea, China, and Vietnam. Anthropogenic lead emissions to the atmosphere in Vietnam were estimated from energy statistics to be 1931 t yr(-1). Calculated BLLs generally agreed with those observed in samples collected in these countries as the error factors were less than 2. The model results revealed that BLLs decreased significantly in Tokyo (by 58%) and Seoul (by 45%) in recent decades and confirmed the effects of efforts to reduce environmental lead in Japan and Korea. The model results also revealed that BLLs in Beijing did not decrease in this decade as much as in Tokyo and Seoul, despite the phasing out of leaded gasoline, and that the contribution from the atmospheric component was increasing (43% in 2009). Finally, we applied EEM to simulate BLLs of children in Hanoi. The probability of children having BLLs greater than 50 μg L(-1) was 7.5%, which was greater than those observed in developed countries.

Previously, we demonstrated that the concentrations of DDTs were greater in breast milk collected from Chinese mothers than from Japanese and Korean mothers. To investigate dicofol as a possible source of the DDTs in human breast milk, we collected breast milk samples from 2007 to 2009 in China (Beijing), Korea (Seoul, Busan) and Japan (Sendai, Takarazuka and Takayama). Using these breast milk samples, we quantified the concentrations of dichlorobenzophenone, a pyrolysis product of dicofol (simply referred to as dicofol hereafter), dichlorodiphenyltrichloroethane and its metabolites (DDTs) using GC–MS. Overall, 12 of 14 pooled breast milk samples from 210 mothers contained detectable levels of dicofol (>0.1 ng g−1 lipid). The geometric mean concentration of dicofol in the Japanese breast milk samples was 0.3 ng g−1 lipid and significantly lower than that in Chinese (9.6 ng g−1 lipid) or Korean breast milk samples (1.9 ng g−1 lipid) (p < 0.05 for each). Furthermore, the ΣDDT levels in breast milk from China were 10-fold higher than those from Korea and Japan. The present results strongly suggest the presence of extensive emission sources of both dicofol and DDTs in China. However, exposure to dicofol cannot explain the large exposure of Chinese mothers to DDTs because of the trace levels of dicofol in the ΣDDTs. In the present study, dicofol was confirmed to be detectable in human breast milk. This is the first report to identify dicofol in human samples.

We previously performed genetic analysis in six unrelated families with infantile limb pain episodes, characterized by cold-induced deterioration and mitigation in adolescence, and reported two new mutations p.R222H/S in SCN11A responsible for these episodes. As no term described this syndrome (familial episodic pain: FEP) in Japanese, we named it as”小児四肢疼痛発作症”. In the current study, we recruited an additional 42 new unrelated Japanese FEP families, between March 2016 and March 2018, and identified a total of 11 mutations in SCN11A: p.R222H in seven families, and p.R225C, p.F814C, p.F1146S, or p.V1184A, in independent families. A founder mutation, SCN11A p.R222H was confirmed to be frequently observed in patients with FEP in the Tohoku region of Japan. We also identified two novel missense variants of SCN11A, p.F814C and p.F1146S. To evaluate the effects of these latter two mutations, we generated knock-in mouse models harboring p.F802C (F802C) and p.F1125S (F1125S), orthologues of the human p.F814C and p.F1146S, respectively. We then performed electrophysiological investigations using dorsal root ganglion neurons dissected from the 6–8 week-old mice. Dissected neurons of F802C and F1125S mice showed increased resting membrane potentials and firing frequency of the action potentials (APs) by high input–current stimulus compared with WT mice. Furthermore, the firing probability of evoked APs increased in low stimulus input in F1125S mice, whereas several AP parameters and current threshold did not differ significantly between either of the mutations and WT mice. These results suggest a higher level of excitability in the F802C or F1125S mice than in WT, and indicate that these novel mutations are gain of function mutations. It can be expected that a considerable number of potential patients with FEP may be the result of gain of function SCN11A mutations.

The increase in the number of chronic kidney disease (CKD) patients from the north central region of Sri Lanka has become a environmental health issue of national concern. Unlike in other countries where long-standing diabetes and hypertension are the leading causes of renal diseases, the majority of CKD patients from this part of Sri Lanka do not show any identifiable cause. As the disease is restricted to a remarkably specific geographical terrain, particularly in the north central dry zone of the country, multidisciplinary in-depth research studies are required to identify possible etiologies and risk factors. During this study, population screening in the prevalent region and outside the region, analysis of geoenvironmental and biochemical samples were carried out. Population screening that was carried out using a multistage sampling technique indicated that the point prevalence of CKD with uncertain etiology is about 2-3% among those above 18 years of age. Drinking water collected from high-prevalent and non-endemic regions was analyzed for their trace and ultratrace element contents, including the nephrotoxic heavy metals Cd and U using ICP-MS. The results indicate that the affected regions contain moderate to high levels of fluoride. The Cd contents in drinking water, rice from affected regions and urine from symptomatic and non-symptomatic patients were much lower indicating that Cd is not a contributing factor for CKD with uncertain etiology in Sri Lanka. Although no single geochemical parameter could be clearly and directly related to the CKD etiology on the basis of the elements determined during this study, it is very likely that the unique hydrogeochemistry of the drinking water is closely associated with the incidence of the disease.

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) have recently received attention owing to their widespread contamination in the environment. One of major manufacturers, 3M Company voluntarily phased out PFOS production in 2002. We measured the PFOS and PFOA concentrations in serum samples from Japan (Sendai, Takayama and Osaka), Korea (Busan and Seoul) and Vietnam (Hanoi) to evaluate the possible effects of the phase-out on the serum levels. There were spatial differences in both the serum PFOS and PFOA concentrations. The serum PFOS concentrations (ngmL(-1)) evaluated as the geometric mean (geometric standard deviation) in 2007-2008 ranged from 4.86 (1.45) in Sendai, Japan, to 9.36 (1.42) in Busan, Korea. The serum PFOA concentrations ranged from 0.575 (2.32) in Hanoi, Vietnam, to 14.2 (1.73) in Osaka, Japan. Historically archived samples collected from Korea in 1994-2008 revealed that the serum PFOA concentrations increased by 1.24-fold in Busan from 2000 to 2008 and 1.41-fold in Seoul from 1994 to 2007. On the other hand, the serum PFOS concentrations did not change from 1994 to 2007/2008. The serum PFOS levels in Japan in 2008 were significantly decreased compared with previously reported values (22.3-66.7% of the values in 2003/2004). However, the serum PFOA levels showed a clear decline from 2003 to 2008 in a high-exposed area, Osaka, but not in low-exposed areas in Japan. The trends toward decreases were not uniformly observed in Asian countries, unlike the case for the United States, suggesting that local factors associated with the production and introduction histories in each country overwhelm the effects of the phase-out.

Background The effects of coffee and green, black and oolong teas and caffeine intake on cardiovascular disease (CVD) mortality have not been well defined in Asian countries. Methods To examine the relationship between the consumption of these beverages and risk of mortality from CVD, 76 979 individuals aged 40–79 years free of stroke, coronary heart disease (CHD) and cancer at entry were prospectively followed. The daily consumption of beverages was assessed by questionnaires. Results 1362 deaths were documented from strokes and 650 deaths from CHD after 1 010 787 person-years of follow-up. Compared with non-drinkers of coffee, the multivariable HR and 95% CI for those drinking 1–6 cups/week, 1–2 cups/day and ≥3 cups/day were 0.78 (0.50 to 1.20), 0.67 (0.47 to 0.96) and 0.45 (0.17 to 0.87) for strokes among men (p=0.009 for trend). Compared with non-drinkers of green tea, the multivariable HR for those drinking 1–6 cups/week, 1–2 cups/day, 3–5 cups/day and ≥6 cups/day were 0.34 (0.06–1.75), 0.28 (0.07–1.11), 0.39 (0.18–0.85) and 0.42 (0.17–0.88) for CHD among women (p=0.038 for trend). As for oolong tea, the multivariable HR of those drinking 1–6 cups/week and ≥1 cups/day were 1.00 (0.65–1.55) and 0.39 (0.17–0.88) for total CVD among men (p=0.049 for trend). Risk reduction for total CVD across categories of caffeine intake was most prominently observed in the second highest quintile, with a 38% lower risk among men and 22% among women. Conclusions Consumption of coffee, green tea and oolong tea and total caffeine intake was associated with a reduced risk of mortality from CVD.