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We investigated the effects of a single and simultaneous intake of allitol and d-allulose on body fat accumulation and cecal short-chain fatty acid (SCFA) production and accurately assessed the contribution of rare sugars to body fat in rats fed a high-fat diet that led to obesity. Thirty-two male 3-week-old Wistar rats were randomly divided into four groups: control, allitol, d-allulose, and allitol + d-allulose. The rats were fed experimental diets and water ad libitum for 11 weeks. High doses of allitol or d-allulose can induce diarrhea in rat; hence, each group of rats was acclimated to 1-5% allitol and d-allulose incrementally for the initial 20 days. After the feeding period, all rats were euthanized and collected tissues. Perirenal, mesenteric, and total intra-abdominal adipose tissue weights were significantly reduced by dietary d-allulose, whereas dietary allitol tended to decrease these adipose tissue weights. Both allitol and d-allulose significantly decreased carcass and total body fat mass. We confirmed that both dietary allitol and d-allulose inhibited body fat accumulation; however, d-allulose did not inhibit hepatic lipogenesis and no synergy was observed between dietary allitol and d-allulose in terms of anti-obesity effects. Dietary allitol significantly increased cecal SCFA levels and these effects were more potent than those of dietary d-allulose. The antiobesity effect of allitol may be due to the action of SCFAs, especially butyric acid, produced by the gut microbiota. Many of the effects of allitol as an alternative sweetener remain unknown, and further research is required.

Introduction: As the incidence of gastric cancer (GC) is increasing in East Asia including Japan, a simple blood test for early GC is needed as an alternative to upper gastrointestinal (UGI) endoscopy. We performed this study to address this issue., Methods: We collected serum samples from 319 participants comprising 225 healthy subjects without GC (control group) and 94 patients with early GC (early GC group). After evaluating copy numbers of serum hTERT and methylated RUNX3 (m-RUNX3) using the Combined Restriction Digital PCR (CORD) assay, which we developed, we assessed the diagnostic performance of hTERT and m-RUNX3 for early GC., Results: Serum levels of hTERT and m-RUNX3 were significantly higher in the early GC group than in the control group. The area under the curve (AUC) was 0.89 for hTERT and 0.78 for m-RUNX3. Multivariate logistic regression analysis revealed age, sex, hTERT copy number, and m-RUNX3 copy number to be independent factors for early GC. We then established a prediction formula and named it the ASTEm-R3 (age, sex, hTERT, and m-RUNX3) index. The AUC of the ASTEm-R3 index was 0.93 with a sensitivity of 79.7% and specificity of 91.1%., Conclusion: We demonstrated excellent performance of the ASTEm-R3 index using the CORD assay to detect early GC. This index might be a promising alternative to UGI endoscopy., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

Abstract: To discharge waste liquid containing radioactive iodine into sewage systems, long-term storage or dilution with a large amount of water may be required until the radioactivity concentration reduces below the standard value. Processing the waste liquid could be easier if radioactive iodine could be separated from the water. This study verified the effectiveness of superabsorbent polymer and α-cyclodextrin as treatment agents to separate radioactive iodine from waste liquids. Sodium iodide (Na 125 I) was added to purified water and artificial urine to prepare simulated waste liquids containing iodine equivalent to the urine of patients treated with radioactive iodine. The as-prepared simulated waste liquid was poured into a container with superabsorbent polymer and left for 90 d. The residual iodine rate in the simulated waste liquid was estimated by measuring 125 I radioactivity. When the water was sufficiently dried, residual iodine rates on day 15 were 0.102 and 0.884 in the simulated waste liquids comprising purified water and artificial urine, respectively. The simulated waste liquid comprising purified water with 5% α-cyclodextrin absorbed by 1 g of superabsorbent polymer had a residual rate of 0.980. Moreover, the residual rate of simulated waste liquid comprising artificial urine with 2% α-cyclodextrin absorbed by 1 g of SAP was 0.949. Superabsorbent polymer combined with α-cyclodextrin was an effective treatment agent for separating radioactive iodine from waste liquids., (Copyright © 2024 Health Physics Society.)

Although the fecal immunochemical test for hemoglobin (FIT) is a widely used screening test for colorectal cancer, it is not sensitive enough to detect advanced colorectal adenoma. To address this issue, we performed this study to investigate whether combining the FIT and fecal DNA testing of methylated somatostatin (SST) could improve diagnostic performance for advanced colorectal adenoma. We collected feces from 79 healthy subjects with negative results on colonoscopy, 43 patients with non-advanced colorectal adenoma, 117 patients with advanced colorectal adenoma, and 126 patients with colorectal cancer. After fecal DNA was incubated with methylation-sensitive restriction enzymes, SST methylation levels were measured by droplet digital PCR. Using logistic multivariate analysis, we established a prediction formula for detecting colorectal neoplasia and named it the FAMS (FIT, age, methylated SST) index. The diagnostic performance of a single use of FIT for advanced colorectal adenoma showed a sensitivity of 29.1% (34/117) and specificity of 89.3% (109/122). In contrast, the FAMS index showed a sensitivity of 56.4% (66/117) at a similar specificity point of 91.0% (111/122). Furthermore, even at the higher specificity point of 94.3% (115/122), the sensitivity was still higher than that of FIT, reaching 42.7% (50/117). As the FAMS index showed better diagnostic performance for advanced colorectal adenoma than a single use of FIT, the FAMS index could be a promising tool for detecting advanced colorectal adenoma., (© 2024 The Author(s). Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

Elevated sperm mitochondrial DNA copy number (mtDNAcn) is associated with damage to sperm and poorer measures of semen quality. Exposure to cadmium (Cd) can increase oxidative stress and damage sperm mitochondria. The adverse effects of Cd can potentially be reduced by sufficient selenium (Se). The objective of this study was to examine the associations between sperm mtDNAcn and urinary concentrations of Cd and Se, as well as the Cd/Se molar ratio. Participants were recruited from patients who sought infertility treatment at two hospitals in Japan. Urine and semen specimens and self-administered questionnaires were collected on the day of recruitment. Sperm mtDNAcn was measured in extracted sperm DNA by multiplex real-time qPCR. Urinary Cd and Se concentrations were measured using inductively coupled plasma mass spectrometry, and their molar weights were calculated to obtain the Cd/Se molar ratio. Linear regression was used to estimate associations after adjusting for age, body mass index, smoking, drinking, exercise, varicocele, and hospital of recruitment. Sperm mtDNAcn showed statistically insignificant associations with creatinine-adjusted concentrations of urinary Cd (β = 0.13, 95% CI -0.18, 0.44) and Se (β = -0.09, 95% CI -0.54, 0.35), and Cd/Se molar ratio (β = 0.12, 95% CI -0.13, 0.37). The current study found no evidence of an association between mtDNAcn and urinary concentrations of Cd or Se, or the Cd/Se molar ratio., (© 2023. The Author(s).)

Adsorption reactions of various cations on clay minerals have different effects on their environmental behaviors depending on the molecular-scale adsorption structure. Some cations form outer-sphere complexes via hydration, while others create inner-sphere complexes through dehydration. This preference dictates their environmental impact. However, the factors controlling these complex formations remain unclear. Furthermore, research on the adsorption preferences of radium (Ra) is lacking. Thus, this study conducted the first EXAFS study of Ra 2+ adsorbed on clay minerals and showed that Ra 2+ forms inner-sphere complexes on vermiculite, which can be surprising because Ra 2+ is a divalent cation and prefers to be hydrated. In order to investigate the factors controlling the complex formations, this study conducted systematic EXAFS measurements and DFT calculations for alkali and alkaline earth metal cations. The results showed the importance of the size-matching effect between the adsorbed cation and the cavity of the tetrahedral sheets and that the complex formation can be estimated by the combination of the ionic radius and hydration enthalpy of the adsorbed cation. Furthermore, this study also analyzed environmental core samples. Their results showed the fixation of Ra 2+ by clay minerals and the controlling factors can effectively predict cation environmental behavior., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Akiko Yamaguchi reports financial support was provided by Japan Society for the Promotion of Science and by Japan Atomic Energy Agency., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

This study aimed to determine the applicability of a tail-attached device in monitoring animal-based indicators (ABI) associated with changes in environmental conditions in calves through investigating the relationship between sensor-derived ABI and the temperature-humidity index (THI). Furthermore, to identify effective ABI indicative of heat stress status, sensor-derived ABI of calves under differing heat stress levels based on rectal temperature (RT) were compared. The tail-attached device, which is capable of measuring skin temperature (ST), activity intensity, and roll angle along the longitudinal axis of the tail at 3-min intervals, was attached to 99 preweaning female Holstein calves for an average of 4 wk (26.4 ± 6.8 d). After selecting data from mild to hot days (daily average THI of ≥55), physiological (daily maximum tail ST) and behavioral (daily average activity intensity, daily total lying time, and daily total body position change) ABI were computed, and their relationship with the daily average THI was determined using piecewise regression analysis. Additionally, during the hot season, RT of 20 randomly selected tested calves were measured thrice a week (every 2.4 ± 0.5 d), and a comparison was conducted between the ABI of calves with normal RT (<39.5°C) and those with high RT (≥39.5°C), utilizing data from days characterized by potential heat stress (daily average THI of ≥75). During the study, ambient temperature (°C) and relative humidity (%) were recorded every 10 min using an automatic digital data logger, from which the daily average THI was calculated. Piecewise regression analysis identified THI breakpoints of 73.6 for tail ST, 79.1 for average activity intensity, 72.3 for lying time, and 79.1 for position change. All the tested ABI tended to increase as THI increased, and this trend was pronounced in tail ST, activity intensity, and position change after the breakpoint. These 3 ABI were higher in calves with high RT compared with those with normal RT, whereas lying time shared similar values between the RT groups. Overall, these findings suggest that the tail-attached device can simultaneously monitor both physiological and behavioral ABI in calves, and among the ABI, tail ST, activity intensity, and position change are the effective ABI indicative of heat stress status., (© 2024.)

Introduction: Patients with hepatitis virus-related hepatocellular carcinoma (viral HCC) are decreasing as hepatitis control improves, but those with non-viral-related HCC (non-viral HCC) are increasing in Japan. No established surveillance system exists for patients with non-viral HCC, so they are often diagnosed at an advanced stage. To address this, we performed this study., Methods: We collected serum samples from 516 participants (154 healthy subjects, 93 chronic liver disease [CLD] patients without HCC, and 269 HCC patients). Participants were divided into a control group comprising healthy subjects and patients with CLD and an HCC group. We evaluated serum methylated HOXA1 (m- HOXA1 ) copy numbers using modified combined restriction digital PCR (CORD) assay (1-step CORD assay). We assessed diagnostic performance of m- HOXA1 compared to HCC tumor markers alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP) and created a novel index to improve HCC prediction., Results: Serum m- HOXA1 level was significantly higher in each HCC stage group versus the control group. Its sensitivity was 69.1% and specificity was 78.5% for diagnosing HCC. The area under the curve (AUC) of m- HOXA1 was superior to that of AFP and equal to that of DCP. Multivariate logistic regression analysis revealed independent contributions of m- HOXA1 , DCP, and AFP, in that order of strength, to diagnose HCC after adjustment for age and sex. We designated the predictive probability of HCC based on the regression model as the ASDAm-H1 (Age, Sex, DCP, AFP, and m- HOXA1 ) index. Its diagnostic accuracy was 0.96 by AUC with a sensitivity of 86.2% and specificity of 93.9%. Sensitivity was identical for viral and non-viral HCCs. When limited to early-stage HCC, sensitivity of the ASDAm-H1 index was 76.3%., Conclusions: We showed distinguished performance of the ASDAm-H1 index to detect viral and non-viral HCC, even at an early stage. This index might have potential as a non-viral HCC surveillance system., Competing Interests: T. Yamasaki received grant support from Eiken Chemical Co., Ltd. All other authors declare no conflicts of interest., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

In this study, we investigated whether monitoring the ventral tail base surface temperature (ST) using a wearable wireless sensor could be effective for fever detection in calves with experimentally induced pneumonia after inoculation with Histophilus somni strain 2336. We found a significant difference in the changes in ST values between the control and H. somni-inoculated groups after 24 h of inoculation and detected fever; however, the rectal temperature showed a significant difference between the groups after 12 h of inoculation. When a significant difference in the ST between the two groups was observed, serum haptoglobin concentration and exacerbation of clinical score increased in the H. somni-inoculated group compared with those in the control group. Pneumonia was observed in the H. somni-inoculated group at necropsy, indicating that the changes in ST may reflect fever with inflammation caused by H. somni infection. Our results demonstrated that monitoring ST using a sensor attached to the ventral tail base can detect fever in calves and may be a useful and labor-saving tool for the health management of calves., (© 2024 Japanese Society of Animal Science.)

Radiocesium released by the Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident still exists in the environment in two forms: adsorbed species on mineral particles in the soil and microparticles containing radiocesium mainly composed of silicate glass (CsMPs). CsMPs are dispersed not only around the FDNPP but also over a wide area of the Kanto region. The behavior and characteristics of CsMPs must be investigated to evaluate the impact of the FDNPP accident. Deposited particles including radiocesium were wiped from metal handrails on balconies and car hoods using tissue papers at six locations in the Kanto region (Tokai village, Ushiku City, Abiko City, Chiba City, Kawaguchi City, and Arakawa Ward) between March 15 and 21, 2011. CsMPs were isolated from the samples, and their characteristics were investigated. In total, 106 CsMPs derived from Unit 2 were successfully separated from 13 tissue paper samples. The radiation images of the two types of CsMPs discovered in Ushiku City demonstrate that CsMPs can easily become susceptible to fragmentation over time, even in the absence of weathering effects., (© 2023. The Author(s).)

Abstract: The plume from Fukushima Dai-ichi Nuclear Power Plant (FDNPP) Unit 2, which reached the Tokyo metropolitan area on 15 March 2011 immediately after the FDNPP disaster, contained insoluble radiocesium-bearing microparticles (CsMPs). This study aimed to reanalyze a mask worn during the disaster to determine the ratio of CsMPs to radiocesium that directly arrived in the Tokyo metropolitan area as a plume immediately after the disaster. CsMPs were isolated from the non-woven masks worn at the Hongo Campus of the University of Tokyo from 15-16 March 2011 that trapped radioactive material from the accident. Twenty-two CsMPs with diameters in the range of 1.9-3.6 μm were found in the mask worn outdoors by one person. The CsMP radioactive ratio to overall radiocesium content of the mask was 9.9%. The CsMPs were collected more efficiently by the mask than by a suspended particulate matter filter. A face mask might trap CsMPs in a plume more efficiently than a suspended particulate matter filter. Determining the amount of CsMPs led to a reassessment of the internal dose by CsMPs that were inhaled by the public immediately after the FDNPP accident., (Copyright © 2023 Health Physics Society.)

Tektins are a group of microtubule-stabilizing proteins necessary for cilia and flagella assembly. TEKTIN1 (TEKT1) is used as a sperm marker for monitoring germ cell differentiation in embryonic stem (ES) and induced pluripotent stem (iPS) cells. Although upregulation of TEKT1 has been reported during spontaneous differentiation of ES and iPS cells, it is unclear which cells express TEKT1. To identify TEKT1-expressing cells, we established an ES cell line derived from cynomolgus monkeys (Macaca fascicularis), which expresses Venus controlled by the TEKT1 promoter. Venus expression was detected at 5 weeks of differentiation on the surface of the embryoid body (EB), and it gradually increased with the concomitant formation of a leash-like structure at the EB periphery. Motile cilia were observed on the surface of the Venus-positive leash-like structure after 8 weeks of differentiation. The expression of cilia markers as well as TEKT1-5 and 9 + 2 microtubule structures, which are characteristic of motile cilia, were detected in Venus-positive cells. These results demonstrated that TEKT1-expressing cells are multiciliated epithelial-like cells that form a leash-like structure during the spontaneous differentiation of ES and iPS cells. These findings will provide a new research strategy for studying cilia biology, including ciliogenesis and ciliopathies., (© 2023 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.)

A decade has passed since the Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident on March 11, 2011. However, radioactive particles have recently been detected in the indoor air of some residences near the FDNPP. Following the recommendations of previous research, we determined the presence of radiocesium-bearing microparticles (CsMPs) and measured the radioactivity of radiocesium that adhered on non-woven face masks worn by six persons during the indoor cleaning of 59 residences in Namie, Futaba, Okuma, and Tomioka towns in Fukushima Prefecture. Of the 284 masks worn in this study, significant 137 Cs radioactivity was detected in 268, and 44 new CsMPs were discovered in 28. The results of this study also suggest the presence of highly concentrated soluble radiocesium particles or soluble radioactive cesium aerosols adhered to house dust. This implies that the CsMPs constituted a large proportion of radioactivity in the indoor air contamination for particles in the 1.0-2.5 µm size range due to the radioactive radiocesium particles. It is desirable to wear masks during cleaning to prevent inhalation of CsMPs., (© 2023. The Author(s).)

It is desirable to attend to the mare at the time of foaling in order to assist fetal delivery and prevent complications. The early detection of the onset of labor is an important issue for the equine industry. The purpose of this study was to examine the applicability of a sensor for foaling detection using the data of surface temperature (ST), roll angle (rotation about the y-axis) and y-axis (long axis of the tail) acceleration which were collected from a multimodal device attached to the ventral tail base of the mare. The data were collected every 3 minutes in 17 pregnant mares. Roll angle differences from the reference values and the mare's posture (standing or recumbent) confirmed by video were compared and associated. Cohen's kappa coefficient was 0.99 when the threshold was set as ± 0.3 radian in roll angle differences. This result clearly showed that the sensor data can accurately distinguish between standing and recumbent postures. The hourly sensor data with a lower ST (LST < 35.5°C), a recumbent posture determined by the roll angle, and tail-raising (TR, decline of 200 mg or more from the reference value in y-axis acceleration) was significantly higher during the last hour prepartum than 2-120 hours before parturition (P < 0.01). The accuracy of foaling detection within one hour was verified using the following three indicators: LST; lying down (LD, change from standing to recumbent posture); and TR. When LST, LD and TR were individually examined, even though all indicators showed that sensitivity was 100%, the precision was 13.1%, 8.1% and 2.8%, respectively. When the data were combined as LST+LD, LST+TR, LD+TR and LST+LD+TR, detection of foaling improved, with precisions of 100%, 32.1%, 56.7% and 100%, respectively. In conclusion, the tail-attached multimodal device examined in this present study is useful for detecting foaling., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Aoki et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Nonylphenol (NP), an endocrine disrupting chemical, is widely used in industrial and agricultural processes, causing NP influx into aquatic environments. NP induces hormonal imbalance, and male feminization, and reduces germ cell production during spermatogenesis; however, the mechanism by which it affects spermatogenesis remains unknown. Here, we investigated the effect of NP on spermatogenesis in honmoroko (Gnathopogon caerulescens), an endangered fish endemic to Lake Biwa, Japan, using an in vitro differentiation system. We collected spermatogonia from the testes of non-spawning G. caerulescens and subjected them to suspension culture. The spermatogonia differentiated into flagellated spermatozoa in 3 weeks, regardless of the presence of NP. NP concentrations as low as 1 nM caused a decrease in the number of germ cells in a dose-dependent manner, whereas the number of somatic cells decreased only at a high concentration of 1 μM. Flow cytometric analysis revealed that the decrease in germ cell number was attributed to haploids (spermatids and spermatozoa); the number of spermatogonia and spermatocytes was not affected by NP treatment. This result is consistent with the hypothesis that NP might repress the second meiosis or induce apoptosis in haploids. This study demonstrated that the combination of in vitro germ cell differentiation and flow cytometric analysis is useful for evaluating the direct effects of NP on germ cell differentiation in endangered endemic fish., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Alpha-cyclodextrin, a six D-glucose cyclic oligosaccharide, has several applications in food and pharmaceuticals, but has also been reported to retain iodine in a stable manner for 16 months. Radioactive iodine, which may cause thyroid cancer and hypofunction, must be properly managed. If the absorption of radioactive iodine is suppressed, it can be expected to lead to a reduction in thyroid exposure. This study clarified the inhibition of radioactive iodine absorption by the oral administration of α-cyclodextrin in a murine model using direct measurement of single photon emission computed tomography. The uptake of radioactive iodine into the thyroid gland in mice administered with radioactive iodine and an α-cyclodextrin solution was approximately 40% lower after 24 h. The finding that oral uptake of α-cyclodextrin has an inhibitory effect on the transfer of radioactive iodine to the thyroid gland has potential for application in many fields such as food, pharmaceuticals, nuclear emergency preparedness, and medicine., (© 2023. The Author(s).)

The present study investigated the applicability of a calving prediction model based on supervised machine learning of ruminal temperature (RT) data in dairy cows. The existence of cow subgroups for prepartum RT changes was also examined, and the predictive performance of the model was compared among these subgroups. RT data were collected from 24 Holstein cows at 10 min intervals using an RT sensor system. The average hourly RT was calculated and data were expressed as residual RTs (rRT = actual RT - mean RT for the same time on the previous three days). The mean rRT decreased beginning at approximately 48 h before calving to a low of -0.5°C at 5 h before calving. However, two cow subgroups were identified: cows with a late and small rRT decrease (Cluster 1, n = 9) and those with an early and large rRT decrease (Cluster 2, n = 15). A calving prediction model was developed using five features extracted from the sensor data (indicative of prepartum rRT changes) through a support vector machine. Cross-validation showed that calving within 24 h was predicted with a sensitivity of 87.5% (21/24) and precision of 77.8% (21/27). A significant difference in sensitivity was observed between Clusters 1 and 2 (66.7 vs. 100%, respectively), while none was observed for precision. Therefore, the model based on RT data with supervised machine learning has the potential to efficiently predict calving, although improvements for specific cow subgroups are required.

A 1.2 m flexible liquid scintillation light guide (LSLG) detector connected to a portable electric device was developed and applied to monitor X-rays scattered from a panoramic dental X-ray imaging apparatus. The X-ray absorption of the LSLG tube was simulated for 20 and 40 keV X-rays. The LSLG detector was calibrated by comparing doses measured by a 3-inch NaI(Tl) detector using Am-241 and Cs-137 sources. A linear relationship was obtained between dose rates (μSv/h) and count rates (cps). The elapsed time profile of scattered radiation from a panoramic dental X-ray imaging apparatus was determined. Local absorbed doses were measured with small OSL dosimeters on the LSLG tube suspended over the shoulder and the eyeglass. Behind the ears and neck (in the center of the LSLG tube) showed high doses. The LSLG detector was useful for real-time monitoring of scattered X-rays., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Whole-body vibration (WBV) is a type of light-resistance exercise that involves exposing the body to rapid and repeated oscillations of a vibrating platform. It has been suggested that long-term WBV can improve bone mass and muscle strength. However, little is known about its effects on body composition, and the safety and efficacy of WBV have not been established. In this study, we investigated the effects of WBV on body fat loss and muscle mass maintenance or improvement in male Wistar rats fed standard or high-fat diets. We also aimed to establish a rat model for future nutritional and physiological studies. We conducted two experiments using male Wistar rats that were 3 weeks old. The rats were randomly divided into two groups: the control group and the vibration group. The rats were fed either a commercial standard diet (Experiment 1) or a high-fat diet (Experiment 2) ad libitum for 8-12 weeks. Our results showed that WBV stimulus dramatically reduced body fat accumulation in rats fed a high-fat diet but not in those fed a standard diet. This suggests that WBV may be particularly effective under dietary conditions that promote obesity. Moreover, WBV increased the mass of several skeletal muscles, which are known to have resistance exercise effects. Our findings indicate that long-term WBV is safe, with no inhibition of growth or feeding. Taken together, our results suggest that WBV may be a promising approach for preventing and treating obesity. Further research is needed to investigate the underlying mechanisms and to determine the optimal WBV for maximum benefits.

Background: Although faecal DNA testing of Fusobacterium nucleatum ( Fn ) is expected to be useful for colorectal neoplasia detection, there is no standardized quantification method of Fn . We performed this study to establish a possible standardized method., Methods: In this study, 322 participants including 71 subjects without colorectal neoplasia (control group), 31 patients with non-advanced colorectal adenoma, 93 patients with advanced colorectal adenoma, and 127 patients with colorectal cancer were enrolled. Faecal Fn were quantified by droplet digital PCR (ddPCR) using two PCR primer-probe sets reported previously that are tentatively named Fn1 and Fn2. Fn1 has been used in ddPCR by us and Fn2 has been widely used in quantitative real-time PCR., Results: The Fn copy number using Fn1 was five times higher than that using Fn2, with a linear relationship shown between them. Receiver operating characteristic curve analysis showed the area under the curve (AUC) to be almost the same between Fn1 and Fn2 in discriminating between the control group and the colorectal cancer group (AUC = 0.81 and 0.81, respectively), and between the control/non-advanced colorectal adenoma group and the advanced colorectal adenoma/colorectal cancer group (AUC = 0.74 and 0.74, respectively)., Conclusions: As the diagnostic performance was quite similar between Fn1 and Fn2, ddPCR-based Fn testing using Fn1 and Fn2 could be a possible standardized method for a colorectal neoplasia screening test, considering that Fn levels quantified by Fn1 are about five times higher than those by Fn2.

In this study, we developed calving prediction models for 24-h and 6-h periods before calving using data on physiological (tail skin temperature) and behavioral (activity intensity, lying time, posture change, and tail raising) parameters obtained using a multimodal tail-attached device (tail sensor). The efficiencies of the models were validated under tethering (tie-stall) and untethering (free-stall and individual pen) conditions. Data were collected from 33 and 30 pregnant cattle under tethering and untethering conditions, respectively, from approximately 15 days before the expected calving date. Based on pre-calving changes, 40 features (8 physiological and 32 behavioral) were extracted from the sensor data, and one non-sensor-based feature (days to the expected calving date) was added to develop models using a support vector machine. Cross-validation showed that calving within the next 24 h under tethering and untethering conditions was predicted with a sensitivity of 97% and 93% and precision of 80% and 76%, respectively, while calving within the next 6 h was predicted with a sensitivity of 91% and 90% and precision of 88% and 90%, respectively. Calving prediction models based on the tail sensor data with supervised machine learning have the potential to achieve effective calving prediction, irrespective of the cattle housing conditions.

Sweetspire (Itea) is the only plant that accumulates rare sugars d-allulose and allitol. However, no reports have indicated that sweetspire has a beneficial physiological activity in mammalians. We have examined the effect of dietary dried sweetspire powder (SP) on body fat accumulation in rats fed with a high-fat diet. Twenty-four male Wistar rats were randomized into three groups, the control (C), SP, and rare sugar (RS) groups. The SP diet contained 5% SP (contained 0.4% d-allulose and 0.6% allitol in the diet), and the RS diet contained the same amount of rare sugars as the SP diet. All rats were given free access to the experimental diets for 8 weeks. The percentages of intra-abdominal adipose tissue and total body fat were significantly lower in the SP group than in the C group, suggesting that SP has an anti-obesity effect. Furthermore, this anti-obesity effect may be attributed to the rare sugars in SP.

Introduction: Production of 99 Mo/ 99m Tc using an electron linear accelerator (linac) and activated carbon (AC)-based 99m Tc generator (linac-AC) is an alternative approach to the conventional fission production of 99 Mo/ 99m Tc. As a preliminary investigation of the clinical applicability of a linac-AC-derived 99m Tc radiopharmaceutical, the biodistribution of linac-AC-derived [ 99m Tc]sodium pertechnetate ([ 99m Tc]NaTcO 4 ) was measured and compared against fission-derived [ 99m Tc]NaTcO 4 at one time point., Methods: 99 Mo was produced by irradiating nonenriched MoO 3 targets with bremsstrahlung photons generated from 55.5-MeV linac electron beams. 99m Tc was then separated and purified from the 99 Mo using an AC-based 99m Tc generator. Subsequently, biodistribution of the linac-AC-derived [ 99m Tc]NaTcO 4 in healthy female Slc:ICR mice (n = 6) was measured by dissection and compared with that of fission-derived [ 99m Tc]NaTcO 4 (n = 4) at 30 min after injection., Results: The two types of [ 99m Tc]NaTcO 4 exhibited similar biodistribution in all the organs and tissues examined: the uptakes of [ 99m Tc]NaTcO 4 prepared from the linac-AC method and those prepared from the fission method were 138.9 ± 69.9%ID/g and 160.6 ± 49.2%ID/g in the thyroids, respectively, 33.4 ± 5.5%ID/g and 29.4 ± 9.1%ID/g in the salivary glands, respectively, and less than 10%ID/g in blood and all the other organs. No adverse effects were observed in the mice administered with either [ 99m Tc]NaTcO 4 ., Conclusion: The clinical applicability of linac-AC-derived [ 99m Tc]NaTcO 4 was suggested by its similar biodistribution with fission-derived [ 99m Tc]NaTcO 4 at one time point. Further biodistribution studies at multiple time points are encouraged to demonstrate the bioequivalence between linac-AC- and fission-derived [ 99m Tc]NaTcO 4 ., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Dementia with Lewy bodies (DLB) is the second most common form of neurodegenerative dementia in elderly people, following Alzheimer's disease. Only three genes, SNCA (α-synuclein), APOE (apolipoprotein E), and GBA (glucosylceramidase), have been convincingly demonstrated to be associated with DLB. Here, we applied whole-genome sequencing to blood samples from 61 DLB patients and 45 cognitively normal controls. We used accumulation of candidate mutations to detect novel DLB-associated genes. Subsequent single nucleotide polymorphism (SNP) genotyping and association studies in a large number of samples from Japanese individuals revealed novel heterozygous variants in MFSD3 (rs143475431, c.888T>A:p.C296*; n = 5,421, p = 0.00063) and MRPL43 (chr10:102746730, c.241A>C:p.N81H; n = 4,782, p = 0.0029). We further found that the MFSD3 variant increased plasma levels of butyrylcholinesterase (n = 1,206, p = 0.029). We believe that our findings will contribute to the understanding of DLB and provide insight into its pathogenic mechanism for future studies., (© 2022 The Authors. American Journal of Medical Genetics Part B: Neuropsychiatric Genetics published by Wiley Periodicals LLC.)

Introduction: We previously developed a novel methylation assay, the combined restriction digital PCR (CORD) assay, consisting of treatment of DNA with methylation-sensitive restriction enzymes and droplet digital PCR., Methods: In this study, we assessed the diagnostic performance of serum methylated Homeobox A1 (mHOXA1) and methylated somatostatin (mSST) using the CORD assay in combination with CA19-9 for pancreatic cancer using serum samples from 82 healthy individuals, 13 patients with benign pancreatic disease, 3 patients with branched-duct intraductal papillary mucinous neoplasm, and 91 patients with pancreatic cancer., Results: For the single marker tests, sensitivity for all stages of pancreatic cancer, stage I cancer, and specificity were, respectively, 71.4%, 50.0%, and 94.9% for CA19-9; 51.6%, 68.8%, and 90.8% for mHOXA1; and 50.1%, 68.8%, and 94.9% for mSST. Those for the combined marker tests were, respectively, 86.8%, 81.3%, and 85.7% for combined mHOXA1 and CA19-9; 86.8%, 87.5%, and 89.8% for combined mSST and CA19-9; and 89.0%, 87.5%, and 85.7% for all three markers combined., Conclusion: The combination of mHOXA1 and mSST with CA19-9 appears to be useful to detect pancreatic cancer even at an early stage., (© 2022 S. Karger AG, Basel.)

Allitol is a rare sugar alcohol obtained by reducing d-allulose (d-psicose). However, information on the effects of long-term dietary allitol intake is limited. This study aimed to investigate the effect of allitol supplementation, as a sugar substitute, on body fat accumulation in rats compared with sucrose, rare sugar d-allulose, or erythritol. Thirty-two male Wistar rats (3 wk old) were fed experimental diets including 5% sucrose, allitol, erythritol, or d-allulose for 8 wk ad libitum. Weight gain, food intake, and food efficiency did not differ among the groups. The total body fat mass and percentage, and intra-abdominal adipose tissue weights were significantly lower in rats fed with the allitol diet than in those fed with the sucrose diet. These body fat indicators tended to be lower in rats fed with the erythritol and d-allulose diets than in those fed with the sucrose diet, but there was no significant difference. The serum glucose-lowering effect obtained in rats fed with the d-allulose diet did not appear in rats fed with the allitol diet. These results suggest that the anti-obesity effect of allitol may be equal to or greater than that of d-allulose.

MicroRNAs (miRNAs) are small non-coding RNAs shown to regulate gene expression by binding to complementary transcripts. Genetic variants, including single-nucleotide polymorphisms and short insertions/deletions, contribute to traits and diseases by influencing miRNA expression. However, the association between genetic variation and miRNA expression remains to be elucidated. Here, by using genotype data and miRNA expression data from 3448 Japanese serum samples, we developed a computational pipeline to systematically identify genome-wide miRNA expression quantitative trait loci (miR-eQTLs). Not only did we identify a total of 2487 cis-miR-eQTLs and 3 155 773 trans-miR-eQTLs at a false discovery rate of <0.05 in six dementia types (Alzheimer's disease, dementia with Lewy bodies, vascular dementia, frontotemporal lobar degeneration, normal-pressure hydrocephalus and mild cognitive impairment) and all samples, including those from patients with other types of dementia, but also we examined the commonality and specificity of miR-eQTLs among dementia types. To enable data searching and downloading of these cis- and trans-eQTLs, we developed a user-friendly database named JAMIR-eQTL, publicly available at https://www.jamir-eqtl.org/. This is the first miR-eQTL database designed for dementia types. Our integrative and comprehensive resource will contribute to understanding the genetic basis of miRNA expression as well as to the discovery of deleterious mutations, particularly in dementia studies. Database URL: https://www.jamir-eqtl.org/., (© The Author(s) 2021. Published by Oxford University Press.)

Fishes expressing a fluorescent protein in germ cells are useful to perform germ cell transfer experiments for conservation study. Nonetheless, no such fish has been generated in endangered endemic fishes. In this study, we tried to produce a fish expressing Venus fluorescent protein in germ cells using Honmoroko (Gnathopogon caerulescens), which is one of the threatened small cyprinid endemic to the ancient Lake Biwa in Japan. To achieve germ cell-specific expression of Venus, we used piwil1 (formally known as ziwi) promoter and Tol2 transposon system. Following the co-injection of the piwil1-Venus expression vector and the Tol2 transposase mRNA into fertilized eggs, presumptive transgenic fish were reared. At 7 months of post-fertilization, about 19% (10/52) of the examined larvae showed Venus fluorescence in their gonad specifically. Immunohistological staining and in vitro spermatogenesis using gonads of the juvenile founder fish revealed that Venus expression was detected in spermatogonia and spermatocyte in male, and oogonia and stage I and II oocytes in female. These results indicate that the Tol2 transposon and zebrafish piwil1 promoter enabled gene transfer and germ cell-specific expression of Venus in G. caerulescens. In addition, in vitro culture of juvenile spermatogonia enables the rapid validation of temporal expression of transgene during spermatogenesis., (© 2021 Fisheries Society of the British Isles.)

In this study, we determined the applicability of the background image subtraction technique to detect estrus in tie-stalled cows. To investigate the impact of the camera shooting direction, webcams were set up to capture the front, top, and rear views of a cow simultaneously. Video recording was performed for a total of ten estrous cycles in six cows. Standing estrus was confirmed by testing at 6 h intervals. From the end of estrus, transrectal ultrasonography was performed every 2 h to confirm ovulation time. Foreground objects (moving objects) were extracted in the videos using the background subtraction technique, and the pixels were counted at each frame of five frames-per-second sequences. After calculating the hourly averaged pixel counts, the change in values was expressed as the pixel ratio (total value during the last 24 h/total value during the last 24 to 48 h). The mean pixel ratio gradually increased at approximately 48 h before ovulation, and the highest value was observed at estrus, regardless of the camera shooting direction. When using front-view videos with an appropriate threshold, estrus was detected with 90% sensitivity and 50% precision. The present method in particular has the potential to be a non-contact estrus detection method for tie-stalled cows.

A part of the radiocaesium from the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident was emitted as glassy, water-resistant caesium-bearing microparticles (CsMPs). Here, we isolated and investigated seven CsMPs from marine particulate matter and sediment. From the elemental composition, the 134 Cs/ 137 Cs activity ratio, and the 137 Cs activity per unit volume results, we inferred that the five CsMPs collected from particulate matter were emitted from Unit 2 of the FDNPP, whereas the two CsMPs collected from marine sediment were possibly emitted from Unit 3, as suggested by (i) the presence of calcium and absence of zinc and (ii) the direction of the atmospheric plume during the radionuclide emission event from Unit 3. The presence of CsMPs can cause overestimation of the solid-water distribution coefficient of Cs in marine sediments and particulate matter and a high apparent radiocaesium concentration factor for marine biota. CsMPs emitted from Unit 2, which were collected from the estuary of a river that flowed through a highly contaminated area, may have been deposited on land and then transported by the river. By contrast, CsMPs emitted from Unit 3 were possibly transported eastward by the wind and deposited directly onto the ocean surface.

Alzheimer's disease (AD) has no cure, but early detection and risk prediction could allow earlier intervention. Genetic risk factors may differ between ethnic populations. To discover novel susceptibility loci of AD in the Japanese population, we conducted a genome-wide association study (GWAS) with 3962 AD cases and 4074 controls. Out of 4,852,957 genetic markers that passed stringent quality control filters, 134 in nine loci, including APOE and SORL1, were convincingly associated with AD. Lead SNPs located in seven novel loci were genotyped in an independent Japanese AD case-control cohort. The novel locus FAM47E reached genome-wide significance in a meta-analysis of association results. This is the first report associating the FAM47E locus with AD in the Japanese population. A trans-ethnic meta-analysis combining the results of the Japanese data sets with summary statistics from stage 1 data of the International Genomics of Alzheimer's Project identified an additional novel susceptibility locus in OR2B2. Our data highlight the importance of performing GWAS in non-European populations.

We aimed to determine the effectiveness of estrus detection based on continuous measurements of the ventral tail base surface temperature (ST) with supervised machine learning in cattle. ST data were obtained through 51 estrus cycles on 11 female cattle (six Holsteins and five Japanese Blacks) using the tail-attached sensor. Three estrus detection models were constructed with the training data (n = 17) using machine learning techniques (random forest, artificial neural network, and support vector machine) based on 13 features extracted from sensing data (indicative of estrus-associated ST changes). Estrus detection abilities of the three models on test data (n = 34) were not statistically different among models in terms of sensitivity and precision (range 50.0% to 58.8% and 60.6% to 73.1%, respectively). The relatively poor performance of the models might indicate the difficulty of separating estrus-associated ST changes from estrus-independent fluctuations in ST.

Background: Mild cognitive impairment (MCI) is a precursor to Alzheimer's disease (AD), but not all MCI patients develop AD. Biomarkers for early detection of individuals at high risk for MCI-to-AD conversion are urgently required., Methods: We used blood-based microRNA expression profiles and genomic data of 197 Japanese MCI patients to construct a prognosis prediction model based on a Cox proportional hazard model. We examined the biological significance of our findings with single nucleotide polymorphism-microRNA pairs (miR-eQTLs) by focusing on the target genes of the miRNAs. We investigated functional modules from the target genes with the occurrence of hub genes though a large-scale protein-protein interaction network analysis. We further examined the expression of the genes in 610 blood samples (271 ADs, 248 MCIs, and 91 cognitively normal elderly subjects [CNs])., Results: The final prediction model, composed of 24 miR-eQTLs and three clinical factors (age, sex, and APOE4 alleles), successfully classified MCI patients into low and high risk of MCI-to-AD conversion (log-rank test P = 3.44 × 10 -4 and achieved a concordance index of 0.702 on an independent test set. Four important hub genes associated with AD pathogenesis (SHC1, FOXO1, GSK3B, and PTEN) were identified in a network-based meta-analysis of miR-eQTL target genes. RNA-seq data from 610 blood samples showed statistically significant differences in PTEN expression between MCI and AD and in SHC1 expression between CN and AD (PTEN, P = 0.023; SHC1, P = 0.049)., Conclusions: Our proposed model was demonstrated to be effective in MCI-to-AD conversion prediction. A network-based meta-analysis of miR-eQTL target genes identified important hub genes associated with AD pathogenesis. Accurate prediction of MCI-to-AD conversion would enable earlier intervention for MCI patients at high risk, potentially reducing conversion to AD.

In this study, we developed a calving prediction model based on continuous measurements of ventral tail base skin temperature (ST) with supervised machine learning and evaluated the predictive ability of the model in 2 dairy farms with distinct cattle management practices. The ST data were collected at 2- or 10-min intervals from 105 and 33 pregnant cattle (mean ± standard deviation: 2.2 ± 1.8 parities) reared in farms A (freestall barn, in a temperate climate) and B (tiestall barn, in a subarctic climate), respectively. After extracting maximum hourly ST, the change in values was expressed as residual ST (rST = actual hourly ST - mean ST for the same hour on the previous 3 d) and analyzed. In both farms, rST decreased in a biphasic manner before calving. Briefly, an ambient temperature-independent gradual decrease occurred from around 36 to 16 h before calving, and an ambient temperature-dependent sharp decrease occurred from around 6 h before until calving. To make a universal calving prediction model, training data were prepared from pregnant cattle under different ambient temperatures (10 data sets were randomly selected from each of the 3 ambient temperature groups: <15°C, ≥15°C to <25°C, and ≥25°C in farm A). An hourly calving prediction model was then constructed with the training data by support vector machine based on 15 features extracted from sensing data (indicative of pre-calving rST changes) and 1 feature from non-sensor-based data (days to expected calving date). When the prediction model was applied to the data that were not part of the training process, calving within the next 24 h was predicted with sensitivities and precisions of 85.3% and 71.9% in farm A (n = 75), and 81.8% and 67.5% in farm B (n = 33), respectively. No differences were observed in means and variances of intervals from the calving alerts to actual calving between farms (12.7 ± 5.8 and 13.0 ± 5.6 h in farms A and B, respectively). Above all, a calving prediction model based on continuous measurement of ST with supervised machine learning has the potential to achieve effective calving prediction, irrespective of the rearing condition in dairy cattle., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

A part of radiocesium emitted during the Fukushima nuclear accident was incorporated in glassy water-resistant microparticles, called Type-A particles, which are spherical with ~ 0.1 to 10 µm diameter and ~ 10 -2 to 10 2  Bq cesium-137 ( 137 Cs) radioactivity; they were emitted from Unit 2 or 3 of the Fukushima Daiichi Nuclear Power Plant. Meanwhile, Type-B particles, having various shapes, 50-400 µm diameter, and 10 1 -10 4  Bq 137 Cs radioactivity, were emitted from Unit 1. The chemical properties of these radioactive particles have been reported in detail, but previous studies investigated only a small number of particles, especially Type-B particles. We tried to understand radioactive particles systematically by analyzing a large number of particles. Micro-X-ray computed tomography combined with X-ray fluorescence analysis revealed the presence of many voids and iron-rich part within Type-B particles. The 137 Cs concentration (Bq mm -3 ) of Type-A particles was ~ 10,000 times higher than that of Type-B particles. Among the Type-B particles, the spherical ones had higher concentration of volatile elements than the non-spherical ones. These differences suggested that Type-A particles were formed through gas condensation, whereas Type-B particles were formed through melt solidification. These findings might contribute to the safe decommissioning of reactors and environmental impact assessment.

Background: With demographic shifts toward older populations, the number of people with dementia is steadily increasing. Alzheimer's disease (AD) is the most common cause of dementia, and no curative treatment is available. The current best strategy is to delay disease progression and to practice early intervention to reduce the number of patients that ultimately develop AD. Therefore, promising novel biomarkers for early diagnosis are urgently required., Methods: To identify blood-based biomarkers for early diagnosis of AD, we performed RNA sequencing (RNA-seq) analysis of 610 blood samples, representing 271 patients with AD, 91 cognitively normal (CN) adults, and 248 subjects with mild cognitive impairment (MCI). We first estimated cell-type proportions among AD, MCI, and CN samples from the bulk RNA-seq data using CIBERSORT and then examined the differentially expressed genes (DEGs) between AD and CN samples. To gain further insight into the biological functions of the DEGs, we performed gene set enrichment analysis (GSEA) and network-based meta-analysis., Results: In the cell-type distribution analysis, we found a significant association between the proportion of neutrophils and AD prognosis at a false discovery rate (FDR) < 0.05. Furthermore, a similar trend emerged in the results of routine blood tests from a large number of samples (n = 3,099: AD, 1,605; MCI, 994; CN, 500). In addition, GSEA and network-based meta-analysis based on DEGs between AD and CN samples revealed functional modules and important hub genes associated with the pathogenesis of AD. The risk prediction model constructed by using the proportion of neutrophils and the most important hub genes (EEF2 and RPL7) achieved a high AUC of 0.878 in a validation cohort; when further applied to a prospective cohort, the model achieved a high accuracy of 0.727., Conclusions: Our model was demonstrated to be effective in prospective AD risk prediction. These findings indicate the discovery of potential biomarkers for early diagnosis of AD, and their further improvement may lead to future practical clinical use.

Introduction: We have reported previously that fecal DNA testing of TWIST1 methylation in combination with the fecal immunochemical test for hemoglobin (FIT) (combination test) is useful for colorectal neoplasia screening. In this study, using larger sample sizes, we studied the clinical performance of the combination test for the detection of colorectal neoplasia and, especially, advanced colorectal adenoma., Methods: We performed a prospective study in which FIT, fecal DNA testing of TWIST1 methylation, and colonoscopy were performed on 372 patients with colorectal neoplasia and 71 subjects without colorectal neoplasia. We assessed the individual clinical performance of each of FIT and fecal DNA testing of TWIST1 methylation and of the combination test for the detection of colorectal neoplasia including advanced adenoma based on morphologic subtypes., Results: The FIT alone had a sensitivity of 7.5% (3/40) for nonadvanced adenoma, 32.3% (41/127) for advanced adenoma, and 93.7% (192/205) for colorectal cancer and a specificity of 87.3% (62/71). The combination test had a sensitivity of 35.0% (14/40) for nonadvanced adenoma, 68.5% (87/127) for advanced adenoma, and 95.6% (196/205) for colorectal cancer and a specificity of 80.3% (57/71). For morphological subtypes of advanced adenoma, the sensitivity of FIT was only 28.2% (20/71) for polypoid type and 16.1% (5/31) for nonpolypoid type, whereas the combination test increased the sensitivities to 64.8% (46/71) and 71.0% (22/31), respectively., Discussion: The combination of the fecal DNA test with FIT seemed to be useful to detect colorectal neoplasia and, especially, advanced adenoma of the nonpolypoid type.

To investigate the publics' internal exposure by inhalation of radiocesium from the Fukushima Daiichi nuclear disaster in 2011, we examined the activity of radiocesium and radiocesium-bearing particles adhering to nonwoven fabric masks worn daily by members of the public in spring 2013 and 2014. We found a maximum cumulative Cs activity of 4.58 ± 0.15 Bq in 4 wk of spring 2013, which is 20.8% of the activity measured for the same subject in spring 2012 using the same method. This decrease was faster than the physical decay of radiocesium. Radiocesium was detected in 21 of 722 masks in 2013; three of these included type A radiocesium-bearing particles. The activity ratio of the radiocesium-bearing particles on the mask to the total radiocesium was at most approximately 20%. The two radiocesium sources, radiocesium-bearing particles and fugitive dust, are both insoluble particles. The largest internal dose from inhalation was 7.6 μSv in spring 2013, which is negligible compared to the dose limit recommended for members of the public by the International Commission on Radiological Protection.

The main modalities for gastric cancer screening are limited to upper gastrointestinal endoscopy and contrast radiography. The former is invasive, and the latter has high false-negative rates. Thus, alternative diagnostic strategies are required. One solution may be a liquid biopsy. Methylated RUNX3 is a well-known biomarker of gastric cancer but it is very difficult to detect with conventional bisulfite-based methylation assays when only a small amount of serum is available. We developed the combined restriction digital PCR (CORD) assay, a new methylation assay allowing for the counting of as little as one copy of a methylated gene in a small sample of DNA without necessitating DNA bisulfite treatment. We evaluated the sensitivity and specificity of the serum DNA testing of methylated RUNX3 by the CORD assay for the detection of early gastric cancer using 50 patients with early gastric cancer and 61 control individuals. The CORD assay had a sensitivity of 50.0% and a specificity of 80.3% for early gastric cancer. Methylated RUNX3 copies were significantly associated with tumor size, massive submucosal invasion, and lymph-vascular invasion. After the treatment, the median number of methylated RUNX3 copies was significantly decreased. The CORD assay may provide an alternative screening strategy to detect even early-stage gastric cancer.

Spherical radioactive caesium (Cs)-bearing microparticles (CsMPs) were emitted during the Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident in March, 2011. The emission source (timing) and formation process of these particles remain unclear. In this study, the isotopic ratios of uranium ( 235 U and 238 U) and caesium ( 133 Cs, 134 Cs, 135 Cs, and 137 Cs) isotopes in the five spherical CsMPs (ca. 2 μm in size) sampled at 50 km west of the FDNPP were determined using secondary ion mass spectrometry and laser ablation-ICPMS, respectively. Results showed that the 235 U/ 238 U ratios of CsMPs were homogeneous (1.93 ± 0.03, N = 4) and close to those estimated for the fuel cores in units 2 and 3, and that the Cs isotopic ratios of CsMP were identical to those of units 2 and 3. These results indicated that U and Cs in the spherical CsMPs originated exclusively from the fuel melt in the reactors. Based on a thorough review of literatures related to the detailed atmospheric releases of radionuclides, the flow of plumes from the FDNPP reactor units during the accident and the U and Cs isotopic ratio results in this study, we hereby suggest that the spherical CsMPs originate only from the fuel in unit 2 on the night of 14 March to the morning of 15 March. The variation range of the analysed 235 U/ 238 U isotopic ratios for the four spherical particles was extremely narrow. Thus, U may have been homogenised in the source through the formation of fuel melt, which ultimately evaporating and taken into CsMPs in the reactor and was released from the unit 2.

Liquid biopsies are not used in practice for hepatocellular carcinoma (HCC). Epi proColon is the first commercial blood-based test for colorectal cancer screening based on methylated DNA testing of the septin 9 gene ( SEPT9 ). However, Epi proColon has some disadvantages, including the requirement of a large amount of blood and lack of quantitative performance. Therefore, we previously developed a novel liquid biopsy test that can quantitatively detect even a single copy of methylated SEPT9 in a small amount of DNA. In the current study, we evaluated the application potential of this assay for diagnosing HCC. Study subjects included 80 healthy volunteers, 45 patients with chronic liver disease (CLD) without HCC, and 136 patients with HCC (stage 0, 12; stage A, 50; stage B, 31; stage C, 41; and stage D, 2), according to the Barcelona Clinic Liver Cancer staging system. For the assay, DNA was treated with methylation-sensitive restriction enzymes in two steps, followed by multiplex droplet digital polymerase chain reaction. The median copy number of methylated SEPT9 was 0.0, 2.0, and 6.4 in the healthy control, CLD, and HCC groups, respectively, with significant differences among the groups (HCC vs. healthy control, P  < 0.001; HCC vs. CLD, P  = 0.002; CLD vs. healthy control, P  = 0.008). Assay sensitivity and specificity were 63.2% and 90.0%, respectively (cutoff value, 4.6 copies), in detecting HCC when compared with healthy subjects. The positive rate of methylated SEPT9 increased with HCC progression (stage 0, 41.7%; stage A, 58.0%; stage B, 61.3%; stage C, 75.6%; and stage D, 100%). Conclusion : We developed a sensitive methylated SEPT9 assay that might serve as a liquid biopsy test for diagnosing HCC., (© 2020 The Authors. Hepatology Communications published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases.)

Effects of endocrine disrupting chemicals (EDCs) on reproduction have not been fully explained comprehensively. In this study, we tried to validate the common effect of Bisphenol A (BPA) and Nonylphenol (NP) on the differentiation of embryonic stem (ES) cells and found that they modify the expression of germ cell specific genes. To elucidate functional significance on biological process, we performed Gene Ontology (GO)-based microarray analysis comparing with published GeneChip data of primordial germ cell development in vivo. Cluster analysis of gene expression profile revealed that EDC treatment and primordial germ cell (PGC) development shared characteristic cluster consists of GO terms related to "germ cell development" and "reproduction". In the GO term "reproduction", meiosis related genes showed high expression level by EDC exposure. These results suggest that BPA and NP affect not only some of the germ cell specific genes, but functionally interferes germ cell development and reproduction., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This paper describes an overview of the radiation protection response to the Plutonium intake accident that occurred at the Plutonium Fuel Facility of the Oarai Research and Development Center of the Japan Atomic Energy Agency on 6 June 2017. In the hood of the analyzing room at the Plutonium Fuel Facility five workers were checking a storage container of fast reactor nuclear fuel material. Around 11:15 a.m., vinyl bags inside the fuel material container containing Plutonium and enriched uranium burst during the inspection work. All the workers heard the bang, which caused misty dust leakage from the container. This event caused significant both skin and nasal α-contamination for three workers and just skin α-contamination for one worker. Decontamination was conducted in the shower room. Then the five workers were transferred to the Nuclear Fuel Cycle Engineering Laboratory to evaluate inhalation intake of Plutonium etc in the lungs. The maximum values of 2.2 × 10 4 Bq for 239 Pu and 2.2 × 10 2 Bq for 241 Am were estimated by the lung monitor. Based on these results, injection of a chelate agent was conducted for prompt excretion of Plutonium etc. The next morning, the five workers were transferred to the National Institute of Radiological Sciences for treatment including decontamination of their skin and measurement by a lung monitor. At that time no obvious energy peak was confirmed for Plutonium. The Japan Health Physics Society launched an ad-hoc working group for Plutonium intake accident around the middle of June to survey issues and to extract lessons for radiological protection. The authors, who are the members of the ad-hoc working group, here report the activity of the working group.

This study aimed to evaluate the effectiveness of estrous detection technique based on continuous measurements of vaginal temperature (VT) and conductivity (VC) with supervised machine learning in cattle. The VT and VC of 17 cows in tie-stalls were measured using our developed wearable vaginal sensor from Day 11 (Day 0 = ovulation day) to Day 11 of the subsequent estrous cycle at 15-min interval. After the maximum VT and VC were extracted hourly, their changes were expressed as residual VT (rVT = actual VT - mean VT for the same time on the previous 3 days) and as VC ratio (VCr = actual VC/mean VC for the same time on Day 11-13), respectively, and were used for analysis. Trans-rectal ultrasonography was performed to monitor ovarian structure changes. The plasma concentrations of reproductive hormones (progesterone: P 4 , estradiol-17β: E 2 , and LH) were measured in the experimental period. Standing estrus was confirmed by testing with herd mates at 3-h interval. The rVT decreased transiently, which coincided with decreasing P 4 a few days before estrus, and a sharp increase was associated with LH surge during estrus. The VCr increased as estrus approached, corresponding with decreasing P 4 and increasing E 2 and LH. After noise reduction, features, possible to follow-up estrus-associated changes in rVT and VCr, were extracted and used for developing estrous detection models; 9 models were developed with 3 feature sets (features extracted from rVT alone, VCr alone, and combination of rVT and VCr) and 3 machine learning algorithms (decision tree: DT, support vector machine: SVM, and artificial neural network: ANN). Cross-validation showed that models using the features from the combination of rVT and VCr showed better performance in terms of sensitivity and precision than those using features from VCr alone, and precision than those of using features from rVT alone. Within the models using the features from the combination of rVT and VCr, sensitivity and precision of the model generated by ANN were numerically, but not statistically, higher than those generated by DT and SVM. Of 17 estruses, 16 were detected, with one false positive, when the best model was used. Furthermore, both mean and variance of the interval from the beginning of the estrous detection alert to ovulation (27.3 ± 6.7 h, mean ± SD of 16 estruses) were not significantly different to those from the onset of standing estrus to ovulation (30.8 ± 5.8 h, n = 17), indicating that the estrus can be detected real-time by the present technique. Hence, the estrous detection technique based on continuous measurements of VT and VC with supervised machine learning has a potential for efficient and accurate estrous detection in cattle., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Purpose: The aim of this study is to determine the efficacy of quantitative real-time PCR (qPCR) and clinical characteristics to diagnose ocular cytomegalovirus (CMV) infections., Methods: The technical factors were assessed by the outcomes of the qPCR assay at five institutions in Japan using the WHO International Standard of cytomegalovirus. The clinical factors were assessed by examining the aqueous humor samples of 197 eyes of 197 consecutive patients suspected of CMV using the receiver operating characteristics (ROCs)., Results: All of the institutions had excellent detection efficacy, although the copy number ranged from 0.82 to 4.66 copies/IU. In the clinical samples, CMV was detected in 51 eyes, and the amount of CMV DNA was significantly higher for CMV retinitis. In corneal diseases, the amount of CMV DNA was significantly associated with frequency of recurrences and IOP elevations. The sensitivity and specificity of qPCR for the diagnosis was 90.0 and 98.7%, respectively. For the corneal and anterior uveitis types of CMV diseases, the area under the curve (AUC) of qPCR was 0.95 and 0.96, followed by frequency of recurrences with AUC of 0.89 and 0.82, and IOP elevations with AUC of 0.78 and 0.76. Unclassified cytomegalovirus detection, which did not meet diagnostic criteria of CMV corneal endotheliitis, anterior uveitis, or retinitis, was 4.6%, and it was significantly associated with corneal diseases and history of corneal transplantation., Conclusions: qPCR with standardization is specific and accurate; however, the inclusion and knowledge of the clinical characteristics improve the diagnostic efficacy.

MiRNA molecules are important post-transcriptional regulators of gene expression in the brain function. Altered miRNA profiles could represent a defensive response against the pathogenesis of neurodegenerative disorders, such as Alzheimer's disease (AD). Endogenous miRNAs have lower toxic effects than other gene silencing methods, thus enhancing the expression of defensive miRNA could be an effective therapy. However, little is known about the potential of targeting miRNAs for the treatment of AD. Here, we examined the function of the miR-200 family (miR-200a, -141, -429, -200b, -200c), identified using miRNA microarray analysis of cortical tissue from Tg2576 transgenic mice. In murine primary neurons, we found that upregulation of miR-200b or -200c was induced by the addition of amyloid beta (Aβ). Neurons transfected with miR-200b or -200c reduced secretion of Aβ in conditioned medium. Moreover, mice infused with miR-200b/c into the brain were relieved of memory impairments induced by intracerebroventricular injection of oligomeric Aβ, and demonstrated proper spatial learning in the Barnes maze. To gain further understanding of the relationship between miR-200b/c and Aβ, we identified target mRNAs via an RNA-binding protein immunoprecipitation-microarray assay. Western blot analysis showed that expression of ribosomal protein S6 kinase B1 (S6K1), a candidate target, was inhibited by miR-200c. S6K1, a downstream effector of mammalian target of rapamycin (mTOR), serves as a negative feedback mediator that phosphorylates insulin receptor substrate 1 at serine residues (IRS-1pSer). S6K1-dependent IRS-1pSer suppresses insulin signaling leading to insulin resistance, which is frequently observed in AD brains. Notably, miR-200b/c transfection of SH-SY5Y cells reduced the levels of IRS-1pSer. This finding indicates that miR-200b/c has the potential to alleviate insulin resistance via modulation of S6K1. Taken together, miR-200b/c may contribute to reduce Aβ secretion and Aβ-induced cognitive impairment by promoting insulin signaling.

We investigated the feasibility of cryopreservation of spermatogonia and oogonia in the critically endangered cyprinid honmoroko Gnathopogon caerulescens using slow-cooling (freezing) and rapid-cooling (vitrification) methods. Initially, we examined the testicular cell toxicities and glass-forming properties of the five cryoprotectants: ethylene glycol (EG), glycerol (GC), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (BG), and we determined cryoprotectant concentrations that are suitable for freezing and vitrification solutions, respectively. Subsequently, we prepared the freezing solutions of EG, GC, DMSO, PG, and BG at 3, 2, 3, 2, and 2 M and vitrification solutions at 7, 6, 5, 5, and 4 M, respectively. Following the cryopreservation of the testicular cells mainly containing early-stage spermatogenic cells (e.g., spermatogonia and primary spermatocytes), cells were cultured for 7 days and immunochemically stained against germ cell marker protein Vasa. Areas occupied by Vasa-positive cells indicated that vitrification led to better survival of germ cells than the freezing method, and the best result was obtained with 5 M PG, about 50% recovery of germ cells following vitrification. In the case of ovarian cells containing oogonia and stage I, II, and IIIa oocytes, vitrification with 5 M DMSO resulted the best survival of oogonia, with equivalent cell numbers to those cultured without vitrification. The present data suggest that male and female gonial cells of the endangered species G. caerulescens can be efficiently cryopreserved using suitable cryoprotectants for spermatogonia and oogonia, respectively.

Although methylated TWIST1 is a biomarker of colorectal neoplasia, its detection from serum samples is very difficult by conventional bisulfite-based methylation assays. Therefore, we have developed a new methylation assay that enables counting of even one copy of a methylated gene in a small DNA sample amount without DNA bisulfite treatment. We performed this study to evaluate the sensitivity and specificity of serum DNA testing by the new methylation assay in combination with and without the fecal immunochemical test for hemoglobin for the detection of colorectal neoplasia. This study comprised 113 patients with colorectal neoplasia and 25 control individuals. For the new methylation assay, DNA was treated in two stages with methylation-sensitive restriction enzymes, followed by measurement of copy numbers of hTERT and methylated TWIST1 by multiplex droplet digital PCR. The fecal immunochemical test had a sensitivity of 8.0% for non-advanced adenoma, 24.3% for advanced adenoma, and 44.4% for colorectal cancer, and a specificity of 88.0%. The new assay had a sensitivity of 36.0% for non-advanced adenoma, 30.0% for advanced adenoma, and 44.4% for colorectal cancer, and a specificity of 92.0%. Combination of the both tests increased the sensitivity to 40.0%, 45.7%, and 72.2% for the detection of non-advanced adenoma, advanced adenoma, and colorectal cancer, respectively, and resulted in a specificity of 84.0%. Combination of both tests may provide an alternative screening strategy for colorectal neoplasia including potentially precancerous lesions and colorectal cancer., Competing Interests: CONFLICTS OF INTEREST The authors have no conflicts of interest.

Background As TWIST1 methylation is specific to colorectal neoplasia, detection of TWIST1 methylation from faeces samples might be useful for colorectal neoplasia screening. However, because the content of human DNA in faeces is very small, it is very difficult to detect TWIST1 methylation by conventional bisulphite-based methylation assays. Therefore, we developed a new methylation assay without bisulphite treatment, the combined restriction digital PCR assay, and evaluated its sensitivity and specificity in combination with and without the faecal immunochemical test for haemoglobin for colorectal neoplasia detection from faeces samples. Methods For the combined restriction digital PCR assay, DNA was treated with three methylation-sensitive restriction enzymes and an exonuclease, followed by measurement of TWIST1 methylation level by droplet digital PCR. Faecal DNA testing and faecal immunochemical test for haemoglobin were performed on 109 patients with colorectal neoplasia and 10 control individuals. Results Basic performance testing showed that the combined restriction digital PCR assay enabled detection of 0.14% of the TWIST1 methylation level for the lymphocyte DNA. The combined restriction digital PCR assay from faeces samples had a sensitivity of 22.2% (95% confidence interval, 2.8-60.0%) for non-advanced adenoma, 47.1% (95% confidence interval, 23.0-72.2%) for advanced adenoma, and 33.7% (95% confidence interval, 23.7-45.0%) for colorectal cancer, and a specificity of 100.0%. Combination of faecal immunochemical test for haemoglobin and faecal combined restriction digital PCR assay increased sensitivity to 82.4% (95% confidence interval, 56.6-96.2%) for the detection of advanced adenoma. Conclusions We developed the combined restriction digital PCR assay, a possible highly sensitive methylation assay. Combination of faecal combined restriction digital PCR assay with faecal immunochemical test for haemoglobin may provide an alternative screening strategy for colorectal neoplasia, especially for potentially precancerous lesions.