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Elite athletes frequently invest in the use of supplements to optimize their dietary regimens and enhance their athletic performance. However, unregulated and unplanned use of supplements can lead to adverse consequences, including anti-doping rule violations or health issues. Thus, athletes should verify their diets, consider scientific evidence, and take necessary precautions regarding supplements before use. To date, no study has explored whether athletes check these factors before using supplements. This study aimed to investigate supplement use using a questionnaire administered to 1,392 athletes (including candidate athletes) who participated in the Tokyo 2020 Olympic/Paralympic and Beijing 2022 Winter Olympic/Paralympic Games. Participants were categorized as follows: 1,040 participants in the Tokyo 2020 Olympic Games, 83 in the Tokyo 2020 Paralympic Games, 239 in the Beijing 2022 Winter Olympic Games, and 30 in the Beijing 2022 Winter Paralympic Games. We collected data on supplement use and gained further knowledge through interviews with the athletes. Approximately 70% of Tokyo 2020 Olympic/Paralympic and Beijing 2022 Winter Olympic athletes and approximately 50% of Beijing 2022 Winter Paralympians used supplements. Over 50% of athletes had not received a doctor's diagnosis or a dietitian's evaluation before supplement use. Moreover, only 50% of the athletes who used dietary supplements reviewed the scientific evidence for the dietary supplements before using them and justified their choice based on their own investigation, while those who did not use dietary supplements cited either a lack of need or fear of an anti-doping rule violation. Considering the holistic health and performance of athletes, as well as the risk associated with unregulated use, such as overdose and anti-doping rule violations, there is a need for nutritional education on supplement use for athletes and their entourages., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Myoenzono, Yasuda, Takai, Shinagawa, Kaneko, Yoshizaki, Namma-Motonaga, Yoshino, Kondo, Nakajima, Hangai, Kamahara, Kamihigashi, Kusano and Kamei.)

The "meal first" strategy is traditionally recommended for athletes' conditioning. However, the importance of the "meal first" principle has not been detailly well documented in athletes' lives. Supplement use has recently become a common part of athletes' diets, but unmonitored supplement use can cause negative consequences, such as anti-doping violations and health issues. Therefore, this review summarizes how the "meal first" strategy and planned supplement use are important for enhancing athletes' health and performance. We believe that the "meal first" strategy is beneficial in terms of the following aspects: (1) consumption of multi-nutrients and other functional components simultaneously; (2) positive effects on psychological well-being; (3) contribution to athletes' health by way of mastication; and (4) less risk for anti-doping violations. Before supplement use, we recommend that athletes first verify their basic factors (e.g., diet, training, and sleep), given that the benefits of supplements are examined and demonstrated with the control of those factors. Otherwise, athletes cannot obtain maximal benefits from the supplements. In contrast, there are situations in which supplements in athletes' lives can be advantageous, such as (1) nutrient deficiency due to ongoing dietary characteristics; (2) interruption of meals due to disease; (3) inaccessibility of quality food during athletic travel; (4) difficulty preparing food due to societal restrictions associated with disasters or infection outbreaks; (5) having a meal before, during, or after exercise is difficult; and (6) achieving targeted intake of performance-enhancing ingredients is not practical. In summary, we emphasize that the "meal first" strategy is recommended for athletes' conditioning, but there are several contexts when supplement use can be more useful in athletes' lives., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Yasuda, Myoenzono, Takai, Toguchi, Tsunezumi, Kondo, Kaizaki, Ode, Ohno, Namma-Motonaga and Kamei.)

Neuroendocrine carcinoma (NEC) is a highly aggressive subtype of the neuroendocrine tumor with an extremely poor prognosis. We have previously conducted a comprehensive genomic analysis of over 100 cases of NEC of the gastrointestinal system (GIS-NEC) and unraveled its unique and organ-specific genomic drivers. However, the epigenomic features of GIS-NEC remain unexplored. In this study, we have described the epigenomic landscape of GIS-NEC and small cell lung carcinoma (SCLC) by integrating motif enrichment analysis from the assay of transposase-accessible chromatin sequencing (ATAC-seq) and enhancer profiling from a novel cleavage under targets and tagmentation (CUT&Tag) assay for H3K27ac and identified ELF3 as one of the super-enhancer-related transcriptional factors in NEC. By combining CUT&Tag and knockdown RNA sequencing for ELF3, we uncovered the transcriptional network regulated by ELF3 and defined its distinctive gene signature, including AURKA, CDC25B, CLDN4, ITGB6, and YWAHB. Furthermore, a loss-of-function assay revealed that ELF3 depletion led to poor cell viability. Finally, using gene expression of clinical samples, we successfully divided GIS-NEC patients into two subgroups according to the ELF3 signature and demonstrated that tumor-promoting pathways were activated in the ELF3 signature-high group. Our findings highlight the transcriptional regulation of ELF3 as an oncogenic transcription factor and its tumor-promoting properties in NEC., (© 2023 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

Neuronal intranuclear inclusion disease (NIID) is a clinically complex neurological disorder that appears sporadically or autosomally. Expansions of intronic GGC trinucleotide repeats in the NOTCH2 N-terminal-like C (NOTCH2NLC) gene cause NIID. In this study, to clarify the clinical characteristics useful for the differential diagnosis of NIID, clinical data of neurological examination, neuroimaging, and nerve conduction studies of six NIID patients diagnosed by pathological or genetic investigations were analyzed. Clinically useful characteristics for diagnosing NIID include general hyporeflexia, episodic disturbance of consciousness, sensory disturbance, miosis, and dementia. Furthermore, neuroimaging findings, such as leukoencephalopathy in T2-weighted magnetic resonance imaging and a linear high intensity of subcortical U-fibers in diffusion-weighted imaging (DWI), as well as decreased motor nerve conduction velocity, are especially important biomarkers for NIID. However, it is necessary to remember that these features may not always be present, as shown in one of the cases who did not have a DWI abnormality in this study. This study also investigated whether expanded GGC repeats were translated into polyglycine. Immunohistochemical analysis using a custom antibody raised against putative C-terminal polypeptides followed by polyglycine of uN2CpolyG revealed that polyglycines were localized in the intranuclear inclusions in skin biopsy specimens from all six patients, suggesting its involvement in the pathogenesis of NIID., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Purpose: Muscle glycogen storage before a race is necessary for endurance athletes to achieve the best performance. Generally, the recommended carbohydrate intake for preparation over 90 min of the race is 10-12 g·kg--1·day--1. However, it remains unclear whether an elite athlete with an already high-carbohydrate diet can further increase muscle glycogen through a very-high-carbohydrate intake. Therefore, we compared the effects of three types of glycogen loading in a 28-year-old male athlete who belongs to the top 50 racewalkers in the world, consuming a daily energy intake of 4507 kcal and a carbohydrate intake of 12.7 g·kg--1·day--1., Methods: The racewalker consumed very-high-carbohydrate diets three times for 2 days each, 13.7 g·kg--1·day--1 for trial 1, 13.9 g·kg--1·day--1 for trial 2, and 15.9 g·kg--1·day-1 for trial 3. Muscle glycogen concentrations in the anterior (vastus lateralis and vastus intermedius) and posterior thighs (semimembranosus, semitendinosus, and biceps femoris) were measured using carbon-13 magnetic resonance spectroscopy., Results: Muscle glycogen concentrations in both the anterior and posterior thighs increased in all trials, particularly in trial 3. Body mass also increased by 1.5 kg in trials 1 and 2 and by 1.8 kg in trial 3 before and after the trials. The participant felt satiated throughout the day and experienced stomach discomfort during trial 3., Conclusion: We found that a 2-day very-high-carbohydrate diet and tapering of training could further increase the muscle glycogen concentration in athletes. However, we speculated that 15.9 g·kg--1·day--1 carbohy.

A highly sensitive and highly multiplexed quantification technique for nucleic acids is necessary to predict and evaluate cancer treatment by liquid biopsy. Digital PCR (dPCR) is a highly sensitive quantification technique, but conventional dPCR discriminates multiple targets by the color of the fluorescent dye of the probe, which limits multiplexing beyond the number of colors of fluorescent dyes. We previously developed a highly multiplexed dPCR technique combined with melting curve analysis. Herein, we improved the detection efficiency and accuracy of multiplexed dPCR with melting curve analysis to detect KRAS mutations in circulating tumor DNA (ctDNA) prepared from clinical samples. The mutation detection efficiency was increased from 25.9% of the input DNA to 45.2% by shortening the amplicon size. The limit of detection of mutation was improved from 0.41 to 0.06% by changing the mutation type determination algorithm for G12A, resulting in a limit of detection of less than 0.2% for all the target mutations. Then, ctDNA in plasma from pancreatic cancer patients was measured and genotyped. The measured mutation frequencies correlated well with those measured by conventional dPCR, which can measure only the total frequency of KRAS mutants. KRAS mutations were detected in 82.3% of patients with liver or lung metastasis, which was consistent with other reports. Accordingly, this study demonstrated the clinical utility of multiplex dPCR with melting curve analysis to detect and genotype ctDNA from plasma with sufficient sensitivity., (© 2023. The Author(s).)

Most of the odors that humans perceive daily are complex odors. It is believed that the modulation, enhancement, and suppression of overall complex odors are caused by interactions between odor molecules. In this study, to understand the interaction between odor molecules at the level of human olfactory receptor responses, the effects of 3-octen-2-one, which has been shown to modulate vanilla flavors, were analyzed using a human olfactory receptor sensor that uses all human olfactory receptors (388 types) as sensing molecules. As a result, the response intensity of 1 common receptor (OR1D2) was synergistically enhanced in vanilla flavor with 3-octen-2-one compared with vanilla flavor, and the response of 1 receptor (OR5K1) to vanilla flavor was completely suppressed. These results strongly suggested that the response of human olfactory receptors to complex odors is enhanced or suppressed by relatively few other odor molecules., (© The Author(s) 2022. Published by Oxford University Press on behalf of Japan Society for Bioscience, Biotechnology, and Agrochemistry.)

Caries sensitivity varies between the two strains of inbred mice, BALB/cA has high sensitivity and C3H/HeN has low sensitivity. One potential reason seems to be a difference in pellicle-forming saliva protein composition. Here, we performed a proteomic analysis in order to identify differences of hydroxyapatite (HAP) adsorbed saliva proteins between these two mouse strains. HAP column chromatography revealed twice the quantity of high-affinity saliva proteins in C3H/HeN compared to BALB/cA. One- and two-dimensional electrophoresis showed 2 bands/spots with deviating migration. They were identified as murine carbonic anhydrase VI (CAVI) by peptide mass fingerprinting and confirmed with western blotting using a specific polyclonal antibody. Total RNA from the salivary glands of both mouse strains, PCR amplification of cDNA with a CAVI specific primer, and sequence analysis revealed one different base in codon 96, resulting in one different amino acid. Glyco-chains of CAVI deviate in one N-glycan, confirmed by mass analysis. CAVI activity was estimated from distinct circular dichroism spectra of the molecules and found higher in C3H/HeN mice. In summary, the CAVI composition of BALB/cA and C3H/HeN differs in one amino acid and a glyco-chain modification. Further, saliva from caries resistant C3H/HeN mice displayed higher CAVI activity and also overall hydroxyapatite adsorption, suggesting a relationship with caries susceptibility., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

The use of dual-energy X-ray absorptiometry (DXA) may be invalid for assessing athletes with larger bodies, larger lean body mass, and thicker trunks. This study compared the athletes' visceral adipose tissue (VAT) assessed using DXA and magnetic resonance imaging (MRI). Eighty-two Japanese male collegiate athletes from 18 sports participated in this study. VAT was assessed using the dual-energy scan that coincided with the 4th lumbar vertebra. The sum of eight magnetic resonance slices corresponding to the region of the dual-energy scan was used for comparison. The VAT volume was higher with the dual-energy scan than with MRI (difference: 35 cm 3 , p<0.01). A significant correlation was noted between the volumes measured using both modalities (r=0.88, p<0.01). Magnetic resonance-derived volumes less than 600 cm 3 showed a stronger significant correlation with dual-energy-derived volumes. However, magnetic resonance-derived VAT volumes exceeding 600 cm 3 were not significantly correlated with dual-energy-derived volumes. In conclusion, VAT volumes derived from DXA were larger and significantly correlated with those derived from MRI across a wide range of values. Methods using DXA for assessing the visceral fat volume may require adjustment to estimate abdominal visceral fat volume in athletes, with care taken when using such methods for heavyweight athletes., Competing Interests: The authors declare that they have no conflict of interest., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

Objective: The primary objective of this study was to identify novel genes that predispose people in the Japanese population to FPC., Summary of Background Data: Familial history of pancreatic cancer is an important risk factor but, to date, few genes predisposing individuals to increased risk of developing FPC have been identified., Methods: We performed whole-exome sequencing of germline DNA from 81 Japanese FPC patients. We also investigated somatic gene alterations in 21 matched tumor tissues through whole-exome sequencing and copy number analysis., Results: Our germline variants identified previously known FPC susceptibility genes such as ATM and BRCA2, and several novel tumor suppressor genes with potentially deleterious variants for FPC. Interestingly, somatic whole-exome analysis demonstrated that most tumor samples with suspicious loss of heterozygosity of candidate genes were KRAS wild-types, implying that these cases may not have required KRAS activation as a driver event for carcinogenesis., Conclusions: Our findings indicate that FPC patients harbor potentially deleterious causative germline variants in tumor suppressor genes, which are known to acquire somatic mutations in pancreatic cancer, and that somatic loss of heterozygosity of some FPC susceptibility genes may contribute to the development of FPC in the absence of somatic KRAS-activating mutation. Genetic testing for a wider variety of FPC-predisposition genes could provide better screening approach for high-risk groups of pancreatic cancer., Competing Interests: The authors report no conflicts of interest., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

The neuroendocrine carcinoma of the gastrointestinal system (GIS-NEC) is a rare but highly malignant neoplasm. We analyzed 115 cases using whole-genome/exome sequencing, transcriptome sequencing, DNA methylation assays, and/or ATAC-seq and found GIS-NECs to be genetically distinct from neuroendocrine tumors (GIS-NET) in the same location. Clear genomic differences were also evident between pancreatic NECs (Panc-NEC) and nonpancreatic GIS-NECs (Nonpanc-NEC). Panc-NECs could be classified into two subgroups (i.e., "ductal-type" and "acinar-type") based on genomic features. Alterations in TP53 and RB1 proved common in GIS-NECs, and most Nonpanc-NECs with intact RB1 demonstrated mutually exclusive amplification of CCNE1 or MYC. Alterations of the Notch gene family were characteristic of Nonpanc-NECs. Transcription factors for neuroendocrine differentiation, especially the SOX2 gene, appeared overexpressed in most GIS-NECs due to hypermethylation of the promoter region. This first comprehensive study of genomic alterations in GIS-NECs uncovered several key biological processes underlying genesis of this very lethal form of cancer., Significance: GIS-NECs are genetically distinct from GIS-NETs. GIS-NECs arising in different organs show similar histopathologic features and share some genomic features, but considerable differences exist between Panc-NECs and Nonpanc-NECs. In addition, Panc-NECs could be classified into two subgroups (i.e., "ductal-type" and "acinar-type") based on genomic and epigenomic features. This article is highlighted in the In This Issue feature, p. 587., (©2021 The Authors; Published by the American Association for Cancer Research.)

Objectives: Olfactory stimulation elicits various physiological responses. However, few reports exist on the changes in gastric motility during olfactory stimulation in humans. In this regard, we carried out electrogastrography (EGG) to non-invasively measure the gastric myoelectrical activity, which regulates gastric motility. Moreover, subjective sensory evaluation was performed to determine which characteristics of vanilla odor at two different concentrations affect the myoelectrical activity., Methods: The participants consisted of eight healthy young males. EGG and electrocardiography (ECG) recordings were obtained approximately 20 min prior to and during olfactory stimulation. Autonomic nervous system activity was evaluated in terms of heart rate variability (HRV) and mean heart rate (HR) from ECG signals. EGG signals were analyzed by spectral analysis. In addition, the translation error was estimated by the Wayland algorithm. Sensory evaluation was performed using the Visual Analog Scale (VAS)., Results: There were no significant differences in HRV and HR values and results of spectral analyses of EGG signals in all sample presentations. The translation error of EGG signals and the rating of perceived odor intensity significantly increased in a concentration-dependent manner. There was a strong positive correlation between translation error and odor intensity., Conclusions: The correlation found between translation error and odor intensity suggests that the higher the vanilla odor intensity was perceived, the greater the randomness of EGG signals was. Our results suggest that the application of the Wayland algorithm to EGG signals can be used as an objective indicator in odor evaluation.

A biomarker that is useful for the detection of human papillomavirus (HPV)-related oropharyngeal cancer (OPC) and cancer of unknown primary (CUP) is indispensable. We evaluated the diagnostic performance of HPV DNA and mRNA in oral gargle samples and circulating tumor HPV16 DNA (ctHPV16DNA) in blood samples. Oral HPV DNA and mRNA were analyzed using commercially available HPV assays of the GENOSEARCH HPV31 and Aptima, respectively. ctHPV16DNA was analyzed using in-house droplet digital polymerase chain reaction. Seventy-four patients with OPC and eight patients with CUP were included. The sensitivity and specificity of oral HPV DNA, oral HPV mRNA, and ctHPV16DNA were 82% (95% confidence interval [CI] = 66-92) and 100% (95% CI = 88-100), 85% (95% CI = 69-94) and 94% (95% CI = 73-100), and 93% (95% CI = 81-99) and 97% (95% CI = 84-100), respectively, for HPV16-related OPC, while those were 20% (95% CI = 1-72) and 100% (95% CI = 3-100), 0% (95% CI = 0-52) and 100% (95% CI = 3-100), and 100% (95% CI = 54-100) and 100% (95% CI = 16-100), respectively, for HPV16-related CUP. The sensitivity of ctHPV16DNA for HPV16-related OPC was higher than that of oral biomarkers, though the difference was not statistically significant. ctHPV16DNA remarkably correlated with the anatomic extent of disease, total metabolic tumor volume and HPV16 copy number per tumor genome in patients with HPV16-related OPC/CUP, whereas oral biomarkers did not. In conclusion, ctHPV16DNA is a potentially promising biomarker for HPV16-related OPC, while further studies are required for HPV16-related CUP., (© 2021 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.)

Assessment of treatment efficacy of immune checkpoint inhibitors in melanoma patients is difficult as the response to these therapies varies among patients or lesions. The clonal evolution of cancer during immune checkpoint blockade therapy could cause treatment resistance. We investigated the potential of liquid biopsy in monitoring the mutational profiles of metastatic melanoma during immunotherapy. Plasma samples collected from 21 Japanese metastatic melanoma patients before immune checkpoint blockade therapy were subjected to whole-exome sequencing (WES). Furthermore, 14 Japanese patients with melanoma were enrolled for longitudinal analysis of circulating tumor DNA (ctDNA). Plasma samples were collected prospectively before and during therapy and sequenced. WES of the pretreatment plasma from Japanese melanoma patients showed detectable ctDNA levels with wide ranges of variant allele frequencies within a sample, suggesting clonal and subclonal mutations in ctDNA. In targeted sequencing using longitudinal samples, ctDNA levels correlated with increased tumor size, while ctDNA content immediately decreased after a surge in a patient exhibiting pseudo-progression, suggesting the potential of ctDNA analysis in discriminating between pseudo- and true progression. Mutant ctDNA levels showed different patterns within the clinical course of specific patients, suggesting that these mutations were derived from different tumor clones with distinct therapeutic responses. During further investigation, WES of plasma samples from 1 patient showed marked differences in the mutational profiles of ctDNA, including expansive tumor evolution during an acute exacerbation. Immunotherapy may induce characteristic clonal evolutions of tumors; longitudinal analysis of ctDNA has the potential of determining these tumor evolution patterns and therapeutic responses., (© 2021 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)