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Yoichi Fukushima,1 Yoshinari Takahashi,2 Yoshimi Kishimoto,3 Chie Taguchi,3 Norie Suzuki,3 Mihoko Yokoyama,4 Kazuo Kondo3,5 1Wellness Communications Section, Nestlé Japan Ltd., Tokyo, Japan; 2Inary Corp., Tokyo, Japan; 3Endowed Research Department “Food for Health”, Ochanomizu University, Tokyo, Japan; 4Yokoyama Skin Clinic, Tokyo, Japan; 5Institute of Life Innovations Studies, Toyo University, Gunma, JapanCorrespondence: Yoichi FukushimaWellness Communications Section, Nestlé Japan Ltd., Tennoz Square, 2-2-20 Higashi-Shinagawa, Shinagawa-ku, Tokyo 140-0002, JapanTel +81 3 5769 6299Fax +81 3 5769 6291Email yoichi.fukushima@jp.nestle.comPurpose: Hyperpigmentation of the skin can occur at any age depending on etiological factors but its intensity increases during adolescence in Japanese females and gradually develops further in adults. The purpose of this study was to characterize factors that influence skin hyperpigmentation, including age, skin type and dietary polyphenol sources.Patients and Methods: A cross-sectional survey of healthy Japanese women aged from 30 to 60 years (n=244) was conducted using food and environmental questionnaires and a VISIA™ facial photoimage analyzer.Results: UV Pigmented Spot (PS) scores correlated negatively with the consumption of total polyphenols (TPs) (R=− 0.224, p< 0.001) and the rate of hyperpigmented spot development (PS score/age after 18 years of age) was suppressed by the consumption of TPs. This trend was independent of the melanin index and the skin type, which indicates the ability of the skin to tan after sun exposure. Consumption of coffee, the largest source of TPs, suppressed the PS score (p< 0.001). Consumption of green tea, the second largest source of TPs, also suppressed the PS score, which was weaker than coffee but was statistically significant (p=0.029). The PS score was suppressed the most in subjects with both a high consumption of coffee and green tea.Conclusion: Higher consumption of TPs may be beneficial to alleviate photoaging of the skin, and coffee as well as green tea contribute to suppress skin hyperpigmentation through adding large amounts of TPs in the diet.Keywords: coffee, green tea, polyphenol, photoaging, skin hyperpigmentation

The aim of this article is to present two case reports for the treatment of patients affected with amelogenesis imperfecta. One case was treated with composite resin and the other case with ceramic veneers. Esthetic and functional results were achieved using both treatments, and a review of advantages and disadvantages is presented. [ABSTRACT FROM AUTHOR]

The aim of this article is to present a case report for the multidisciplinary treatment of anterior tooth fractures with invasion of the biologic width and pulpectomy. Successful esthetic and functional results were achieved by bonding the crown fragment, without any form of preparation or the utilization of intracanal posts. [ABSTRACT FROM AUTHOR]

Objective: NMK36 is a novel PET tracer containing a synthetic amino acid analogue anti-[(18)F]FACBC as the active ingredient, and is under development for the use of tumor diagnosis. A Phase I clinical study of NMK36 was conducted to evaluate its safety, biodistribution, and radiation dosimetry in healthy volunteers.Methods: Six healthy volunteers (Japanese male) received a bolus injection of NMK36 (174.4-201.4 MBq) intravenously. The safety of NMK36 was evaluated by monitoring signs/symptoms, electrocardiography, recording vital signs, and laboratory examinations at baseline and several time points in 6 days after injection. A total of 11 whole-body PET-CT scans were acquired up to 4 h post-injection, and venous blood and urine samples were also collected for 6 and 24 h post-injection, respectively. Based on the results of the biodistribution study, absorbed radiation dose was estimated by the MIRD method.Results: Although two adverse events occurred after the injection of NMK36, they were mild and disappeared without any specific treatment. NMK36 preferentially accumulated in the pancreas and liver early after injection, followed by rapid clearance from the pancreas. Persistent uptake was observed in the skeletal muscle. NMK36 showed low uptake in the brain, and its urinary excretion was limited (5.40 ± 1.43% of the injected dose at 24 h post-injection). The liver was the critical organ, with a mean absorbed dose of 40.6 μGy/MBq. The estimated effective dose of NMK36 was 13.8 μSv/MBq, which was similar to or lower than those of radiotracers approved for clinical use including [(18)F]FDG.Conclusions: The findings of this study indicate that NMK36 is well tolerated. NMK36 has favorable characteristics for imaging brain and pelvic tumors, such as low brain uptake, slow urinary excretion, and high in vivo stability. [ABSTRACT FROM AUTHOR]

Gingival overgrowth caused by cyclosporine A is due to increased fibroblast proliferation in gingival tissues. Cell cycle system balances proliferation and anti-proliferation of gingival fibroblasts and plays a role in the maintenance of its population in gingival tissues. When cells detect and respond to abnormalities (e.g. DNA damage), cell cycle progression is arrested in the G1 phase until the completion of damage restoration. In this study, we investigated the effects of cyclosporine A on G1 cell cycle arrest and on its regulators in gingival fibroblasts to clarify the mechanism of cyclosporine A-induced gingival overgrowth. Human gingival fibroblasts from healthy donors were cultured to semi-confluence and were then treated with or without 200 ng/mL (166 nM) cyclosporine A in D-MEM with 2% fetal bovine serum. Cell proliferation was assessed by counting total cell numbers. The distribution of cell cycle phases was assessed using flow cytometric analysis. The levels of mRNA and protein expression for cell cycle regulators were quantified using reverse transcription-quantitative PCR and western blot analysis, respectively. Treatment with cyclosporine A markedly increased cell proliferation, inhibited G1 cell cycle arrest, significantly increased CDC25A and CYCLIN E1 mRNA expression levels, significantly decreased P21, SMAD3 and SMAD4 mRNA expression levels, significantly upregulated the protein expression levels of CDC25A, CYCLIN E1, pCDK2 and pRB1 and significantly downregulated the protein expression levels of P21, SMAD3 and SMAD4. Treatment with cyclosporine A also increased MYC and ATM mRNA expression levels and decreased CDK2, ATR, P27, P53 and RB1 mRNA expression levels but not significantly. These results demonstrate that cyclosporine A causes gingival overgrowth due to the following mechanism in gingival fibroblasts: cyclosporine A increases levels of phospho-CDK2 and CYCLIN E1 by upregulating CDC25A and downregulating P21 with the downregulation of SMAD3 and SMAD4, which results in the inhibition of G1 cell cycle arrest., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Takeuchi 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.)

Objectives: Cyclosporine A promotes gingival fibrosis by enhancing the proliferation of gingival fibroblasts, leading to gingival overgrowth. The population of gingival fibroblasts is regulated by cell cycle machinery, which balances cell growth and inhibition. Cells that detect DNA damage pause at the G1/S checkpoint to repair the damage instead of progressing to the S phase. Previous studies have linked drug-induced gingival overgrowth to the response of fibroblasts to lipopolysaccharide (LPS) and cyclosporine A. This research investigates the effects of cyclosporine A on the G1/S checkpoint and its mediators in LPS-treated gingival fibroblasts to clarify the mechanisms behind cyclosporine-A-induced gingival overgrowth., Methods: Semi-confluent human gingival fibroblasts were treated with LPS or cyclosporine A in DMEM. Cell proliferation was evaluated by counting the total number of cells. The distribution of the cell cycle phases was analyzed using flow cytometry. Additionally, the expression levels of mRNAs and proteins related to cell cycle regulators were quantified by reverse-transcription quantitative PCR and Western blotting, respectively., Results: Cyclosporine A treatment significantly enhanced cell proliferation and the G1-S cell cycle transition. It increased the mRNA levels of CDC25A and CYCLIN D while decreasing those of RB1 , SMAD3 , and SMAD4 . Additionally, it upregulated the protein levels of CDC25A, CYCLIN D, CDK4, CDK6, and pRB and downregulated the protein levels of SMAD3 and SMAD4., Conclusions: Gingival overgrowth induced by cyclosporine A could be attributed to these alterations.

Cells regularly repair numerous mutations. However, the effect of CRISPR/Cas9-induced dsDNA breaks on the repair processes of naturally occurring genome-wide mutations is unclear. In this study, we used TSCE5 cells with the heterozygous thymidine kinase genotype (TK +/- ) to examine these effects. We strategically inserted the target sites for guide RNA (gRNA)/Cas9 and I-SceI into the functional allele and designed the experiment such that deletions of > 81 bp or base substitutions within exon five disrupted the TK gene, resulting in a TK -/- genotype. TSCE5 cells in the resting state exhibited 16 genome-wide mutations that affected cellular functions. After gRNA/Cas9 editing, these cells produced 859 mutations, including 67 high-impact variants that severely affected cellular functions under standard culture conditions. Mutation profile analysis indicated a significant accumulation of C to A substitutions, underscoring the widespread induction of characteristic mutations by gRNA/Cas9. In contrast, gRNA/Cas9-edited cells under conditions of S∼G2/M arrest and cyclin-dependent kinase 1 inhibition showed only five mutations. Transcriptomic analysis revealed the downregulation of DNA replication genes and upregulation of alternative DNA repair genes, such as zinc finger protein 384 (ZNF384) and dual specificity phosphatase, under S∼G2/M conditions. Additionally, activation of nucleotide and base excision repair gene, including O-6-methylguanine-DNA methyltransferase and xeroderma pigmentosum complementation group C, was observed. This study highlights the profound impact of CRISPR/Cas9 editing on genome-wide mutation processes and underscores the emergence of novel DNA repair pathways. Finally, our findings provide significant insights into the maintenance of genome integrity during genome editing., Competing Interests: Conflicts of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Phenytoin (PHT)-induced gingival overgrowth is caused by the increased proliferation and reduced apoptosis of gingival fibroblasts in inflammatory gingiva. Licorice has long been used as a component of therapeutic preparations. It inhibits cell proliferation, induces cell apoptosis and has anti-inflammatory effects. 18-α-glycyrrhetinic acid (18α-GA), the active compound in licorice, promotes apoptosis in various types of cells. The present study determined whether 18α-GA affects apoptosis in gingival fibroblasts exposed to PHT. The present study aimed to establish a basis for the therapeutic application of 18α-GA to treat the gingival overgrowth induced by PHT. Human gingival fibroblasts from healthy donors were cultured to semi-confluence and then stimulated in serum-free DMEM containing PHT with or without 18α-GA for subsequent experiments. Apoptotic cells were detected by ELISA. Analysis of the distribution of cell cycle phases and the apoptotic cell population was performed by flow cytometry. The expression levels of mRNAs and proteins of apoptotic regulators were measured using reverse transcription-quantitative PCR and western blotting, respectively. Caspase (CASP) activities were assessed by an ELISA. Treatment with 18α-GA markedly increased the number of apoptotic cells, reduced BCL2 mRNA expression, increased CASP2 and receptor (TNFRSF)-interacting serine-threonine kinase 1 (RIPK1) domain containing adaptor with death domain, Fas (TNFRSF6)-associated via death domain, RIPK1, tumor necrosis factor receptor superfamily; member 1A, TNF receptor-associated factor 2, CASP2, CASP3 and CASP9 mRNA expression, and also upregulated the protein expression levels and activities of caspase-2, caspase-3 and caspase-9. These results demonstrated that 18α-GA induced apoptosis through the activation of the Fas and TNF pathways in the death receptor signaling pathway in gingival fibroblasts treated with PHT. 18α-GA exhibited therapeutic potential for the treatment of PHT-induced gingival overgrowth., Competing Interests: The authors declare that they have no competing interests., (Copyright: © 2024 Takeuchi et al.)

In the Japanese official detection method for unauthorized genetically modified (GM) papayas, one of two types of real-time PCR reagents with DNA polymerase (TaqMan Gene Master Mix [TaqMan Gene] or FastGene QPCR Probe Mastermix w/ROX [FastGene]) is primarily used for measurement. In 2022, we conducted a laboratory performance study on the unauthorized GM papaya line PRSV-YK, and the results revealed that high threshold cycle (Cq) values for the PRSV-YK detection test were obtained using TaqMan Gene with the 7500 Fast & 7500 Real-Time PCR System (ABI7500) and QuantStudio 12K Flex (QS12K), indicating the possibility of false negatives. The possibility of similar problems with all unauthorized GM papaya lines detection tests needs to be evaluated. In this study, we performed detection tests on unauthorized GM papaya lines (PRSV-YK, PRSV-SC, and PRSV-HN), the cauliflower mosaic virus 35S promotor (CaM), and a papaya positive control (Chy), and examined how the limits of detection (LOD) for each test are affected by two types of DNA polymerases (TaqMan Gene and FastGene) and three types of real-time PCR instruments (ABI7500, QS12K, and LightCycler 480 Instrument II [LC480]). In the PRSV-YK and PRSV-SC detection tests using ABI7500 and QS12K, measurement with TaqMan Gene showed a higher LOD than FastGene. In this case, an exponential amplification curve was confirmed on the amplification plot; however, the amplification curve did not cross the ΔRn threshold line and the correct Cq value was not obtained with a threshold line=0.2. The other tests (PRSV-HN, CaM, and Chy with ABI7500 and QS12K, and all detection tests with LC480) showed no important differences in the LOD for each test using either DNA polymerase. Therefore, when performing PRSV-YK and PRSV-SC detection tests with the ABI7500 or QS12K, FastGene should be used to avoid false negatives for foods containing GM papaya lines PRSV-YK and PRSV-SC at low mixing levels.

Since the establishment of procedures for the safety assessment of food products that use recombinant DNA technology, the manufacture, import, and sale of genetically modified (GM) foods that have not undergone safety assessment are prohibited under the Food Sanitation Act. Therefore, a performance study to confirm the GM food testing operations of each laboratory is very important to ensure the reliability of the GM food monitoring system. In 2022, GM papaya line PRSV-YK-which has not yet been authorized in Japan-was selected for testing, and a papaya paste and a DNA solution were used as the test samples. With these samples, a laboratory performance study of the DNA extraction and real-time PCR operations was conducted. This confirmed that the 18 participating laboratories were generally performing the DNA extraction and real-time PCR operations correctly. However, some laboratories using certain DNA amplification reagent with some real-time PCR instruments were not able to determine the PRSV-YK detection test. This suggests that the PRSV-YK detection test may not be able to correctly detect samples containing GM papaya when performed with these combinations of instruments and reagent. In order to ensure the reliability of the PRSV-YK detection test, it is necessary to examine in detail how the combination of DNA polymerase reagents and real-time PCR instruments affects the detection limit, and to implement an appropriate solution.

Although the safety of genome-edited foods in Japan has been confirmed through pre-submission consultation under the notification process, public perception of safety confirmation methodology has not been investigated to date. Therefore, we created three media to provide information on the safety assurance of genome-edited foods and surveyed the perception of current safety confirmation. In addition, we examined the opinions of researchers in health science on current safety confirmation methods. As a result, 62% of general consumers and 68% of researchers in health science recognized that safety is ensured. Acceptance of genome-edited foods improved when they realized that safety was ensured. Researchers in health science who felt that safety confirmation was insufficient were concerned about the third-party verification. Therefore, it was suggested that in order to boost public understanding of genome-edited foods, it would be useful to inform the public by communicating in an easy-to-understand way the safety assurance approaches being made in pre-submission consultation.

The Japanese Health Ministry recently granted permission for the market distribution of genome-edited (GE) foods, yet there remains a lack of full understanding among consumers regarding this technology. In this study, we conducted a survey to assess the acceptability of GE foods among Japanese consumers and examined the impact of providing information about GE foods on their acceptability. We conducted a web-based survey among 3,408 consumers aged 20-69 years, focusing on three aspects: (1) the commercial availability of GE foods, (2) the consumption of GE foods by others, and (3) your own consumption of GE foods. The survey findings revealed that participants were most accepting of the consumption of GE foods by others, followed by their acceptance of GE foods being commercially available. Notably, participants' acceptance of GE foods increased in all three aspects after they viewed an informative video. The video had a particularly strong impact on participants who fully or partially understood its content, compared to those who did not. Furthermore, regression analyses showed that participants' understanding of two key areas, namely "Why are GE foods important" and "What procedures are in place to ensure the safety of GE foods," played a crucial role in increasing acceptability. Overall, these results indicate that providing information about GE foods to Japanese consumers can effectively enhance their acceptance of such foods. The findings highlight the importance of understanding the benefits and safety measures associated with GE foods in influencing consumer attitudes.

Strawberry contains many bioactive compounds such as vitamin C and polyphenols as well as folate, a vitamin that is especially important for women of childbearing age. We investigated the effects of the acute consumption of strawberry on the serum levels of vitamin C and folate, and on the antioxidant potential of low-density lipoprotein (LDL). In a randomised, placebo-controlled, double-blind, crossover study, twenty-three healthy female volunteers (age 22⋅5 ± 1⋅4 years) ingested 500 g of a strawberry purée beverage or a sugar content-matched placebo beverage. Blood samples were collected at fasting and at 0⋅5, 1, 2 and 4 h post-ingestion. The serum concentrations of vitamin C and folate were significantly elevated from 0⋅5 to 4 h after the strawberry beverage ingestion ( P < 0⋅001); the levels peaked at 2 h, with peak levels of 15⋅0 ± 2⋅5 μg/ml for vitamin C and 14⋅4 ± 7⋅0 ng/ml for folate. Notably, at 1 h after the strawberry beverage ingestion, the LDL oxidation lag time was significantly prolonged ( P < 0⋅05), suggesting that the antioxidant potential of LDL was increased. After the ingestion of either beverage, the serum levels of glucose and insulin reached a peak at 0⋅5 h and then quickly returned to baseline levels. These results suggest that strawberries are a useful source of vitamin C and folate and may help enhance the antioxidant potential of LDL in healthy young women., (© The Author(s) 2023.)

Carotenoid intake is associated with low mortality and cancer risks; data on non-provitamin carotenoid intake is limited especially in Asians. We aimed to estimate carotenoid intake in Japanese adult women. Carotenoid content database comprises 196 food items, including 39 fruits, 87 vegetables and mushrooms, and 11 seaweeds, and was established using data from the literature and analyses of foods available in Japan. We surveyed the intake of these foods in Japanese women aged 21-56 years (n=109). Total intake of 7 carotenoids (mean±SD [range]) was 7,450±3,840 (1,160-21,300) μg/day; α-carotene, β-carotene, β-cryptoxanthin, lutein, zeaxanthin, lycopene, and fucoxanthin represented 4.3%, 23%, 3.4%, 15%, 2.0%, 39%, and 13% of total intake, respectively. Lutein intake was 1,132±686 (294-3,490) μg/day; its best sources were spinach, cucumber, chicken egg, green onion, and Chinese chives, representing 51% of total intake. Lutein can be obtained from a variety of sources. Thus, lutein intake levels did not vary widely among individuals and very few individuals consumed insufficient levels of lutein. Intake of zeaxanthin, lycopene, and fucoxanthin was 149±93 (2-479), 2,890±2,970 (0-17,100), and 980±1,230 (0-5,660) μg/day, respectively. Their intake required rich sources including chicken egg for zeaxanthin (52%); tomato products for lycopene (98%), and wakame seaweed for fucoxanthin (76%). The carotenoid content database including all food items consumed in Japan will be helpful for further investigations on carotenoid intake and its health benefits.

Given that the number of genetically modified (GM) maize events that have been announced as having undergone safety assessment procedures in Japan is increasing yearly, more information is needed about their actual recent domestic distribution in Japan. In this study, we investigated whether current Japanese official qualitative and quantitative methods (the current official methods) for GM maize can comprehensively target events in domestically distributed maize. For samples with the identity-preserved (IP) handling system and non-IP samples from the United States (US) and non-IP samples from Brazil, we performed event-specific real-time PCR targeting 25 authorized single GM maize events in addition to the current official methods. According to our results, 15 events targeted by the current official methods were detected, but insect-resistance (IR) Event5307 and herbicide-tolerant (HT) DAS40278, not targeted by the current official methods, were detected in the US (one out of 5 lots) and Brazilian (four out of 5 lots) non-IP samples, respectively. Nevertheless, a survey of recent GM maize acreage in recent years has revealed that more than 95% of the acreage in US maize is occupied by HT or IR/HT stacked events, and that more than 95% of the acreage in Brazilian maize is occupied by IR or IR/HT stacked events. Because the current official methods can target all stacked events related to Event5307 and DAS40278, the only undetectable events are the single Event5307 and DAS40278, whose production is estimated to be less than 5% of the total production in the producing country. Therefore, we conclude that the current official methods for the labelling of GM maize should be maintained in view of practicability.

The food supply system is facing important challenges and its sustainability has to be considered. Genome-editing technology, which accelerates the development of new variety, could be used to achieve sustainable development goals, thereby protecting the environment and ensuring the stable production of food for an increasing global population. The most widely used genome-editing tool, CRISPR/Cas9, is easy to use, affordable, and versatile. Foods produced by genome-editing technologies have been developed worldwide to create novel traits. In the first half of the review, the latest scientific findings on genome-editing technologies are summarized, and the technical challenge in genome sequence analysis are clarified. CRISPR/Cas9 has versatile alternative techniques, such as base editor and prime editor. Genome sequencing technology has developed rapidly in recent years. However, it is still difficult to detect large deletions and structural variations. Long-read sequencing technology would solve this challenge. In the second part, regulatory framework and approach for genome-edited foods is introduced. The four government ministries, including the Ministry of Environment, the Ministry of Agriculture, Forestry and Fisheries, and the Ministry of Health, Labour and Welfare (MHLW), started to discuss how the regulation should be implemented in 2019. The SDN-1 technique is excluded from the current genetically modified organism (GMO) regulation. The Japanese regulatory framework includes pre-submission consultation and submission of notification form. In the last part of this review, transparency of regulatory framework and consumer confidence were described. Since maintaining consumer trust is vital, transparency of regulatory framework is a key to consumers. The information of notification process on approved genome-edited foods is made public immediately. This review will help regulators build regulatory frameworks, and lead to harmonization of the framework between the countries., Competing Interests: Conflict of interest: The author declares there are no conflicts of interest associated with this manuscript., (©2022 Food Safety Commission, Cabinet Office, Government of Japan.)

Introduction: Appendicolith causes acute appendicitis. However, surgical indications for appendicolith-related acute appendicitis have not been established. We aimed to clarify the clinical features of appendicolith-associated appendicitis and determine an appropriate treatment strategy based on the initial presentation., Materials and Methods: We retrospectively reviewed the records of 479 consecutive patients with acute appendicitis and verified the therapeutic strategy as per the appendicolith and clinical status., Results: Appendicoliths were identified in 214 of 479 patients (44.6%) using computed tomography. Surgery was more frequently required in patients with appendicolith than in patients without appendicolith (82.7 versus 64.9%; P < 0.001). The stones were smaller and serum C-reactive protein (CRP) concentration was lower among patients with appendicoliths treated with medication alone than among those surgically treated (both P < 0.001). An appendicolith measuring ≤5 mm in diameter and CRP concentration ≤5.36 mg/dL were predictive of completion of nonsurgical therapy. CRP concentration >10 mg/dL and stone diameter of 10 mm were significantly associated with appendiceal perforation., Conclusions: Nonsurgical therapy could be considered for patients with appendicoliths measuring ≤5 mm in diameter and in cases where the serum CRP concentration is ≤5 mg/dL. An appendicolith measuring >10 mm in diameter or CRP concentration >10 mg/dL is an indication for surgery., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Purpose: A water fluoridation program launched in the United States in 1945 has become a worldwide application for caries prevention. Although such a program is important in Japan, water fluoridation in large communities has not been established. This study aimed to develop safe small-scale water fluoride equipment that can be easily used to carry out water fluoridation programs in areas with children in long-term care facilities, and in developing countries without water supply facilities., Methods: Batch-type NaF addition adjustment equipment was manufactured as small-scale water fluoride equipment. The fluoride concentration of the adjusted water with this equipment was measured using an ion meter and a fluoride composite electrode. All 51 water quality standards set by the Ministry of Health, Labour, and Welfare of Japan were tested., Results: The fluoride ion concentration of the adjusted water was 0.7 mg/L and it was constant and stable. The adjusted water conformed to the water quality standard values of the Japanese Water Supply Law., Conclusion: Water produced with small-scale water fluoridation equipment had a fluoride concentration of 0.7 mg/L, which is the recommended concentration for caries prevention. The fluoride concentration was stable.

Background/objectives: Japan has responded to the spread of COVID-19 through declaration of a state of emergency to regulate human mobility. Although the declaration was enforced by the government for prefectures, there is limited evidence as to whether the public complied with requests for voluntary stay at home. In this study, we evaluated the impact of declaring a state of emergency on human mobility during the COVID-19 pandemic in Japan., Methods: We utilized daily human mobility data for 47 prefectures in Japan. Data were collected via mobile phone from February 1, 2020 to April 30, 2021. Difference-in-difference analysis was utilized to estimate the effects of the declaration of a state of emergency on prefectures in the Tokyo Metropolitan Area (Tokyo, Kanagawa, Saitama, and Chiba) in comparison to other prefectures where the state of emergency was first lifted (Osaka, Hyogo, Fukuoka, and Aichi)., Results: Human mobility was suppressed during the second state of emergency, from January 8 to March 21, 2021. However, the impact was weaker for the second state of emergency compared to the first., Conclusion: In Japan, government requests for stay at home, such as the declaration of a state of emergency, were temporarily able to control human mobility. However, the second state of emergency was not as effective as the first. If additional need to regulate human mobility arises, self-restraint with stronger enforcement should be considered., (© 2022 The Author(s).)

Dental professionals are at increased risk of being infected with airborne pathogens such as SARS-CoV-2 because they are often exposed to droplets/aerosols production during dental treatment. To scientifically clear the effects of extraoral and oral suctions on the droplets and aerosols produced by dental treatments using an ultrasonic scaler was analyzed. The adenosine triphosphate and bacteria in droplets and aerosols produced during simulated scaling were quantitatively observed by reactions with luciferin/luciferase and incubation in culture plates to grow bacteria, respectively. The protection against spreading droplets and aerosols by oral and extraoral suctions was recognized, and the areas were limited to the left and posterior sides of the dental chair head when a right-handed dentist and dental hygienist performed scaling. Extraoral suction is a very useful tool for reducing the infection risk of COVID-19 in dental care, but the effective area is limited depending on physical characteristics of dentist and dental hygienist.

Estimations of individuals' polyphenol intake contributes to the understanding of the health benefits of dietary polyphenol. We developed a food frequency questionnaire for polyphenol intake (FFQ) and a short-form FFQ for polyphenol intake (SF-FFQ) for assessing the polyphenol intake of Japanese. The aim of this study was to compare the relative validity of polyphenol intake derived from the FFQ with that from the SF-FFQ, using a 4-consecutive-d dietary record (DR) as the reference. Sixty Japanese subjects aged 30-69 y completed the 4-d DR and the two FFQs regarding their polyphenol intake in November 2019. The polyphenol intake values estimated by the DR, FFQ, and SF-FFQ were 1,057±524 mg/d, 1,061±537 mg/d and 1,015±491 mg/d, respectively. No significant differences were present in the estimated polyphenol intake between the 4-d DR and both FFQs. The correlation coefficient with the DR was 0.779 for the FFQ and 0.814 for the SF-FFQ. These results indicate that the total polyphenol intake in a Japanese population were accurately estimated by the FFQ and SF-FFQ.

Mortality is obviously intended for epidemiological studies of community-dwelling older adults. There are several health-related factors associated with nutritional status and mortality. The aim of this study was to elucidate the risk factor for mortality in community-dwelling oldest-older adults at the age of 90 and clarify the structure of health-related factors associated with mortality. A 10-year follow-up study was performed for 93 subjects at the age of 90. The mean and median of their survival days were 2373 and 2581 days for women, and 1694 and 1793 days for men. By Cox's proportional hazards model, health-related factors associated with mortality were self-assessed for chewing ability, activities of daily living (ADLs), serum albumin, total cholesterol, serum creatinine, and gripping power for women but not for men. These factors interacted with each other, and the association of these factors was different in women and men. Self-assessed chewing ability was a powerful risk factor for mortality in women at the age of 90. It acted independently from nutritional status. For older adults, addressing healthy food choices together with improved oral functions is useful. However, risk factors for mortality may depend on the life stage of subjects. To investigate the risk factor for the mortality, the life course approach is necessary., Competing Interests: The authors declare no conflict of interest.

Quality of life (QOL) and mortality are true endpoints of epidemiological or medical research, especially for community-dwelling older adults. Nutritional status and activities of daily living (ADLs) are associated with QOL and mortality. Good oral health status supports a good nutritional status. The aim of this study was to elucidate the complex structure of these important health-related factors. We surveyed 354 healthy older adults at the age of 85. Nutritional status was evaluated by the serum level of albumin. QOL, ADLs, self-assessed chewing ability, serum albumin level, and mortality during the 15 year follow up period were analyzed. Self-assessed chewing ability was associated with QOL and ADLs. Self-assessed chewing ability for slight-hard foods was associated with mortality in men. However, it was not associated with the serum albumin level. The serum albumin level was associated with mortality in women. These results indicate that maintaining good oral function is not enough. Nutritional instruction in accordance with oral function is indispensable for health promotion in older adults. When planning health promotion strategies for older adults, different strategies are needed for men and women.

Autophagy is a highly conserved intracellular digestion process that degrades damaged proteins and organelles but the biological roles of autophagy in pathological aspects of oral tissues remain largely unknown. We sought to elucidate the function of autophagy, especially its interplay with apoptosis and oxidative stress, in the oral toxicity induced by exposure to 5 mM sodium fluoride (NaF). Human cementoblasts (HCEM-2) in culture were exposed to 5 mM NaF for 5 min, after which cell viability and cell apoptosis were assessed using the MTS assay and an Annexin V-FITC/PI apoptosis detection kit, respectively. Quantitative RT-PCR and Western blotting were performed to characterize the expression levels of markers for autophagy, apoptosis, and oxidative stress. Senescence-resistant (SAMR1) mice were exposed to 5 mM NaF in their drinking water from 12 to 58 weeks. Micro-computed tomography was used to measure changes in their alveolar bone while immunohistochemistry and immunofluorescence staining was used to evaluate protein expression levels. HCEM-2 cells exposed to 5 mM NaF had decreased levels of autophagy, as shown by reduced expression levels of ATG5, Beclin-1 and LC3-II, elicited apoptosis, which in turn induced oxidative stress and inflammation, as manifested by elevated levels of Bax, cleaved caspase-3, SOD1 and phospho NF-κB. Treatment of mice with 5 mM NaF resulted in histological abnormalities in periodontal tissues, induced excessive oxidative stress and apoptosis, and reduced autophagy. Micro-computed tomography analysis demonstrated that 5 mM NaF caused a decrease in bone areas of mice compared with controls. Exposure to 5 mM NaF induced RANKL (receptor activator of nuclear factor κB ligand) and cathepsin K expression in periodontal tissues, while ATG5 and Beclin-1 expression was abrogated by 5 mM NaF. Taken together, our findings suggest that 5 mM NaF elicits oral toxicity that contributes to excessive apoptosis, oxidative stress, and defective autophagy, which aggravates periodontal tissue damage.

Purpose: To report a case of primary vitreoretinal lymphoma in which a macular hole developed after a diagnostic pars plana vitrectomy., Methods: A retrospective interventional case report., Results: A 65-year-old woman presented with worsening vision in the left eye. Fundus examination showed vitreous haze and multifocal, yellow-white infiltrates in the retina and under the retinal pigment epithelium in the left eye. She underwent a diagnostic pars plana vitrectomy in that eye. Undiluted vitreous specimen showed an increased interleukin-10 level (1,470 pg/mL) with an elevated interleukin-10 to interleukin-6 ratio of 15.1; cytologic analysis of the vitreous showed atypical lymphoid cells with large irregular nuclei and scanty cytoplasm. The retinal and sub-retinal pigment epithelial infiltrates responded well to intravitreal methotrexate injections, but a macular hole developed in the left eye. The second pars plana vitrectomy with internal limiting membrane peeling and 20% sulfur hexafluoride gas tamponade successfully closed the macular hole., Conclusion: Macular hole closure can be accomplished in eyes receiving intravitreal methotrexate injections for treating primary vitreoretinal lymphoma.

Background/aims: Lacerations of the oral mucosa and fractures of alveolar processes commonly occur in traumatic dental injuries (TDIs). Impaired wound healing and tissue regeneration have severe consequences on the quality of life. Bone marrow mesenchymal stem cells (BMMSCs) possess the ability of self-renewal and multipotential differentiation. Treatment with low-level sodium fluoride (NaF) has emerged as a promising approach to enhance wound repair. The aim of this study was to assess the effects of low-level NaF on soft tissue healing and on the proliferation, migration and extracellular matrix synthesis of BMMSCs., Material and Methods: BMMSCs derived from mice were treated with 50 μM, 500 μM, or 5 mM NaF for 12, 24, and 48 hours, and cell proliferation was assessed by the MTS assay. Cell motility was detected at 12 and 24 hours by a wound healing assay, and osteoblastic differentiation for 21 days by 1% Alizarin Red S staining in 50 μM NaF-treated BMMSCs. Gene expression of Runx2 and Osteocalcin was evaluated by quantitative real-time PCR. An experimental rat skin wound model was employed, and levels of c-Myc, Ki67, fibronectin, and vimentin were assessed by immunohistochemistry., Results: There was a significant induction in the proliferation and migration of BMMSCs treated with 50 μM NaF. The expression of Ki67 and c-Myc protein was increased in tissues treated with 50 μM NaF, and the expression of fibronectin and vimentin in the 50 μM NaF-treated tissues was stimulated. Alizarin Red staining revealed enhanced mineralization in 50 μM NaF-treated BMMSCs with increased expression of Runx2 and Osteocalcin, indicating their upregulated osteogenic differentiation., Conclusion: Low-level NaF could promote soft tissue healing and hard tissue regeneration., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The association between dental status and mortality in community-dwelling older adults has been documented by several studies. The aim of this study was to analyze the contribution of self-assessed chewing ability, number of remaining teeth and serum albumin levels to mortality and the interactions between the three factors., Methods: A 20-year follow-up study was conducted with 666 subjects aged 80 years (from 1996 to 2017) who resided in the 8 areas served by one health center in Iwate Prefecture. Health check-ups including physical fitness measurements were conducted at a meeting place or gymnasium. Medical interview and blood sampling were conducted by physician. Oral examination was examined by dentist. The number of remaining teeth, serum albumin levels, and self-assessed chewing ability were used as predictors of mortality., Results: Among the 608 subjects (233 men and 375 women) included in this study, only 12 subjects (1.97%) survived after 20 years of follow-up. For men, dental status and serum levels of albumin were significantly associated with mortality. The hazard ratios of self-assessed chewing ability calculated by item response theory analysis and the inability to chew at least one food adjusted for serum albumin and tooth conditions were statistically significant in men. When adjusted by health status evaluated by blood tests, self-assessed chewing ability was statistically significant in men. According to path analysis, self-assessed chewing ability and serum albumin independently affected mortality in men., Conclusion: Masticatory dysfunction may be an important risk factor for mortality in men, even though it was self-assessed. Retaining chewing ability might be a useful predictor of longevity in older male adults.

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Purpose: Several epidemiological studies have demonstrated the health benefits of polyphenols, but the associations between polyphenol intake and mortality including total and major causes of death remain unclear. We investigated the associations between subjects' total polyphenol intake and their mortality from all causes, cardiovascular disease (CVD), cancer, and other causes of death in a population-based cohort study in Japan., Methods: A total of 29,079 residents of Takayama City, Japan were analyzed. Their dietary intake was assessed using a validated semi-quantitative food-frequency questionnaire (FFQ) in 1992. Mortality was ascertained over the subsequent 16 years. The dietary polyphenol intake was calculated by matching the subjects' food consumption data with our original polyphenol content database., Results: A total of 5339 deaths occurred during the follow-up. After multivariable adjustment, the highest quartile of total polyphenol intake compared with the lowest quartile was significantly associated with a lower risk of all-cause mortality (HR: 0.93, 95% CI: 0.82-0.99, p trend = 0.003). The subjects in the highest quartile showed significantly lower CVD mortality compared to those in the lowest quartile, and among the types of CVD mortality, a strong inverse association was observed for stroke mortality. Inverse associations were also observed for mortality from other causes, specifically digestive disease. The total polyphenol intake was not significantly associated with the risk of cancer mortality., Conclusions: The results of this prospective study indicate that dietary total polyphenol intake in Japanese is inversely associated with all-cause mortality and mortality from cardiovascular and digestive diseases.

Green tea and coffee contain various bioactive compounds (e.g., polyphenols), and their consumption has been proposed to decrease the risk of cardiovascular diseases. Here, we investigated the associations between the consumption of green tea and that of coffee and the prevalence of coronary artery disease (CAD) in Japanese patients. The study group was 612 patients who underwent coronary angiography at Tokyo Medical Center between July 2008 and February 2017. CAD was confirmed in 388 of the patients: one-vessel disease (1-VD, n=166); two-vessel disease (2-VD, n=112); three-vessel disease (3-VD, n=110). Myocardial infarction (MI) was found in 138 patients. After adjustment for well-known atherosclerotic risk factors and other dietary habits, greater green tea consumption was significantly inversely associated with CAD prevalence (p for trend=0.044), and the patients who drank >3 cups/d had a lower prevalence of CAD compared to those who drank <1 cup/d (odds ratio [OR]: 0.54, 95% CI: 0.30-0.98). Greater green tea consumption (>3 cups/d) was also associated with a decreased prevalence of 3-VD (OR: 0.49, 95% CI: 0.24-0.98, p-trend=0.047) and MI (OR: 0.51, 95% CI: 0.27-0.97, p-trend=0.037). In contrast, coffee consumption was not associated with CAD or MI. In subgroup analyses, the inverse association between green tea consumption and CAD or MI was found in the high intake groups of vegetables or fruits but not in the low intake groups of vegetables or fruits. These results suggest a beneficial effect of green tea consumption, especially with a diet rich in vegetables and fruits, against coronary atherosclerosis in Japanese.

Background: Inflammation in endothelial cells induces production of inflammatory cytokines and monocytes adhesion, which are crucial events in the initiation of atherosclerosis. Aronia berry ( Aronia meranocalpa ), also called black chokeberry, contains abundant anthocyanins that have received considerable interest for their possible relations to vascular health., Objective: The aim of this study was to investigate whether an anthocyanin-rich extract obtained from aronia berry can attenuate inflammatory responses in vascular endothelial cells., Methods: As a model of vascular endothelial inflammation, human umbilical vein endothelial cells (HUVECs) pretreated with aronia berry extract were stimulated with tumor necrosis factor-alpha (TNF-α). The expression levels of cytokines and adhesion molecules were analyzed. To investigate the effects of aronia berry extract on the adhesion of THP-1 monocytic cell, the static adhesion assay was carried out. The possible molecular mechanisms by which aronia berry extract regulated vascular inflammatory responses were explored., Results: The mRNA expressions of interleukins (IL-1β, IL-6, and IL-8) and monocyte chemoattractant protein-1 (MCP-1) upregulated by TNF-α were significantly suppressed by pretreatment with aronia berry extract. Aronia berry extract decreased TNF-α-induced monocyte/endothelial adhesion and suppressed vascular cell adhesion molecule-1 (VCAM-1) expression, but did not affect intercellular adhesion molecule-1 (ICAM-1) expression. Moreover, aronia berry extract decreased the phosphorylation of signal transducer and activator of transcription 3 (STAT3) and the nuclear levels of STAT3 and interferon regulatory transcription factor-1 (IRF1). The nuclear translocation of nuclear factor-kappa B (NF-κB) was not inhibited by aronia berry extract., Conclusion: Aronia berry extract could exert anti-atherosclerotic effects on TNF-α-induced inflammation through inhibition of STAT3/IRF1 pathway in vascular endothelial cells., Competing Interests: This work was partly supported by JSPS KAKENHI (Grant nos. 15H02895, 16H03033, and 17K00847) from the Japan Society for the Promotion of Science. The authors declare no potential conflicts of interest.