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Background: Phrenic nerve injury (PNI) is one of the common complications in atrial fibrillation (AF) ablation, which often recovers spontaneously. However, the course of its recovery has not been examined fully, especially in regard to the different ablation methods. We sought to compare the recovery course of PNI in cryoballoon, laser balloon, and radiofrequency ablation., Methods: This multicenter retrospective study analyzed 355 patients who suffered from PNI during AF ablation. PNI occurred during cryoballoon ablation (CB group) and laser balloon ablation (LB group) for a pulmonary vein isolation in 288 and 20 patients, and radiofrequency ablation for a superior vena cava (SVC) isolation (RF-SVC group) in 47 patients, respectively RESULTS: There was a significant difference in the estimated probability of PNI recovery after the procedure between the methods (p = 0.01). PNI recovered significantly earlier in the CB group, especially within 24 h and 3 months post-procedure (the percentage of the recovery within 24 h and 3 months: 49.7% and 71.5% in the CB group, 15.0% and 22.2% in the LB group, and 23.4% and 41.9% in the RF-SVC group, respectively). Persistent PNI after 12 months was observed in only seven patients in the CB group, one in the LB group, and four in the RF-SVC group, respectively., Conclusion: PNI rarely persists over 12 months after AF ablation; however, there is a difference in the timing of its recovery. PNI recovers quicker with cryoballoon ablation than with laser balloon ablation or radiofrequency ablation of the SVC., Competing Interests: Declarations. Conflict of interest: Dr. Miyazaki belongs to the endowed departments of Medtronic, Boston Scientific, and Japan Lifeline, and has received speaker honoraria from Medtronic and Boston Scientific. The other authors declare that they have no conflict of interest. Ethical standards: The study protocol was approved by the institutional review board of the Tokyo Medical and Dental University Hospital. The need for patient consent was waived due to the anonymized and retrospective nature of the study (opt-out method). The study complied with the Declaration of Helsinki., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Background: QRS morphology can change during ventricular arrhythmias (VAs) with the appearance of bundle branch block (BBB)., Methods: We retrospectively investigated 195 consecutive patients who underwent an initial ablation of VA. The study inclusion criteria were VAs that were successfully ablated in the outflow tract (OT) and in whom right bundle branch block (RBBB) was induced by catheter manipulation close to the His bundle area during sinus rhythm, before any radiofrequency application. We analyzed the QRS morphology of the VAs with and without RBBB during sinus beats., Results: Twenty-four patients (age 59 ± 17 years, female 14) developed RBBB at some point during their procedure. The successful ablation sites of the VAs were the right ventricular outflow tract (RVOT) in 12 patients, pulmonary artery in one, left coronary cusp in five, right coronary cusp in three, right-left cusp junction in two, and great cardiac vein in two. QRS-morphology change was observed in five (20%) cases. The successful ablation sites in that group were the left coronary cusp in three cases, right coronary cusp in one, and RVOT septum in one. The QRS duration of the VAs increased during RBBB., Conclusions: There are some cases of OT-VAs in which the QRS waveform changes with the appearance of catheter induced RBBB. We need to be aware that when QRS morphology changes during an OT-VA ablation, it does not necessarily mean that the origin or exit of the VA has changed., (© 2024 Wiley Periodicals LLC.)

Gingival wound healing plays a critical role in maintaining oral health. However, this process can be delayed by oxidative stress and excessive inflammatory responses. In this study, we established a human inflammatory gingival tissue equivalent (iGTE) to investigate the inhibitory effects of hydrogen-rich water (HW), enzyme-digested edible bird's nest (EBND) and sialic acid (SA) on PMA (an inducer of oxidative free radicals)- and LPS (an inflammatory stimulus)-impaired wound healing. The iGTE was constructed by human gingival fibroblasts (hGFs), keratinocytes and macrophages under three-dimensional conditions. Wounds in the iGTE and hGF/keratinocyte monolayers were created by mechanical injury. Tissues and cells were pretreated with HW, EBND, and SA, and then exposed to the inflammatory and oxidative environment induced by PMA (10 ng/mL) and LPS (250 ng/mL). The inflammatory cytokines IL-6 and IL-8 were quantitatively analyzed by ELISA. Histopathological image analysis was performed by HE and immunofluorescence staining. In the iGTE, PMA/LPS significantly reduced the epithelial thickness while causing a decrease in K8/18, E-cadherin, laminin and elastin expression and an increase in COX-2 expression along with ulcer-like lesions. In mechanically scratched hGFs and keratinocyte monolayers, PMA/LPS significantly impaired wound healing, and promoted the secretion of IL-6 and IL-8. Pretreatment of HW, EBND, and SA significantly suppressed PMA/LPS-induced wound healing delay and inflammatory responses in cell monolayers, as well as in the iGTE. Remarkably, the combined use of HW and EBND exhibited particularly robust results. Combined use of HW and EBND may be applied for the prevention and treatment of wound healing delay., (© 2024. The Author(s) under exclusive licence to Japan Human Cell Society.)

Background/aims: Extracellular acidic conditions impair cellular activities; however, some cancer cells drive cellular signaling to adapt to the acidic environment. It remains unclear how ovarian cancer cells sense changes in extracellular pH. This study was aimed at characterizing acid-inducible currents in an ovarian cancer cell line and evaluating the involvement of these currents in cell viability., Methods: The biophysical and pharmacological properties of membrane currents in OV2944, a mouse ovarian cancer cell line, were studied using the whole-cell configuration of the patch-clamp technique. Viability of this cell type in acidic medium was evaluated using the MTT assay., Results: OV2944 had significant acid-sensitive outwardly rectifying (ASOR) Cl - currents at a pH 50 of 5.3. The ASOR current was blocked by pregnenolone sulfate (PS), a steroid ion channel modulator that blocks the ASOR channel as one of its targets. The viability of the cells was reduced after exposure to an acidic medium (pH 5.3) but was slightly restored upon PS administration., Conclusion: These results offer first evidence for the presence of ASOR Cl - channel in ovarian cancer cells and indicate its involvement in cell viability under acidic environment., Competing Interests: The authors have no conflicts of interest to declare., (© Copyright by the Author(s). Published by Cell Physiol Biochem Press.)

Objective: Pancreatic cancer (PC) originates and progresses with genetic mutations in various oncogenes and suppressor genes, notably KRAS, CDKN2A, TP53, and SMAD4, prevalent across diverse PC cells. In addition to genetic mutations/deletions, persistent exposure to high-risk factors, including obesity, induces whole-genome scale epigenetic alterations contributing to malignancy. However, the impact of obesity on DNA methylation in the presymptomatic stage, particularly in genes prone to PC mutation, remains uncharacterized., Results: We analyzed the methylation levels of 197 loci in six genes (KRAS, CDKN2A, TP53, SMAD4, GNAS and RNF43) using Illumina Mouse Methylation BeadChip array (280 K) data from pancreatic exocrine cells obtained from high-fat-diet (HFD) induced obese mice. Results revealed no significant differences in methylation levels in loci between HFD- and normal-fat-diet (NFD)-fed mice, except for RNF43, a negative regulator of Wnt signaling, which showed hypermethylation in three loci. These findings indicate that, in mouse pancreatic exocrine cells, high-fat dietary obesity induced aberrant DNA methylation in RNF43 but not in other frequently mutated PC-related genes., (© 2024. The Author(s).)

Background: The increasing number of patients with miliary tuberculosis (MTB) is a concern in an aging society because of its high mortality rate. Several prognostic biomarkers for MTB have been identified; however, the predictive ability of monocytes as biomarkers remains unknown. This study demonstrates the usefulness of monocytes as prognostic biomarkers for MTB., Materials and Methods: We retrospectively compared the clinical findings of 52 patients with MTB hospitalized between April 2013 and October 2021. The predictive ability of biomarkers for 3-month prognosis and their cutoff values were calculated. Survival times and longitudinal changes in monocytes after initiating treatment were compared., Results: A smaller number of monocytes (#M), higher lymphocyte-monocyte ratio (LMR), higher neutrophil-monocyte ratio, and poorer performance status were associated with death within 3 months. #M was an independent prognostic factor. #M and LMR exhibited the highest predictive performance compared to others using receiver operating characteristic curve analysis (area under the curve = 0.86 and 0.85, respectively). Survival time was shorter in patients with #M ≤ 200 cells/μL and LMR > 2.5. Rapidly increasing #M after treatment was related to better prognosis in patients with #M ≤ 200 cells/μL at diagnosis., Conclusions: #M at diagnosis and longitudinal changes in monocytes are related to MTB prognosis., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dr. Shima received personal fees from AstraZeneca and Ono Pharmaceutical Co., Ltd. outside of the submitted work during the study period. The other authors declare that they have no conflicts of interest.., (© 2024 The Authors.)

Background: The concept of ablation index (AI) was introduced to evaluate radiofrequency (RF) ablation lesions. It is calculated from power, contact force (CF), and RF duration. However, other factors may also affect the quality of ablation lesions. To examine the difference in RF lesions made during sinus rhythm (SR) and atrial fibrillation (AF)., Methods: Sixty patients underwent index pulmonary vein isolation during SR ( n  = 30, SR group) or AF ( n  = 30, AF group). All ablations were performed with a power of 50 W, a targeted CF of 5-15 g, and AI of 400-450 using Thermocool Smarttouch SF. The CF, AI, RF duration, temperature rise (Δtemp), impedance drop (Δimp), and the CF stability of each ablation point quantified as the standard deviation of the CF (CF-SD) were compared between the two groups., Results: A total of 3579 ablation points were analyzed, which included 1618 and 1961 points in the SR and the AF groups, respectively. Power, average CF, RF duration per point, and the resultant AI (389 ± 59 vs. 388 ± 57) were similar for the two rhythms. However, differences were seen in the CF-SD (3.5 ± 2.2 vs. 3.8 ± 2.1 g, p  < .01), Δtemp (3.8 ± 1.3 vs. 4.0 ± 1.3°C, p  < .005), and Δimp (10.3 ± 5.8 vs. 9.4 ± 5.4 Ω, p  < .005)., Conclusions: Despite similar AI, various RF parameters differed according to the underlying atrial rhythm. Ablation delivered during SR demonstrated less CF variability and temperature increase and greater impedance drop than during AF., Competing Interests: The authors declare no conflicts of interest., (© 2024 The Authors. Journal of Arrhythmia published by John Wiley & Sons Australia, Ltd on behalf of Japanese Heart Rhythm Society.)

Background: A well-functioning vascular access (VA) is crucial for the patients on maintenance hemodialysis (HD). Central venous stenosis (CVS) is a common, yet, overlooked complication of VA as its diagnosis is challenging. Moreover, its effect on the flow volume (FV) and the peripheral resistive index (RI) was not well discussed before. Despite the availability of doppler ultrasound (DUS) in most centers, direct visualization of central veins using DUS is quite difficult., Methods: This is a retrospective single-center self-controlled cohort study that was conducted in a specialized vascular access management tertiary center in Japan and included all patients with CVS who underwent percutaneous transluminal angiography (PTA) with or without vascular stenting in the period from January 2014 to September 2022. The patients were divided according to their VA type into arteriovenous fistula (AVF), and arteriovenous graft (AVG) groups, then subdivided, according to the type of stenosis, into two subgroups: CVS, and mixed central and peripheral venous stenosis (MVS) groups. The FV and RI of the feeding artery were compared in the same procedure before and after PTA to assess the impact of the procedure itself., Results: Data of 485 percutaneous transluminal angiography procedures (PTA), performed in 95 patients during the period from January 2014 to September 2022, were collected. FV and RI were significantly affected in the patients with MVS than patients with CVS. After PTA, both FV and RI were significantly improved. The improvement rate of FV and RI after PTA were significantly higher in patients with MVS than patients with CVS. However, it was difficult to determine the cut-off value to diagnose or to assess the improvement of CVS., Conclusion: Our findings suggest that FV and RI measurement by DUS are good tools, along with the clinical findings for assessment of CVS in certain situations., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: Skin disorders are the most common side effect associated with epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) therapy. It is important to manage skin lesions. Adapalene has been used to treat skin lesions caused by EGFR-TKIs in some cases. The aim of this study was to investigate the functional mechanism of adapalene in erlotinib-induced skin disorder., Methods: To analyze the effect of adapalene on skin rash, afatinib and adapalene were administered to mice. The relationship between the concentration of adapalene and skin disorders was also examined by analyzing AQP3 expression. A skin lesion model was experimentally established in human skin keratinocytes (HaCaT) by using erlotinib with TNF-α and IL-1β. We used qRT-PCR to analyze chemokine-induced inflammation and western blotting to analyze the effects of adapalene on the NF-κB signaling pathway. Antimicrobial peptides and adhesion factors were also examined using qRT-PCR., Results: Mice administered 0.01% adapalene had less skin inflammation than mice treated with afatinib alone. The expression level of AQP3 decreased in an adapalene concentration-dependent manner. The mRNA levels of proinflammatory cytokines such as CCL2 and CCL27 in HaCaT cells were significantly reduced by adapalene. The expression of an antimicrobial peptide, hBD3, was upregulated after adapalene treatment. Adhesion factors, such as E-cadherin, were significantly downregulated by EGFR-TKI and significantly upregulated by adapalene treatment. Western blot analysis suggested that erlotinib-induced phosphorylation of p65 was decreased by adapalene., Conclusion: We suggest that adapalene may be a possible treatment option for skin disorders induced by EGFR-TKIs., (© 2024 The Authors. Thoracic Cancer published by John Wiley & Sons Australia, Ltd.)

Introduction: It would be helpful in determining ablation strategy if the occurrence of perimitral atrial tachycardia (PMAT) could be predicted in advance. We investigated whether estimated perimitral conduction time (E-PMCT), namely, twice the time between coronary sinus (CS) pacing and the ensuing wave-front collision at the opposite side of the mitral annulus, correlated with the cycle length of PMAT and could predict future PMAT., Methods and Results: We retrospectively (retrospective cohort) and prospectively (validation cohort) investigated atrial fibrillation patients who had received pulmonary vein isolation (PVI) and in whom left atrial maps had been created during CS pacing. We calculated their E-PMCT. PMAT was observed either by provocation or during follow-up in 25, AT other than PMAT was observed in 24 (non-PMAT AT group), and 53 patients never displayed any AT (no-AT group) in the retrospective cohort. In the PMAT group of the retrospective cohort, a strong positive correlation was observed between the PMAT CL and E-PMCT (r = .85, p < 0.001). PMAT was never induced nor observed in patients with E-PMCT less than 176 ms, and the best cut-off value for PMAT was 180 ms by receiver-operating characteristic curve analysis. In the validation cohort of 76 patients, the cut-off value of the E-PMAT less than 180 ms predicted noninducibility of PMAT, with a sensitivity of 78.6%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 25.0%., Conclusion: Short E-PMCT may predict noninducibility of PMAT and guide a less invasive ablation strategy., (© 2024 Wiley Periodicals LLC.)

Background: Symptomatic gastric hypomotility (SGH) is a rare but major complication of atrial fibrillation (AF) ablation, but data on this are scarce., Objective: We compared the clinical course of SGH occurring with different energy sources., Methods: This multicenter study retrospectively collected the characteristics and clinical outcomes of patients with SGH after AF ablation., Results: The data of 93 patients (67.0 ± 11.2 years, 68 men, 52 paroxysmal AF) with SGH after AF ablation were collected from 23 cardiovascular centers. Left atrial (LA) ablation sets included pulmonary vein isolation (PVI) alone, a PVI plus a roof-line, and an LA posterior wall isolation in 42 (45.2%), 11 (11.8%), and 40 (43.0%) patients, respectively. LA ablation was performed by radiofrequency ablation, cryoballoon ablation, or both in 38 (40.8%), 38 (40.8%), and 17 (18.3%) patients, respectively. SGH diagnoses were confirmed at 2 (1-4) days post-procedure, and 28 (30.1%) patients required re-hospitalizations. Fasting was required in 81 (92.0%) patients for 4 (2.5-5) days; the total hospitalization duration was 11 [7-19.8] days. After conservative treatment, symptoms disappeared in 22.3% of patients at 1 month, 48.9% at 2 months, 57.6% at 3 months, 84.6% at 6 months, and 89.7% at 12 months, however, one patient required surgery after radiofrequency ablation. Symptoms persisted for >1-year post-procedure in 7 patients. The outcomes were similar regardless of the energy source and LA lesion set., Conclusions: The clinical course of SGH was similar regardless of the energy source. The diagnosis was often delayed, and most recovered within 6 months, yet could persist for over 1 year in 10%., 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. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (© 2023 Miyazaki, Kobori, Jo, Keida, Yoshitani, Mukai, Sagawa, Asakawa, Sato, Yamao, Horie, Manita, Fukaya, Hayashi, Tanimoto, Iwayama, Chiba, Sato, Sekiguchi, Sugiura, Iwai, Isonaga, Miwa, Kato, Inaba, Hirota, Nagata, Ono, Hachiya, Yamauchi, Goya, Nitta, Tada and Sasano.)

Metastasis, a multistep process including cancer cell migration and invasion, is the major cause of mortality in patients with cancer. Here, we investigated the effect of dicalcin, a Ca 2+ -binding protein, on the invasion and metastasis of ovarian cancer (OC) cells. Extracellularly administered dicalcin bound to the membrane of OV2944 cells, mouse OC cells, and suppressed their migration in vitro; however, cell viability or proliferation were unaffected. Repeated intraperitoneal injection of a partial peptide of dicalcin (P6) prolonged the survival, and reduced the number of microcolonies in the livers of cancer-bearing mice. P6 bound to the ganglioside GM1b in a solid-phase assay; treatment with P6 inhibited the constitutive activation of Erk1/2 in OC cells, whereas excess administration of GM1b augmented Erk activity and cancer cell migration in vitro. Thus, dicalcin, a novel suppressor of invasion and metastasis of OC cells, acts via the GM1b-Erk1/2 axis to regulate their migration., (© 2023. Springer Nature Limited.)

Background: Although atrial flutter (AFL) is a common arrhythmia that is based on a macro-reentrant tachycardia around the tricuspid annulus, the factors giving rise to typical AFL (t-AFL) versus reverse typical AFL (rt-AFL) are unknown. To investigate the difference between t-AFL and rt-AFL circuits using ultrahigh resolution mapping of the right atrium., Methods: We investigated 30 isthmus-dependent AFL patients (mean age 71, 28 male) who underwent first-time cavo-tricuspid isthmus (CTI) ablation guided by Boston Scientific's Rhythmia mapping system and divided them into two groups: t-AFL (22 patients) and rt-AFL (8 patients). We compared the anatomy and electrophysiology of their reentrant circuits., Results: Baseline patient characteristics, use of antiarrhythmic drugs, prevalence of atrial fibrillation, AFL cycle length (227.1 ± 21.4 vs. 245.5 ± 36.0 ms, p = .10), and CTI length (31.9 ± 8.3 vs. 31.1 ± 5.2 mm, p = .80) did not differ between the two groups. Functional block was observed at the crista terminalis in 16 patients and at the sinus venosus in 11. No functional block was observed in three patients, all of whom belonged to the rt-AFL group. That is, functional block was observed in 100% of the t-AFL group as opposed to 5/8 (62.5%) of the rt-AFL (p < .05). Slow conduction zones were frequently observed at the intra-atrial septum in the t-AFL group and at the CTI in the rt-AFL group., Conclusion: Mapping with ultrahigh-resolution mapping showed differences between t-AFL and rt-AFL in conduction properties in the right atrium and around the tricuspid valve, which suggested directional mechanisms., (© 2023 Wiley Periodicals LLC.)

Background and Aims: First-year students encounter substantial stress when they enter university. Their mental health often depends upon how well they cope with the stress of university life. Salivary components are well known to reflect the stress status of the students; however, the relationship between salivary components and coping styles remains unknown., Methods: In this study, 54 healthy first-year students voluntarily completed a questionnaire that addressed three different coping styles: problem-focused, emotion-focused, and escape-focused. We simultaneously collected salivary samples from students in the classroom and measured concentrations of salivary cortisol and α-amylase by enzyme-linked immunosorbent assays over 4 months., Results: We examined the relationship between coping style and salivary cortisol concentrations and found that the mean salivary cortisol concentrations were significantly lower in students who had a higher Likert-type score for the problem-focused coping style than in students who had a lower score. The difference in the mean cortisol concentrations between the two groups increased over time. However, we observed no apparent correlation between α-amylase concentrations and Likert scores of the three coping styles., Conclusion: These results suggest that salivary cortisol concentrations might reflect the stress-coping status, particularly involving the problem-focused coping style., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors. Health Science Reports published by Wiley Periodicals LLC.)

Background: Mfa1 fimbriae of the periodontal pathogen Porphyromonas gingivalis are responsible for biofilm formation and comprise five proteins: Mfa1-5. Two major genotypes, mfa1 70 and mfa1 53 , encode major fimbrillin. The mfa1 70 genotype is further divided into the mfa1 70A and mfa1 70B subtypes. The properties of the novel mfa1 70B remain unclear., Methods: Fimbriae were purified from P. gingivalis strains JI-1 (mfa1 70A ), 1439 (mfa1 70B ), and Ando (mfa1 53 ), and their components and their structures were analyzed. Protein expression and variability in the antigenic specificity of fimbrillins were compared using Coomassie staining and western blotting using polyclonal antibodies against Mfa1 70A , Mfa1 70B , and Mfa1 53 proteins. Cell surface expression levels of fimbriae were analyzed by filtration enzyme-linked immunosorbent assays., Results: The composition and structures of the purified Mfa1 fimbriae of 1439 was similar to that of JI-1. However, each Mfa1 protein of differential subtype/genotype was specifically detected by western blotting. Mfa1 70B fimbriae were expressed in several strains such as 1439, JKG9, B42, 1436, and Kyudai-3. Differential protein expression and antigenic heterogeneities were detected in Mfa2-5 between strains., Conclusion: Mfa1 fimbriae from the mfa170A and mfa170B genotypes indicated an antigenic difference suggesting the mfa170B, is to be utilized for the novel classification of P. gingivalis., Competing Interests: No potential conflict of interest was reported by the authors., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

We report a rare case of a mobile ectopic calcification in the left atrium requiring surgical excision 9 years after multiple atrial fibrillation ablations. ( Level of Difficulty: Intermediate. )., Competing Interests: The authors have reported that they have no relationships relevant to the contents of this paper to disclose., (© 2023 The Authors.)

Background: Cardiac manifest of COVID-19 infection was widely reported. The pathophysiology is thought the combination of direct damage caused by viruses and myocardial inflammation caused by immune responses. We tracked the inflammatory process of fulminant myocarditis associated with COVID-19 infection using multi-modality imaging., Case Summary: A 49-year-old male with COVID-19 went into cardiac arrest from severe left ventricular dysfunction and cardiac tamponade. He was treated with steroids, remdesivir, and tocilizumab but failed to maintain circulation. He recovered with pericardiocentesis and veno-arterial extracorporeal membrane oxygenation in addition to the immune suppression treatment. In this case, a series of chest computed tomography (CT) was performed on Days 4, 7, and 18 and cardiac magnetic resonance (MR) on Days 21, 53, and 145., Discussion: Analysis of the inflammatory findings on CT in this case showed that intense inflammation around the pericardial space was observed at an early stage of the disease. Although inflammatory findings in the pericardial space and chemical markers had improved according to non-magnetic resonance imaging (MRI) tests, the MRI revealed a notable long inflammatory period more than 50 days., Competing Interests: Conflict of interest: None declared., (© The Author(s) 2023. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Aberrant DNA methylation is associated with oncogenesis of various human cancers, including pancreatic cancer (PC). PC is the seventh most common cancer, and obesity is a known high-risk factor. However, whether obesity influences DNA methylation in pancreatic exocrine cells and if this influences PC development remain unclear. Here, we performed an epigenome-wide analysis of isolated pancreatic exocrine cells obtained from mice with high-fat-diet-induced obesity (DIO). Using the Illumina Mouse Methylation BeadChip array (280K), we identified 316 differentially methylated regions (DMRs) that were enriched for cellular processes, such as DNA repair, transcription regulation, and cell proliferation, which confirmed obesity-related dysregulation of certain metabolic processes in the pancreatic cells in DIO mice. Comparing the DMRs with those in stage IB PC helped identify 82 overlapping DMRs. Three pathways including the cell hypertrophy pathway involving PLC, PKC, SMAD2/3, and TRKA; the metabolic control pathway involving CREB and AMPK; and the potassium regulation pathway involving K + -channels, were shared between the pancreatic exocrine cells from DIO mice and stage IB PC. Enhanced alteration in the methylation level was observed in PC compared to that in DIO mice. These findings indicated that obesity influences DNA methylation in pancreatic exocrine cells of DIO mice, and persistent dysregulation of DNA methylation in individuals with obesity may result in PC development., 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 © 2022 Elsevier Inc. All rights reserved.)

Aims: To describe the performance of machine learning (ML) applied to predict future metabolic syndrome (MS), and to estimate lifestyle changes effects in MS predictions., Methods: We analyzed data from 17,182 adults attending a checkup program sequentially (37,999 visit pairs) over 17 years. Variables on sociodemographic attributes, clinical, laboratory, and lifestyle characteristics were used to develop ML models to predict MS [logistic regression, linear discriminant analysis, k-nearest neighbors, decision trees, Light Gradient Boosting Machine (LGBM), Extreme Gradient Boosting]. We have tested the effects of changes in lifestyle in MS prediction at individual levels., Results: All models showed adequate calibration and good discrimination, but the LGBM showed better performance (Sensitivity = 87.8 %, Specificity = 70.2 %, AUC-ROC = 0.86). Causal inference analysis showed that increasing physical activity level and reducing BMI by at least 2 % had an effect of reducing the predicted probability of MS by 3.8 % (95 % CI = -4.8 %; -2.7 %)., Conclusion: ML models based on data from a checkup program showed good performance to predict MS and allowed testing for effects of lifestyle changes in this prediction. External validation is recommended to verify models' ability to identify at-risk individuals, and potentially increase their engagement in preventive measures., 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 © 2022 Elsevier B.V. All rights reserved.)

Introduction: Mitral valve surgery employing a superior transseptal approach (STA) is associated with arrhythmogenicity and intra-atrial conduction delay, despite being optimal for visualization of the surgical field. It is sometimes difficult to treat atrial tachycardias (AT) that arise after STA. To investigate AT circuits that arise after STA in detail in order to identify the optimal ablation line, using ultra-high-resolution mapping (UHRM)., Methods: We retrospectively analyzed 12 AT from 10 patients (median age 70 years, nine males) who had undergone STA surgery. The tachycardias were mapped using the Rhythmia mapping system (Boston Scientific, Natick, Massachusetts)., Results: The 12 STA-related AT (STA-AT) circuits were classifiable as follows according to location of the optimal ablation line: (1) peri-septal incision STA-AT (n = 3), (2) cavotricuspid isthmus (CTI) dependent STA-AT (n = 7), and (3) biatrial tachycardia (n = 2). Radiofrequency (RF) application terminated 11 of the 12 STA-AT. We found that difference in STA-AT circuit type was due to characteristics of the septal incision line made for STA. UHRM was important in identifying optimal ablation sites that did not create additional conduction disturbances in the right atrium (RA)., Conclusions: ATs after STA involve complex arrhythmia circuits due to multiple and long incision lines in the RA. Accurate understanding of the arrhythmia circuit and sinus conduction in the RA after STA is recommended for treating post-surgical tachycardia in a minimally invasive manner., (© 2022 Wiley Periodicals LLC.)

Background The association between alcohol consumption, atrial substrate, and outcomes after atrial fibrillation (AF) ablation remains controversial. This study evaluated the impacts of drinking on left atrial substrate and AF recurrence after ablation. Methods and Results We prospectively enrolled 110 patients with AF without structural heart disease (64±12 years) from 2 institutions. High-density left atrial electroanatomic mapping was performed using a high-density grid multipolar catheter. We investigated the impact of alcohol consumption on left atrial voltage, left atrial conduction velocity, and AF ablation outcome. Patients were classified as abstainers (<1 drink/wk), mild drinkers (1-7 drinks/wk), or moderate-heavy drinkers (>7 drinks/wk). High-density mapping (mean 2287±600 points/patient) was performed on 49 abstainers, 27 mild drinkers, and 34 moderate-heavy drinkers. Low-voltage zone and slow-conduction zone were identified in 39 (35%) and 54 (49%) patients, respectively. There was no significant difference in the proportions of low-voltage zone and slow-conduction zone among the 3 groups. The success rate after a single ablation was significantly lower in drinkers than in abstainers (79.3% versus 95.9% at 12 months; mean follow-up, 18±8 months; P =0.013). The success rate after a single or multiple ablations was not significantly different among abstainers and drinkers. In multivariate analysis, alcohol consumption ( P =0.02) and the presence of a low-voltage zone ( P =0.032) and slow-conduction zone ( P =0.02) were associated with AF recurrence after a single ablation, while low-voltage zone ( P =0.023) and slow-conduction zone ( P =0.024) were associated with AF recurrence after a single or multiple ablations. Conclusions Alcohol consumption was associated with AF recurrence after a single ablation but not changes in atrial substrate.

This study compared the effects of hydrogen-water (HW) bath on the oxygen radical absorption-based antioxidant capacity and the inflammatory indicator, C-reactive protein (CRP), in serum between healthy volunteers and inflammatory/collagen disease-patients. The HW bath apparatus supplied nano-bubbles with a diameter of 110 ± 10 nm and 338-682 μg/L of dissolved hydrogen after 120 minutes electrolysis, and nano-bubbles increased to 9.91 × 10 7 /mL along with the increase of correlative dissolved hydrogen. Ten-minute HW bath increased the oxygen radical absorption-based antioxidant capacity to 110.9 ± 9.2% at post-bathing 120 minutes, although unaltered with 10-minute normal water bath at 40°C in healthy subjects. The CRP level was repressed to 70.2 ± 12.1% at 120 minutes after HW bath, although rather increased for normal water bath. In the patients with connective tissue diseases, the CRP level was repressed to 3-24% upon 9 days to 4 months of HW bathing. In another six patients with diverse autoimmune-related diseases, upon daily HW bathing as long as 2-25 months, the pre-bathing CRP level of 5.31 mg/dL decreased to 0.24 mg/dL being within the standard-range, with relief of visible inflammatory symptoms for some cases. Thus, the HW bath with high-density nano-bubbles has beneficial effects on serum antioxidant capacity, inflammation, and the skin appearance. The study was approved by the Committee of Ethics, Japanese Center of Anti-Aging Medical Sciences (Authorization No. H-15-03-2, on January 15, 2019), which was a non-profitable organization officially authenticated by the Hiroshima Prefecture Government of Japan., Competing Interests: None

The physiological practice course at Saitama Medical University provides students with the opportunity to learn physiological principles through wet labs and discussions. To develop a more effective method for maximizing learning outcomes, we extended the course's schedule from one day (1d) to two days (2d) per theme, evaluated self-administered questionnaires between two different years (pre and post-change), and examined whether the increased course length affected learning outcomes. Within the 2018 curriculum year, every theme of the course was completed in a day, including experiments in the wet lab and discussions. In 2019, each theme was assessed for two days. The second-year undergraduate medical students anonymously submitted the self-assessment questionnaire that addressed several aspects, such as understanding of the theme, through a 5-point Likert scale. The average Likert scores varied from 4 to 4.5 point for all questions, and significant differences were not found between the 1d and 2d courses. However, the ratio of students with the highest points increased for one question of the 2d course: 43.6% (1d) to 53.4% (2d) for understanding. Further, the standard deviation (SD) values decreased in the 2d course for every question: 0.29 (1d) to 0.15 (2d) for interest, 0.33 (1d) to 0.19 (2d) for understanding, 0.30 (d) to 0.17 (d) for communication, 0.34 (1d) to 0.19 (2d) for general evaluation. This reduction in the SD values indicated that the educational content was imparted more efficiently to students in the 2d course. Thus, we concluded that extending the course time facilitated dissemination of educational content for every theme., Supplementary Information: The online version contains supplementary material available at 10.1007/s40670-022-01563-4., Competing Interests: Conflict of InterestThe authors declare no competing interests., (© The Author(s) 2022.)

Background: Ischemic compression is a manual therapy technique for myofascial pain. This study aimed to verify the effect of ischemic compression performed by family caregivers on myofascial pain syndrome (MPS) in patients and on the family's care burden., Methods: This multicenter, open-label, randomized, comparative study included patients with myofascial pain and their family caregivers who were randomized into the following groups: ischemia compression (performed by a family caregiver), sham ischemia compression, or untreated control. The effectiveness and safety of ischemic compression and the burden on family caregivers were evaluated. The primary endpoint was the rate of 50% or more improvement in the patient's mean numerical rating scale pain score in the previous 24 hours, 14 days after starting the intervention. The secondary endpoint was the rate of change in the family caregivers' reaction assessments., Results: A total of 75 patients and caregivers (70 patients with cancer and family caregivers) who received home medical care were enrolled at three facilities. The study completion rate was 94.7%, and there were no adverse events. The rate of 50% or more improvement in the numerical rating scale score was 64.0% in the ischemic compression group, 16.0% in the sham ischemic compression group, and 4.0% in the control group (P<0.001). Caregivers' self-esteem was significantly lower in the ischemic compression and sham ischemic compression groups than in the control group. However, there was no significant difference between the two groups (P=0.370)., Conclusions: Ischemic compression for myofascial pain in patients performed by family caregivers can increase the analgesic effect in patients and self-esteem in family caregivers., Trial Registration: The University Hospital Medical Information Network Clinical Trials Registry (approval number: UMIN000036605).

Introduction: Catheter ablation for perimitral atrial tachycardia (PMAT) that persists despite lateral mitral isthmus (LMI) ablation is challenging. The aim of this study was to identify the role of the ligament of Marshall (LOM) in PMATs that persist after LMI conduction block has been created, and evaluate the validity of ethanol infusion into the vein of Marshall (VOM) as treatment., Methods and Results: Sixteen consecutive PMATs in 13 patients that persisted despite apparent LMI conduction block, which was confirmed by ultrahigh-resolution mapping and entrainment pacing along the mitral annulus, were analyzed. PMATs were classified into two types based on the location of the endocardial breakthrough site: those utilizing the LOM (n = 13), which had a breakthrough site along with the LOM, and those not utilizing the LOM (n = 3), which had a breakthrough site at an anterior or posterior side of the LOM. Of the 16 PMATs, 5 PMATs (31%) were not suitable for ethanol infusion into the VOM because the LOM was not involved in the tachycardia circuit or because of the anatomy of the VOM. Fourteen PMATs (88%) were successfully terminated solely by breakthrough site ablation. At a mean follow-up period of 12 ± 9 months, 10 (77%) patients have remained free from atrial tachyarrhythmias., Conclusion: In cases of PMAT following LMI ablation, epicardial conduction over the LMI can occur independently of the LOM. Ethanol infusion into the VOM in such cases would not abolish residual epicardial conduction. The anatomy of the VOM can also preclude the use of this method., (© 2021 Wiley Periodicals LLC.)

Objectives: Previous studies reported the association between inflammation and atrial fibrillation (AF). Pericoronary adipose tissue (PCAT) attenuation, PCATA, on cardiac CT angiography (CTA) reflects pericoronary inflammation. We hypothesized that the PCATA predicts AF recurrence after cryoballoon ablation (CBA) for paroxysmal and persistent AF., Methods: We studied 364 patients (median age, 65 years) with persistent ( n = 41) and paroxysmal ( n = 323) AF undergoing successful first-session second-generation CBA with pre-ablation cardiac CTA. Three-vessel (3V)-PCATA was defined as the mean CT attenuation value of PCAT of all three major coronary arteries. Predictors of AF recurrence during follow-up were evaluated., Results: AF recurrence after the 3-month blanking period was detected in 90 patients (24.7%) during the median follow-up of 26 (interquartile range, 19-42) months. AF recurrence was associated with prior stroke and statin use, NT-proBNP and high-sensitivity cardiac troponin-I levels, left ventricular dimension, left atrial volume index (LAVI), 3V-PCATA, and early AF recurrence during the blanking period. On multivariable Cox proportional hazard analysis, prior stroke (hazard ratio [HR], 2.208, 95% confidence interval [CI], 1.166-4.180, p = 0.015), LAVI (HR, 1.030, 95% CI, 1.010-1.051, p = 0.003), 3V-PCATA (HR, 1.034, 95% CI, 1.001-1.069, p = 0.046), and early AF recurrence (HR, 2.858, 95% CI, 1.855-4.405, p < 0.001) remained statistically significant., Conclusion: Pre-ablation CTA-derived 3V-PCATA, representing pericoronary inflammation, was an independent predictor of recurrence after first-session AF ablation using a second-generation cryoballoon., Advances in Knowledge: Assessment of 3V-PCATA may identify patients at high risk of AF recurrence after CBA for AF.

Implantation is a highly organized process that involves an interaction between a competent blastocyst and a receptive uterus. Despite significant research efforts, the molecular mechanisms governing this complex process remain elusive. Here, we investigated the effect of dicalcin, an S100-like Ca 2+ -binding protein, on the attachment of choriocarcinoma cells (BeWo cells) onto a monolayer of endometrial carcinoma cells (Ishikawa cells). Extracellularly administered dicalcin bound to both BeWo and Ishikawa cells. Pretreatment of BeWo spheroids with dicalcin reduced the attachment ratio of the spheroids onto the monolayer, whereas that of Ishikawa cells showed no apparent change. We identified the partial amino acid sequence of human dicalcin that exhibited maximum suppression for BeWo spheroid attachment. Transmission electron microscopy analysis revealed that the dicalcin-derived peptide caused a dilation of the intercellular junction between BeWo and ISK cells. Peptide treatment of BeWo spheroids downregulated the expression of integrinαvβ3 in BeWo cells, and induced alterations in their phalloidin-staining pattern, as measured by the length of each F-actin fiber and the thickness of the cortical stress fiber. Thus, dicalcin affects reorganization of the intracellular actin meshwork and subsequently the intensity of attachment, functioning as a novel suppressor of implantation., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Radiofrequency catheter ablation (RFCA) for pulmonary artery ventricular arrhythmia (PAVA) can be difficult because of the occasional existence of PAVA with preferential conduction.This study described the characteristics of PAVA that demonstrate preferential conduction.We analyzed electrocardiographic and electrophysiological data from 8 patients found to have PAVAs with preferential conduction out of 183 patients (4.4%) with right ventricular outflow tract (RVOT) arrhythmias who underwent RFCA at our hospitals. The PAVA with preferential conduction were classified into two types. In type 1 PAVA, successful ablation sites (success-sites) exhibited discrete prepotentials with an isoelectric line, in which the activation time (AT) was ≥ 50 milliseconds. In type 2 PAVA, excellent pace mapping was achieved at two sites separated by ≥ 20 mm: one in the RVOT free wall and the other at the success-site in the pulmonary artery. Type 1 and 2 PAVA features were considered signs of a short and long preferential conduction pathway, respectively.There were four patients each with type 1 and 2 PAVA. Type 1 PAVA was distinguished by the isoelectric line at success-sites with the mean AT of 78 ± 25.1 milliseconds. In type 2 PAVAs, although the AT at RVOT sites was very short (18.5 ± 10.1 milliseconds), the AT at success-sites was longer than that at the RVOT by 42.3 ± 36.2 milliseconds. Type 2 PAVAs displayed distinct electrocardiogram (ECG) features (R wave in lead I, RR' in inferior leads, and transitional zone in V4) not found in typical PAVA ECGs.PAVA with preferential conduction can manifest in distinct ways on the ECG and intracardiac mapping. Knowledge of these features may facilitate successful RFCA of such PAVA cases.

Introduction: The optimal ablation strategy is unknown regarding a superior vena cava isolation (SVCI). This study aimed to examine the feasibility and safety and to analyze the lesion characteristics of the SVCI using high-power, short-duration (HPSD) ablation., Methods and Results: A total of 100 patients underwent an index SVCI using HPSD (n = 50, HPSD group) or conventional lower-power and longer-duration (n = 50, LPLD group) ablation, using the Thermocool Smarttouch SF. In the HPSD group, ablation was performed with a power of 50 W for 7 s, and was limited to 4 s at the lateral segment close to the right phrenic nerve. The ablation setting used in the LPLD group was 20-25 W for 20-30 s and was limited to 10-20 W for 15-30 s at the lateral segment when diaphragmatic capture was seen. An electrical SVCI was achieved in all patients. The HPSD group required a significantly shorter procedure time (10.8 ± 3.2 vs. 14.8 ± 6.4 min; p < .01), shorter radiofrequency duration (49 ± 16 vs. 282 ± 124 s; p < .01), fewer lesions (8.3 ± 2.5 vs. 10.4 ± 4.4; p < .01), and lower ablation index (316 ± 38 vs. 356 ± 62; p < .001) than the LPLD group. The incidence of a postprocedural asymptomatic mild diaphragmatic elevation was comparable (2% in the HPSD group vs. 6% in the LPLD group; p = .61)., Conclusion: The 50-W HPSD ablation strategy allowed for a successful, fast, and safe SVCI with the fewer ablation lesions and the lower ablation index., (© 2021 Wiley Periodicals LLC.)

Background: Pacemaker positioning on the right ventricular (RV) septum during implantation is conventionally conducted utilizing two fixed fluoroscopy angles, a 45° left anterior oblique (LAO) and 35° right anterior oblique projection. However, placement location can be suboptimal, especially for leadless pacemakers (LPMs)., Objective: To evaluate the safety and ease of LPM implantation using individualized LAO projection., Methods: Consecutive patients undergoing LPM implantation were prospectively included. The angle of the RV septum was recorded for each patient by studying the angle at which an RV pigtail catheter (RV-PC) could be seen edge on. This was then used as the preferred LAO projection angle for that patient. We evaluated the success rate and safety of this method. We also compared the RV septum angle as measured by this method versus that measured by chest CT., Results: Of the 31 patients (mean age 80.6 ± 7.0 years, 15 females), LPM implantation was successful in 30. The pacemaker was implanted on the RV septum in 29 and on the free wall in one. LPM implantation was abandoned for anatomical reasons in one. Complications were limited to a groin arteriovenous fistula and one deep vein thrombosis. The angle of RV septum as measured by pigtail catheter and chest CT was not significantly different (CT: 54.8 ± 6.0°, RV pigtail catheter: 52.9 ± 6.1°, P  = .07)., Conclusions: Using an RV-PC to determine the preferred angle of LAO projection facilitates differentiation between the RV septum and free wall, which in turn facilitates optimal LPM placement., Competing Interests: All authors have no disclosure associated with this manuscript., (© 2021 The Authors. Journal of Arrhythmia published by John Wiley & Sons Australia, Ltd on behalf of Japanese Heart Rhythm Society.)

Introduction: Various types of inhalation devices have been released, and it is necessary to acquire the skills for using each of them. The factors that have been previously associated with poor inhalator usage include gender, duration of disease, age, and the type of device. However, it is unclear whether these factors also apply to the Japanese population. The number of education sessions needed to acquire inhaler usage skills is also not established., Patients and Methods: We performed a retrospective review of the medical records of selected patients and their subjective assessments of their inhaler usage skills between January 2016 and March 2018. The primary outcome was the effect of inhaler education for each inhaler device. The secondary outcomes were the factors affecting the effectiveness of inhaler education, the effects of inhalation education stratified by age, and the number of inhaler education sessions needed to improve inhaler usage skills., Results: Data from 399 patients were analyzed. Age and the type of delivery device affected the mastery of inhaler usage skills. Approximately half of the patients had acquired inhaler usage skills during baseline evaluation. Approximately 90% of patients acquired inhalation usage skills after two education sessions, regardless of the type of inhalation device. Among the older patients, 35.0% had acquired inhaler usage skills during the baseline evaluation, and 86.8% acquired them after two education sessions., Conclusions: Inhaler usage skills significantly improved, regardless of the device, after inhalation education, and this was also observed in elderly patients after two education sessions., Competing Interests: Conflict of Interest Tatsuya Nagano has received research funding from AstraZeneca, United State, Grant ID: NCR-18-14138. Other authors deny conflicts of interest or honoraria., (Copyright © 2020 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.)

Hydrogen molecules have attracted attention as a new antioxidant, but are left to be confirmedly verified whether the oral administration is highly safe or not, concurrently with retention of abundant hydrogen. When electrolysis was performed for 10 minutes using a direct-current electrolytic hydrogen-water generating bottle with tap water, "residual free chlorine" concurrently upon the production of molecular hydrogen (444 μg/L) could be appreciably decreased from 0.18 mg/L to 0.12 mg/L as quantified by a N,N-diethyl-p-phenylenediamine-dye colorimetric method. Moreover, the total chlorine concentration (residual bound chlorine plus free chlorine) was estimated to be decreased from 0.17 mg/L to 0.11 mg/L. Although a merit of electrolytic hydrogen-generating bottles exists in electrolysis for periods as short as 10 minutes, the 30-minute electrolysis brought about the more abundant hydrogen (479 μg/L) together with an oxidation-reduction potential of -245 mV; even upon this long-term electrolysis, the gross amounts of chlorine, hypochlorous acid and chloramine were shown not to be increased (0.09-0.10 mg/L from 0.11 mg/L for tap water) as detected by orthotolidine colorimetry. Above-mentioned levels of diverse-type chlorines might fulfill the World Health Organization guideline for drinking water below 5 mg/L. In addition, the dissolved ozone upon electrolytic generation of hydrogen-water was below the detection limit (< 0.05 mg/L) or undetectable, which fulfilled the official safety standards in Japan and the USA for drinking water below 0.1 mg/L, as evaluated by three methods such as an electrode-type ozone checker, indigo dye-utilizing ozone detector capillaries and potassium iodide-based colorimetry. Importantly, even when half the amount of tap water was poured into the tank of the apparatus and electrolyzed, both the residual chlorine and ozone concentrations measured were also below the safety standard. Thus, major potently harmful substances, such as residual free/bound chlorine, or hypochlorous-acid/chloramine, respectively, and dissolved ozone, as the drinking hydrogen-water was direct-current-electrolytically generated, were estimated to be repressed within safety concentration ranges with achievements of abundant hydrogen generation.

Reactive oxygen species (ROS)-induced oxidative stress in adipose tissue is associated with inflammation and the development of obesity-related metabolic disorders. The aim of this study is to investigate the effects of hydrogen nano-bubble water (HW) on ROS generation, adipogenesis, and interleukin-6 (IL-6) secretion in hydrogen peroxide (H 2 O 2 ) or phorbol 12-myristate 13-acetate (PMA)-stimulated OP9 adipocytes, and three-dimensional (3D) subcutaneous adipose equivalents. Nanoparticle tracking analysis showed that fresh HW contains 1.17 × 10 8 /mL of nano-sized hydrogen bubbles. Even after 8 to 13 months of storage, approximately half of the bubbles still remained in the water. CellROX ® staining showed that HW could diminish H 2 O 2 - or PMA-induced intracellular ROS generation in human keratinocytes HaCaT and OP9 cells. We discovered that PMA could markedly increase lipid accumulation to 180% and IL-6 secretion 2.7-fold in OP9 adipocytes. Similarly, H 2 O 2 (5 µM) also significantly stimulated lipid accumulation in OP9 cells and the 3D adipose equivalents. HW treatment significantly repressed H 2 O 2 - or PMA-induced lipid accumulation and IL-6 secretion in OP9 adipocytes and the 3D adipose equivalents. In conclusion, HW showed a possibility of repressing oxidative stress, inflammatory response, and adipogenesis at cellular/tissue levels. It can be used for preventing the development of metabolic disorders amongst obese people.

Carcinostatic effects of combined use of ascorbic acid (Asc), 2-O-phospho- or 6-O-palmitoyl ascorbate (Asc2Phos, Asc6Palm) or diverse alkanoyl Asc, and nano-sized platinum-poly(N-vinyl-pyrrolidone) colloid (PVP-Pt; 2-nm diameter) were examined on human esophagus carcinoma-derived cells KYSE70. Cell viability was repressed by 'Asc6Palm + PVP-Pt' mixture more markedly than by Asc6Palm or PVP-Pt alone, together with cell shrinkage and fragmentation, in contrast to no additive carcinostatic effect of 'Asc + PVP-Pt' or 'Asc2Phos + PVP-Pt'. The effects might be partly due to efficiency for intracellular uptake of PVP-Pt, as previously shown by our studies that Pt atoms composed of PVP-Pt were incorporated into human tongue carcinoma cells by 9.6-fold compared to normal human tongue epitheliocytes. Asc6Palm was advantageous for intracellular uptake, in terms of the proper balance for molecular hydrophilicity-lipophilicity (BMHL), whereas 6-O-stearoyl (C 18 ) Asc or 2,6-O-dipalmitoyl (2 × C 16 ) was demonstrated to be less carcinostatic owing to a lower BMHL. Although esterolytically converted from Asc6Palm, Asc was necessitated to be retained for efficient carcinostasis, and demonstrated by HPLC-coulometric ECD analysis to be appreciably stabilized in electrolytically generated hydrogen (dissolved hydrogen: 0.575 mg/L)-water, but scarcely in hydrogen-gas-bubbled water (0.427 mg/L), Mg stick-derived hydrogen (0.044 mg/L) water, or tap water, suggesting that hydrogen-rich water suppresses oxidative decomposition of Asc. Thus, Asc6Palm plus PVP-Pt with hydrogen-rich water supplement might be applicable for carcinostatic therapy.

The network mechanisms underlying how inhibitory circuits regulate ON- and OFF-responses (the b- and d-waves) in the electroretinogram (ERG) remain unclear. The purpose of this study was to investigate the contribution of inhibitory circuits to the emergence of the b- and d-waves in the full-field ERG in the newt retina. To this end, we investigated the effects of several synaptic transmission blockers on the amplitudes of the b- and d-waves in the ERG obtained from newt eyecup preparations. Our results demonstrated that (a) L-APB blocked the b-wave, indicating that the b-wave arises from the activity of ON-bipolar cells (BCs) expressing type six metabotropic glutamate receptors; (b) the combined administration of UBP310/GYKI 53655 blocked the d-wave, indicating that the d-wave arises from the activity of OFF-BCs expressing kainate-/AMPA-receptors; (c) SR 95531 augmented both the b- and the d-wave, indicating that GABAergic lateral inhibitory circuits inhibit both ON- and OFF-BC pathways; (d) the administration of strychnine in the presence of SR 95531 attenuated the d-wave, and this attenuation was prevented by blocking ON-pathways with L-APB, which indicated that the glycinergic inhibition of OFF-BC pathway is downstream of the GABAergic inhibition of the ON-system; and (e) the glycinergic inhibition from the ON- to the OFF-system widens the response range of OFF-BC pathways, specifically in the absence of GABAergic lateral inhibition. Based on these results, we proposed a circuitry mechanism for the regulation of the d-wave and offered a tentative explanation of the circuitry mechanisms underlying ERG formation., (© 2020 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

Ultraviolet-A ( UVA) irradiation induces harmful effects on skin cells and accelerates skin aging through oxidative stress. In this study, the effects of a hydrogen-generating silica material named ULH-002 against UVA injuries in human cells and 3D skin equivalents were investigated. The oxygen radical absorption capacity (ORAC) assay showed that both freshly prepared ULH-002 solutions and 7-day-old solutions exhibited equal peroxyl radical (ROO·) scavenging activities concentration-dependently. CellROX ® green/orange staining showed that ULH-002 could reduce UVA-induced oxidative stress in human keratinocytes HaCaT and human gingival fibroblasts (HGFs). ULH-002 significantly prevented UVA-induced apoptotic/necrotic cell death and cell-viability decline in HGFs and keratinocytes, as shown by Annexin V/PI apoptosis assay and PrestoBlue assay, respectively. Immunostaining showed that ULH-002 prevented the UVA-induced deterioration of expression of both type IV and I collagens in the 3D skin equivalents, and similarly in monolayer HGFs. UVA-enhanced melanogenesis was observed in human melanocytes HMV-II and HMV-II cell-containing 3D skin equivalents, but markedly prevented by ULH-002 as demonstrated by Fontana-Masson's staining. In conclusion, our data suggested that ULH-002 could protect human keratinocytes and fibroblasts from UVA-induced injuries, prevent the loss of type IV and I collagens, as well as reduce melanogenesis. ULH-002 might be developed as a skin care reagent in the cosmetic industry.

Hydrogen-rich water bath devices are commercially available, but have been scarcely clarified for heat-retention effects. In this study, heat-retention effects of hydrogen-rich water bath were assessed by thermographic clinical trials, which employed twenty-four healthy subjects. The thermograms indicated that, under the same conditions (41 °C, 10-min bathing), hydrogen-rich water bath (hydrogen concentrations: 185-548 μg/L; oxidation-reduction potentials: -167 to -91 mV, versus 0.8 μg/L and +479 mV for normal bath, respectively) brought about the heat-retention being more marked than those of normal water bath for several body-parts in the order as follows: abdomen > upper legs > arms > hands > feet, for 30- and 60-min post-bathing, being in contrast to scarce heat-retention for head, armpits and lower legs. Then, as reflection to promotive effects on blood stream, we also examined the thickness of fingertip-capillary in hands. The thickness was expanded in the hydrogen-rich water bath more markedly than that in the normal water bath, suggesting that the hydrogen-rich water bath may have the hydrogen-based promotive effect, exceeding over mere heat retention-based effects, on blood circulation of the whole body. Meanwhile, the heat-retention in hydrogen-rich water bath weakly or moderately correlated with contents of the subcutaneous fat, whole body fat and body mass index, and inversely correlated with skeletal muscle rates, although their correlation degrees did not obviously exceed over normal water bath, with a poor relation with the basal metabolism rate. Thus, the hydrogen-rich water bath was suggested to exert heat-retention effects exceeding over normal water bath, in diverse body-parts such as abdomen, upper legs, arms and hands, via promotion to blood flow which was reflected by expanding the thickness of capillary. The heat-retention after bathing can be noted as effects of the hydrogen-rich water bath, which is applicable for most of people widespread regardless of their body composition index., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

As an aging-associated degenerative disease, Alzheimer's disease is characterized by the deposition of amyloid beta (Aβ), oxidative stress, inflammation, dysfunction and loss of cholinergic neurons. Colla Corii Asini (CCA) is a traditional Chinese medicine which has been used for feebleness-related diseases and anti-aging. CCA might delay aging-induced degenerative changes in neurons. In the present study, we evaluated antioxidant activity, cytoprotective effects, and Aβ removability of enzyme-digested Colla Corii Asini (CCAD). Oxygen radical absorbance capacity (ORAC) activity assay showed that, as compared to gelatins from the skin of porcine, bovine and cold water fish, CCA exhibited the highest ORAC activity. The ORAC activity of CCA and CCAD was increased gradually by the length of time in storage. Ultrastructure analysis by scanning electron microscopy showed that among CCA manufactured in 2008, 2013, 2017 and gelatin from cold water fish skin, CCA manufactured in 2008 presented the smoothest surface structure. We further tested the protective effects of CCAD (manufactured in 2008) and enzyme-digested gelatin from cold water fish skin (FGD) on hydrogen peroxide (H 2 O 2 )-induced cell death in nerve growth factor-differentiated neuronal-like PC12 cells. Presto blue assay showed that both FGD and CCAD at 0.5 mg/mL increased cell viability in H 2 O 2 -treated neuronal-like PC12 cells. The protection of CCAD was significantly superior to that of FGD. Acetylcholinesterase (AchE) assay showed that both FGD and CCAD inhibited AchE activity in nerve growth factor-differentiated neuronal-like PC12 cells to 89.1% and 74.5% of that in non-treated cells, respectively. The data suggest that CCAD might be able to increase the neurotransmitter acetylcholine. Although CCAD inhibited AchE activity in neuronal-like PC12 cells, CCAD prevented H 2 O 2 -induced abnormal deterioration of AchE. ELISA and neprilysin activity assay results indicated that CCAD reduced amyloid beta accumulation and increased neprilysin activity in Aβ 1-42 -treated neuronal-like PC12 cells, suggesting that CCAD can enhance Aβ clearance. Our results suggest that CCA might be useful for preventing and treating Alzheimer's disease., Competing Interests: None

Cutaneous wound healing delay, collagen synthesis decline and wrinkle formation are the common features of skin aging. The aim of this study is to investigate repressive effects of Colla Corii Asini (CCA) (a traditional Chinese medicine which has been used for anti-aging) on hydrogen peroxide (300 µM, 2 h) and ultraviolet A (UVA) (3.2 mJ/cm 2 )-induced skin aging in vitro. To simulate the in vivo condition of CCA, CCA was digested by gastrointestinal enzymes and added to human gingival fibroblasts (HGF) and three dimensional (3D) skin equivalents at different concentrations. Cell viability assay showed that the enzyme-digested CCA (CCAD) exhibited significant preventive effects on hydrogen peroxide- and UVA-induced cell death. The in vitro scratch assay showed that CCAD was able to prevent hydrogen peroxide-induced wound healing delay in HGF cell sheets. Immunostaining and imaging analysis showed that CCAD could suppress UVA-reduced expression of type IV collagen and elastin in both HGF cells and the 3D skin equivalents. Using a tissue stretching system, wrinkles were formed on UVA-irradiated 3D skin equivalents. Without CCAD-treatment, the wrinkles on the skin were deep, whereas CCAD markedly reduced the depth of wrinkles. In conclusion, CCAD could protect skin cells from oxidative stress and UVA-induced harmful effects, accelerate wound healing, promote synthesis of collagen and elastin, and reduce wrinkles formation. CCAD might be developed as an anti-skin aging reagent in the cosmetic industry.

Carcinostatic effects of combined use of hydrogen nano-bubbles (nano-H) and platinum-povidone (PVP--Pt) were examined. Hydrogen-dissolved medium was prepared by hydrogen-gas bubbling with a microporous gas-emittance-terminal into a medium in the absence or presence of PVP-Pt (nano-H, nano-H/PVP-Pt). Human esophagus-derived carcinoma cells KYSE70 were repressed for cell proliferation with nano-H/PVP-Pt more markedly than with nano-H, indicating the hydrogen-intensification for PVP-Pt-alone-carcinostasis. However, the intensified carcinostasis required co-administration of nano-H and PVP-Pt, and no intensified carcinostasis was shown in two-step separate administration of nano-H and PVP-Pt. Furthermore, hydrogen bubbling into PVP-Pt-containing medium achieved more appreciable carcinostasis than mere addition of PVP-Pt into nano-H-containing medium, indicating the potent interaction of hydrogen and PVP-Pt. The nano-H/PVP-Pt-administered human tongue-derived carcinoma cells HSC-4 were repressed for cell proliferation more markedly than pre-malignant human tongue-derived epitheliocytes DOK, concurrently with more abundant intracellular Pt-intake into HSC-4 cells than DOK as analyzed by ICP-MS. Thus, PVP-Pt is able to adsorb hydrogen nano-bubbles on Pt and applicable for cancer therapy by diminishing the side-effects to normal cells.

Purpose: Caspase recruitment domain-containing protein 9 (CARD9) is expressed at high levels in bone marrow cells and has a crucial role in innate immunity. Current studies indicate that CARD9 also plays a key role in tumor progression, but there are few reports on the role of CARD9 in lung cancer. The aim of this study was to clarify the role of CARD9 in lung adenocarcinoma., Patients and Methods: Lung adenocarcinoma tumor samples from 74 patients who underwent complete resection at Kobe University Hospital from January 2014 to December 2014 were analyzed by immunohistochemistry. The role of CARD9 in cancer cells was analyzed using lung cancer cell lines treated with CARD9 siRNA., Results: High expression of CARD9 was observed in 32.4% of tumors, and compared to low expression of CARD9, high expression was associated with poorer overall survival (P = 0.0365). Univariate and multivariate analyses showed that high expression of CARD9 was an independent prognostic factor. Knockdown of CARD9 in lung adenocarcinoma cells inhibited proliferation but did not increase apoptosis. In addition, CARD9 activated the NF-κB pathway in a lung adenocarcinoma cell line., Conclusion: CARD9 was shown to be an independent prognostic factor of poor outcome for lung cancer and may represent a molecular target for treatment., Competing Interests: This work was supported by grant 17K09614 from JSPS KAKENHI to Tatsuya Nagano. The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors report no other potential conflicts of interest for this work., (© 2020 Miwa et al.)

Hydrogen-rich water is conventionally prepared by direct current-electrolysis, but has been not or scarcely prepared by alternating current (AC)-electrolysis. The AC preparations from tap water for 20-30 minutes exhibit a dissolved hydrogen concentration of 1.55 mg/L, which was close to the theoretical maximum value of 1.6 mg/L. These preparations also displayed an oxidation-reduction potential of -270 mV (tap water: +576 mV) and pH of 7.7-7.8, being closer to physiological values of body fluids than general types of direct current-electrolytic hydrogen-rich water. We examined whether AC-electrolytic hydrogen-water is retained for hydrogen-abundance after boiling or for antioxidant abilities, and whether the oral administration of this water is clinically effective for diabetes and prevention against systemic DNA-oxidative injuries. 5,5-Dimethyl-1-pyrroline-N-oxide spin trapping and electron spin resonance revealed that the hydrogen-rich water generated by AC-electrolysis exhibited hydroxyl-radical-scavenging activities. Laser nanoparticle tracking method revealed that nanoparticle suspensions as abundant as 5.4 × 10 7 /mL were efficiently retained (up to 3.5 × 10 7 /mL) even after boiling for 10 minutes, being thermodynamically contrary to Henry's law. Oral intake of hydrogen-rich water, 1500 mL per day, lasted for 8 weeks in nine people with the diabetes-related serum markers beyond the normal ranges. The subjects exhibited significant tendencies for the decreased fasting blood glucose and fructosamine, and for the increased 1,5-anhydro-D-glucitol, concomitantly with significant decreases in urinary 8-hydroxy-2-deoxyguanosine contents and its rate of generation. Hydrogen-rich water prepared by AC-electrolysis may be effective in improving diverse diabetes-related markers and systemic DNA oxidative injuries through the formation of abundant heat-resistant nanobubbles and the increased hydrogen concentrations. The study protocol was officially approved by the Medical Ethics Committee of the Japanese Center for Anti-Aging Medical Sciences (approval No. 01S02) on September 15, 2009., Competing Interests: None

Background: Sneezes produce many pathogen-containing micro-droplets with high velocities of 4.5-50.0 m/s. Face masks are believed to protect people from infection by blocking those droplets. However, current filtration efficiency tests can't evaluate masks under sneeze-like pressure. The goal of this study was to establish a method to evaluate the filtration efficiency of mask materials under extreme conditions., Materials and Methods: Efficiency of surgical masks, gauze masks, gauze, cotton, silk, linen and tissue paper on blocking micro-droplet sized starch particles (average 8.2 μm) and latex microspheres (0.75 μm) with a velocity of 44.4 m/s created by centrifugation was qualitatively analyzed by using imaging-based analysis., Results: The 4 layers of silk could block 93.8% of microspheres and 88.9% of starch particles, followed by the gauze mask (78.5% of microspheres and 90.4% of starch particles) and the 2 layers of cotton (74.6% of microspheres and 87.5-89.0% of particles). Other materials also blocked 53.2-66.5% of microspheres and 76.4%-87.9% of particles except the 8 layers of gauze which only blocked 36.7% of particles. The filtration efficiency was improved by the increased layers of materials., Conclusion: Centrifugation-based filtration efficiency test not only compensates shortcomings of current tests for masks, but also offers a simple way to explore new mask materials during pandemics. Common mask materials can potentially provide protection against respiratory droplet transmission., (Copyright© 2020, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Transglutaminase (TG) catalyzes cross-linking between the γ-carboxyamide groups of glutamine residues and the ε-amino groups of lysine residues in polypeptide chains, yielding ε- (γ-glutamyl) lysine (G-L) bonds. By forming a network structure in the protein via G-L bonds, it is possible to increase the viscosity of protein solutions or to cause gelation. Nearly thirty years have passed since microbial TG (MTG) appeared in the food enzyme market. Since the start of research and development, MTG has been used in fishery products such as kamaboko (boiled fish paste), meat products such as sausages, milk products such as yogurt, processed-soybean products such as tofu, and wheat products such as bread and noodles. MTG has provided effects such as adding new functions and reducing waste in food applications. The purpose of this review is to describe not only the history of research and development of TG but also the key aspects that have facilitated the great success of this process as a technology for enzymatically modifying protein-containing foods., Competing Interests: Declaration of competing interest The author declares no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Recently, the SARS-CoV-2 induced disease COVID-19 has spread all over the world. Nearly 20% of the patients have severe or critical conditions. SARS-CoV-2 exploits ACE2 for host cell entry. ACE2 plays an essential role in the renin-angiotensin-aldosterone system (RAAS), which regulates blood pressure and fluid balance. ACE2 also protects organs from inflammatory injuries and regulates intestinal functions. ACE2 can be shed by two proteases, ADAM17 and TMPRSS2. TMPRSS2-cleaved ACE2 allows SARS-CoV-2 cell entry, whereas ADAM17-cleaved ACE2 offers protection to organs. SARS-CoV-2 infection-caused ACE2 dysfunction worsens COVID-19 and could initiate multi-organ failure. Here, we will explain the role of ACE2 in the pathogenesis of severe and critical conditions of COVID-19 and discuss auspicious strategies for controlling the disease.

Widely distributed electrolytic-generators for hydrogen-water are not fully considered for the dependencies of post-electrolytic values of the dissolved hydrogen concentration (DH) and the oxidation-reduction potential (ORP) on the properties of the pre-electrolytic water. We investigated the dependencies of DH and ORP on mineral-based hardness, temperatures and the container materials, and effects on the oral cavity by oral washing or drinking. Along with an increase in mineral-based water-hardness, DH decreased from 960 to 870 μg/L and the ORP unexpectedly increased from -460 to -320 mV. Purified water of almost zero hardness, however, caused a post-electrolytic DH as low as 80 μg/L and an ORP as high as +20 mV. Post-electrolytic DHs were not significantly changed (780-900 μg/L) upon electrolysis at 1.5-30°C and decreased at 40-50°C. The diffusion of hydrogen from the inside to the outside of the container was extremely small even after 12 hours for an aluminum- or stainless steel-made container, but not for containers made of diverse plastics. The ORP of the intact saliva was +136 mV, and decreased to +90 mV at 20 minutes after 1-minute oral-cramming of hydrogen-water, but returned to +135 mV after 60-minute leaving, showing a transient ORP-decrease in the saliva. Drinking-pause for 4 weeks after drinking hydrogen-water, however, saliva ORP, gradually but not instantly, increased to +60 to +80 mV, but upon drinking-resumption and 2 weeks thereafter, decreased again to -100 to -110 mV, suggesting that several-week hydrogen-water drinking caused a certain decrease in the saliva ORP. Thus, the present study provided the appropriate conditions such as hardness and temperatures for hydrogen-water production by the electrolytic generator, and the container materials suitable for hydrogen-water preservation. Furthermore, we clarified ORP changes of human saliva, being an indicator for human oxidative stress. The study was approved by the Medical Ethics Committee of the NPO (Non-Profitable Organization)-Corporate Japanese Center for Anti-Aging Medical Sciences (approval No. 09S02) on May 2, 2012.

Patients with arrhythmogenic right ventricular cardiomyopathy (ARVC) and hypertrophic cardiomyopathy (HCM) implanted with implantable cardioverter-defibrillators (ICDs) may show a large decrease in R-wave amplitude during long-term follow-up. However, it is unclear whether this decrease is higher in these patients than in those without structural heart disease. This study investigated ICD-lead intracardiac parameters over a long duration in patients with ARVC and HCM and compared these parameters with those of a control group. We included 50 patients (mean age, 55.2 ± 17.2 years; 26% female) with ICD leads in the right ventricular apex, and compared 7 ARVC and 14 HCM patients with 29 control patients without structural heart disease. ICD-lead parameters, including R-wave amplitude, pacing threshold, and impedance during follow-up, were compared. The difference in these parameters between the time of implantation and year 5 were also compared. There were no significant differences in R-wave amplitude at implantation among the 3 groups. The change in R-wave amplitude between the time of implantation and year 5 was significantly greater in the ARVC group (-3.3 ± 5.4 mV, P = 0.012) in comparison to the control group (1.3 ± 2.8 mV); the HCM group showed no significant difference (-0.4 ± 2.3 mV, P = 0.06). Thus, in the ARVC group, R-wave amplitude at year 5 was significantly lower than that in the control group (5.7 ± 4.8 mV versus 12.5 ± 4.5 mV, P = 0.001). In ARVC patients with ICDs, ventricular sensing is likely to deteriorate during long-term follow-up; however, in HCM patients, sensing may not deteriorate.