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Abstract Background Epigenetic modifications have a central role in transcriptional regulation. While several studies using next-generation sequencing have revealed genome-wide associations between epigenetic modifications and transcriptional states, a direct causal relationship at specific genomic loci has not been fully demonstrated, due to a lack of technology for targeted manipulation of epigenetic modifications. Recently, epigenome editing techniques based on the CRISPR-Cas9 system have been reported to directly manipulate specific modifications at precise genomic regions. However, the number of editable modifications as well as studies applying these techniques in vivo is still limited. Results Here, we report direct modification of the epigenome in medaka (Japanese killifish, Oryzias latipes) embryos. Specifically, we developed a method to ectopically induce the repressive histone modification, H3K27me3 in a locus-specific manner, using a fusion construct of Oryzias latipes H3K27 methyltransferase Ezh2 (olEzh2) and dCas9 (dCas9-olEzh2). Co-injection of dCas9-olEzh2 mRNA with single guide RNAs (sgRNAs) into one-cell-stage embryos induced specific H3K27me3 accumulation at the targeted loci and induced downregulation of gene expression. Conclusion In this study, we established the in vivo epigenome editing of H3K27me3 using medaka embryos. The locus-specific manipulation of the epigenome in living organisms will lead to a previously inaccessible understanding of the role of epigenetic modifications in development and disease. more...

The heavily methylated vertebrate genomes are punctuated by stretches of poorly methylated DNA sequences that usually mark gene regulatory regions. It is known that the methylation state of these regions confers transcriptional control over their associated genes. Given its governance on the transcriptome, cellular functions and identity, genome-wide DNA methylation pattern is tightly regulated and evidently predefined. However, how is the methylation pattern determined in vivo remains enigmatic. Based on in silico and in vitro evidence, recent studies proposed that the regional hypomethylated state is primarily determined by local DNA sequence, e.g., high CpG density and presence of specific transcription factor binding sites. Nonetheless, the dependency of DNA methylation on nucleotide sequence has not been carefully validated in vertebrates in vivo. Herein, with the use of medaka (Oryzias latipes) as a model, the sequence dependency of DNA methylation was intensively tested in vivo. Our statistical modeling confirmed the strong statistical association between nucleotide sequence pattern and methylation state in the medaka genome. However, by manipulating the methylation state of a number of genomic sequences and reintegrating them into medaka embryos, we demonstrated that artificially conferred DNA methylation states were predominantly and robustly maintained in vivo, regardless of their sequences and endogenous states. This feature was also observed in the medaka transgene that had passed across generations. Thus, despite the observed statistical association, nucleotide sequence was unable to autonomously determine its own methylation state in medaka in vivo. Our results apparently argue against the notion of the governance on the DNA methylation by nucleotide sequence, but instead suggest the involvement of other epigenetic factors in defining and maintaining the DNA methylation landscape. Further investigation in other vertebrate models in vivo will be needed for the generalization of our observations made in medaka. more...

Background and Objective: Embolic stroke of undetermined source (ESUS) patients undergoing long-term rhythm monitoring with Implantable Loop Recorder (ILR) have an atrial fibrillation (AF) detection rate of approximately 12 % at 1 year and 30 % with extended follow-up over 3 years. However, research specifically focusing on the majority of patients in whom AF is not detected through implantable cardiac monitors remains limited. Abnormal lipid profiles may be associated with embolic risks from non-AF sources. This study aimed to develop a model to predict the absence of AF in patients with ESUS using multiple variables, including lipid profiles. Methods: A retrospective, multicenter cohort study was conducted across four institutions, involving 99 ESUS patients. Patients were categorized based on AF detection via ILR. Patient characteristics, blood test results, and echocardiographic findings were assessed through univariate and multivariate logistic regression analyses. ROC curve analysis was performed to evaluate the biomarkers’ predictive accuracy. Results: AF was detected in 30.3 % of patients over a median follow-up of 25.5 months. Multivariate analysis confirmed elevated D-dimer (OR: 2.77, p = 0.002), low LDL/HDL ratio (OR: 2.0, p = 0.01), and CHA2DS2-VASc score (OR: 1.4, p = 0.04) as independent predictors of AF detection. The CHA2DS2-VASc score was excluded due to multicollinearity, and patients with D-dimer 1.98 had significantly lower AF detection rates (6.8 %, P more...

Walking patterns can differ between children and adults, both kinematically and kinetically. However, the detailed nature of the ankle pattern has not been clarified. We investigated musculature, biomechanics, and muscle activation strategies and their relevance to walking performance in preschool (PS) and school children (SC), with adults (AD) as reference. Twenty-six PS (3-5 yr), 20 SC (6-8 yr), and 17 AD (18-30 yr) participated. Tibialis anterior (TA) and medial gastrocnemius (MG) thicknesses, fascicle lengths, and maximal voluntary dorsi- and plantar flexion isometric torques were measured. Hip, knee, and ankle kinematics, ground reaction forces (GRFs), and TA and MG electromyographic activities were recorded during shod walking at each participant's preferred speed. Walking speed, step length, and cadence were correlated with age in PS. These walking performance measures were also correlated with muscle thickness and fascicle length, showing higher speed in individuals with thicker muscles and longer TA and MG fascicles (conversely, higher cadence with thinner muscles and shorter fascicles). AD demonstrated the largest values for muscle thickness (p more...

Abstract Some hardwoods have a higher density and superior structural performance than softwoods, which are generally used as construction materials in buildings. Therefore, the characteristics of hardwoods are advantageous in their use for wooden joints. However, the effects of the dimensions of hardwood specimens and wood species on the partial compression performance perpendicular to the grain are unclear. In this study, we conducted partial compression tests perpendicular to the grain of hardwoods to clarify the effects of the edge distance and wood species on the characteristic values. The relationships between the partial compression performance perpendicular to the grain and the edge distance perpendicular to the grain of hardwoods were similar to those of softwoods. A significant correlation was observed between the density and the characteristic values of partial compression perpendicular to the grain. There were differences in the fracture morphologies and stress–strain curves between diffuse and ring-porous woods. However, factors other than density and whether the wood is diffuse- or ring-porous are responsible for the differences in the effect of the edge distance on the characteristic values between wood species. more...

Abstract Aim Photopharmacology is a new technique for modulating biological phenomena through the photoconversion of substances in a specific target region at precise times. Caged compounds are thought to be compatible with photopharmacology as uncaged ligands are released and function in a light irradiation‐dependent manner. Here, we investigated whether a microscale light‐emitting diode (MicroLED) probe is applicable for the photoconversion of caged‐glutamate (caged‐Glu) in vivo. Methods A needle‐shaped MicroLED probe was fabricated and inserted into the mouse hippocampal dentate gyrus (DG) with a cannula for drug injection and a recording electrode for measuring the local field potential (LFP). Artificial cerebrospinal fluid (ACSF) or caged‐Glu was infused into the DG and illuminated with light from a MicroLED probe. Results In the caged‐Glu‐injected DG, the LFP changed in the 10–20 Hz frequency ranges after light illumination, whereas there was no change in the ACSF control condition. Conclusion The MicroLED probe is applicable for photopharmacological experiments to modulate LFP with caged‐Glu in vivo. more...

Abstract Motor contagions refer to implicit effects induced by the observation of actions made by others on one’s own actions. A plethora of studies conducted over the last two decades have demonstrated that both observed and predicted actions can induce various kinds of motor contagions in a human observer. However, motor contagions have always been investigated with regard to different features of an observed action, and it remains unclear whether the background environment in which an observed action takes place modulates motor contagions as well. Here, we investigated participant movements in an empirical hand steering task during which the participants were required to move a cursor through a visual channel after being presented with videos of an actor performing the same task. We manipulated the congruency between the actions shown in the video and the background channels and examined whether and how they affected the participants’ own movements. We observed a clear interaction between the observed action and its background. The movement time of the participants’ actions tended to increase or decrease depending on whether they observed a faster or slower movement, respectively, and these changes were amplified if the background was not congruent with the action contained within it. These results suggest that background information can modulate motor contagions in humans. more...

Abstract Vitamin C plays important roles as a cofactor in many enzymatic reactions and as an antioxidant against oxidative stress. As some mammals including humans cannot synthesize vitamin C de novo from glucose, its uptake from dietary sources is essential, and is mediated by the sodium-dependent vitamin C transporter 1 (SVCT1). Despite its physiological significance in maintaining vitamin C homeostasis, the structural basis of the substrate transport mechanism remained unclear. Here, we report the cryo-EM structures of human SVCT1 in different states at 2.5–3.5 Å resolutions. The binding manner of vitamin C together with two sodium ions reveals the counter ion-dependent substrate recognition mechanism. Furthermore, comparisons of the inward-open and occluded structures support a transport mechanism combining elevator and distinct rotational motions. Our results demonstrate the molecular mechanism of vitamin C transport with its underlying conformational cycle, potentially leading to future industrial and medical applications. more...

We report a case in which a 74-year-old man suffering from esophageal achalasia complicated with epiphrenic esophageal diverticulum was successfully treated with a simultaneous laparoscopic surgery. The gentleman was referred with symptoms suggestive of a passage disorder in the lower esophagus for the past 5 years. Esophagogastroduodenoscopy demonstrated an epiphrenic diverticulum at the left wall of the lower esophagus, and esophagography led to the suspicion of a combined esophageal achalasia. A simultaneous laparoscopic surgery with an abdominal approach was performed in which, following the opening of the esophageal hiatus, the diverticular wall was separated from the mediastinal organs and diverticulectomy was performed with linear staplers. After Heller’s myotomy, Dor’s fundoplication was subsequently performed in which both the incisional line of muscle layer and the suturing line of diverticulectomy were wrapped by the fornix of the stomach to make up for the wall strength and avoid the suture leakage. It was theoretically considered logical and effective to reinforce this vulnerable site with Dor’s fundoplication. He had an uneventful recovery and a rapid relief from symptoms following surgery. more...

Anticipating the onset, location and severity of radiation dermatitis before radiotherapy can aid in dermatological care. This study developed a method for creation of a prediction diagram for dermatitis and conducted a comparative verification between the prediction diagram and actual patient condition. The prediction diagram involved converting skin doses into 2 Gy fractionated equivalent doses using α/β of 10.0, defining regions of interest (ROIs) from 20-50 Gy at 10 Gy intervals. Overlaps between these ROIs at each dose level and the skin (external) was sequentially color-coded as blue, yellow, red, and purple. The study included four patients: two underwent head and neck treatment and two received neck and chest treatment. This approach involved a visual assessment comparing the prediction diagram with skin photographs captured at the end of treatment. Nurses marked skin sites corresponding to grades 1 and 2 on photographs, assessing their correlation with the predicted range. Visual assessment results were largely positive, although one patient exhibited slightly lower scores. Results revealed a correlation between grade 1 skin reactions and the 20 Gy regions. Grade 2 reactions were observed in regions near 30 and 40 Gy. Although discrepancies between prediction diagram and actual skin symptoms were observed in patients, a broad agreement was evident. The prediction diagram cannot accurately predict radiation dermatitis, as it does not account for skin symptoms unrelated to the dose. However, the diagram is significant as it provides physicians, nurses, and patients with concise and visually comprehensible information regarding the location of dermatitis. more...

Key Clinical Message In addition to the reentrant map, the wave speed map can be helpful in accurately identifying the CTI gap during radiofrequency application for atrial flutter(AFL). However, in complex cases involving extensive scarring and multiple low‐velocity local areas, this technique may not be useful. Abstract A 73‐year‐old male patient with a history of pulmonary vein isolation and cavotricuspid isthmus ablation underwent a second catheter ablation owing to recurrent atrial flutter (AFL). The AFL was diagnosed as cavotricuspid isthmus‐dependent AFL caused by the reconnection of the previous cavotricuspid isthmus ablation. Wave speed mapping was performed at the same site, and results comprehensively revealed a low‐velocity local area. The AFL was terminated after the first radiofrequency application, and the block line was easily completed. Therefore, this technique could be an adjunctive tool for cavotricuspid isthmus gap identification and minimal radiofrequency application. more...

Purpose: Muscle quality is explained by the ratio between muscle size and strength. Conventionally, muscle size is evaluated without considering the composition of contractile and non-contractile tissues in muscle, hence the influence of non-contractile tissues on muscle quality is not fully understood, especially within aging muscle. This study investigated the differences in intramuscular non-contractile tissues between different age and sex groups, and investigated their influence on muscle quality. Methods: Eighty-two older and 64 young females and males participated. Muscle cross-sectional area (quadriceps and hamstrings), separating contractile and non-contractile areas, was calculated from the magnetic resonance image of the right mid-thigh. Maximal voluntary isometric knee extension and flexion torque was measured. Torque/muscle area and torque/contractile area were calculated for each age and sex group. Results: Non-contractile/muscle area was higher in older than in young individuals in both muscle groups (p females) (p more...

Introduction: The peribiliary gland is an accessory bile duct gland. Hyperplasia of these tissues may lead to elevation of the mucosa in the bile ducts and bile duct stenosis. We herein report a case of peribiliary gland hyperplasia that required preoperative differentiation from bile duct cancer, with a discussion of the literature. Case Presentation: The patient had an adenomatous lesion in the ascending colon that was difficult to treat endoscopically; therefore, surgery was planned. Preoperative abdominal ultrasonography revealed a bile duct tumor, and endoscopic ultrasonography revealed a mass lesion around the confluence of the cystic duct. Computed tomography revealed localized wall thickening in the middle bile duct, and the upstream bile ducts were slightly dilated. In addition, continuous thickening of the bile duct wall from the gallbladder to the confluence of the cystic duct was observed. No distant metastases, such as liver metastases, or nearby enlarged lymph nodes were observed. Endoscopic retrograde cholangiopancreatography and magnetic resonance cholangiopancreatography revealed a papillary-like elevated lesion in the bile duct near the confluence of the cystic duct, and a biopsy and bile cytology from the same area showed no malignant findings. As a result, the possibility that the patient had gallbladder or bile duct cancer could not be ruled out; therefore, a policy of surgery together with the ascending colon tumor was decided after receiving sufficient informed consent. During surgery, the patient underwent extrahepatic bile duct resection, reconstruction of the biliary tract using the Roux-en-Y method, and right hemicolectomy. Both duodenal and hilar bile duct transects were subjected to a fast frozen section analysis during surgery, and the results were negative for cancer. A histopathological examination of the resected specimen revealed no malignant findings in the bile duct lesions, and a diagnosis of peribiliary gland hyperplasia with chronic inflammatory cell infiltration and fibrosis of the extrahepatic bile duct wall was made. Conclusions: We encountered a case of peribiliary gland hyperplasia that was difficult to distinguish from bile duct cancer. more...

Optogenetics enables precise neural control but is limited by conventional optical fibers in complex networks. We present a hybrid device integrating multi-point micro-light-emitting diodes (MicroLEDs) with neural electrodes for localized light stimulation and simultaneous neural recording. Fabricated via direct bonding, it ensures optimal alignment for high spatial-temporal resolution. The thin MicroLED probes minimize invasiveness while maintaining optical performance. Validated in mouse brain models, the system achieves selective neural activation and recording with minimal thermal effects. This scalable, flexible tool overcomes integration challenges, advancing optogenetic research and opening avenues for exploring neural dynamics and targeted neurological therapies. more...

We investigated a newly developed digitized Trail Making Test using an iPad (iTMT) as a brief cognitive function screening test. We found that the iTMT part-A (iTMT-A) can estimate generalized cognitive function in rehabilitation inpatients examined using the Mini-Mental State Examination (MMSE). Forty-two hospitalized participants undergoing rehabilitation (rehab participants), 30 of whom had cerebral infarction/hemorrhage (stroke participants), performed the iTMT five times (first three times: iTMT-A; fourth: paper version of TMT-A; fifth: the inverse version of iTMT-A) and the MMSE once. Each iTMT-A trial's completion time was divided into the move and dwell times. A linear mixed model following post-hoc tests revealed that the completion time of the third and fourth iTMT-A was faster compared to that of the first iTMT-A, suggesting the presence of a learning effect. In the partial least squares (PLS) regression analysis, the coefficient of determination for estimating the MMSE score was increased by using the dwell and move times extracted from the repeated iTMT-A and the availability of TMT-B, even for subjects with low MMSE scores. These findings indicate that the dwell time of iTMT-A may be important for estimating cognitive function. The iTMT-A extracts significant factors temporally and spatially, and by incorporating the learning effect of repeated trials, it may be possible to screen cognitive and physical functions for rehabilitation patients. more...

The Japan Atomic Energy Agency (JAEA) is developing an evaluation method for a two-phase flow in the reactor core using simulation codes based on the volume of fluid (VOF) method. However, it is impossible to simulate boiling on the heating surface in large-scale domains, such as fuel assemblies, using this simulation method because simulating boiling based on the VOF method requires fine meshes to resolve the initiation of boiling. Therefore, the JAEA started developing a simplified boiling model (SBM) for the two-phase flow simulation in fuel assemblies. In the SBM, the motion and growth equations of a bubble are solved to obtain their diameter and time length at the detachment, the size scale of which is within/around the calculation grid size of the numerical simulation. This information is given as the vapor injection flow rate for the boundary condition on the heating surface. JUPITER calculates the bubble behavior with a scale of more than several millimeters. Using the developed SBM, this study simulates convection boiling on a vertical heating surface. A comparison between the simulation and experimental results showed good reproducibility of the heat flux and velocity dependency on the passage period of the bubble. more...

Abstract The Higgs potentials of extended Higgs sectors exhibit a complex and interesting vacuum structure. When travelling back in time, i.e. going to higher temperatures, the structure may change and exhibit interesting phase patterns and sequences of phases related to the respective minima of the potential. The investigation of the vacuum structure can give us indirect insights in beyond-Standard-Model physics and the evolution of the Universe. In this paper, we investigate the possibility of an intermediate charge-breaking (CB) phase in the 2-Higgs-Doublet Model (2HDM) type I. The existence has been reported previously by using a simple potential setup. We here confirm that the intermediate CB phase can still exist when using the one-loop corrected effective potential including thermal masses. We discuss its features and the relation with SU(2) symmetry (non-)restoration as well as its consistency with the current experimental data. Lastly, we show for some selected benchmark points the rich and interesting phase patterns and sequences that the 2HDM can undergo during its evolution from the early Universe to today’s electroweak vacuum. more...

LoRaWAN (Long Range Wide Area Network), a type of LPWAN (Low Power Wide Area Network), has gained popularity for enabling long-distance data transmission with low-power consumption. However, single-hop LoRaWAN may not be able to provide a sufficiently wide communication range in a practical environment. Although multi-hop communication can offer a wider communication range in LoRaWAN, it has its distinct challenges: the hidden node problem, synchronization misalignment, throughput degradation, and increased power consumption due to noncentralized operation. This paper aims to solve those challenges by proposing an autonomous distributed adaptive resource allocation method and a synchronization misalignment compensation method for multi-hop LoRaWAN systems. In the proposed method, each device selects the radio resource, i.e., time slot and frequency channel, for packet transmission through a mapping rule shared among the devices in advance. Since each device can calculate the timing of the current packet transmission from its preceding device, it can compensate for the misalignment of synchronization. Furthermore, predicting the timing of the next packet from the preceding device enables the device to open the receive window during that time only. Thus, this resource selection not only helps avoid packet collisions but also reduces power consumption by preventing unnecessary reception window openings, thereby improving the efficiency of LoRaWAN communication. The proposed methods are implemented in commercially available private LoRa systems to show their potential to reduce device power consumption by up to 50% reduction. more...

Introduction: The intentional perforation of the pulp chamber floor before tooth replantation promotes pulpal healing by facilitating the revascularization of the pulp cavity. This study aimed to elucidate the effects of this method on the dynamics of quiescent dental pulp stem cells (DPSCs). Methods: The right and left maxillary first molars of Crlj:CD1 mice and TetOP-histone 2B (H2B)-green fluorescent protein (GFP) mice were extracted. The left molars were immediately replanted as the control group (CG), whereas the pulp chamber floor of the right molars were perforated before the tooth was replanted as the experimental group (EG). Immunohistochemistry for Nestin and GFP, and quantitative RT-PCR for Nestin, Opn, CD11c, and Oct3/4 mRNA were performed. Results: The rate of Nestin-positive perimeter along the pulp–dentin border in the EG tended to be higher than that of the CG at days 5 and 7 and was significantly increased between days 3 and 7. The rate of GFP-positive cells in the EG was significantly higher than that of the CG at days 5 and/or 7 in the mesial and middle coronal pulp. CD11c mRNA in the EG at day 5 was significantly higher than that of the CG and tended to be higher than that of the CG during the observation period. Oct3/4 mRNA expression in the EG was significantly higher than that of the CG at day 7. Conclusions: The current experimental model demonstrated the promotion of the survival of DPSCs and their differentiation into odontoblast-like cells (OBLCs). Thus, the use of this model is expected to clarify the crosstalk mechanism between immune cells, including macrophages and dendritic cells, and DPSCs with regards to pulpal healing after tooth replantation. It also provides insight into the differentiation process of DPSCs into OBLCs. more...

Abstract Many governments and non-state actors have pledged to achieve net-zero greenhouse gas emissions, raising questions about the feasibility of these decarbonization goals. The existing literature, however, mostly relied on technoeconomic assessments and lack broad contextual considerations such as national conditions and local sociocultural characteristics. Here, we present a framework for assessing perceived feasibility and multi-dimensional barriers for net-zero transition that can complement existing methods of technoeconomic traditions. We applied this framework to the Japanese net-zero goal by surveying more than 100 experts from diverse fields with a shared national context. Most of the experts supported the desirability of the net-zero goal and chose a probability of 33–66% for its feasibility. However, the distribution of feasibility assessments differs between groups of integrated assessment modelers and the Intergovernmental Panel on Climate Change authors and other researchers, suggesting opportunities for further exploration within and between communities. Identified barriers reflect a unique national condition of Japan and include the limitations of national strategies and clean energy supply. The present framework can be extended to non-experts, data-scarce geographies and sectors. more...

Introduction: Leucine-rich alpha-2-glycoprotein (LRG) is a potential biomarker for disease activity and reflects mucosal healing in patients with ulcerative colitis (UC). However, only a few studies have described a detailed sensitivity analysis of LRG in predicting mucosal healing in patients. This study aimed to evaluate the association between LRG and the endoscopic activity of UC and its predictability for mucosal healing and explore the utility and clinical application of LRG. Methods: The diagnostic accuracy of biomarkers, including LRG, in predicting the endoscopic activity of UC was evaluated. All consecutive patients who underwent total colonoscopy between April 2021 and September 2022 were included. The Mayo endoscopic subscore (MES) was used for assessing endoscopic activity. Furthermore, endoscopic remission was defined as an MES of ≤1. Clinical activity was evaluated based on stool frequency and bloody stool. Receiver operating characteristic curve analysis and binary logistic regression were performed to assess the diagnostic accuracy of the biomarkers. We evaluated LRG trends and treatment response in patients with MES ≥2 who underwent induction therapy. Results: This study comprised 214 patients. The proportions of endoscopically and clinically active patients were 33.6% and 49.1%, respectively. LRG had an area under the curve (AUC) of 0.856, with a higher diagnostic accuracy than other biomarkers, such as C-reactive protein, leukocyte, neutrophil, platelet, and albumin. The cutoff value for LRG was 15.6 μg/mL (sensitivity, 72.2%; specificity, 86.6%). Using the MES, patients with higher scores had higher LRG levels than those with lower scores. The cutoff value, AUC, sensitivity, and specificity varied with a higher AUC for left-sided colitis and pancolitis than for proctitis. Logistic regression analysis showed that LRG was an independent predictor of endoscopic remission using multivariate analysis, even with the factor of clinical activity. The change ratio of LRG pre- and post-treatment was statistically significant in the higher LRG group. Conclusion: LRG reflected endoscopic activity independently, regardless of clinical symptoms. An LRG below the cutoff value could indicate a significantly low probability of endoscopic activity in asymptomatic patients, and follow-up endoscopy (not for cancer screening) may be unnecessary. Furthermore, a higher LRG level might be more useful as an indicator of treatment efficacy. more...