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Given that the hydration water of polymer matrices may differ from that of outermost polymer surfaces, processes at biomaterial-biofluid interfaces and role of hydration water therein cannot be adequately examined using most conventional characterization methods. To bridge this gap, a gold substrate was herein modified with linear and cyclic poly(2-methoxyethyl acrylate) to prepare gl -PMEA and gc -PMEA surfaces, respectively, as models for the outermost surfaces of blood-contacting medical devices. Both surfaces suppressed the adhesion of human platelets but differed in the adhesion behaviors of normal and tumor cells despite having the same areal density of fixed-end units. The surfaces were analyzed using quartz crystal microbalance (QCM), frequency modulation atomic force microscopy (FM-AFM), and X-ray emission spectroscopy (XES) measurements under wet conditions to clarify the relationship between bioresponsivity and hydration water. QCM measurements provided evidence that both grafted-PMEA were hydrated. FM-AFM observations revealed that the swelling layer was thicker for gc -PMEA. To rationalize the differences in the surface hydration states, we performed XES measurements under conditions enabling control over the number of hydration water molecules. In the low-water-content region, hydrogen bonds or interactions between water molecules developed in the vicinity of gl -PMEA but not gc -PMEA. Thus, the initial hydration behavior of the gc -PMEA surface, which promoted intermediate water formation, was different from that of the gl -PMEA surface. The results suggested that the adjustment and optimization of the hydration state of outermost biomaterial surfaces enable the control of bioresponsivity, including the selective isolation of tumor cells.

Infection by enterotoxigenic Escherichia coli (ETEC) causes severe watery diarrhea and dehydration in humans. Heat-labile enterotoxin (LT) is a major virulence factor produced by ETEC. LT is one of AB 5 -type toxins, such as Shiga toxin (Stx) and cholera toxin (Ctx), and the B-subunit pentamer is responsible for high affinity binding to the LT-receptor, ganglioside GM1, through multivalent interaction. In this report, we found that Glu51 of the B-subunit plays an essential role in receptor binding compared with other amino acids, such as Glu11, Arg13, and Lys91, all of which were previously shown to be involved in the binding. By targeting Glu51, we identified four tetravalent peptides that specifically bind to the B-subunit pentamer with high affinity by screening tetravalent random-peptide libraries, which were tailored to bind to the B-subunit through multivalent interaction. One of these peptides, GGR-tet, efficiently inhibited the cell-elongation phenotype and the elevation of cellular cAMP levels, both induced by LT. Furthermore, GGR-tet markedly inhibited LT-induced fluid accumulation in the mouse ileum. Thus, GGR-tet represents a novel therapeutic agent against ETEC infection., 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 © 2024 Elsevier Inc. All rights reserved.)

Adipose-derived stem cell (ASC)-released exosomes (ASCexos) have multiple biological activities. We examined the effect of ASCexos derived from the inguinal adipose tissue of exercise-trained rats (EX-ASCexos) on adipogenic conversion of 3T3-L1 cells and analyzed their microRNA (miRNA) expression profiles. Differentiation of 3T3-L1 cells into adipocytes was performed for 9 d with EX-ASCexos or ASCexos from sedentary control rats (SED-ASCexos), and the expression of proteins and miRNA involved in adipogenic differentiation were determined. EX-ASCexos but not SED-ASCexos attenuated 3T3-L1 adipocyte differentiation with increased phosph-Ser112PPARγ expression, the inactive form of PPARγ. These differentiated adipocytes were also accompanied by increased phosph-Thr202/Tyr204ERK and decreased dual-specificity phosphatase 3 (DUSP3) levels. The exosomal miRNAs miR-323-5p, miR-433-3p, and miR-874-3p were identified specifically in EX-ASCexos. Of these, miR-323-5p mimic replicated the EX-ASCexo-induced suppression of 3T3-L1 adipocyte differentiation and altered adipogenesis-related factor expression. In conclusion, exercise training-driven exosomal miR-323-5p suppressed 3T3-L1 adipogenesis by increasing phosph-Ser112PPARγ expression, while phosph-Thr202/Tyr204ERK accumulation inhibited DUSP3 expression., Competing Interests: Declaration of competing interest There are no conflicts of interest to declare., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Because bats emit ultrasound and listen to echoes to perceive their surroundings, presenting phantom echoes is useful for understanding their perception of echolocation. Here, we present a protocol for the real-time playback of phantom echoes, especially from approaching objects. We describe steps for presenting virtual objects using a microphone, digital signal processor, and loudspeaker to induce the escape behavior of bats. We then detail procedures for clarifying their detection algorithm for approaching objects. For complete details on the use and execution of this protocol, please refer to Yoshida et al. 1 ., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Purpose: Acoustic radiation force impulse (ARFI) elastography and contrast-enhanced ultrasonography (CEUS) are emerging techniques that are becoming common in ultrasound examinations. We previously reported that ARFI (push pulse) induced lung hemorrhage in rabbits, indicating that greater risks are associated with ARFI than with conventional ultrasound. In this study, we assessed the risk of lung hemorrhage under a combination of ARFI elastography and CEUS, considering potential exacerbation of ARFI-induced lung hemorrhage as a result of the ultrasound contrast agent (UCA) used in CEUS., Methods: Twenty-three rabbits were divided into non-UCA and UCA groups. ARFI exposure parameters were set at six mechanical index (MI) levels (0.29, 0.45, 0.60, 0.88, 1.0, 1.39) in non-UCA groups and five MI levels (0.29, 0.66, 0.88, 0.97, 1.25) in UCA groups. Lung exposure was performed bilaterally through the intercostal space in each rabbit. Lung damage was assessed through macroscopic and microscopic observation post euthanasia., Results: Lung hemorrhage was detected at MI 0.3 levels of 0.88 or higher. Logistic regression analyses showed that MI 0.3 was a statistically significant factor for occurrence of lung hemorrhage in both non-UCA and UCA groups, and the MI 0.3 threshold (ED 05 ) for inducing lung hemorrhage was 0.68 and 0.71, respectively. However, multivariate logistic regression and linear regression analyses across all samples indicated that UCA did not significantly affect the occurrence or area of lung hemorrhage., Conclusion: This study demonstrates that UCA does not significantly worsen ARFI-induced lung hemorrhage in terms of occurrence or severity. However, risks and benefits of ARFI elastography on the lung should be considered, irrespective of UCA administration., Competing Interests: Declarations Conflict of interest The authors declare that they do not have any conflict of interest. Ethical approval All institutional and national guidelines for the care and use of laboratory animals were followed., (© 2024. The Author(s), under exclusive licence to The Japan Society of Ultrasonics in Medicine.)

The association between subjective walking speed and metabolic diseases has received limited attention, particularly in individuals with obesity. We aimed to clarify this association using comprehensive health checkup data of participants with obesity. In total, 8578 individuals with a body mass index ≥ 25.0 kg/m 2 , 9626 individuals with waist circumference ≥ 85 cm in men and ≥ 90 cm in women, and 6742 individuals who met both criteria of body mass index and waist circumference were included in this cross-sectional analysis. Subjective walking speed was investigated using the question "Is your walking speed faster than the speed of those of your age and sex?" in a health examination questionnaire. Metabolic diseases were defined according to the guidelines for each disease, and modified Poisson regression analyses were performed. In the model adjusted for age and sex, individuals with obesity based on body mass index and fast subjective walking speed showed significantly lower risk of diabetes mellitus (risk ratio [RR] 0.70; 95% CI 0.63-0.77) and dyslipidemia (RR 0.97; 95% CI 0.94-1.00). Similarly, among those with obesity based on waist circumference and both body mass index and waist circumference, fast subjective walking speed showed a significant negative association with hypertension (RR 0.94; 95% CI 0.90-0.97 and RR 0.95; 95% CI 0.92-0.99, respectively), diabetes mellitus (RR 0.70; 95% CI 0.64-0.77 and RR 0.70; 95% CI 0.63-0.77, respectively), and dyslipidemia (RR 0.96; 95% CI 0.94-0.99 and RR 0.96; 95% CI 0.94-0.99, respectively). Thus, among individuals with obesity, the odds of metabolic diseases were lower if their subjective walking speed was fast. This study contributes to earlier prevention of the cascade of diseases that begin with obesity., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: School-based cognitive behavioral therapy (CBT) programs could contribute toward preventing anxiety disorders in children. However, setting aside class time for such programs is difficult. Internet-based CBT (I-CBT) is an efficient way to provide CBT. However, studies on I-CBT for anxiety prevention remain scarce, including I-CBT for school-based universal prevention programs. Therefore, we developed an e-learning version of a school-based anxiety-prevention CBT program, "Journey of the Brave," which was effective in Japan, to make it more flexible and accessible. This study evaluated its feasibility., Methods: We conducted a single-arm study based on 44 parents and children. Children aged 10-12 years took the e-learning program at home. Parents and children responded to a questionnaire three times: before the learning (Pre), after the learning (Post), and three months later (FU: Follow-up). Feasibility was comprehensively evaluated through dropout rates, satisfaction and learning records, and changes in scores on psychological scales assessing anxiety symptoms, emotion-regulation skills, and children's emotional and behavioral problems., Results: Of the 44 children, 42 started the e-learning, and 32 continued it (dropout rate of 23.8%). Furthermore, 83.9% of the children and 96.8% of the parents responded "agree" or "somewhat agree" regarding overall satisfaction. The parent-rated Spence Children's Anxiety Scale(SCAS) (SCAS-P) showed a significant decrease between Pre and FU (p = 0.014, 95% CI = (-9.22, -0.84)); however, the child-rated SCAS (SCAS-C) reduction was not significant (p = 0.08). The Emotion‑Regulation Skills Questionnaire (ERSQ) also increased significantly between Pre and FU (p = 0.045, 95% CI = (0.18, 18.31)). The Total Difficulties Score of the Strengths and Difficulties Questionnaire(SDQ) decreased significantly from Pre to Post (p = 0.025, 95% CI = (-3.62, -0.19)); however, it was not significantly different between Pre and FU (p = 0.67)., Conclusion: The e-learning version showed low dropout rates and high satisfaction ratings from parents and children. Moreover, this study did not rule out the possibility that the program reduced children's anxiety and improved their emotional-regulation skills. Therefore, its potential and feasibility were indicated., Trial Registration: UMIN, UMIN000049182, Registered 11 October 2022., Competing Interests: Declarations Ethics approval and consent to participate The protocol was approved by the Ethics Review Committee of Chiba University School of Medicine (Registration number: M10352) in accordance with the Ethical Guidelines for Medical and Health Research Involving Human Subjects as well as the Declaration of Helsinki. Informed consent forms were available on the web which were filled out by the parents. A plain-language children’s assent document was also available on the web to ensure that both the parents and the children were willing to participate. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Oxide-based all-solid-state batteries are ideal next-generation batteries that combine high energy density and high safety, but their realization requires the development of interface bonding technology between the stiff solid electrolyte and electrode. Even if the interface could be bonded, it is difficult to hold the interface, because only the electrode expands/contracts unilaterally during charge/discharge reactions. In particular, silicon (Si), which has eagerly awaited as a next-generation negative-electrode material for many years, changes in volume by several hundred percent. To solve these problems, in this work, highly porous silicon oxide (SiO x ) electrodes with different porous structures were fabricated on a stiff garnet-type Li 7 La 3 Zr 2 O 12 solid electrolyte, the three-dimensional nanoporous structure was analyzed quantitatively, and the charge/discharge characteristics were investigated. The microscopic observation and electrochemical analysis revealed how we should control the porous structure, such as sizes of pores and SiO x , size distribution, and porosity, for repeated and stable charge/discharge cycles. In addition, the resultant porous SiO x electrodes demonstrated superior charge/discharge cycle performance even when it thickened to 5 μm, whereas non-porous SiO x easily peeled off from the solid electrolyte when its thickness exceeded 0.1 μm. The thick SiO x films greatly improved the energy density per unit area (mAh cm -2 ). Nanosized fine pores with an interconnected open-pore architecture effectively mitigated the internal and interfacial stress upon expansion (charge)/contraction (discharge) of Si, and as a result, the thick and porous SiO x electrode maintained the interfacial joint with the stiff solid electrolyte after repeated charge/discharge cycles. These results will provide useful insights for effectively designing more practical porous SiO x powder effectively.

Recently, providing smartphone-based health-improving applications to employees has emerged as a promising strategy for sustaining their well-being. This study estimates the impact of the routine use of an application, introduced in 2020 by a Japanese manufacturing company, on various health-related behaviours and outcomes among employees by exploiting a distinctive large-scale longitudinal dataset and personnel records. The analysis addresses potential selection biases arising from the non-random nature of application usage by employing the instrumental variable approach. Regular application use generates significant positive impacts on health-related habits, including moderate alcohol consumption, regular breakfast intake and refraining from eating two hours before bedtime. Furthermore, regarding physical and psychological stress, noteworthy reductions in physical burden and less frequent experiences of annoyance are observed. Employees also report a lower frequency of dizziness, headaches and palpitations, albeit an increase in the frequency of strained eyes is noted. Additionally, application use is associated with lower systolic and diastolic blood pressure, as well as decreased levels of triglycerides and gamma-GTP., 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 © 2024 Elsevier B.V. All rights reserved.)

Recent advancements in microelectromechanical system technology have significantly enhanced our ability to monitor neuronal activity in free-swimming fish without disrupting their natural movement, thereby greatly improving the capabilities of neural logging using "neurologger" technology. In this review, we compiled the findings from studies applying neurologgers to teleost fish, emphasizing the discovery of various spatial-cognition cells in regions of the telencephalon analogous to the mammalian hippocampus that are deeply involved in spatial navigation. We detailed how different fish species, such as goldfish and salmonids, correlate their neural activity with environmental boundaries, head direction, speed, and other navigational cues for spatial memory and navigation strategies. We critically analyzed the similarities and differences in these mechanisms to provide insights into the evolutionary aspects of spatial cognition. We also identified gaps in current methodologies and suggest directions for future research, emphasizing the need for further exploration of spatial encoding in aquatic environments. The insights gained herein suggest the existence of a complex and evolutionarily conserved substrate for navigation and memory in vertebrates, highlighting the potential of neurologgers to expand our understanding of spatial cognition., 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., (Copyright © 2024 Takahashi, Sawatani, Ide, Abe, Kitagawa and Makiguchi.)

In the 21st-century era of globalization, language proficiency is a pivotal connector across cultures, with artificial intelligence (AI) revolutionizing educational paradigms through Robot-Assisted Language Learning (RALL). This systematic review examines the role of RALL in adult second language acquisition, focusing on its pedagogical strategies and learner engagement. Unlike the previous systematic reviews that explore the multifaceted roles of robots in language learning, including as teachers, tutors, assistants, and peer learners, we identify explicit and implicit instructional strategies within RALL, highlighting the unique learning landscape of adult learners characterized by self-regulation and self-direction. We assess the latest advancements in RALL for adult learners through three research questions, compare the effectiveness of explicit versus implicit instructions, and investigate affective factors enhancing RALL performance. Our review contributes a comprehensive status analysis, in-depth exploration of interaction modes, and insights for future research directions, providing a roadmap for academic research and practical guidance for educators and robot developers. This study aims to optimize RALL strategies to better meet the needs of adult learners, fostering a more efficient and engaging language learning experience., 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., (Copyright © 2024 Deng, Fu, Ban and Iio.)

Glaucoma is an optic degenerative neuropathy that is driven by a vicious cycle of oxidative stress and mechanical stress. Current clinical treatments aim exclusively at alleviating mechanical stress by reducing the intraocular pressure (IOP). With the unattended oxidative stress, recurrence and deterioration of mechanical stress are inevitable. Nitric oxide (NO) and carbon monoxide (CO) are endogenous gaseous signaling molecules for vasodilation and anti-inflammation, respectively. Mounting evidence suggests an intricate interplay between NO and CO to mediate their biological roles, like how it takes two to dance a waltz. This leads to the concept of "gas waltz therapy" for glaucoma, in which NO is released to reduce IOP and stoichiometric CO is coreleased to suppress oxidative stress. CND570 is the first phototriggered cascade NO/CO donor, to the best of our knowledge. Notably, the release of NO/CO is accompanied by the concomitant release of a rhodamine dye whose bright fluorescence is harnessed as a convenient calibration mechanism of the gas release profile. CND570 exhibits excellent transcorneal permeability and reaches the target aqueous humor outflow pathway. Further, green-light irradiation triggers release of CO and NO in the eye tissue of glaucoma mice. NO and CO could promote the upregulation of soluble guanylate cyclase (sGC) in both in vitro and in vivo models. Notably, CND570 treatment significantly reduces the oxidative stress associated with glaucoma. NO/CO-based gas waltz therapy is a promising new avenue for glaucoma treatment.

The homochirality of life remains one of the most enigmatic issues in the study of the origin of life. A proposed mechanism for symmetry breaking involves irradiation by circularly polarized light (CPL). To investigate the photoreaction of amino acids under CPL irradiation, a vacuum ultraviolet (VUV) CPL irradiation system was developed at the synchrotron light source UVSOR-III. Hydrogen Lyman-α CPL (121.6 nm) is considered a potential asymmetric source in space. Therefore, racemic alanine film samples were irradiated with Lyman-α CPL to explore the photoreaction of biomolecules. Circular dichroism (CD) spectra measurements revealed that irradiation with right- (left-) handed CPL induced a positive (negative) anisotropy factor g in the wavelength range of 180-240 nm. However, the spectra differed from those of enantiopure alanine, exhibiting broad wavelength ranges and no sign change. Liquid chromatography-mass spectrometry (LC-MS) measurements indicate formation of larger molecules, such as oligomeric alanine adducts or modified oligomers after the Lyman-α CPL irradiation. Additionally, CPL irradiation considerably changes the microstructure of the alanine film surface, leading to the formation of circular network aggregates on the scale of 100 nm. The morphology changes in the alanine film and/or the formation of the larger molecules could be possible causes of the modified anisotropy factor spectra compared to those of enantiopure alanine. These findings highlight the need for further research on the photoreaction of biomolecules in solid states under VUV CPL irradiation, particularly in the photoionization energy range, to validate the cosmic scenario of homochirality., (© 2024 The Author(s). Chirality published by Wiley Periodicals LLC.)

It is established that neurogenesis of dentate gyrus is increased after ischemic insult, although the regulatory mechanisms have not yet been elucidated. In this study, we focused on Ezh2 which suppresses gene expression through catalyzing trimethylation of lysine 27 of histone 3. Male gerbils were injected with adeno-associated virus (AAV) carrying shRNA targeting to Ezh2 into right dentate gyrus 2 weeks prior to forebrain ischemia. One week after ischemia, animals were injected with thymidine analogue to label proliferating cells. Three weeks after ischemia, animals were killed for histological analysis. AAV-mediated knockdown of Ezh2 significantly decreased the ischemia-induced increment of proliferating cells, and the proliferated cells after ischemia showed significantly longer migration from subgranular zone (SGZ), compared to the control group. Furthermore, the number of neural stem cells in SGZ significantly decreased after ischemia with Ezh2 knockdown group. Of note, Ezh2 knockdown did not affect the number of proliferating cells or the migration from SGZ in the non-ischemic condition. Our data showed that, specifically after ischemia, Ezh2 knockdown shifted the balance between self-renewal and differentiation toward differentiation in adult dentate gyrus., (© 2024. The Author(s).)

We have developed a HPLC system where phase-separation multiphase flow works in the separation column as an eluent. We call the novel separation mechanism a phase-separation mode. The ternary mixed solution of water/acetonitrile/ethyl acetate, which is one of the two-phase-separation mixed solutions, caused the phase-separation multiphase flow via phase change from homogeneous to heterogeneous with the temperature effect (from 20 to 0 °C). In this study, we tried to perform phase-separation multiphase flow with the pressure effect instead of the temperature one. The fused-silica capillary tube (50 cm length and 50 µm inner diameter) was allied to the downstream of the column to apply the pressure of 5.5 MPa to the system. Model analytes of 2,6-naphthalenedisulfonic acid (2,6-NDS) and 1-naphthol (1-NA) were examined. For example, solutions (the volume% of water/acetonitrile/ethyl acetate; 20:55:25, organic-rich) and (60:30:10, water-rich) were used as eluent. The model analytes were not separated with both solutions at the pressure of 1.5 MPa and 20 °C. But with the organic-rich solution, 1-NA and 2,6-NDS were separated in this order and with the water-rich solution, they were separated in the reverse order at the pressure of 5.5 MPa and 20 °C. The phase-separation mode could be performed at the high pressure even at the room temperature., (© 2024. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

A ferrofluid droplet on a plate undergoes interesting fission under a magnetic field. In this article, a recently developed simplified multiphase lattice Boltzmann method coupled with a self-correction solution for the magnetic potential equation is employed to explore this complex flow phenomenon. It is found that the droplet fission follows a four-pattern process under a nonuniform magnetic field. The mechanisms behind this behavior are influenced by the strength and gradient of the magnetic field, surface tension, droplet size, and the wetting boundary condition. These findings provide valuable insights into the application of ferrofluids in droplet microfluidics and contribute to advancements in this field.

This study investigated a technique for controlling the orientation of C2C12-derived myotube cells using ultrasonication for future clinical applications of cultured skeletal muscle tissues. An ultrasonicating cell culture dish, comprising a plastic-bottomed culture dish and a circular glass plate (diameter, 35 mm; thickness, 1.1 mm) attached to an annular piezoelectric ultrasonic transducer (inner diameter, 10 mm; outer diameter, 20 mm; thickness, 1 mm), was constructed. A concentric resonant vibrational mode at 89 kHz was generated on the bottom of the dish, and the orientations of myotube cells were quantitatively evaluated using two-dimensional Fourier transform analysis of phase contrast microscopy images captured over a 14 × 10 mm 2 area at the center of the dish. Unsonicated myotube cells grew in random directions, but ultrasonication aligned them circumferentially in the culture dish. The timing of treatment was important, with ultrasonication for 48 h before differentiation having a greater impact on myotube orientation than ultrasonication after differentiation. A larger ultrasonic vibration, with an amplitude of over 20 V pp , resulted in significantly smaller angles of deviation in the circumferential direction than the control. Ultrasonication enhanced the expression of differentiation-related genes and the formation of aligned myotubes, suggesting that it promotes differentiation of C2C12 cells into myotubes., (© 2024. The Author(s).)

Infrared laser stimulation of the cochlea has been proposed as a possible alternative to conventional auditory prostheses. Whereas previous studies have focused primarily on the short-term effects of laser stimulation, the practical application of this technics requires an investigation into whether prolonged laser exposure can induce neural responses and safely. This study assessed the effect of laser-induced damage to the cochlea on auditory perception using Mongolian gerbils (Meriones unguiculatus) trained with a classical conditioning task. The broadband noise was presented as a conditioned stimulus, and reward licking was recorded as a conditioned response. After training, the subject's cochlea was exposed to a continuous pulsed laser for 15 h. Broadband noise of various intensities was presented without pairing it with water before and after laser exposure to assess the decrease in auditory perception due to laser-induced injury. The licking rate did not change after laser exposure of 6.6 W/cm 2 or weaker but drastically decreased after 26.4 W/cm 2 or higher. These findings showed, for the first time, that the safety margin of long-term, at least several hours, cochlear laser stimulation exists and will contribute to the appropriate delimitation of the safe and effective laser stimulation parameters in future research., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Synaptic-vesicle (SV) recruitment is thought to maintain reliable neurotransmitter release during high-frequency signaling. However, the mechanism underlying the SV reloading for sustained neurotransmission at central synapses remains unknown. To elucidate this, we performed direct observations of SV reloading and mobility at a single-vesicle level near the plasma membrane in cerebellar mossy fiber terminals using total internal reflection fluorescence microscopy, together with simultaneous recordings of membrane fusion by capacitance measurements. We found that actin disruption abolished the rapid SV recruitment and reduced sustained release. In contrast, induction of actin polymerization and stabilization did not affect vesicle recruitment and release, suggesting that the presence of actin filaments, rather than actin dynamics, was required for the rapid recruitment. Single-particle tracking experiments of quantum dot-labeled vesicles, which allows nanoscale resolution of vesicle mobility, revealed that actin disruption caused vesicles to diffuse more rapidly. Hidden Markov modeling with Bayesian inference revealed that SVs had two diffusion states under normal conditions: free-diffusing and trapped. After disruption of the actin filament, vesicles tended to have only the free-diffusing state. F-actin staining showed that actin filaments were localized outside the active zones (AZs) and surrounded some SV trajectories. Perturbation of SV mobility, possibly through interference with biomolecular condensates, also suggested that the restricted diffusion state determined the rate of SV recruitment. We propose that actin filaments confined SVs near the AZ to achieve rapid and efficient recruitment followed by priming and sustained synaptic transmission., Competing Interests: Competing interests statement:The authors declare no competing interest.

Hydroxyapatite was crystallized in an alkaline dispersion of mechanically fibrillated cellulose to prepare their composites with hydroxyapatite contents of 26-86 wt %. The composite powder was uniaxially pressed at 120 °C and 300 MPa to obtain the compacts. Three-point bending tests revealed that the bending strengths of the compacts were 40-100 MPa, and the elastic moduli were 4-9 GPa. The composite containing 43% hydroxyapatite showed the largest bending strength and the largest work of fracture, and the composite containing 62% hydroxyapatite showed the largest elastic modulus. The composites, derived from the bioderived eco-friendly materials, showed the mechanical properties comparable to those of engineering plastics such as polyamide-6. Scanning electron microscopic observation of the fracture surface showed that the organic phase was discontinuous when the hydroxyapatite weight fraction was increased from 43% to 62%, and the compacts with a discontinuous organic phase lost toughness., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Laccase, a multi-copper oxidase, is limited by its optimal temperature range and isolation costs. To overcome these challenges, we synthesized copper-doped zeolitic imidazolate framework-67 (Cu-doped ZIF-67) with 16 mol % Cu as an artificial laccase catalyst. The introduced Cu site acts as the phenol oxidation site, and Co-based ZIF-67 is the four-electron oxygen reduction site. Laccase also employs this division of oxidation and reduction sites. Cu-doped ZIF-67 demonstrated significant catalytic activity, superior to natural laccase, especially at elevated temperatures, and maintained stability across multiple reaction cycles. These findings suggest that Cu-doped ZIF-67 is a robust, reusable alternative for industrial applications requiring high thermal stability and efficient catalysis., (© 2024 Wiley-VCH GmbH.)

The universe began 13.8 billion years ago, and Earth was born 4.6 billion years ago. Early traces of life were found as soon as 4.1 billion years ago; then, ~200,000 years ago, the human being was born. The evolution of life on earth was to become individual rather than cellular life. The birth of mitochondria made this possible to be the individual life. Since then, individuals have had a limited time of life. It was 1.4 billion years ago that a bacterial cell began living inside an archaeal host cell, a form of endosymbiosis that is the development of eukaryotic cells, which contain a nucleus and other membrane-bound compartments. The bacterium started to provide its host cell with additional energy, and the interaction eventually resulted in a eukaryotic cell, with both archaeal (the host cell) and bacterial (mitochondrial) origins still having genomes. The cells survived high concentrations of oxygen producing more energy inside the cell. Further, the roles of mitochondria in human being's life and aging will be discussed.

Study Design: A cadaveric study., Objective: Measuring the pressure exerted on lumbar lateral interbody fusion (LLIF) cages and examining its correlation with qualitative assessment of tactile feedback related to pullout strength during cage extraction to determine the optimal cage height., Summary of Background Data: No data exists on the relationship between the surgeon's tactile feedback related to pullout strength during the LLIF cage extraction and the cage pressures measured during surgery., Methods: We used four cadavers to insert the original stress-measuring cages at the L2/3 to L4/5 levels (12 intervertebral spaces) using the oblique lumbar lateral approach. The intervertebral disc height (IDH) was measured before cage insertion. These stainless-steel stress-measuring cages were designed with the same shape as standard cages and included a built-in load cell to measure the stress applied. The cage heights were set at 8, 10, and 12 mm and they were inserted sequentially from 8mm, with stress measurements taken at each step. Four spine surgeons qualitatively evaluated the extraction strength using three rating categories ("loose," "appropriate," and "hard to extract"). After cage removal, endplate damage (ED) and anterior longitudinal ligament injury (ALLI) were checked. The anterior part of the vertebral body was dissected to expose the ALL and it is checked for obvious injuries. Additionally, a surgical probe is used to check for ED and ALLI., Results: The measured cage pressures were 45, 120, and 191 N for the "loose," "appropriate," and "hard to extract" ratings, respectively (P < 0.001). ED was observed in five intervertebral spaces, whereas no ALLI was noted. The average maximum pressure when ED occurred was 127 N. The primary risk factor for ED was identified as the difference between the height of the inserted cage and IDH, with a cut-off value of 4.5 mm., Conclusion: The appropriate extraction strength corresponded to a cage pressure of 120 N. A cage pressure ≥127 N posed a risk for ED. ED incidence was high when the difference between the IDH and the height of the inserted cage exceeded 4.5 mm., Level of Evidence: Level Three., Competing Interests: Conflicts of Interest and Sources of Funding: This study received funding from Globus Medical Research grant: No.22-277 and JSPS KAKENHI Grant number JP24K19575., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Pea (Pisum sativum L.), a widely cultivated legumes globally, is attracting interest as a functional food owing to its antioxidant properties derived from nutritional components such as polyphenols. We previously reported that yellow pea-based pasta (YPP) aids in controlling blood glucose and enhances the sensitivity to saltiness. This study examined the antioxidant effect of YPP and its effects on the salt intake and postprandial blood glucose levels by simulating a real-life scenario. In this open, parallel-group, randomized controlled trial, 40 healthy adult men and women aged 20-65 years, whose salt intake exceeded the target salt equivalent level of the Japanese dietary intake standard, were allocated to the following groups (n = 20): the group consuming one serving of YPP per day and the group maintaining their regular daily dietary habits. The participants who were allocated to the YPP group showed significantly improved oxidative stress markers (BAP/d-ROMs ratio change: control =  - 0.11, YPP = 0.27, p = 0.044; lipid peroxide change: control = 0.11, YPP =  - 0.25, p < 0.001) than control participants. The effects on salt intake and blood glucose levels were limited. In conclusion, YPP may serve as a functional staple food that improves oxidative stress., (© 2024. The Author(s).)

Objective: This study quantifies the change in collision avoidance performance of autonomous emergency braking (AEB) systems for traffic accidents in Japan since 2015., Method: This study used data on Japanese traffic accidents compiled by Japan's National Police Agency. The data included only accidents involving loss of or injury to human life; accidents involving only property damage were excluded. We restricted our analysis to collisions between two 4-wheel vehicles and considered only collisions for which we could determine whether the primary party's car was equipped with an AEB system. Both Poisson and negative binomial mixed-effects regression analyses were conducted using the data for 2021 and 2022 to measure the collision avoidance performance of first registered cars in 2015 to 2020 equipped with AEB systems compared with cars without AEB systems first registered in 2015. Collision avoidance performance was measured for 2 types of intervehicle collisions: rear-end collisions and right-turn collisions. Collision avoidance performance for rear-end collisions was also measured for each of the 3 car types-Standard, small, and light cars., Results: The collision reduction rate for rear-end collisions increases with the year of first registration and for cars equipped with AEB systems first registered in 2020 compared with non-AEB-equipped cars first registered in 2015 is 69.2% (95% confidence interval [CI] 67.0%-71.1%), indicating that the performance of AEB systems has dramatically improved in terms of preventing rear-end collisions. For right-turn collisions, the rate increased to 20.4% (95% CI 5.9%-32.6%) for cars equipped with AEB systems first registered in 2019. However, no clear trend is observed., Conclusions: This study evaluated a time series of the collision reduction performance of AEB systems using an original methodology. Japan's New Car Assessment Program (JNCAP) has included AEB's effectiveness in reducing damage from traffic collisions as an evaluation item since FY2014. The results could demonstrate the effectiveness of JNCAP.

Purpose: To determine whether the clinical and paraclinical course of Fuchs endothelial corneal dystrophy (FECD) over 1 year is related to the extent of triplet repetition in the transcription factor 4 (TCF4) gene., Methods: A prospective study with a 1-year follow-up was conducted. A total of 104 patients (160 eyes) with FECD and an equivalent number of age- and sex-matched control subjects without FECD were included. At inclusion, the corneas were graded using the modified Krachmer grade (KG) and patients were genotyped for the number of trinucleotide repeats (TNRs) in the TCF4 gene by the short tandem repeat assay. Visual acuity, Scheimpflug tomographic features, and the Visual Function and Corneal Health Status using a visual disability instrument were measured on 2 visits at 1-year intervals., Results: KGs ranged from 1 to 6, and 46% of eyes had grades 1 to 4. 71% of the patients harbored TNR expansion (>40) versus 13% in control subjects ( P < 0.001). Severity at inclusion was higher in the presence of TNR expansion when considering eyes independently (mean grade ±SD, 4.08 ± 1.42) without TNR expansion and 4.66 ± 1.27 with TNR expansion ( P = 0.024). In 1 year, the ETDRS score significantly decreased by -2.97 (95% confidence interval -4.69 to -1.26, P = 0.001) and the ETDRS score with glare by -4.25 (95% confidence interval -6.22 to -2.27, P < 10 -5 ). There was no relationship between the rate of decline and TNR expansion or KG. Central corneal thickness and Visual Function and Corneal Health Status scores did not significantly vary., Conclusions: It is possible to measure a subtle progression of FECD over a period as short as 1 year. We did not find a relationship between the presence of TNR expansion and the speed of deterioration over 1 year. This work should facilitate the design of future clinical trials on FECD., Competing Interests: Conflicts of interest statement: P. Gain, N. Koizumi, N. Okumura, and G. Thuret were shown using the ICMJE disclosures form. The remaining authors have no funding or conflicts of interest to disclose., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Sarcopenic obesity is the co-existence of obesity and sarcopenia in individuals aged 40-75 years. The Japanese Working Group on Sarcopenic Obesity has developed diagnostic criteria tailored for the Japanese population, considering their unique characteristics compared with European populations. Our algorithm consists of two steps: screening and diagnosis. The screening of obesity mandates using waist circumference and/or body mass index (BMI) based on national standards, while screening for sarcopenia involves the "finger ring test" in addition to the Asian Working Group for Sarcopenia 2019 criteria. The final diagnosis of sarcopenia involves handgrip strength for low muscle strength, the five-times chair stand test for low physical function, and limb skeletal muscle mass (corrected for BMI) for low muscle mass. Obesity is assessed by visceral fat area or body fat percentage. Sarcopenic obesity is then categorized into Stage I, with muscle weakness/loss of function, loss of muscle mass, and obesity; or Stage II, which includes complications. Further clinical validation is needed to refine the consensus and age range. Geriatr Gerontol Int 2024; 24: 997-1000., (© 2024 The Author(s). Geriatrics & Gerontology International published by John Wiley & Sons Australia, Ltd on behalf of Japan Geriatrics Society.)

Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive form of nonalcoholic fatty liver disease characterised by fat accumulation, inflammation, oxidative stress, fibrosis, and impaired liver regeneration. In this study, we found that heme oxygenase-1 (HO-1) is induced in both MASH patients and in a MASH mouse model. Further, hepatic carbon monoxide (CO) levels in MASH model mice were >2-fold higher than in healthy mice, suggesting that liver HO-1 is activated as MASH progresses. Based on these findings, we used CO-loaded red blood cells (CO-RBCs) as a CO donor in the liver, and evaluated their therapeutic effect in methionine-choline deficient diet (MCDD)-induced and high-fat-diet (HFD)-induced MASH model mice. Intravenously administered CO-RBCs effectively delivered CO to the MASH liver, where they prevented fat accumulation by promoting fatty acid oxidation via AMP-activated protein kinase (AMPK) activation and peroxisome proliferator-activated receptor induction. They also markedly suppressed Kupffer cell activation and their corresponding anti-inflammatory and antioxidative stress activities in MASH mice. CO-RBCs also helped to restore liver regeneration in mice with HFD-induced MASH by activating AMPK. We confirmed the underlying mechanisms by performing in vitro experiments in RAW264.7 cells and palmitate-stimulated HepG2 cells. Taken together, CO-RBCs show potential as a promising cellular treatment for MASH., 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 © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background & Aims: Caloric overfeeding combined with adequate protein intake increases not only body fat mass but also fat-free mass. However, it remains unclear whether the increase in fat-free mass due to overfeeding is associated with an increase in total body protein mass. We evaluated the hypothesis that overfeeding would promote an increase in total body protein mass., Methods: In our randomized controlled trial, 23 healthy young men were fed a diet equivalent to their energy requirements with a +10 % energy surplus from protein alone or a +40 % energy surplus (+10 % from protein, +30 % from carbohydrate) for 6 weeks. We estimated total body protein mass by a four-compartment model using dual-energy X-ray absorptiometry, deuterium dilution, and hydrostatic underwater weighing., Results: The 40 % energy surplus over 6 weeks significantly increased body protein mass compared to baseline by 3.7 % (0.44 kg; 95 % confidence interval [CI], 0.21-0.67 kg; P = 0.003); however, the 10 % energy surplus did not result in a significant change (0.00 kg; 95 % CI, -0.38-0.39 kg; P = 0.980). A significant interaction between intervention duration (time) and energy surplus (group) was observed for total body protein mass (P = 0.035, linear mixed-effects model), with a trend toward a significant difference in total body protein mass gain between groups (P = 0.059, Wilcoxon rank sum test). The increase in body protein mass due to the energy surplus was correlated with an increase in fat mass (r = 0.820, p = 0.002)., Conclusions: A higher energy intake was found to promote an increase in body protein mass in healthy men consuming excess protein, suggesting the importance of energy surplus in body protein accumulation. This effect of energy surplus may be related to factors such as increased body fat mass and the associated secretion of adipokines., Trial Registration: The trial was registered with the University Hospital Medical Information Network Clinical Trial Registry as UMIN000034158., Competing Interests: Conflict of interest KK reports receiving grants and research funds from JSPS Grants-in-Aid for Scientific Research and Meiji Co., Ltd. during the conduct of this study. KK also reports receiving lecture and manuscript fees from the Japanese Society of Anti-Aging Medicine and Bunkodo Co., Ltd. RT, KU, and CS were employed by Meiji Co., Ltd. during the study period. The other authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This review discusses the prevention and treatment of coronavirus disease 2019 (COVID-19) caused by infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Mutations in its spike glycoprotein have driven the emergence of variants with high transmissibility and immune escape capabilities. Some antiviral drugs are ineffective against the BA.2 subvariant at the authorized dose. Recently, 150 natural metabolites have been identified as potential candidates for development of new anti-COVID-19 drugs with higher efficacy and lower toxicity than those of existing therapeutic agents. Botanical drug-derived bioactive molecules have shown promise in dampening the COVID-19 cytokine storm and thus preventing pulmonary fibrosis, as they exert a strong binding affinity for viral proteins and inhibit their activity. The Health Ministry of Thailand has approved Andrographis paniculata (Jap. Senshinren) extracts to treat COVID-19. In China, over 85% of patients infected with SARS-CoV-2 receive treatments based on traditional Chinese medicine. A comprehensive map of the stages and pathogenetic mechanisms related to the disease and effective natural products to treat and prevent COVID-19 are presented. Approximately 10% of patients with COVID-19 are affected by long COVID, and COVID-19 infection impairs mitochondrial DNA. As the number of agents to treat COVID-19 is limited, adjuvant botanical drug treatments including vitamin C and E supplementation may reduce COVID-19 symptoms and inhibit progression to long COVID., 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., (Copyright © 2024 Chatatikun, Indo, Imai, Kawakami, Kubo, Kitagawa, Ichikawa, Udomwech, Phongphithakchai, Sarakul, Sukati, Somsak, Ichikawa, Klangbud, Nissapatorn, Tangpong and Majima.)

Objective: Water is an essential nutrient for all organisms and is important for maintaining life and health. We aimed to develop a biomarker-calibrated equation for predicting water turnover (WT) and pre-formed water (PW) using the doubly labelled water (DLW) method., Design: Cross-sectional study., Setting: General older population from the Kyoto-Kameoka Study, Japan., Participants: The 141 participants aged ≥ 65 years were divided into a model developing ( n 71) and a validation cohort group ( n 70) using a random number generation. WT and PW was measured using the DLW method in May-June of 2012. In developing the cohort, equations for predicting WT and PW were developed by multivariate stepwise regression using all data from the questionnaires in the Kyoto-Kameoka study (including factors such as dietary intake and personal characteristics). WT and PW measured using the DLW method were compared with the estimates from the regression equations developed using the Wilcoxon signed-rank test and correlation analysis in validation cohort., Results: The median WT and PW for 141 participants were 2·81 and 2·28 l/d, respectively. In the multivariate model, WT ( R 2 = 0·652) and PW ( R 2 = 0·623) were moderately predicted using variables, such as height, weight and fluid intake from beverages based on questionnaire data. WT ( r = 0·527) and PW ( r = 0·477) predicted that using this model was positively correlated with the values measured by the DLW method., Conclusions: Our results showed factors associated with water requirement and indicated a methodological approach of calibrating the self-reported dietary intake data using biomarkers of water consumption.

The purpose of this study was to determine the effect of the COVID-19 pandemic on infant development. The study investigated the development of infants at 10-11 months of age between 2020 and 2023 by using the Kyoto Scale of Psychological Development-2020 (KSPD2020), an individualized developmental scale, and the Kinder Infant Developmental Scale (KIDS), a developmental questionnaire. We compared the results of the KSPD2020 with those of a pre-pandemic developmental research and compared the developmental age (DA) of KIDS with children's chronological age (CA). Moreover, the same developmental research was conducted again on the same children at 18-24 months of age. DA for receptive language and expressive language was lower in the KIDS compared to CA in the investigation at 10-11 months. However, in the investigation at 18-24 months, there were no areas where KIDS' DA was lower than CA, and DA in the areas of manipulation, receptive language, social relationship with adults, discipline, and eating was higher than CA. On the other hand, using the KSPD2020, there were no differences when compared to pre-pandemic data in the investigation at 10-11 months. Furthermore, the investigation at 18-24 months showed that developmental quotient (DQ) was lower in the Language-Social (L-S) areas than in the investigation at 10-11 months. The lower DQ of L-S in this study was also evident in comparison to the 18-24 months pre-pandemic data. These results suggested that to investigate the medium- and long-term effects of the COVID-19 pandemic on children's development, it is necessary to use not only parent-filled questionnaires but also individualized developmental scales. In addition, the finding that results may differ depending on the method of developmental assessment is considered important not only for developmental researchers but also for professionals involved in supporting children's development., 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., (Copyright © 2024 Otani, Kato, Haraguchi and Goma.)

Two novel diarylethene-fused subporphyrinoids were prepared and characterized. A mono diarylethene derivative was obtained via a statistical condensation reaction with 2 eq. of 1,2-dicyanobenzene and 1 eq. of thiophene-disubstituted butenedinitrile. The symmetric triply diarylethene-fused subporphyrazine was synthesized via a cyclotrimerization reaction of the thiophene-disubstituted butenedinitrile derivative. These compounds were characterized by NMR spectroscopy and high-resolution mass spectrometry. The spectroscopic properties have been measured in hexane and in chloroform. The mono diarylethene-fused-type compound showed photochromism at 580 nm and >700 nm wavelength, accompanied by degradation. According to DFT calculations, photoreactivity likely depends on the contribution of aromatic feature of pyrrole ring bonded to two thiophene rings., (© 2024 Wiley-VCH GmbH.)

A comprehensive analysis of cross-country skiing races is a pivotal step in establishing effective training objectives and tactical strategies. This study aimed to develop a method of classifying sub-techniques and analyzing skiing characteristics during cross-country skiing skating style timed races on snow using high-precision kinematic GNSS devices. The study involved attaching GNSS devices to the heads of two athletes during skating style timed races on cross-country ski courses. These devices provided precise positional data and recorded vertical and horizontal head movements and velocity over ground (VOG). Based on these data, sub-techniques were classified by defining waveform patterns for G2, G3, G4, and G6P (G6 with poling action). The validity of the classification was verified by comparing the GNSS data with video analysis, a process that yielded classification accuracies ranging from 95.0% to 98.8% for G2, G3, G4, and G6P. Notably, G4 emerged as the fastest technique, with sub-technique selection varying among skiers and being influenced by skiing velocity and course inclination. The study's findings have practical implications for athletes and coaches as they demonstrate that high-precision kinematic GNSS devices can accurately classify sub-techniques and detect skiing characteristics during skating style cross-country skiing races, thereby providing valuable insights for training and strategy development.

Bioimaging such as magnetic resonance is used to monitor atherosclerotic plaques consisting of foam cells, which are derived from macrophages that have ingested oxidized low-density lipoprotein (oxLDL). However, the current bioimaging techniques are not highly specific and sensitive in detecting foam cells, calling for the development of higher precision foam cell detection probes. Here, we investigated the utility of iodine-125-labeled oxLDL ( 125 I-oxLDL) as a prototype radiotracer in the radioimaging of foam cells infiltrating atherosclerotic plaques. Mouse bone marrow-derived macrophages (BMDMs) were used to analyze oxLDL uptake. Atherosclerosis mouse model was injected with 125 I-oxLDL and DiI-labeled oxLDL (DiI-oxLDL). Accumulation of 125 I-oxLDL and DiI-oxLDL in foam cells infiltrating atherosclerotic plaques was examined using Oil Red O (ORO) staining, autoradiography, and fluorescent immunohistochemistry. BMDMs phagocytosed oxLDL/ 125 I-oxLDL via CD36, but not LDL/ 125 I-LDL. The radioactive signal from 125 I-oxLDL phagocytosed by the BMDMs could be detected for at least 3 days. In atherosclerosis mouse model, atherosclerotic plaques formed in the aortic arches and valves. The radioactive signal of the injected 125 I-oxLDL was detected in atherosclerotic plaques of the aortic arch, and its intensity was positively correlated with the lesion size. Furthermore, the DiI-oxLDL fluorescent signals were detected in foam cells accumulating in atherosclerotic plaques. Thus, we found that 125 I-oxLDL can be used as a radiotracer in the radioimaging of foam cells in atherosclerotic plaques by autoradiography, suggesting its potential future applications in bioimaging methods such as single-photon emission computed tomography., (© 2024. The Author(s), under exclusive licence to World Molecular Imaging Society.)

Cinema, a modern titan of entertainment, holds power to move people with the artful manipulation of auditory and visual stimuli. Despite this, the mechanisms behind how sensory stimuli elicit emotional responses are unknown. Thus, this study evaluated which brain regions were involved when sensory stimuli evoke auditory- or visual-driven emotions during film viewing. Using functional magnetic resonance imaging (fMRI) decoding techniques, we found that brain activities in the auditory area and insula represent the stimuli that evoke emotional response. The observation of brain activities in these regions could provide further insights to these mechanisms for the improvement of film-making, as well as the development of novel neural techniques in neuroscience. In near feature, such a "neuro-designed" products/ applications might gain in popularity., (© 2024. The Author(s).)

Genetically encoded fluorescent sensors continue to be developed and improved. If they could be expressed across multiple cortical areas in non-human primates, it would be possible to measure a variety of spatiotemporal dynamics of primate-specific cortical activity. Here, we develop an Automated Robotic Virus injection System (ARViS) for broad expression of a biosensor. ARViS consists of two technologies: image recognition of vasculature structures on the cortical surface to determine multiple injection sites without hitting them, and robotic control of micropipette insertion perpendicular to the cortical surface with 50 μm precision. In mouse cortex, ARViS sequentially injected virus solution into 100 sites over a duration of 100 min with a bleeding probability of only 0.1% per site. Furthermore, ARViS successfully achieved 266-site injections over the frontoparietal cortex of a female common marmoset. We demonstrate one-photon and two-photon calcium imaging in the marmoset frontoparietal cortex, illustrating the effective expression of biosensors delivered by ARViS., (© 2024. The Author(s).)

Understanding the complex formation of metal ions in room-temperature ionic liquids (ILs) is essential for the application of ILs in solvent extraction. Nevertheless, the research on metal complex formation in ILs lags behind other applications. The complex formation equilibria may be influenced by specific interactions among the metal ion, ligand molecule, and IL cation and anion. In the present investigation, the complex formation of Ni 2+ with ethanol (EtOH) and methanol (MeOH) molecules in ILs, 1-alkyl-3-methylimidazolium bis(trifluoromethylsulfonyl)amide ([C N mim][TFSA], where N represents the alkyl chain lengths of 2 and 8) was discussed in terms of the microscopic interactions among alcohol molecules, [C N mim] + and [TFSA] - , and the mesoscopic mixing states of alcohols in [C N mim][TFSA], with N = 2-12. The microscopic interaction of alcohol molecules with the imidazolium ring H atoms in the ILs was evaluated by using ATR-IR and 1 H and 13 C NMR spectroscopies. The self-hydrogen bonding of alcohol molecules was clarified from the O-H stretching vibration of alcohol molecules. MeOH molecules can be more strongly hydrogen-bonded with themselves than EtOH molecules due to the less steric hindrance and the weaker dispersion force of MeOH with the IL cation's alkyl chain. In fact, small-angle neutron scattering (SANS) experiments revealed the more heterogeneous mixing of MeOH with the ILs by the self-hydrogen bonding among MeOH molecules than EtOH. The longer the IL cation's alkyl chain, the more the MeOH clusters significantly form. In contrast, the formation of EtOH clusters becomes weaker with elongating the alkyl chain. Ultraviolet (UV)-visible spectroscopic measurements on Ni 2+ -[C N mim][TFSA]-alcohol solutions with N = 2 and 8 revealed that di-, tetra-, and hexa-alcohol-Ni 2+ complexes are formed in both the ILs. With N = 2, the stabilities of Ni 2+ -EtOH and Ni 2+ -MeOH complexes are comparable in the IL. However, with N = 8, the complexes are more stable in the EtOH solutions than in the MeOH solutions. This is because the less heterogeneous mixing of EtOH molecules with the IL results in the larger enthalpic contribution in the complex formation, as shown by the thermodynamic parameters estimated by the van't Hoff plots on the stability constants at several temperatures.

Drinking alcohol is considered one of the risk factors for development of diabetes mellitus. Recently, it was reported that selenoprotein P levels in blood are increased by ethanol intake. However, the mechanism by which ethanol increases selenoprotein P has not been elucidated. The expression of selenoprotein P protein and its mRNA were increased in a concentration- and time-dependent manner when human liver-derived HepG2 cells were treated with ethanol. Levels of AMPK and JNK proteins, which have been known to regulate selenoprotein P transcription, were unchanged by ethanol treatment. However, the amount of nuclear FoxO3a, a transcription factor of SeP, was increased. This was associated with dephosphorylation of ERK1 but not ERK2. It was found that ERK1 was dephosphorylated by activation of dual-specific phosphatase 5 and dual-specific phosphatase 6. However, the phosphorylation of MEK by ERK phosphokinase was not affected by ethanol treatment. These results suggest that the ethanol-induced increase in SeP levels occurs by enhanced transcription of SeP mRNA via the DUSP5/6-ERK1-FoxO3a pathway., Competing Interests: No potential conflicts of interest were disclosed., (Copyright © 2024 JCBN.)

Numerous cross-sectional studies have attempted to identify the muscle morphology required to achieve high sprint velocity. Our longitudinal study addressed an unanswered question of cross-sectional studies: whether hypertrophy of the individual trunk and thigh muscles induced by daily training (e.g., sprint, jump, and resistance training) is linked to an improvement in sprint performance within well-trained sprinters. Twenty-three collegiate male sprinters (100-m best time of 11.36 ± 0.44 s) completed their daily training for 1 year without our intervention. Before and after the observation period, the sprint velocities at 0-100 m, 0-10 m, and 50-60 m intervals were measured using timing gates. The volumes of 14 trunk and thigh muscles were measured using magnetic resonance imaging. Muscle volumes were normalized to the participants' body mass at each time point. Sprint velocities increased at the 0-100 m (p < 0.001), 0-10 m (p = 0.019), and 50-60 m (p = 0.018) intervals after the observation period. The relative volumes of the tensor fasciae latae, sartorius, biceps femoris long head, biceps femoris short head, semitendinosus, and iliacus were increased (all p < 0.050). Among the hypertrophied muscles, only the change in the relative volume of the semitendinosus was positively correlated with the change in sprint velocity at the 50-60 m interval (p = 0.018 and ρ = 0.591). These findings suggest that semitendinosus hypertrophy seems to be associated with sprint performance improvement within well-trained sprinters during the maximal velocity phase., (© 2024 The Author(s). European Journal of Sport Science published by Wiley‐VCH GmbH on behalf of European College of Sport Science.)

Importance: Whether guttae in Fuchs endothelial corneal dystrophy (FECD) can be removed by polishing without Descemet stripping and whether postoperative maintenance of reduced guttae can be achieved through cultured corneal endothelial cell (CEC) transplant therapy are critical issues to be addressed., Objective: To investigate the decrease of guttae through polishing degenerated CECs and abnormal extracellular matrix (ECM) without Descemet stripping and to observe the behavior of guttae following cultured CEC transplant., Design, Setting, and Participants: This case series prospective observational study was conducted in a hospital outpatient clinic setting. Between December 2013 and January 2019, 22 eyes with corneal endothelial failure caused by FECD received cultured CEC transplant therapy at Kyoto Prefectural University Hospital. Of these, 15 eyes were consistently monitored at the same central corneal area during the preoperative phase, as well as in the early (within 1 year) and late (after 3 years) postoperative phases. The images from these phases were categorized into 3 groups: typical guttae, atypical guttae, and no guttae., Exposures: Cultured CEC transplant therapy., Main Outcomes: Proportion of guttae in the observable area was measured, comparing the early and late postoperative phases for each group., Results: The mean age of the patients at the time of surgery was 69 years (range, 49-79 years). All 15 eyes exhibited the presence of confluent guttae preoperatively (100%). Among these, 3 of 15 eyes belonged to male patients. The early postoperative phase of guttae morphologies was classified into 3 groups: 5 eyes with typical guttae, 7 with atypical guttae, and 3 with no guttae. The decrease in the number of these guttae was achieved by surgical procedures. The median percentage of guttae in the typical guttae, atypical guttae, and no guttae groups was 41.8%, 44.4%, and 16.2%, respectively, in the early phase, and 42.2%, 38.2%, and 18.8%, respectively, in the late phase., Conclusions and Relevance: The findings demonstrate that in some cases of FECD, guttae can be removed by scraping and polishing abnormal ECM and degenerated CECs, while preserving the Descemet membrane. Furthermore, cultured CEC transplant resulted in no increase in guttae for up to 3 years, providing insights into surgically eliminating guttae.

Background and Aim: Autotaxin (ATX) is an extracellular lysophospholipase D that catalyzes the hydrolysis of lysophosphatidylcholine into lysophosphatidic acid (LPA). Recent accumulating evidence indicates the biological roles of ATX in malignant tumors. However, the expression and clinical implications of ATX in human cholangiocarcinoma (CCA) remain elusive., Methods: In this study, the expression of ATX in 97 human CCA tissues was evaluated by immunohistochemistry. Serum ATX levels were determined in CCA patients (n = 26) and healthy subjects (n = 8). Autotaxin expression in cell types within the tumor microenvironment was characterized by immunofluorescence staining., Results: High ATX expression in CCA tissue was significantly associated with a higher frequency of lymph node metastasis (p = 0.050). High ATX expression was correlated with shorter overall survival (p = 0.032) and recurrence-free survival (RFS) (p = 0.001) than low ATX expression. In multivariate Cox analysis, high ATX expression (p = 0.019) was an independent factor for shorter RFS. Compared with low ATX expression, high ATX expression was significantly associated with higher Ki-67-positive cell counts (p < 0.001). Serum ATX levels were significantly higher in male CCA patients than in healthy male subjects (p = 0.030). In the tumor microenvironment of CCA, ATX protein was predominantly expressed in tumor cells, cancer-associated fibroblasts, plasma cells, and biliary epithelial cells., Conclusions: Our study highlights the clinical evidence and independent prognostic value of ATX in human CCA., (© 2024 Japan Society of Hepatology.)

We developed a novel HPLC device where the phase-separation multiphase flow worked as the eluent in the separation column by using a water/acetonitrile/ethyl acetate triple mixed solution as a dual-phase-separation solution. Dual-phase-separation solutions form a phase-separation multiphase flow in a microscopic space. The new separation mechanism in the HPLC is called phase-separation mode. In this study, we used water and acetonitrile with NaCl mixed solution as a dual-phase-separation solution instead of the triple mixed solution. Octadecylsilyl (ODS)-modified particle- and porous silica particle-packed separation columns were united with the HPLC device for phase-separation mode caused by phase-separation multiphase flow. NA (1-naphthol) and NDS (2,6-naphthalenedisulfonic acid) were analyzed by the device as model sample. Using the water and acetonitrile with NaCl mixed solution at the solvent volume ratio of 5:5, NA and NDS were not separated on either column at 25 °C. On the other hand, they were separated with the order NDS and NA on the ODS column and separated with the order NA and NDS on the silica column in phase-separation mode at 0 °C. We discuss the separation mechanism of phase-separation mode using the water and acetonitrile with NaCl mixed solution at 0 °C., (© 2024. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

Humans exhibit unique skeletal muscle morphologies that are known to matter in upright bipedalism. However, their relevance to the ease of leg swing, which limits locomotion performance, remains unclear. Here, we aimed to examine muscle mass distribution within the human leg and the effect of each muscle on the ease of leg swing. We calculated the mass, center of mass position, and moment of inertia around the hip extension-flexion axis for all leg muscles by using a publicly available dataset of the 3D reconstruction of the musculoskeletal components in human male and female legs. The leg muscles showed a top-heavy-bottom-light tapering trend; muscles far from the hip joint tended to have smaller masses. Interestingly, however, the soleus exhibited sizable mass for its location. Consequently, the moment of inertia of the soleus was exceptionally greatest, accounting for approximately one-quarter of that of all muscles. These results indicate that compared to the other muscles the soleus muscle has a much larger effect on the leg moment of inertia and uniquely makes humans difficult to swing the leg, although the leg muscles basically show the top-heavy bottom-light tapering trend favoring the leg swing. Our findings highlight a novel functional consequence of human body evolution, suggesting that muscular enlargement for postural stability and endurance capacity has compromised the locomotion speed during the adaptation to bipedalism., 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 © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Previous cross-cultural research has described two different attention styles: a holistic style, characterized by context-sensitive processing, generally associated with interdependent cultural contexts, and an analytic style, a higher focus on salient objects, generally found in independent cultural contexts. Though a general assumption in the field is that attention styles are gradually socialized in culture-specific interactions in childhood, empirical evidence for the proximal mechanisms underlying this development is scarce. This study aimed to document the emergence of cross-cultural differences in attention styles in three cultural contexts differing in social orientations, namely in urban middle-class families from Münster, Germany (i.e., more independent context), and Kyoto, Japan, and Indigenous-heritage families from Cotacachi, Ecuador (i.e., more interdependent contexts). Furthermore, to test the assumption that caregivers' attention guidance is one of the forces driving differential development, we investigated how caregivers guide children's attention. In total, 270 children between 4 and 9 years of age and their mothers participated in three tasks: an eye-tracking task, a picture description task and a forced-choice recognition task. Results indicate a mixed pattern of findings: While some tasks revealed the expected cultural differences, namely a higher object focus in Münster compared to Kyoto and Cotacachi, others did not. Regarding caregivers' attention guidance, we found that mothers in Münster more strongly emphasized the focal object than mothers in Kyoto and Cotacachi. The results are discussed in terms of culture-specific developmental trajectories and the generalizability of attentional processes across tasks and cultural contexts. RESEARCH HIGHLIGHTS: We investigated visual attention styles in 4- to 9-year-old children and their mothers from urban Germany, urban Japan, and rural Ecuador in three different tasks. Special emphasis lied on mothers' verbal attention guidance toward their children as a proximal mechanism underlying the emergence of culture-specific attention styles. Mothers from urban Germany guided their children's attention in more analytic ways than mothers from urban Japan and rural Ecuador. The relevance of verbal attention guidance in the development of culture-specific attention styles has been demonstrated beyond the East-West dichotomy., (© 2023 The Authors. Developmental Science published by John Wiley & Sons Ltd.)

Purpose: To evaluate the pharmacokinetics of ganciclovir eye drops by comparing solutions prepared from ganciclovir for intravenous infusion and ganciclovir gel and to assess the impact of systemic administration on drug levels in ocular tissues and serum., Study Design: Experimental study design., Methods: Ganciclovir solutions (0.5% and 1.0%) prepared by diluting DENOSINE ® IV Infusion in saline and 0.15% ganciclovir gel (Virgan ® ) were topically administered in rabbit eyes, with and without concomitant systemic administration of ganciclovir. The concentrations of ganciclovir in the corneal epithelium, stroma, and endothelium, aqueous humor; and blood plasma were analyzed by high-performance liquid chromatography (HPLC)., Results: The ganciclovir solutions (0.5% and 1.0%) maintained therapeutic ganciclovir levels in the corneal endothelium above the effective dose required for 50% inhibition (ED50) up to 6 h, albeit with a swift decline thereafter. The 0.15% ganciclovir gel maintained higher therapeutic concentrations in the corneal endothelium for up to 12 h, exceeding the ED50. Serum concentrations of ganciclovir were significantly elevated in the groups receiving combined systemic administration., Conclusion: Topical application of 0.15% ganciclovir gel maintained high endothelial concentrations, well above the therapeutic threshold, with or without systemic administration. Furthermore, the observed increase in ganciclovir levels within the plasma and aqueous humor following systemic administration posits it as a viable strategy for severe cases of cytomegalovirus corneal endotheliitis or those inadequately managed by local treatments alone., (© 2024. Japanese Ophthalmological Society.)

Background/objectives: While some dietary guidelines have established daily water requirements for adults, those for older adults are not well defined. Factors associated with water turnover (WT), indicating water requirements and preformed water (PW), referring to fluids consumed from food and beverages, remain poorly understood. Therefore, we aimed to investigate these factors in Japanese adults., Subjects/methods: This cross-sectional study included 57 participants aged 54-85 years, living in Fukuoka City, Japan. The WT and PW were measured using the doubly labelled water (DLW) methods from August 2015 to September 2015. Dietary intake and physical activity were evaluated using a 3-day dietary record (DR) and a validated triaxial accelerometer, respectively. Fasting blood and urine samples, as well as environmental conditions, were assessed using a validated technique. We evaluated the variables for predicting water consumption by using a multivariate generalised linear model, with forward stepwise selection using these covariates., Results: The median WT and PW were 2842 ml/day and 2227 ml/day, respectively. In the multivariate model, WT (R 2  = 0.629) and PW (R 2  = 0.621) were moderately predicted by variables such as sex, body weight, moderate-to-vigorous physical activity, water consumption estimated by DR, urinary aldosterone and osmolality, creatinine clearance and wet-bulb globe temperature. The PW estimates from DR were ~40% lower than those from DLW method, despite a significant correlation between the values., Conclusions: WT and PW are associated not only with lifestyle and environmental factors but also with urinary markers and kidney function. These findings provide useful insights into the differences in each individual's water requirement., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

We synthesized a phenolic hydroxy group-bearing version (1) of a simplified analog of aplysiatoxin comprising a carvone-based conformation-controlling unit. Thereafter, we evaluated its antiproliferative activity against human cancer cell lines and its binding affinity to protein kinase C (PKC) isozymes. The antiproliferative activity and PKC-binding ability increased with the introduction of the phenolic hydroxy group. The results of molecular dynamics simulations and subsequent relative binding free-energy calculations conducted using an alchemical transformation procedure showed that the phenolic hydroxy group in 1 could form a hydrogen bond with a phospholipid and the PKC. The former hydrogen bonding formation facilitated the partitioning of the compound from water to the phospholipid membrane and the latter compensated for the loss of hydrogen bond with the phospholipid upon binding to the PKC. This information may facilitate the development of rational design methods for PKC ligands with additional hydrogen bonding groups., (© The Author(s) 2024. Published by Oxford University Press on behalf of Japan Society for Bioscience, Biotechnology, and Agrochemistry.)

It has generally been accepted that vitamin E refers to a group of tocochromanols, α-, β-, γ-, and δ-tocopherols and the corresponding four tocotrienols. Recently, Azzi and colleagues proposed to restrict the term vitamin E only to RRR-α-tocopherol, not to other tocopherols and tocotrienols (Azzi A et al. Free Radic Biol Med. 2023; 207:178-180. doi: 10.1016/j.freeradbiomed.2023.06.029). The aim of this paper is to express our opinion on the nomenclature of vitamin E based on available scientific data. In our opinion, it would be inappropriate to exclude all the tocochromanols other than RRR-α-tocopherol from the vitamin E group at this stage when the molecular mechanisms showing how vitamin E deficiency causes diseases such as ataxia and how vitamin E prevents/reverses such diseases are not elucidated. Understanding of whole functions of tocochromanols including underlying mechanisms and dynamics is essential before revision of currently accepted definition of vitamin E. The potential roles of γ-tocopherol and tocotrienols are discussed despite whether they are vitamin function should be clarified in the future studies., Competing Interests: Declaration of competing interest There is no declaration of competing interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Sensorimotor learning requires reorganization of neuronal activity in the premotor cortex (PM) and primary motor cortex (M1). To reveal PM- and M1-specific reorganization in a primate, we conducted calcium imaging in common marmosets while they learned a two-target reaching (pull/push) task after mastering a one-target reaching (pull) task. Throughout learning of the two-target reaching task, the dorsorostral PM (PMdr) showed peak activity earlier than the dorsocaudal PM (PMdc) and M1. During learning, the reaction time in pull trials increased and correlated strongly with the peak timing of PMdr activity. PMdr showed decreasing representation of newly introduced (push) movement, whereas PMdc and M1 maintained high representation of pull and push movements. Many task-related neurons in PMdc and M1 exhibited a strong preference to either movement direction. PMdc neurons dynamically switched their preferred direction depending on their performance in push trials in the early learning stage, whereas M1 neurons stably retained their preferred direction and high similarity of preferred direction between neighbors. These results suggest that in primate sensorimotor learning, dynamic directional motor tuning in PMdc converts the sensorimotor association formed in PMdr to the stable and specific motor representation of M1., (© 2024. The Author(s).)