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Tungsten-containing formate dehydrogenase from Methylorubrum extroquens AM1 (FoDH1) was investigated from structural biology and bioelectrochemistry. The electron transfer (ET) pathways were investigated by structural information, electrostatic interactions between the electrode and the enzyme, and the differences in the substrates. Two electrode-active sites and multiple ET pathways in FoDH1 were discovered.

A plasma nitriding-assisted 3D printing method was developed to build up the micro-punch and micro-die systems. Two dimensional punch head and core-die cavity geometries were ink-jet printed or screen-printed onto the AISI316 and SKD11 tool substrate surfaces in following their two-dimensional computer-aided design (CAD) data. The low-temperature plasma nitriding process was utilized to make nitrogen supersaturation only into the unprinted substrates. The sand-blasting and chemical etching were utilized to mechanically or chemically remove the printed parts from punch and die substrate. As sand-blasted and chemically etched AISI316 and SKD11 punches and core-dies were simply finished and used as a die set for micro-embossing, micro-piercing and micro-punching processes. In particular, a micro-pump was selected as a miniature mechanical element. Its 3D CAD geometry was sliced to 2D CAD data for each functional AISI304 stainless steel sheet. A pair of punch and die for each 2D CAD geometry for constituent sheet was prepared by the plasma nitriding-assisted 3D printing. Each sheet was punched out by using this set of punch and die to functionalize each sheet unit in correspondence to the sliced CAD data. These constituent sheets were assembled and joined to a structural unit of micro-pump.

With a rapidly aging society, there is increasing interest in the health of female workers in the field of care services for older adults due to increasing demands to maintain 24-h care and to support older adults without errors or accidents. Therefore, the purpose of this cross-sectional study was to examine the association between sleep-disordered breathing (SDB) and sustained attention in women caring for older adults in Japan. The study was conducted in women aged 18–67 years old working in care service facilities for older adults in Japan. The sustained attention of participants was measured by the 10-min psychomotor vigilance task (PVT). SDB was assessed based on the respiratory disturbance index (RDI), which was measured using an ambulatory airflow monitor with a polyvinylidene fluoride (PVDF) film sensor to monitor the respiratory airflow of nasal and oral breathing. The participants wore the monitor to record the breathing status while asleep at home. The severity of SDB was categorized as follows: normal, RDI

The high-density plasma nitriding at 673 K and 623 K was employed to make 10% of nitrogen supersaturation on AISI316 base austenitic stainless steels. The processing parameters and nitrogen-hydrogen gas flow ratio were optimized to increase the yield of N2+ ion and NH-radical for efficient nitriding. The nitrided AISI316 specimens were prepared for multidimensional analysis to describe the fundamental features of low-temperature plasma nitriding. First, macroscopic evaluation revealed that nitrogen supersaturation induced the γ-lattice expansion and the higher nitrogen content than 4% of mass in depth. The mesoscopic analysis describes the holding temperature and initial grain-size effects on the microstructure changes. Plastic straining, grain-size refinement, and nitrogen zone-boundary diffusion processes advance with nitrogen supersaturation to drive the inner nitriding behavior. The microscopic analysis explains the microstructure refinement, the two-phase structuring, and the microstructure modification. Through this multi-dimensional analysis, the essential characteristics of the low-temperature plasma nitriding of 316 austenitic stainless steels were precisely understood to extend the engineering treatise on the bulk nitrogen stainless steels for surface modification and treatment of stainless steels by nitriding. This plasma nitriding was applied to strengthen and harden the AISI316 wire surfaces toward its application on surgery wires.

A femtosecond laser micro−/nano-texturing was proposed to fabricate the coated and surface treated dies with the tailored textures for surface decoration and surface property control of metal, polymer and glass products. The polygonal model for microtextures with nanotextures by the LIPSS-effect was utilized to fabricate a DLC-coated SKD11 die with a star-shaped emblem. This die was set up into the cassette die set for directly imprinting this emblem into aluminum alloy and PET sheets. The periodic surface structure was synthesized as a surface geometry model to build up the super-hydrophobic surface on the nitrogen supersaturated AISI316 die. This die was also set up into a hot stamping system to directly imprint the hydrophobic surface onto the phosphorous glass products. Through the femtosecond laser micro−/nano-texturing and CNC-imprinting, the metal, polymer and glass product surfaces were optically decorated to have color grating and plasmonic brilliance and functionally controlled to be hydrophobic.

Purpose: To investigate whether the topical administration of ripasudil ophthalmic solution enhances aqueous outflow in the episcleral vein of the human eye.Design: Two-sequence, prospective, randomized, double-blind, crossover trial.Participants: Sixteen eyes of 16 healthy participants were recruited in this study.Methods: Participants were randomized into one of the two crossover sequences to the instillation of ripasudil or the control drug, latanoprost, followed by a washout period of more than 2 days, and crossed over to the alternative instillation. The aqueous columns in the episcleral veins were recorded using a video capture system connected to a slit-light microscope (hemoglobin video imaging) before and 2 and 8 h after the instillation.Primary Outcome Measure: Comparisons between ripasudil and latanoprost for the changes of the aqueous column width after the instillation.Results: Two hours after the instillation, the ripasudil group had significantly greater dilation of the aqueous column width than the latanoprost group. Eight hours after the instillation, the ripasudil group had significantly greater dilation of the aqueous column width than the latanoprost group.Conclusions: Hemoglobin video imaging revealed the dilation of the aqueous column width in the episcleral veins of the human eye after the instillation of ripasudil ophthalmic solution.

Objective Based on the increasing incidence of smell and taste dysfunction among coronavirus disease 2019 (COVID-19) patients, such issues have been considered an early symptom of infection. However, few studies have investigated the type of taste components that are most frequently affected in COVID-19 patients. This study investigated the difference in frequencies of the types of taste component disorders among hospitalized COVID-19 patients. Methods In this retrospective, single-center, observational study, patients' background characteristics, clinical course, laboratory and radiological findings, and details on taste and/or smell disorders were collected and analyzed from medical records. Patients A total of 227 COVID-19 patients were enrolled, among whom 92 (40.5%) complained of taste disorders. Results Multiple types of taste disorders (hypogeusia/ageusia and hypersensitivity, or hypersensitivity and changing tastes) were reported in 10 patients. In particular, 23 patients reported hypersensitivity to at least 1 type of taste, and 2 patients complained of a bitter taste on consuming sweet foods. Impairment of all taste components was found in 48 patients (52.2%). The most frequent taste disorder was salty taste disorder (81 patients, 89.0%). Hypersensitivity to salty taste was most frequently observed (19 patients, 20.9%). Conclusion Patients with COVID-19 develop multiple types of taste disorders, among which salty taste disorder was the most frequent, with many patients developing hypersensitivity to salty taste. As smell and taste are subjective senses, further studies with the combined use of objective examinations will be required to confirm the findings.

Pulmonary emphysema is predominantly caused by chronic exposure to cigarette smoke (CS). Novel tobacco substitutes, such as heated tobacco products (HTPs), have emerged as healthier alternatives to cigarettes. IQOS, the most popular HTP in Japan, is advertised as harmless compared with conventional cigarettes. Although some studies have reported its toxicity, few in vivo studies have been conducted. Here, 12-wk-old C57BL6/J male mice were divided into three groups and exposed to air (as control), IQOS aerosol, or CS for 6 mo. After exposure, the weight gain was significantly suppressed in the IQOS and CS groups compared with the control (−4.93 g; IQOS vs. air and −5.504 g; CS vs. air). The serum cotinine level was significantly higher in the IQOS group than in the control group. The neutrophils and lymphocyte count increased in the bronchoalveolar lavage fluid of the IQOS and CS groups compared with those in the control group. Chronic IQOS exposure induced pulmonary emphysema similar to that observed in the CS group. Furthermore, expression levels of the genes involved in the apoptosis-related pathways were significantly upregulated in the lungs of the IQOS-exposed mice. Cytochrome c, cleaved caspase-3, and cleaved poly (ADP-ribose) polymerase-1 were overexpressed in the IQOS group compared with the control. Single-stranded DNA and TdT-mediated dUTP nick-end labeling-positive alveolar septal cell count significantly increased in the IQOS group compared with the control. In conclusion, chronic exposure to IQOS aerosol induces pulmonary emphysema predominantly via apoptosis-related pathways. This suggests that HTPs are not completely safe tobacco products.

Flavin adenine dinucleotide (FAD)-dependent fructose dehydrogenase (FDH) from Gluconobacter japonicus NBRC3260, a membrane-bound flavohemoprotein capable of direct electron transfer (DET)-type bioelectrocatalysis, was investigated from the viewpoints of structural biology and bioelectrochemistry. As FDH provides a strong DET-type catalytic signal, extensive research has been conducted. Structural analysis using cryo-electron microscopy (cryo-EM) and single-particle analysis revealed the entire FDH structure. The electron transfer (ET) pathway during the catalytic oxidation of D-fructose was investigated using thermodynamic and kinetic approaches in bioelectrochemistry, as well as structural information. Key amino acid residues that play important roles in substrate specificity and ET acceleration have also been proposed.

Bilirubin oxidase from Myrothecium verrucaria (mBOD) is a promising enzyme for catalyzing the four-electron reduction of dioxygen into water and realizes direct electron transfer (DET)-type bioelectrocatalysis. It has two N-linked glycans (N-glycans), and N472 and N482 are known as binding sites. Both binding sites located on opposite side of the type I (T1) Cu, which is the electrode-active site of BOD. We investigated the effect of N-glycans on DET-type bioelectrocatalysis by performing electrochemical measurements using electrodes with controlled surface charges. Two types of BODs with different N-glycans, mBOD and recombinant BOD overexpressed in Pichia pastoris (pBOD), and their deglycosylated forms (dg-mBOD and dg-pBOD) were used in this study. Kinetic analysis of the steady-state catalytic waves revealed that both size and composition of N-glycans affected the orientation of adsorbed BODs on the electrodes. Interestingly, the most favorable orientation was achieved with pBOD, which has the largest N-glycans. Furthermore, the effect of the orientation control by the N-glycans is cooperative with electrostatic interaction.

We aimed to evaluate the association between hyperactivity/anxiety and obesity among elementary school students in Matsuyama City, Japan. We conducted a cross-sectional study of all 46 elementary school students (24, 296 students) in Matsuyama City. The questionnaire included question items from the Strengths and Difficulties Questionnaire (SDQ), as well as questions about height and weight. The students were classified into two groups according to Rohrer index. After separating the data by gender, we examined the association between (1) obesity and hyperactivity, (2) obesity and anxiety, and (3) obesity and combination of hyperactivity and anxiety, by estimating relative risk using Poisson regression model. We also conducted stratified analyses to examine the effect modification by age groups and unbalance of diet on those associations. Moreover, we calculated relative excess risk due to the interaction (RERI) to examine whether there was an additive interaction between hyperactivity and anxiety. Sixteen thousand forty-eight students were included in the present analysis. The prevalence ratio (PR) of being obese in girls who had both hyperactivity and anxiety was higher compared to girls without those symptoms (PR = 1.80; 95% CI 1.04–3.13). There was no significant difference in the prevalence ratio for obesity in boys, whether they were hyperactive, anxious, or neither. RERI was 0.00 for boys and 0.18 for girls. We found a significant association between obesity and co-existence of hyperactivity and anxiety among elementary school girls. Our findings strengthen the need to further explore the association between childhood obesity, hyperactivity, and anxiety.

A CVD (Chemical Vapor Deposition) diamond coated tungsten carbide (WC) and cobalt (Co) sintered alloy punch was trimmed by the femtosecond laser machining to sharpen its edge with about 2 μm and to simultaneously make nanostructuring to its side surface. In addition to the sharpened edge, its edge profile was formed to be homogeneous enough to reduce the damage layer width by piercing the electrical amorphous steel sheet stack. Each brittle sheet in the stacked work was damaged to have three kinds of defects by piercing, e.g., the droop-like cracking in the thickness and at the vicinity of hole, the wrinkling in peak-to-valley with partial cracking on the peaks, and the circumferential cracking. When using the WC (Co) punch with the inhomogeneous edge profile in the sharpened edge width, these three damages were induced into each sheet and the maximum damage width exceeded 80 μm. When using the punch with the sharpened edge and homogeneous edge profile, the wrinkling mode was saved and the total affected layer width was significantly reduced to less than 20 μm. Through the precise embossing experiments, this effect of punch edge profile condition to the induced damages was discussed with a statement on the nanostructuring effect on the reduction of damaged width in electrical amorphous steel sheets. The developed tool with the sharpened edge and homogenous edge condition contributes to the realization of a low iron loss motor with a reduced affected layer width.

CVD-diamond coated special tools have been widely utilized to prolong their tool life in practical production lines. WC (Co) punch for fine piercing of metallic sheets required for high wear-toughness to be free from chipping and damages and for high product quality to punch out the holes with sufficient dimensional accuracy. The laser trimming process was developed to reduce the surface roughness of diamond coating down to submicron level and to adjust its diamond layer dimensions with a sharp punch edge for accurate piercing. The pulsed laser irradiation was employed to demonstrate that micro-groove was accurately formed into the diamond coating. Less deterioration in the worked diamond film by this laser treatment was proved by the Raman spectroscopy. The femtosecond laser trimming was proposed to sharpen the punch edge down to 2 μm and to form the nano-textured punch side surfaces with the LIPSS (Laser Induced Periodic Surface Structuring)-period of 300 nm. Fine piercing experiments were performed to demonstrate that punch life was significantly extended to continuous punching in more than 10,000 shots and that mirror-shining hole surfaces were attained in every shot by regularly coining the nanotextures. The sharp punch edge with homogeneous edge profile was responsible for reduction of the induced damages into work sheet by piercing. The punch life was extended by the ejection mechanism of debris particles through the nanotextures on the punch side surface. The present laser treatment was useful in trimming and nanostructuring the complex-shaped punch edge for industrial application.

With a rapidly aging society, there is increasing interest in the health of female workers in the field of care services for older adults due to increasing demands to maintain 24-h care and to support older adults without errors or accidents. Therefore, the purpose of this cross-sectional study was to examine the association between sleep-disordered breathing (SDB) and sustained attention in women caring for older adults in Japan.The study was conducted in women aged 18-67 years old working in care service facilities for older adults in Japan. The sustained attention of participants was measured by the 10-min psychomotor vigilance task (PVT). SDB was assessed based on the respiratory disturbance index (RDI), which was measured using an ambulatory airflow monitor with a polyvinylidene fluoride (PVDF) film sensor to monitor the respiratory airflow of nasal and oral breathing. The participants wore the monitor to record the breathing status while asleep at home. The severity of SDB was categorized as follows: normal, RDI 5 events/h; mild SDB, RDI 5-10 events/h; and moderate-to-severe SDB, RDI ≥ 10 events/h.Of 688 women enrolled, medians of age, body mass index (BMI), sleep duration, and prevalence of hypertension tended to be higher with increasing RDI. No significant association was found between RDI and PVT parameters. However, when we limited the analysis to women with BMI ≥ 22 kg/mOur findings suggest that SDB is associated with reduced sustained attention in participants with BMI ≥ 22 kg/m

BACKGROUND How the time sequence of cardiopulmonary resuscitation (CPR) procedures is related to clinical outcomes in patients with out-of-hospital cardiac arrest (OHCA) remains unclear. This study examined the impact of the time interval from collapse to start of CPR (no-flow time, NF time) and the time interval from start of CPR to implementation of extracorporeal CPR (ECPR) (low-flow time, LF time) on neurological outcomes.Methods and Results:During the period from 2010 to 2015, we enrolled 85 patients who received ECPR. Fourteen patients (16.5%) showed favorable 30-day neurological recovery. NF time was shorter in the favorable neurological recovery group than in the unfavorable recovery group (1.4±3.0 vs. 5.2±5.8 min, P

To understand the role of forest ecosystems in the global carbon cycle, it is important to clarify the factors affecting the carbon balance of forest ecosystems. However, little is known about the direct effect of forest types, especially dominant species, on their different carbon dynamics. To clarify the effect of difference in forest types, an experiment was conducted in three forests, which were located in the same place and exposed to the same climate conditions. These forests were middle-aged (40 - 45 years) and dominated by Quercus serrata (Q forest), Larix kaempferi (L forest) and Pinus densiflora (P forest). Net primary production (NPP), heterotrophic respiration (HR) and net ecosystem production (NEP) were estimated in each forest, using a biometric method over one year. For NPP estimated from the annual growth of tree biomass (ΔB) and amount of litter (LF), P forest NPP (5.3 MgC·ha-1·yr-1) was higher than Q and L forest NPP (4.6 and 3.2 MgC·ha-1·yr-1). The difference was affected by a significant difference in ΔB (p = 0.032) and LF (p -1·yr-1) was higher than L and P forest (2.3 and 2.1 MgC·ha-1·yr-1). This difference could result from the amount of litter (respiration substrate) and chemical properties of litter (lability of decomposition). The NEP, which was calculated from the difference between NPP and HR, varied widely among the forest types (0.5, 0.9 and 3.2 MgC·ha-1·yr-1 in Q, L and P forests, respectively). The range of values among the forest types was comparable to those among age sequences and climate zones in previous studies. These results suggest that the difference in forest types (especially dominant species) can potentially lead to a large variation in carbon dynamics, in ecosystems located in the same place.