Clozapine Induces Neuronal Activation in the Medial Prefrontal Cortex in a Projection Target-Biased Manner.

The medial prefrontal cortex (mPFC) is associated with various behavioral controls via diverse projections to cortical and subcortical areas of the brain. Dysfunctions and modulations of this circuitry are related to the pathophysiology of schizophrenia and its pharmacotherapy, respectively. Clozapine is an atypical antipsychotic drug used for treatment-resistant schizophrenia and is known to modulate neuronal activity in the mPFC. However, it remains unclear which prefrontal cortical projections are activated by clozapine among the various projection targets. To identify the anatomical characteristics of neurons activated by clozapine at the mesoscale level, we investigated the brain-wide projection patterns of neurons with clozapine-induced c-Fos expression in the mPFC. Using a whole-brain imaging and virus-mediated genetic tagging of activated neurons, we found that clozapine-responsive neurons in the mPFC had a wide range of projections to the mesolimbic, amygdala and thalamic areas, especially the mediodorsal thalamus. These results may provide key insights into the neuronal basis of the therapeutic action of clozapine.

A multi-institutional, observational study of outcomes after catheter placement for peritoneal dialysis in Japan.

BACKGROUND: This multi-institutional, observational study examined whether the outcomes after peritoneal dialysis (PD) catheter placement in Japan meet the audit criteria of the International Society for Peritoneal Dialysis (ISPD) guideline and identified factors affecting technique survival and perioperative complications. METHODS: Adult patients who underwent first PD catheter placement for end-stage kidney disease between April 2019 and March 2021 were followed until PD withdrawal, kidney transplantation, transfer to other facilities, death, 1 year after PD start or March 2022, whichever came first. Primary outcomes were time to catheter patency failure and technique failure, and perioperative infectious complications within 30 days of catheter placement. Secondary outcomes were perioperative complications. Appropriate statistical analyses were performed to identify factors associated with the outcomes of interest. RESULTS: Of the total 409 patients, 8 who underwent the embedded catheter technique did not have externalised catheters. Of the 401 remaining patients, catheter patency failure occurred in 25 (6.2%). Technical failure at 12 months after PD catheter placement calculated from cumulative incidence function was 15.3%. On Cox proportional hazards model analysis, serum albumin (hazard ratio (HR) 0.44; 95% confidence interval (CI) 0.27-0.70) and straight type catheter (HR 2.14; 95% CI 1.24-3.69) were the independent risk factors for technique failure. On logistic regression analysis, diabetes mellitus was the only independent risk factor for perioperative infectious complications (odds ratio 2.70, 95% CI 1.30-5.58). The occurrence rate of perioperative complications generally met the audit criteria of the ISPD guidelines. CONCLUSION: PD catheter placement in Japan was proven to be safe and appropriate.

Midterm outcomes after surgical aortic valve replacement with the INSPIRIS RESILIA from a multicenter AVR registry.

BACKGROUND: There have been few reports on the mid- to long-term safety and efficacy validation of the INSPIRIS RESILIA aortic bioprosthesis (Edwards Lifesciences LLC, Irvine, CA, USA) in Japan. Herein, we report the mid-term results of surgical aortic valve replacement (AVR) for aortic stenosis using INSPIRIS and evaluate the hemodynamics compared to the CEP Magna series from the multicenter AVR registry (the ACTIVIST registry). METHODS: Of the 1967 patients who underwent surgical or transcatheter AVR from the ACTIVIST registry, 66 patients who underwent isolated surgical AVR with INSPIRIS by December 2020 were included in this study, and the early and mid-term results were evaluated. Hemodynamics were evaluated by comparing 272 patients undergoing isolated surgical AVR with the Magna group using propensity score matching. RESULTS: The mean age was 74.0 ±â€¯7.8 years, and 48.5 % were women. In-hospital mortality was 1.5 %, and the survival rates at 1- and 2-years were 95.2 % and 95.2 %, respectively. After propensity score matching, echocardiographic findings at discharge demonstrated that peak velocity and mean pressure gradient in the INSPIRIS group were comparable, while the effective orifice area in the INSPIRIS group was significantly larger than those in the Magna group (p = 0.048). A patient-prosthesis mismatch at discharge was significantly lower in the INSPIRIS group (11.8 %) than in the Magna group (36.4 %) (p = 0.004). CONCLUSIONS: Surgical AVR with INSPIRIS was performed safely and the mid-term results were satisfactory. The hemodynamics of INSPIRIS were comparable to those of Magna.

Mechanism for exercise-mediated prevention against muscle wasting on extensor digitorum longus muscle in Spontaneously Diabetic Torii fatty rats.

We previously reported the significant increase in limb muscle strength and cross-sectional area of the type IIb muscle fibers in the extensor digitorum longus (EDL) muscle in a type 2 diabetic animal model, with Spontaneously Diabetic Torii (SDT) fatty rats (n = 6) undergoing regular treadmill exercise from 8 to 16 weeks of age compared with sedentary SDT fatty rats (n = 6). This study investigated the mechanism by which exercise training prevented skeletal muscle wasting in the EDL muscle of the SDT fatty rats. The endurance exercise for 8 weeks downregulated the expression of muscle RING-finger protein-1 (an E3 ubiquitin ligase) and upregulated the expression of CD31, insulin receptor substrate-2, and phosphorylated endothelial nitric oxide synthase in the EDL muscle of 16-week-old SDT fatty rats.Endurance exercise training might reduce muscle wasting by preventing muscle degradation and increasing the angiogenic response in the EDL muscle in type 2 diabetes.

A high level of vaccine knowledge increases people's behavioral risks for contracting COVID-19 in Japan.

BACKGROUND: There has been no study in which the association between levels of vaccine knowledge and preventive behaviors was examined during the COVID-19 pandemic. We examined the transition to risky (transmission) behavior according to level of vaccine knowledge over a seven-month period when vaccines became widely available in Japan. METHODS: A series of cross-sectional surveys were conducted using rapid online surveys of residents in Iwate Prefecture from December 4 to 7 in 2020 (the first survey) and from July 2 to 4 in 2021 (the fourth survey). We calculated each individual's risk of acquiring SARS-CoV-2 infection using a quantitative assessment tool (the microCOVID calculator). The respondents' level of knowledge regarding the COVID-19 vaccine was assessed by a questionnaire and was divided into four groups: very low level, low level, moderate level, and high level of vaccine knowledge. RESULTS: People with a high-level knowledge about the vaccine had significantly higher odds ratios (ORs) of transitioning to high-risk behavior compared to people with a low level of vaccine knowledge (OR [95% confidence interval (CI)], 1.50 [1.17-1.93]; P = 0.001). There was a dose-response association according to the four levels of vaccine knowledge, while engagement in preventive measures in the first survey was not associated with high-risk behavior in the follow-up survey. CONCLUSIONS: Since new variants of SARS-CoV-2 have evolved, policy makers should continue to communicate strong messages to keep a high level of consciousness and maintain basic preventive measures even after widespread vaccination.

Glucagon-like peptide-1 receptor agonist, liraglutide, attenuated retinal thickening in spontaneously diabetic Torii fatty rats.

BACKGROUND: This study aims to investigate the effect of the glucagon-like peptide-1 (GLP-1) receptor agonist (GLP-1RA) liraglutide on retinal pathological findings as compared with insulin and hydralazine using an animal model of type 2 diabetes with obesity, hypertension, and hyperlipidemia. METHODS: Male spontaneously diabetic Torii (SDT) fatty rats at 8 weeks of age were randomly assigned to three groups: the liraglutide group (SDT-lira, n = 6) received a subcutaneous injection of liraglutide from the age of 8 to 16 weeks, the SDT-ins-hyd group (n = 6) was provided both insulin against hyperglycemia and hydralazine against hypertension to match levels of both blood glucose and blood pressure to those of the liraglutide group, and the control group of SDT fatty rats (SDT-vehicle, n = 7) and a nondiabetic control group of Sprague-Dawley rats (SD, n = 7) were injected with vehicle only. Both eyeballs of all groups were collected at the age of 16 weeks. RESULTS: Retinal thickness, which was found in the SDT-vehicle group, was significantly prevented to similar levels in both the SDT-lira and SDT-ins-hyd groups. Immunohistological analysis revealed that GLP-1 receptor was not expressed in the retina of all rats. The ocular protein expression of monocyte chemoattractant protein-1, which causes a proinflammatory situation, was significantly upregulated in all SDT fatty rats as compared to SD rats, but the expression levels were similar between all SDT fatty rats. With regard to neovascularization in the eyes, there were no significant differences in protein expressions of vascular endothelial growth factor, CD31, or endothelial nitric oxide synthase in all rats. CONCLUSIONS: The present study indicates that liraglutide prevents retinal thickening, dependent on blood glucose and blood pressure levels in SDT fatty rats without ocular neovascularization. However, the effects did not improve the ocular proinflammatory state.

Increase in people's behavioural risks for contracting COVID-19 during the 2021 New Year holiday season: longitudinal survey of the general population in Japan.

OBJECTIVES: There has been no study in Japan on the predictors of risk for acquiring SARS-CoV-2 infection based on people's behaviour during the COVID-19 pandemic. The aim of this study was to document changes in risk behaviour during the New Year's holiday season in 2021 and to identify factors associated with high-risk behaviour for infection using a quantitative assessment tool. DESIGN: A longitudinal survey. SETTING: Multiphasic health check-ups for the general population in Iwate Prefecture. PARTICIPANTS: Serial cross-sectional data were obtained using rapid online surveys of residents in Iwate Prefecture from 4 to 7 December 2020 (baseline survey) and from 5 to 7 February 2021 (follow-up survey). The data in those two surveys were available for a total of 9741 participants. MAIN OUTCOME MEASURES: We estimated each individual's risk of acquiring SARS-CoV-2 infection based on the microCOVID calculator. We defined four trajectories of individual risk behaviours based on the probabilities of remaining at low risk, increasing to high risk, improving to low risk and persistence of high risk. RESULTS: Among people in the low-risk group in the first survey, 3.6% increased to high risk, while high risk persisted in 80.0% of people who were in the high-risk group at baseline. While healthcare workers were significantly more likely to be represented in both the increasing risk and persistently high-risk group, workers in the education setting were also associated with persistence of high risk (OR 2.58, 95% CI 1.52 to 4.39; p<0.001). CONCLUSIONS: In determining countermeasures against COVID-19 (as well as future outbreaks), health officials should take into account population changes in behaviour during large-scale public events.

Endurance Exercise Training-Attenuated Diabetic Kidney Disease with Muscle Weakness in Spontaneously Diabetic Torii Fatty Rats.

BACKGROUND: The aim of this study was to evaluate protective effects of endurance exercise training against diabetic kidney disease (DKD) with muscle weakness by using male spontaneously diabetic Torii (SDT) fatty rats as type 2 diabetic animal models with obesity, hypertension, and hyperlipidemia. METHODS: Eight-week-old SDT fatty rats (n = 12) and Sprague-Dawley (SD) rats (n = 10) were randomly divided into exercise (Ex; SDT-Ex: n = 6, SD-Ex: n = 5) and sedentary groups (SDT-Cont: n = 6, SD-Cont: n = 5), respectively. Each group underwent regular treadmill exercise 4 times a week from ages 8-16 weeks. RESULTS: The exercise attenuated hypertension and hyperlipidemia and prevented increases in renal parameter levels without affecting blood glucose levels. In the SDT fatty rats, it prevented induction of renal morphological abnormalities in the interstitium of the superficial and intermediate layers of the cortex. Downregulated expression of endothelial nitric oxide synthase in the glomerulus of the SDT fatty rats was significantly upregulated by the exercise. The exercise upregulated the renal expressions of both medium-chain acyl-CoA dehydrogenase and peroxisome proliferator-activated receptor γ coactivator-1α related to fatty acid metabolism. It increased muscle strength and both muscle weight and cross-sectional area of type IIb muscle fibers in the extensor digitorum longus muscle in the SDT fatty rats. CONCLUSION: Endurance exercise training in type 2 diabetes ameliorates DKD by improving endothelial abnormality and enhancing fatty acid metabolism in addition to attenuated hypertension, hyperlipidemia, and muscle weakness independently of blood glucose levels.

U-Net Convolutional Neural Network for Segmenting the Corneal Endothelium in a Mouse Model of Fuchs Endothelial Corneal Dystrophy.

PURPOSE: The purpose of this study was to assess the U-Net-based convolutional neural network performance for segmenting corneal endothelium and guttae of Fuchs endothelial corneal dystrophy. METHODS: Twenty-eight images of corneal endothelial cells and guttae of Col8a2L450W/L450W knock-in mice were obtained by specular microscopy. We used 20 images as training data to develop the U-Net for analyzing guttae and cell borders. The proposed network was validated using independent test data of 8 images. Cell density, hexagonality, and coefficient of variation were calculated from the predicted cell borders and compared with ground truth. RESULTS: U-Net allowed the prediction of cell borders and guttae, and overlays of those segmentations on specular microscopy images highly corresponded to ground truth. The average number of guttae per field was 6.25 ± 8.07 for ground truth and 6.25 ± 7.87 when predicted by the network (Pearson correlation coefficient 0.989, P = 3.25 × 10 -6 ). The guttae areas were 1.60% ± 1.79% by manual determination and 1.90% ± 2.02% determined by the network (Pearson correlation coefficient 0.970, P = 6.72 × 10 -5 ). Cell density, hexagonality, and coefficient of variation analyzed by the proposed network for cell borders showed very strong correlations with ground truth (Pearson correlation coefficient 0.989, P = 3.23 × 10 -6 , Pearson correlation coefficient 0.978, P = 2.66 × 10 -5 , and Pearson correlation coefficient 0.936, P = 6.20 × 10 -4 , respectively). CONCLUSIONS: We demonstrated proof of concept for application of U-Net for objective analysis of corneal endothelial cells and guttae in Fuchs endothelial corneal dystrophy, based on limited ground truth data.

Development of a CRISPR interference system for selective gene knockdown in Acidithiobacillus ferrooxidans.

Acidithiobacillus ferrooxidans is an iron-oxidizing chemolithotroph used for bioleaching of precious metals and is also regarded as a potential host for bioelectrochemical production of value-added chemicals. Despite its industrial utility, however, it is difficult to genetically engineer A. ferrooxidans due to low transformation and recombination efficiencies. Here, we developed a clustered regularly interspaced short palindromic repeats interference (CRISPRi) system that can selectively repress the expression of a target gene in A. ferrooxidans. The mutated gene encoding a nuclease-deactivated Cas9 protein was cloned into the broad-host-range plasmid pBBR1-MCS2, and the applicability of the CRISPRi system was examined using the nitrogenase nifH gene as a knockdown target. Introduction of the CRISPRi plasmid into A. ferrooxidans resulted in decreased nifH transcription and retarded cell growth in the absence of nitrogen sources, demonstrating that the CRISPRi system altered the phenotype of this bacterium via selective gene knockdown. We suggest that the CRISPRi system developed in this study provides an efficient technique for constructing A. ferrooxidans knockdown mutants that are useful for the genetic dissection of this bacterium.

Effect of GLP-1 receptor agonist, liraglutide, on muscle in spontaneously diabetic torii fatty rats.

This study investigated the effect of liraglutide, a glucagon-like peptide-1 receptor agonist, on skeletal muscles in rats with type 2 diabetes. Male SDT fatty rats (8-week-old) were provided liraglutide, or insulin-hydralazine for 8 weeks; control SDT fatty rats and SD rats were administered a vehicle. At 16 weeks of age, muscle strength of limbs was significantly lower in all SDT fatty rats compared to SD rats. While cross-sectional areas of type IIb muscle fibers in extensor digitorum longus muscle were significantly lower in SDT fatty rats than in SD rats, those of type I muscle fibers in soleus were similar in all rats. In the soleus of SDT fatty rats, liraglutide led to greater citrate synthase activity and cytochrome c oxidase subunit 5 B protein expression, independently of blood glucose and blood pressure levels. Liraglutide may contribute to preservation of mitochondrial content on soleus muscle in type 2 diabetes.

Renoprotective effect of GLP-1 receptor agonist, liraglutide, in early-phase diabetic kidney disease in spontaneously diabetic Torii fatty rats.

BACKGROUND: The aim of this study is to investigate the renoprotective effect of the GLP-1 receptor agonist, liraglutide, in early-phase diabetic kidney disease (DKD) using an animal model of type 2 diabetes with several metabolic disorders. METHODS: Male 8-week-old spontaneously diabetic Torii (SDT) fatty rats (n = 19) were randomly assigned to three groups. The liraglutide group (n = 6) was injected subcutaneously with liraglutide. Another treatment group (n = 6) received subcutaneous insulin against hyperglycemia and hydralazine against hypertension for matching blood glucose levels and blood pressure with the liraglutide group. The control groups of SDT fatty (n = 7) and non-diabetic Sprague-Dawley rats (n = 7) were injected only with a vehicle. RESULTS: The control group of SDT fatty rats exhibited hyperglycemia, obesity, hypertension, hyperlipidemia, glomerular sclerosis, and tubulointerstitial injury with high urinary albumin and L-FABP levels. Liraglutide treatment reduced body weight, food intake, blood glucose and blood pressure levels, as well as ameliorated renal pathologic findings with lower urinary albumin and L-FABP levels. Liraglutide increased expressions of phosphorylated (p)-eNOS and p-AMPK in glomeruli, downregulated renal expression of p-mTOR, and increased renal expressions of LC3B-II, suggesting activation of autophagy. However, these effects were not caused by the treatments with insulin and hydralazine, despite comparable levels of hyperglycemia and hypertension to those achieved with liraglutide treatment. CONCLUSIONS: Liraglutide may exert a renoprotective effect via prevention of glomerular endothelial abnormality and preservation of autophagy in early-phase DKD, independent of blood glucose, and blood pressure levels.

Towards Application of Electro-Fermentation for the Production of Value-Added Chemicals From Biomass Feedstocks.

According to recent social demands for sustainable developments, the value of biomass as feedstocks for chemical industry is increasing. With the aid of metabolic engineering and genome editing, microbial fermentation has been developed for producing value-added chemicals from biomass feedstocks, while further improvements are desired for producing more diverse chemicals and increasing the production efficiency. The major intrinsic limitation in conventional fermentation technologies is associated with the need for balancing the net redox equivalents between substrates and products, resulting in limited repertories of fermentation products. One solution for this limitation would be "electro-fermentation (EF)" that utilizes bioelectrochemical systems for modifying the intracellular redox state of electrochemically active bacteria, thereby overcoming the redox constraint of fermentation. Recent studies have attempted the production of chemicals based on the concept of EF, while its utility has not been sufficiently demonstrated in terms of low production efficiencies. Here we discuss EF in terms of its concept, current status and future directions, which help us develop its practical applications to sustainable chemical industries.

Seasonality of peritoneal dialysis-related peritonitis in Japan: a single-center, 10-year study.

BACKGROUND: Peritonitis is one of the major complications of peritoneal dialysis (PD). Although several reports have indicated seasonality of peritonitis, the observation periods were short, and there were no reports from Japan. Therefore, the purpose of this study was to investigate the long-term seasonality of peritonitis in a single institution in Japan. METHODS: Of 126 patients who started PD in our hospital between January 1, 2009, and December 31, 2018, 25 patients (15 men, 10 patients with diabetes) developed peritonitis with a total of 42 episodes. The median age at onset was 63 years, and the median duration from the start of PD to the onset of peritonitis was 22 months. RESULTS: The 10-year incidence of peritonitis was 0.12 episodes per patient-year. Compared with the reference season of winter (December-February), the incidence rate ratios (95% confidence interval) for spring (March-May), summer (June-August), and autumn (September-November) were 1.75 (0.65-4.75), 1.56 (0.57-4.31), and 2.42 (0.94-6.23), respectively. In addition, no seasonality of Gram-positive and Gram-negative organisms was observed. CONCLUSION: No seasonality was evident in the incidence of PD-related peritonitis in our hospital over a 10-year period. These findings suggest that the development of peritonitis in Japanese PD patients is not affected by seasonality.

Enhanced electricity generation in rice paddy-field microbial fuel cells supplemented with iron powders.

Microbial fuel cells installed in rice paddy fields (RP-MFCs) are able to serve as on-site batteries for operating low-power environmental sensors. In order to increase the utility and reliability of RP-MFCs, however, further research is necessary for boosting the power output. Here we examined several powdered iron species, including zero valent iron (ZVI), goethite, and magnetite, for their application to increasing power outputs from RP-MFCs. Soil around anodes was supplemented with either of these iron species, and RP-MFCs were operated for several months during the transplanting and harvesting. It was found that power outputs from RP-MFCs supplemented with ZVI were more than double the outputs from control (not supplemented with iron species) and other RP-MFCs, even after iron corrosion was ceased, and the maximum power density reached 130 mW/m2 (per projected area of the anode). Metabarcoding of 16S rRNA gene amplicons suggested that several taxa represented by fermentative and exoelectrogenic bacteria were substantially increased in MFCs supplemented with ZVI. Results suggest that ZVI lowers oxidation/reduction potential around anodes, activates anaerobic microbes involved in the conversion of organic matter into electricity and increases power output from RP-MFCs.