Accumulation of Phosphorylated SnRK2 Substrate 1 Promotes Drought Escape in Arabidopsis.

Plants adopt optimal tolerance strategies depending on the intensity and duration of stress. Retaining water is a priority under short-term drought conditions, whereas maintaining growth and reproduction processes takes precedence over survival under conditions of prolonged drought. However, the mechanism underlying changes in the stress response depending on the degree of drought is unclear. Here, we report that SNF1-related protein kinase 2 (SnRK2) substrate 1 (SNS1) is involved in this growth regulation under conditions of drought stress. SNS1 is phosphorylated and stabilized by SnRK2 protein kinases reflecting drought conditions. It contributes to the maintenance of growth and promotion of flowering as drought escape by repressing stress-responsive genes and inducing FLOWERING LOCUS T (FT) expression, respectively. SNS1 interacts with the histone methylation reader proteins MORF-related gene 1 (MRG1) and MRG2, and the SNS1-MRG1/2 module cooperatively regulates abscisic acid response. Taken together, these observations suggest that the phosphorylation and accumulation of SNS1 in plants reflect the intensity and duration of stress and can serve as a molecular scale for maintaining growth and adopting optimal drought tolerance strategies under stress conditions.

Progesterone increases the success of in vitro fertilization via enhanced sperm hyperactivation in mice.

Progesterone (P) enhances spermatozoal hyperactivation, a capacitation event. Hyperactivation is associated with successful in vitro fertilization (IVF). In this study, we examined the effects of P on hyperactivation and IVF in mice. P enhanced spermatozoal hyperactivation and increased IVF success rate in a dose-dependent manner. Moreover, P affected spermatozoal hyperactivation and IVF through the membrane progesterone receptor of the spermatozoal head. These results show that P regulates spermatozoal capacitation and fertilization in mice. The concentration of P changes during the estrous cycle, indicating that spermatozoa are capacitated in response to the oviductal environment and subsequently fertilize the oocyte.

Characteristics of gaze tracking during movement analysis by therapists.

[Purpose] Visual assessment of the quality of movement is a common and important component of physiotherapy. The purpose of this study is to quantify the level of proficiency of therapists and to obtain a new index of proficiency by measuring the coordinates of the gaze tracking trajectories of therapists with years of experience. [Participants and Methods] Eighteen voluntary physiotherapists (1st year (n=4), 7th year (n=1), 9th year (n=4), 10th year (n=3), 11th year (n=4), 13th year (n=1), and 21st year (n=1)) were recruited for this study. [Results] Discriminant analysis according to the size of the deviation between the X-axis and Y-axis of the range of gaze tracking during movement analysis measured from each therapist showed that the percentage of classification accuracy in the 10th year or less was 72.2%. Cluster analysis showed that two clusters were formed. Thirteen therapists in Cluster 2 were in their 9th year or more. Eye tracking trajectories can be classified by the 10th year of experience as a therapist. [Conclusion] It was shown that full-fledged therapists with 10 years of experience also expanded the range of eye tracking. The trajectory in the Y-axis direction tends to be extended with their 9th year or more of experience. In this point, quantitative judgments of eye-tracking results can serve as indicators of proficiency. The eye movements are important as a tool to objectively measure skills from experience.

Thrombin-deficient mutant of medaka, a model fish, displays serious retardation in blood coagulation.

At the last stage of the blood coagulation cascade, thrombin plays a central role in the processing of fibrinogen for the polymerization and in the additional activation of Factor XIII for the stable cross-linking of fibrin. In addition, thrombin carries out possible multiple roles via processing or interaction with various functional proteins. Several studies conducted in order to elucidate additional physiological significance are ongoing. To clarify further significance of thrombin and to establish an associated disease model, we characterized the orthologue gene for medaka (Oryzias latipes), a research model fish. Tissue distribution of medaka prothrombin has been immunotechnically analyzed. Furthermore, thrombin-deficient medaka mutants were viably established by utilizing a genome-editing method. The established gene-deficient mutants exhibited retarded blood coagulation even in the heterozygous fish. Taking advantage of their ease of handling, this specific model is useful for further investigation in medical research areas on human coagulation diseases.

Transglutaminase orthologues in medaka fish - biochemical characterization and establishment of gene-deficient mutants.

By genome analysis, seven homologous genes (orthologues) of human transglutaminases (TGases) have been identified in medaka fish (Oryzias latipes), some of which clearly corresponded to Factor XIII, TG1, and TG2. The enzymatically active-recombinant proteins for these medaka TGases have been successfully produced in bacteria or baculovirus-infected insect cell systems. Specific antibodies have been prepared and used in immunohistochemical analyses to reveal tissue distribution. Furthermore, gene-deficient medaka mutants for the genes encoding Factor XIII and TG1 have been established together with analysis of their phenotypes. Retarded cross-linking of fibrin and higher sensitivity to osmolality are observed when each gene is knocked-out. In this review, we summarize these biochemical features and the phenotypes of these gene-deficient fish.

Integrated volitional control electrical stimulation for the paretic hand: a case report.

[Purpose] Integrated volitional control electrical stimulation (IVES) is a type of electrical stimulation therapy that promotes agonist muscle contraction in limbs with motion paralysis. This case study describes the improvement in the paretic hand with stroke hemiplegia, eight years after the onset, with IVES for one month in the extrinsic and intrinsic muscles, including change of mode of stimulation based on the degree of improvement. [Participant and Methods] A 76 year-old male with hemiplegia for eight years. The patient was evaluated for two weeks and performed IVES in the right flexor pollicis brevis, abductor pollicis brevis, and extensor carpi ulnaris with the change of mode of IVES. [Results] The upper limb function improved in a short period of time. The hemiplegia test showed Brunnstrom stages II-III and II-IV for the right upper limb and right hand and fingers, respectively, 28 days after IVES initiation. [Conclusion] After one month of undergoing IVES, the patient showed improvement in hand and finger motor function, which was maintained even after IVES was completed. In this case, there was improvement with a short-term intervention using appropriately combined IVES modes.

Anti-HMGB1 Neutralizing Antibody Attenuates Periodontal Inflammation and Bone Resorption in a Murine Periodontitis Model.

High mobility group box 1 (HMGB1) is a non-histone DNA-binding protein that is secreted into the extracellular milieu in response to inflammatory stimuli. The secreted HMGB1 mediates various inflammatory diseases, including periodontitis; however, the underlying mechanisms of HMGB1-induced periodontal inflammation are not completely understood. Here, we examined whether anti-HMGB1 neutralizing antibody inhibits periodontal progression and investigated the molecular pathology of HMGB1 in vitro and in vivo. In vitro analysis indicated that HMGB1, granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-1ß (IL-1ß) were secreted in response to tumor necrosis factor-α (TNF-α) stimuli in human gingival epithelial cells (HGECs) and human monocytic leukemia cells (THP-1) treated with phorbol myristate acetate. Increased levels of GM-CSF and IL-1ß were observed in the conditioned media from TNF-α-stimulated HGECs and THP-1 in vitro Simultaneous stimulation with TNF-α and anti-HMGB1 antibody significantly decreased TNF-α-induced inflammatory cytokine secretion. Experimental periodontitis was induced in mice using Porphyromonas gingivalis-soaked ligatures. The extracellular translocation was confirmed in gingival epithelia in the periodontitis model mice by immunofluorescence analysis. Systemic administration of anti-HMGB1 neutralizing antibody significantly inhibited translocation of HMGB1. The anti-HMGB1 antibody inhibited periodontal inflammation, expression of IL-1ß and C-X-C motif chemokine ligand 1 (CXCL1), migration of neutrophils, and bone resorption, shown by bioluminescence imaging of myeloperoxidase activity, quantitative reverse transcription-PCR (RT-PCR), and micro-computed tomography analysis. These findings indicate that HMGB1 is secreted in response to inflammatory stimuli caused by periodontal infection, which is crucial for the initiation of periodontitis, and the anti-HMGB1 antibody attenuates the secretion of a series of inflammatory cytokines, consequently suppressing the progression of periodontitis.

Development of a low-cost EMG biofeedback device kit as an educational tool for physical therapy students.

[Purpose] Electromyography biofeedback therapy is applied to various diseases during physical therapy for motor learning. Our aim was to develop a low-cost electromyography biofeedback device kit that students could build by themselves in class and to evaluate whether this kit was an adequate educational tool for physical therapy students. [Subjects and Methods] This study included 34 volunteers from a 3 year physical therapy vocational school and a 4 year university course. All students were tasked with: 1) completing a pre-questionnaire; 2) attending basic lectures on electromyography biofeedback therapy and how to build the device; 3) building the device; and 4) completing a post-questionnaire. [Results] Mean time required to build the device was 50.8 ± 34.1 minutes for the vocational course students and 27.4 ± 8.8 minutes for university students. For both groups, there was a significant increase in the number of students stating "have an interest in," "be involved with," and "have knowledge of electromyography biofeedback" when pre- and post-questionnaires were compared. In the post questionnaire, the frequency of the "knowledge increased" response was higher. [Conclusion] Physical therapy students can easily build our new device in a classroom, and this kit is a useful educational tool to motivate learning in electromyography biofeedback.

Immobilization of a Bacterial Cytochrome P450 Monooxygenase System on a Solid Support.

Bacterial cytochrome P450s (P450s), which catalyze regio- and stereoselective oxidations of hydrocarbons with high turnover rates, are attractive biocatalysts for fine chemical production. Enzyme immobilization is needed for cost-effective industrial manufacturing. However, immobilization of P450s is difficult because electron-transfer proteins are involved in catalysis and anchoring these can prevent them from functioning as shuttle molecules for carrying electrons. We studied a heterotrimeric protein-mediated co-immobilization of a bacterial P450, and its electron-transfer protein and reductase. Fusion with subunits of a heterotrimeric Sulfolobus solfataricus proliferating cell nuclear antigen (PCNA) enabled immobilization of the three proteins on a solid support. The co-immobilized enzymes catalyzed monooxygenation because the electron-transfer protein fused to PCNA via a single peptide linker retained its electron-transport function.

Interprofessional Team Collaboration as a Mediator Between Workplace Social Capital and Patient Safety Climate: A Cross-Sectional Study.

BACKGROUND AND OBJECTIVES: Patient-safety climate is one of the most important organizational factors contributing to health care quality. We hypothesized that a patient safety climate is fostered by the willingness to collaborate and trust among members as well as by daily collaborative practices. This study aimed to clarify the effect of workplace social capital on patient safety climate. We also sought to investigate the mediating effect of interprofessional team collaboration on the relationship between workplace social capital and patient safety climate. METHODS: This cross-sectional survey was conducted from November 2021 to January 2022 using anonymous web-based questionnaires. The survey was distributed to 1495 employees working in a hospital in Tokyo, Japan. The questionnaire included the patient safety climate scale, workplace social capital scale, Japanese version of the Assessment of Interprofessional Team Collaboration Scale-II (AITCS-II-J), and demographic items. Structural equation modeling was performed to verify the associations among the 3 variables. In addition, a significance test for indirect effects was conducted using the bootstrap method to confirm the mediating effect of AITCS-II-J. RESULTS: A total of 725 employees participated in this survey, and 632 data items were analyzed. Nurses were the highest number of respondents (68.2%), followed by physicians (13.3%). Workplace social capital and patient safety were directly and significantly associated (ß = .309, P < .01). Furthermore, the partially indirect effect of the AITCS-II-J on the association between workplace social capital and patient safety climate was also significant (ß = .430, P < .01). CONCLUSIONS: Workplace social capital was significantly and directly related to patient safety climate and was also significantly related to patient safety climate partially mediated by interprofessional team collaboration. Our findings suggest the importance of workplace social capital and routine multidisciplinary collaboration for a patient safety climate to manage health care quality.

Characterizing medaka visual features using a high-throughput optomotor response assay.

Medaka fish (Oryzias latipes) is a powerful model to study genetics underlying the developmental and functional traits of the vertebrate visual system. We established a simple and high-throughput optomotor response (OMR) assay utilizing medaka larvae to study visual functions including visual acuity and contrast sensitivity. Our assay presents multiple adjustable stripes in motion to individual fish in a linear arena. For that the OMR assay employs a tablet display and the Fish Stripes software to adjust speed, width, color, and contrast of the stripes. Our results demonstrated that optomotor responses were robustly induced by black and white stripes presented from below in the linear-pool-arena. We detected robust strain specific differences in the OMR when comparing long established medaka inbred strains. We observed an interesting training effect upon the initial exposure of larvae to thick stripes, which allowed them to better respond to narrower stripes. The OMR setup and protocol presented here provide an efficient tool for quantitative phenotype mapping, addressing visual acuity, trainability of cortical neurons, color sensitivity, locomotor response, retinal regeneration and others. Our open-source setup presented here provides a crucial prerequisite for ultimately addressing the genetic basis of those processes.

Applicability of supercritical fluid chromatography for oligonucleotide analysis: A proof-of-concept study.

We evaluated the suitability of supercritical fluid chromatography (SFC) for oligonucleotide analysis using 4-mer oligonucleotides with various phosphorothioate (PS) contents as model compounds. Column screening showed that the diol-modified column was able to separate sequences with different PS contents. Optimization of the column body and additives allowed us to analyze polar oligonucleotides using SFC. Various sequences were also analyzed using the optimized method. A good peak shape was obtained when the guanine plus cytosine content of the analyte was two or less in the 4-mer oligonucleotides. Furthermore, we found that the retention times of the selected sequences were positively correlated with polar surface areas, indicating that oligonucleotides interact with polar stationary phases. In contrast, more hydrophobic full PS sequences were retained more strongly in the diol column than the full phosphodiester (PO) sequences. This suggests that the diol column has unique selectivity for PO and PS linkages. These results indicate that SFC is potentially applicable to oligonucleotide analysis with a separation mechanism that is different from that of ion-pair reversed-phase liquid chromatography.

Circadian Rhythm of Subspecies of Low-Density Lipoprotein-Cholesterol and High-Density Lipoprotein-Cholesterol in Healthy Subjects and Patients with Type 2 Diabetes.

AIMS: We established automated assay kits for quantifying small dense low-density lipoprotein (sdLDL)-cholesterol (C), LDL-triglyceride (TG), and high-density lipoprotein (HDL)3-C, and apolipoprotein (apo)E-rich HDL-C, and these have been recognized as sensitive biomarkers for predicting coronary artery disease. We investigated the circadian rhythms of these novel lipids to determine if fasting is required to determine basal levels. METHODS: Forty-eight inpatients with type 2 diabetes and 19 healthy volunteers were studied. Blood samples were collected at seven time points, which were obtained after an overnight fast, before and 2 h after each meal, and before the next breakfast. sdLDL-C, LDL-TG, remnant-like particle (RLP)-C, TG-rich lipoprotein (TRL-C), HDL3-C, and apoE-rich HDL-C were measured by the homogeneous methods. NonHDL-C, large buoyant (lb)LDL-C and HDL2-C were calculated by subtracting sdLDL-C from LDL-C or HDL3-C from HDL-C, respectively. RESULTS: Serum TG levels were significantly increased after meals in both healthy participants and patients with diabetes. RLP-C and TRL-C were also increased postprandially. LDL-TG, LDL-C, nonHDL-C, HDL2,3-C, and apoE-rich HDL-C did not exhibit significant fluctuation during the day in healthy participants and patients with diabetes. sdLDL-C was slightly increased postprandially in subjects with diabetes (1-2 mg/dl, 3%-9%), though its increase was not significant compared to the baseline (fasting) level. Significant postprandial reduction was observed with LDL-C and lbLDL-C. There was no influence of statin therapy or oral anti-diabetes drugs on the circadian rhythm of LDL-C subspecies. CONCLUSIONS: Subtle postprandial increase in sdLDL-C is considered a negligible level in general clinical practice. Fasting is not mandatory to measure basal concentrations of LDL and HDL subspecies.

The early neutrophil-committed progenitors aberrantly differentiate into immunoregulatory monocytes during emergency myelopoiesis.

Inflammatory stimuli cause a state of emergency myelopoiesis leading to neutrophil-like monocyte expansion. However, their function, the committed precursors, or growth factors remain elusive. In this study we find that Ym1+Ly6Chi monocytes, an immunoregulatory entity of neutrophil-like monocytes, arise from progenitors of neutrophil 1 (proNeu1). Granulocyte-colony stimulating factor (G-CSF) favors the production of neutrophil-like monocytes through previously unknown CD81+CX3CR1lo monocyte precursors. GFI1 promotes the differentiation of proNeu2 from proNeu1 at the cost of producing neutrophil-like monocytes. The human counterpart of neutrophil-like monocytes that also expands in response to G-CSF is found in CD14+CD16- monocyte fraction. The human neutrophil-like monocytes are discriminated from CD14+CD16- classical monocytes by CXCR1 expression and the capacity to suppress T cell proliferation. Collectively, our findings suggest that the aberrant expansion of neutrophil-like monocytes under inflammatory conditions is a process conserved between mouse and human, which may be beneficial for the resolution of inflammation.

Involvement of cholesterol membrane transporter Niemann-Pick C1-like 1 in the intestinal absorption of lutein.

PURPOSE: Lutein is a carotenoid mainly found in green leafy vegetables and is located in the macula lutea in the human eye. Since humans cannot synthesize lutein de novo, it must be digested as food. The physiological importance of an orally administered compound depends on its interaction with target tissues. It is therefore important to clarify the absorption mechanism in the intestine. Cholesterol membrane transporters Niemann-Pick C1 Like 1 (NPC1L1) and scavenger receptor class B type 1 (SR-B1) are involved in the intestinal absorption of highly lipophilic compounds including cholesterol. Ezetimibe, a selective inhibitor of intestinal NPC1L1, is the widespread lipid-lowering agent. It is important to investigate the possibility of food-drug interactions in order to prevent undesirable and harmful clinical consequences. The aim of this work was to determine whether NPC1L1, SR-B1 and other transporters are involved in absorption of lutein. METHODS: Caco-2 cells were used for accumulation and permeability study of lutein. Lutein concentration was determined by an HPLC system. The cDNA of transporters was isolated from total RNA of Caco-2 cells, and the expression of these transporters was confirmed by RT-PCR (reverse transcription-polymerase chain reaction). RESULTS: Ezetimibe inhibited up to 40% of lutein accumulation by Caco-2 cell monolayers. Block lipid transport 1 (BLT-1), a selective chemical inhibitor of SR-B1, also inhibited lutein accumulation by Caco-2 cells. On the other hand, ATP-depletion reagents (sodium fluoride and sodium azide or carbonyl cyanide m-chlorophenylhydrazone) did not influence the accumulation or permeation of lutein significantly. CONCLUSIONS: The results show that lutein absorption is, at least in part, mediated by influx transporters NPC1L1 and SR-B1 rather than mediated by efflux transporters such as ABC (ATP-binding cassette) transporters.

Association of body mass index with Clostridioides difficile infection among older patients with pneumonia in Japan.

AIM: Obesity is reported to be a risk factor for Clostridioides difficile infection. However, obesity rarely occurs in older Asian patients, and the effects of obesity on health and disease are different in Asian and Western countries. This study aimed to assess the association between body mass index and C. difficile infection risk among older patients with pneumonia in Japan. METHODS: This retrospective observational cohort study used data from the nationwide database of acute hospital inpatients' data in Japan between July 2014 and March 2016. All patients aged ≥65 years admitted with a primary diagnosis of pneumonia were enrolled. Risk factors for C. difficile infection were determined by logistic regression analysis, including known risks as covariates. RESULTS: Among 221 242 pneumonia patients, 611 developed C. difficile infection. Underweight patients (body mass index <18.5 kg/m2 ) showed higher odds for C. difficile infection (odds ratio 1.38, 95% confidence interval 1.17-1.62, P < 0.001) than normal weight patients (body mass index 18.5-24.9 kg/m2 ), whereas overweight patients (body mass index ≥25 kg/m2 ) showed lower odds (odds ratio 0.63, 95% confidence interval 0.45-0.89, P < 0.01). CONCLUSIONS: Body mass index was associated with C. difficile infection in older pneumonia patients in Japan. Underweight was a risk factor, whereas overweight was a protective factor for C. difficile infection. Geriatr Gerontol Int 2022; 22: 63-67.

IgG4-Related Hypertrophic Pachymeningitis with Skull Base Involvement Presenting with Isolated Glossopharyngeal and Vagus Nerve Palsy.

We herein report a 70-year-old man diagnosed with IgG4-related hypertrophic pachymeningitis with skull base involvement, who presented with isolated glossopharyngeal and vagus nerve palsy. Contrast-enhanced magnetic resonance imaging (MRI) showed enhanced dural thickening of the posterior clivus and skull base involvement. When a patient with hypertrophic pachymeningitis presents with isolated cranial neuropathy without systemic manifestations or definite MRI abnormalities, it is difficult to make a diagnosis, and the patient may be misdiagnosed. This case suggests that a detailed radiological evaluation including contrast enhancement of the skull base is very important in patients with isolated glossopharyngeal and vagus nerve palsy.

The Medaka Inbred Kiyosu-Karlsruhe (MIKK) panel.

BACKGROUND: Unraveling the relationship between genetic variation and phenotypic traits remains a fundamental challenge in biology. Mapping variants underlying complex traits while controlling for confounding environmental factors is often problematic. To address this, we establish a vertebrate genetic resource specifically to allow for robust genotype-to-phenotype investigations. The teleost medaka (Oryzias latipes) is an established genetic model system with a long history of genetic research and a high tolerance to inbreeding from the wild. RESULTS: Here we present the Medaka Inbred Kiyosu-Karlsruhe (MIKK) panel: the first near-isogenic panel of 80 inbred lines in a vertebrate model derived from a wild founder population. Inbred lines provide fixed genomes that are a prerequisite for the replication of studies, studies which vary both the genetics and environment in a controlled manner, and functional testing. The MIKK panel will therefore enable phenotype-to-genotype association studies of complex genetic traits while allowing for careful control of interacting factors, with numerous applications in genetic research, human health, drug development, and fundamental biology. CONCLUSIONS: Here we present a detailed characterization of the genetic variation across the MIKK panel, which provides a rich and unique genetic resource to the community by enabling large-scale experiments for mapping complex traits.

Genomic variations and epigenomic landscape of the Medaka Inbred Kiyosu-Karlsruhe (MIKK) panel.

BACKGROUND: The teleost medaka (Oryzias latipes) is a well-established vertebrate model system, with a long history of genetic research, and multiple high-quality reference genomes available for several inbred strains. Medaka has a high tolerance to inbreeding from the wild, thus allowing one to establish inbred lines from wild founder individuals. RESULTS: We exploit this feature to create an inbred panel resource: the Medaka Inbred Kiyosu-Karlsruhe (MIKK) panel. This panel of 80 near-isogenic inbred lines contains a large amount of genetic variation inherited from the original wild population. We use Oxford Nanopore Technologies (ONT) long read data to further investigate the genomic and epigenomic landscapes of a subset of the MIKK panel. Nanopore sequencing allows us to identify a large variety of high-quality structural variants, and we present results and methods using a pan-genome graph representation of 12 individual medaka lines. This graph-based reference MIKK panel genome reveals novel differences between the MIKK panel lines and standard linear reference genomes. We find additional MIKK panel-specific genomic content that would be missing from linear reference alignment approaches. We are also able to identify and quantify the presence of repeat elements in each of the lines. Finally, we investigate line-specific CpG methylation and performed differential DNA methylation analysis across these 12 lines. CONCLUSIONS: We present a detailed analysis of the MIKK panel genomes using long and short read sequence technologies, creating a MIKK panel-specific pan genome reference dataset allowing for investigation of novel variation types that would be elusive using standard approaches.