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BACKGROUND The 30-item Questionnaire for Triage and Assessment (QTA30) is a standardized triage and assessment tool for assessing pediatric psychosomatic disorders. It is estimated that one in 10 children in Japan experience difficulties in their school life. Using the QTA30 we evaluated mental health in children at an outpatient clinic in a local hospital. METHODS All elementary and junior high school students (≥9 years) who visited our institution between December 1 2019 and March 31 2020 were asked to complete the QTA30. RESULT A total of 372 children responded. Half of the children with a psychosomatic disorder and 9% of children with other chronic disorders were suspected to have poor mental health. Suspected poor mental health was associated with higher odds of female gender (odds ratio [OR]: 1.89, 95% confidence interval [CI]: 1.07-3.39), junior high school students (OR: 3.73, 95% CI: 2.11-6.73), and not enjoying exercise (OR: 2.13, 95% CI: 1.16-3.9). The mean ± standard deviation total QTA30 score was significantly worse in children with psychosomatic disorders (38.0 ± 19.1) among children with other chronic diseases; however, only the score in children with central precocious puberty (27.4 ± 13.7) showed no difference. CONCLUSION Based on our survey, the percentage of children suspected to have mental health problems manifesting as non-psychosomatic chronic disorders was similar to the proportion of children suspected to experience difficulties with their school life. Pediatricians should carefully consider the possibility of mental health problems when children are seen in regular visits to the outpatient clinic.

Background High-dose intravenous immunoglobulin (IVIG) is the mainstay of treatment for Kawasaki disease (KD). Usually, 2 g/kg of IVIG is administered over 10–24 h, depending on the institution or physician, but the association between infusion speed and effectiveness has not been reported. In this study, we evaluated the differences in efficacy and safety between two different IVIG administration speeds. Methods This was a multicenter, unblinded, randomized controlled study. Patients newly diagnosed with KD were randomized into two groups: one who received IVIG over 12 h (12H group, double speed), and one that received IVIG over 24 h (24H group, reference speed). The endpoints included the duration of fever, incidence of coronary artery abnormalities (CAAs) and of adverse events. Laboratory data were evaluated before and after IVIG administration. Results A total of 39 patients were enrolled. There was no difference between groups in fever duration after the initiation of IVIG (21 h vs. 21.5 h, p = 0.325), and no patient experienced CAAs. Two adverse events were observed in the 12H group (elevation of aspartate aminotransferase and vomiting), however no severe adverse events requiring treatments or extension of hospital stay were observed in either group. After initial IVIG administration, the change ratio of inflammatory markers, such as white blood cell counts, neutrophils, C-reactive protein, and albumin, did not show significant differences between the two groups. On the other hand, a greater increase of serum immunoglobulin G from its baseline level was observed in the 24H group compared to the 12H group (3037 ± 648 mg/dl vs. 2414 ± 248 mg/dl, p Conclusion The efficacy and safety of IVIG administered over 12 h (double speed) were similar to those administered over 24 h (reference speed). Trial registration University Hospital Medical Information Network (UMIN000014665). Registered 27 July 2014 – Prospectively registered, https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000017058

Assessing phenotypic changes in both cancer cells and surrounding cells, which construct the tumour microenvironment, is essential for understanding the role of bi-directional communication among cells in the tumorigenic process. Here, a 3D in vitro cancer-stroma co-culture system, a co-culture disc, was reported for the spatiotemporal image analysis of cancer-stromal cell interactions. Due to their centre-open disc structure, the lung cancer A549 spheroids could be co-cultured with a high concentration of fibroblasts, without gel shrinkage in the long term (>1 month). In the co-culture disc, some populations of applied normal human lung fibroblasts showed morphological and phenotypic changes into activated myofibroblasts (AMFs) with high expression of myo-fibrotic α-smooth muscle actin fibre in the cell, which is a well-known feature of cancer-associated fibroblasts. The AMFs appeared heterogeneously at the boundary of cancer spheroids, which could not be detected by standard mass analysis using a quantitative RT-qPCR system, and they led to A549 cancer cell migration. In addition, the effects of oncogenic or medicinal additives were quantitatively assessed by combining co-culture discs with image analysis. This system provides a new potential technique to analyse the complicated crosstalk among cancer tissue-constructing cells during the tumorigenic process and provides insight into applications for the quantitative evaluation of substances inducing tumorigenesis as well as drugs to prevent and inhibit cancer progression.

Rehabilitation is an important treatment for spinocerebellar ataxia (SCA). The lack of improvement in ataxia, deficit of motor learning, and unstable balance causes disability for activities of daily living and restricts participation in social activities, further resulting in a disturbance of the restoration of quality of life. This narrative review describes physical rehabilitation, including measurement of movement disorder, associated with ataxia and possible interventions. Several lines of evidence suggest that high-intensity individualized physical rehabilitation programs, especially for gait and balance training, improve motor function. Continuous exercise at home contributes to the maintenance of the gait and balance function. Moreover, videography and mechanical technology contribute to the evaluation of ataxia and motor learning ability, and assistive robotic systems may improve gait stability. Neuromodulation montages, such as repetitive transcranial magnetic stimulation and transcranial electrical stimulation, can enhance the effect of physical rehabilitation. Further research aimed at developing a more-effective physical rehabilitation for these patients is expected.

CpG methylation of genomic DNA is a well-known repressive epigenetic marker in eukaryotic transcription, and DNA methylation of promoter regions is correlated with gene silencing. In contrast to the promoter regions, the function of DNA methylation during transcription termination remains to be elucidated. A recent study revealed that mouse DNA methyltransferase 3a (Dnmt3a) mainly functions in de novo methylation in the promoter and gene body regions, including transcription termination sites (TTSs), during development. To investigate the relationship between DNA methylation overlapping the TTSs and transcription termination, we performed bioinformatics analysis using six pre-existing Dnmt

Since the current melanoma clinicopathological staging system remains restricted to predicting survival outcomes, establishing precise prognostic targets is needed. Here, we used gene expression signature (GES) classification and Cox regression analyses to biologically characterize melanoma cells at the single-cell level and construct a prognosis-related gene signature for melanoma. By analyzing publicly available scRNA-seq data, we identified six distinct GESs (named: “Anti-apoptosis”, “Immune cell interactions”, “Melanogenesis”, “Ribosomal biogenesis”, “Extracellular structure organization”, and “Epithelial-Mesenchymal Transition (EMT)”). We verified these GESs in the bulk RNA-seq data of patients with skin cutaneous melanoma (SKCM) from The Cancer Genome Atlas (TCGA). Four GESs (“Immune cell interactions”, “Melanogenesis”, “Ribosomal biogenesis”, and “Extracellular structure organization”) were significantly correlated with prognosis (p = 1.08 × 10−5, p = 0.042, p = 0.001, and p = 0.031, respectively). We identified a prognostic signature of melanoma composed of 45 genes (MPS_45). MPS_45 was validated in TCGA-SKCM (HR = 1.82, p = 9.08 × 10−6) and three other melanoma datasets (GSE65904: HR = 1.73, p = 0.006; GSE19234: HR = 3.83, p = 0.002; and GSE53118: HR = 1.85, p = 0.037). MPS_45 was independently associated with survival (p = 0.002) and was proved to have a high potential for predicting prognosis in melanoma patients.

Aspergillus section Versicolores species, except Aspergillus sydowii, produce a carcinogenic mycotoxin sterigmatocystin (STC). Since these fungi are found in varied environmental milieu including indoor dust and food products, our aim was to develop a sensitive and convenient assay to detect STC producing fungal strains. We made use of a high discrimination DNA polymerase (HiDi DNA polymerase), for single nucleotide polymorphism (SNP)-based PCR amplification. Using specific primer pairs based on the SNPs between A. sydowii and other strains of Aspergillus section Versicolores, we succeeded in amplifying the genomic DNA all target strains except A. sydowii. These results confirm that the SNP-based PCR amplification technique, using a high discrimination DNA polymerase, was a reliable and robust screening method for target fungal strains.

【Objective】 To predict the recurrence of advanced cervical cancer patients treated with radiotherapy from radiomics features on pre-treatment T1- and T2-weighted MRI images. 【Methods】 A total of 90 patients were split into two sets: 67 patients for model training and 23 patients for model testing. The patient outcome was classified into two groups; patients with a recurrence (group I) and without a recurrence (group II). The predictors were selected using the least absolute shrinkage and selection operator (LASSO) regression. The machine learning for the predictive models was sued neural network classifiers. The accuracy, sensitivity, specificity, and the area under the curve (AUC) from the receiver operating characteristic were evaluated. 【Results】 By the LASSO analysis, we found 25 radiomics features from the T1-weighted MRI image and 4 radiomics features from the T2-weighted MRI image. The accuracy of the prediction model was highest with the combination of T1- and T2-weighted MRI images. The model performances with T1-weighted MRI image and T2-weighted MRI image were 86.4% and 89.4% of accuracy, 74.9% and 38.1% of sensitivity, 81.8% and 72.2% of specificity, and 0.89 and 0.69 of AUC. The model performance was improved with the combination of T1- and T2-weighted MRI images, which was 93.1% of accuracy, 81.6% of sensitivity, 88.7% of specificity, and 0.94 of AUC. 【Conclusions】 The radiomics analysis with T1- and T2-weighted MRI images could highly predict the recurrence of the cervix cancer after radiotherapy. The variation of the distribution and the difference of the pixel number at the peripheral and the center were important predictors.

CpG methylation in genomic DNA is well known as a repressive epigenetic marker in eukaryotic transcription, and DNA methylation of the promoter regions is correlated with silencing of gene expression. In contrast to the promoter regions, the function of DNA methylation during transcription termination remains to be elucidated. A recent study has revealed that mouse DNA methyltransferase 3a (Dnmt3a) mainly functions inde novomethylation in the promoter and gene body regions (including transcription termination sites (TTSs)) during development. To investigate the relationship between DNA methylation overlapping the TTSs and transcription termination, we employed two strategies: informatic analysis using already deposited datasets ofDnmt3a-/-mouse cells and the zebrafish model system. Bioinformatic analysis using methylome and transcriptome data showed that hypomethylated differentially methylated regions overlapping the TTSs were associated with increased read counts and chimeric transcripts downstream of TTSs inDnmt3a-/-Agouti-related protein neurons, but not inDnmt3a-/-embryonic stem cells and mouse embryonic fibroblasts. We experimentally detected increased read-through and chimeric transcripts downstream of hypomethylated TTSs in zebrafish maternal-zygoticdnmt3aa-/-mutant. This study is the first to identify transcription termination defects in DNA hypomethylated TTSs inDnmt3a-/-vertebrates.

The discovery of anticancer therapeutics effective in eliminating dormant cells is a significant challenge in cancer biology. Here, we describe new synthetic polymer-based anticancer agents that mimic the mode of action of anticancer peptides. These anticancer polymers developed here are designed to capture the cationic, amphiphilic traits of anticancer peptides. The anticancer polymers are designed to target anionic lipids exposed on the cancer cell surfaces and act by disrupting the cancer cell membranes. Because the polymer mechanism is not dependent on cell proliferation, we hypothesized that the polymers were active against dormant cancer cells. The polymers exhibited cytotoxicity to proliferating prostate cancer. Importantly, the polymer killed dormant prostate cancer cells that were resistant to docetaxel. This study demonstrates a new approach to discover novel anticancer therapeutics.

Breast cancer is the most common cancer affecting women worldwide. Although many analyses and treatments have traditionally targeted the breast cancer cells themselves, recent studies have focused on investigating entire cancer tissues, including breast cancer cells. To understand the structure of breast cancer tissues, including breast cancer cells, it is necessary to investigate the three-dimensional location of the cells and/or proteins comprising the tissues and to clarify the relationship between the three-dimensional structure and malignant transformation or metastasis of breast cancers. In this review, we aim to summarize the methods for analyzing the three-dimensional structure of breast cancer tissue, paying particular attention to the recent technological advances in the combination of the tissue-clearing method and optical three-dimensional imaging. We also aimed to identify the latest methods for exploring the relationship between the three-dimensional cell arrangement in breast cancer tissues and the gene expression of each cell. Finally, we aimed to describe the three-dimensional imaging features of breast cancer tissues using noninvasive photoacoustic imaging methods.

Assessing phenotypic changes in both cancer cells and surrounding cells, which construct the tumour microenvironment, is essential for understanding the role of bi-directional communication among cells in the tumorigenic process. Here, a 3D in vitro cancer-stroma co-culture system, a co-culture disc, was reported for the spatiotemporal image analysis of cancer-stromal cell interactions. Due to their centre-open disc structure, the lung cancer A549 spheroids could be co-cultured with a high concentration of fibroblasts, without gel shrinkage in the long term (1 month). In the co-culture disc, some populations of applied normal human lung fibroblasts showed morphological and phenotypic changes into activated myofibroblasts (AMFs) with high expression of myo-fibrotic α-smooth muscle actin fibre in the cell, which is a well-known feature of cancer-associated fibroblasts. The AMFs appeared heterogeneously at the boundary of cancer spheroids, which could not be detected by standard mass analysis using a quantitative RT-qPCR system, and they led to A549 cancer cell migration. In addition, the effects of oncogenic or medicinal additives were quantitatively assessed by combining co-culture discs with image analysis. This system provides a new potential technique to analyse the complicated crosstalk among cancer tissue-constructing cells during the tumorigenic process and provides insight into applications for the quantitative evaluation of substances inducing tumorigenesis as well as drugs to prevent and inhibit cancer progression.

Background Changes in relationships, sleep rhythms, and physical activity caused by school closures instituted to curb the spread of COVID‐19 influenced children’s mental health. We explored changes in children’s daily life and effects on their mental health during school closures. Methods Participants included elementary and junior high school students 9 years of age and older seen in the outpatient clinic during school closures and were required to complete the Japanese version of WHO Five Well‐Being Index (WHO‐5‐J). The results were compared with those of students seen after schools reopened. Results Participants included 78 students in the school closure group and 113 in the school reopening group. Although those in the closure group devoted more time to family and sleep, their sleep rhythms, eating habits, and physical activities were disrupted. Although there were no significant differences between the two groups in total WHO‐5‐J scores, single WHO‐5‐J items such as activity and vigor and interest were significantly worse and rest was significantly better in the school closure group. Conclusion Although school closures resulted in elementary and junior high school students spending more time with family and sleeping, their sleep rhythms, eating habits, and physical activities were disrupted. As the children’s living environment changed, they felt less active and vigorous and had difficulty finding things that interested them. However, their sleep improved and overall, the number of children with potential mental health problems did not change.

BACKGROUND Transcranial magnetic stimulation (TMS), when applied over the primary motor cortex, elicits a motor-evoked potential (MEP) in electromyograms measured from peripheral muscles. MEP amplitude has often been observed to fluctuate trial to trial, even with a constant stimulus. Many factors cause MEP fluctuations in TMS. One of the primary factors is the weak stationarity and instability of cortical activity in the brain, from which we assumed MEP fluctuations originate. We hypothesized that MEP fluctuations are suppressed when TMS is delivered to the primary motor cortex at a time when several electroencephalogram (EEG) channels measured on the scalp are highly similar in the frequency domain. OBJECTIVE We developed a TMS triggering system to suppress MEP fluctuations using EEG coherence analysis, which was performed to detect the EEG signal similarity between the 2 channels in the frequency domain. METHODS Seven healthy adults participated in the experiment to confirm whether the TMS trigger system works adequately, and the mean amplitude and coefficient of the MEP variation were recorded and compared with the values obtained during the control task. We also determined the experimental time under each condition and verified whether it was within the predicted time. RESULTS The coefficient of variation of MEP amplitude decreased in 5 of the 7 participants, and significant differences (P=.02) were confirmed in 2 of the participants according to an F test. The coefficient of variation of the experimental time required for each stimulus after threshold modification was less than that without threshold modification, and a significant difference (PF test. CONCLUSIONS We found that MEP could be suppressed using the system developed in this study and that the TMS trigger system could also stabilize the experimental time by changing the triggering threshold automatically.

This study evaluated the performance of a new O3 /H2 O2 mixed gas sterilization instrument for killing microorganisms and inactivating bacterial endotoxin at low temperatures. Sterility assurance level was achieved by an over 6-log reduction of Geobacillus stearothermophilus ATCC 12980, and the decimal reduction value was 0.77 min in sterilization mode. A reduction of over 3 logs in Limulus amebocyte lysate coagulation activity of purified endotoxin from Escherichia coli was observed after treatment in endotoxin-inactivation mode. The same inactivation ability was observed when treating dried bacterial cells. Biomaterials made of polymer or metal did not exhibit cytotoxicity after gas exposure at O3 concentrations below 200 ppm. As the results of human cell-based pyrogen testing, significant amounts of endotoxin that were over the limit for medical devices contacting cerebrospinal fluid (2.15 EU/device) were detected on scissors washed with a washer-disinfector and sterilized with ethylene oxide or autoclaving. In contrast, endotoxin decreased to 0.29 ± 0.05 EU/device after O3 /H2 O2 mixed gas sterilization in endotoxin-inactivation mode. Compared to conventional gas sterilization methods, O3 /H2 O2 mixed gas has high sterilization ability and a strong capacity to inactivate endotoxin. It is expected that this sterilization technology will improve the safety of reusable medical devices and utensils for regenerative medicine.

Prions are infectious agents causing prion diseases, which include Creutzfeldt-Jacob disease (CJD) in humans. Several cases have been reported to be transmitted through medical instruments that were used for preclinical CJD patients, raising public health concerns on iatrogenic transmissions of the disease. Since preclinical CJD patients are currently difficult to identify, medical instruments need to be adequately sterilized not to transmit the disease. In this study, we investigated the sterilizing activity of two oxidizing gases, ozone gas and hydrogen peroxide gas, against prions fixed on stainless steel wires using a mouse bioassay. Mice intracerebrally implanted with prion-contaminated stainless steel wires treated with ozone gas or hydrogen peroxide gas developed prion disease later than those implanted with control prion-contaminated stainless steel wires, indicating that ozone gas and hydrogen peroxide gas could reduce prion infectivity on wires. Incubation times were further elongated in mice implanted with prion-contaminated stainless steel wires treated with ozone and hydrogen peroxide-mixed gas, indicating that ozone-mixed hydrogen peroxide gas inactivates prions on these wires more potently than ozone gas or hydrogen peroxide gas. Taken together, these results suggest that ozone-mixed hydrogen peroxide gas may be useful for prion sterilization of medical instruments.

This study aimed to investigate whether cerebellar transcranial magnetic stimulation (C-TMS) affected the cortical silent period (cSP) induced by TMS over the primary motor cortex (M1) and the effect of interstimulus interval (ISI) on cerebellar conditioning and TMS to the left M1 (M1-TMS). Fourteen healthy adult participants were instructed to control the abduction force of the right index finger to 20% of the maximum voluntary contraction. M1-TMS was delivered during this to induce cSP on electromyograph of the right first dorsal interosseous muscle. TMS over the right cerebellum (C-TMS) was conducted prior to M1-TMS. In the first experiment, M1-TMS intensity was set to 1 or 1.3 &times, resting motor threshold (rMT) with 20-ms ISI. In the second experiment, the intensity was set to 1 &times, rMT with ISI of 0, 10, 20, 30, 40, 50, 60, 70, or 80 ms, and no-C-TMS trials were inserted. In results, cSP was significantly shorter in 1 &times, rMT condition than in 1.3 &times, rMT by C-TMS, and cSP was significantly shorter for ISI of 20&ndash, 40 ms than for the no-C-TMS condition. Further, motor evoked potential for ISI40-60 ms were significantly reduced than that for ISI0. Thus, C-TMS may reduce cSP induced by M1-TMS with ISI of 20&ndash, 40 ms.

Objective To verify the effects of structured home-based exercises without supervision by a physical therapist in patients with early-stage amyotrophic lateral sclerosis (ALS). Design A historical controlled study that is part of a multicenter collaborative study. Setting Rehabilitation departments at general hospitals and outpatient clinics with a neurology department. Participants Patients (N=21) with ALS were enrolled and designated as the home-based exercise (Home-EX) group, and they performed unsupervised home-based exercises. As a control group, 84 patients with ALS who underwent supervised exercise with a physical therapist for 6 months were extracted from a database of patients with ALS and matched with the Home-EX group in terms of their basic attributes and clinical features. Intervention The Home-EX group was instructed to perform structured home-based exercises without supervision by a physical therapist that consisted of muscle stretching, muscle training, and functional training for 6 months. Main Outcome Measures The primary outcome was the score on the ALS Functional Rating Scale–Revised (ALSFRS-R), which is composed of 3 domains: bulbar function, limb function, and respiratory function. The score ranges from 0 to 48 points, with a higher score indicating better function. Results In the Home-EX group, 15 patients completed the home-based exercises for 6 months, and 6 patients dropped out because of medical reasons or disease progression. No adverse events were reported. The Home-EX group was found to have a significantly higher respiratory function subscore and total score on the ALSFRS-R than the control group at follow-up (P Conclusions Structured home-based exercises without supervision by a physical therapist could be used to alleviate functional deterioration in patients with early-stage ALS.

Isolation and culture of Xenopus laevis neural tubes resulted in differentiation of melanophores and iridophores from neural crest cells; the differentiated melanophores and iridophores were then maintained in culture for more than 6 months. Guanosine has been reported to promote reflecting platelet formation in melanin-producing pigment cells; however, the process of pigment organellogenesis is still unclear. In the present study, unusual light-reflecting pigment cells were observed upon addition of guanosine to the neural tube culture system, which contained melanosomes specific to melanophores, and reflecting platelets specific to iridophores. Ultrastructural studies suggested that irregularly shaped reflecting platelets were formed from stage II melanosomes (the early stage of melanosome formation) in these unusual pigment cells.

n animal regeneration, control of position-dependent cell proliferation is crucial for the complete restoration of patterned appendages in terms of both, shape and size. However, detailed mechanisms of this process are largely unknown. In this study, we identified leucine/glutamine and v-ATPase/lysosomal acidification, via mechanistic target of rapamycin complex 1 (mTORC1) activation, as effectors of amputation plane-dependent zebrafish caudal fin regeneration. mTORC1 activation, which functions in cell proliferation, was regulated by lysosomal acidification possibly via v-ATPase activity at 3 h post amputation (hpa). Inhibition of lysosomal acidification resulted in reduced growth factor-related gene expression and suppression of blastema formation at 24 and 48 hpa, respectively. Along the proximal-distal axis, position-dependent lysosomal acidification and mTORC1 activation were observed from 3 hpa. We also report that Slc7a5 (L-type amino acid transporter), whose gene expression is position-dependent, is necessary for mTORC1 activation upstream of lysosomal acidification during fin regeneration. Furthermore, treatment with leucine and glutamine, for both proximal and distal fin stumps, led to an up-regulation in cell proliferation via mTORC1 activation, indicating that leucine/glutamine signaling possesses the ability to change the position-dependent regeneration. Our findings reveal that leucine/glutamine and v-ATPase/lysosomal acidification via mTORC1 activation are required for position-dependent zebrafish fin regeneration., This study was supported by grants from Grant-in-Aid for Scientific Research from the JSPS (KAKENHI JP15J05983) to K.T.