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Introduction: Gerstmann-Sträussler-Scheinker disease (GSS) is an autosomal-dominant inherited prion disease most often associated with the human prion protein gene (PRNP)-P102L mutation. Although patients manifest considerable phenotypic heterogeneity, the involvement of the nigrostriatal system has not been well-studied., Methods: We performed dopamine transporter single-photon emission computed tomography (DAT-SPECT) using 123 I-ioflupane to investigate the nigrostriatal system function in nine patients with the PRNP-P102L mutation. We also examined the pathological findings in another patient whose predominant feature was ataxia and who died 5 years after disease onset., Results: Striatum uptake of 123 I-ioflupane indicated by specific binding ratio (SBR) values was significantly reduced in two patients. The DAT-SPECT examination was performed 6 months after disease onset in one of these patients who manifested rapidly developing cognitive decline mimicking Creutzfeldt-Jakob disease. DAT-SPECT was also performed 9 years after disease onset in another patient who manifested the conventional features of GSS involving ataxia and dementia in the initial phase but showed akinetic mutism at the examination time. Another patient examined 2 years after disease onset who predominantly manifested ataxia showed marginally abnormal SBR values. An autopsy case showed moderate neuronal loss in the substantia nigra, and the degree of neuronal loss was similar in most other parts of the brain., Conclusion: Nigrostriatal system involvement may occur in patients with GSS associated with the PRNP-P102L mutation, even though parkinsonism is not the predominant feature., Competing Interests: Declaration of competing interest The authors declare that they have no financial and personal relationships with other people or organizations that could inappropriately influence (bias) their work., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Deep brain stimulation (DBS) has emerged as an important therapeutic option for several movement disorders; however, the management of acute complications, such as acute subdural hematoma (ASDH), remains challenging. This is the case of a 71-year-old woman with Parkinson's disease who developed ASDH 12 years after bilateral DBS placement. On admission with altered consciousness, imaging revealed significant displacement of the DBS electrodes because of the hematoma. Emergent craniotomy with endoscopic evacuation was performed with preservation of the DBS system. Postoperatively, complete evacuation of the hematoma was confirmed, and the patient experienced significant clinical improvement. ASDH causes significant electrode displacement in patients undergoing DBS. After hematoma evacuation, the electrodes were observed to return to their proper position, and the patient exhibited a favorable clinical response to stimulation. To preserve the DBS electrodes, endoscopic hematoma evacuation via a small craniotomy may be useful., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Tanaka et al.)

Human parainfluenza virus type 3 (HPIV-3, human respirovirus 3) is the second most frequently detected virus in lower respiratory tract infections in children after human respiratory syncytial virus (HRSV). HPIV-3, similar to related respiratory viruses such as HRSV and influenza virus, may cause encephalopathy; however, the relevance of HPIV-3 as a pathogenic factor in encephalopathy is unknown. We attempted to detect HPIV-1, HPIV-2, HPIV-3, HPIV-4, HRSV, and human metapneumovirus (HMPV) in 136 patients with encephalitis/encephalopathy or suspected encephalitis/encephalopathy during a 6-year period from 2014 to 2019. HPIV-3 was detected in 6 patients, followed by HRSV in 3 patients. The HPIV-3 strains detected were closely related to those detected in a patient with respiratory disease during the same period. Although HPIV-3 is less widely recognized than HRSV as a triggering virus of encephalopathy, our results suggest that HPIV-3 is as important as HRSV. Surveillance of the causative viruses of encephalopathy, including HPIV-3, would help clarify the causes of encephalopathy in Japan, as the cause is currently reported in less than half of cases in Japan.

Background and Purpose: The coast of Kyushu Island on Ariake Sea in Japan is known to be an accumulation area for patients with a proline-to-leucine substitution mutation at residue 102 (P102L) of the human prion protein gene ( PRNP ), which is associated with Gerstmann-Sträussler-Scheinker disease. We designated this geographical distribution as the "Ariake PRNP P102L variant." The purpose of this study was to characterize the clinical features of this variant., Methods: We enrolled patients with the PRNP P102L variant who were followed up at the Saga University Hospital from April 2002 to November 2019. The clinical information of patients were obtained from medical records, including clinical histories, brain magnetic resonance imaging (MRI), and electroencephalography (EEG). A brain autopsy was performed on one of the participants., Results: We enrolled 24 patients from 19 family lines, including 12 males. The mean age at symptom onset was 60.6 years (range, 41-77 years). The incidence rate of the Ariake PRNP P102L variant was 3.32/1,000,000 people per year in Saga city. The initial symptoms were ataxia (ataxic gait or dysarthria) in 19 patients (79.2%), cognitive impairment in 3 (12.5%), and leg paresthesia in 2 (8.3%). The median survival time from symptom onset among the 18 fatal cases was 63 months (range, 23-105 months). Brain MRI revealed no localized cerebellar atrophy, but sparse diffusion-weighted imaging abnormalities were detected in 16.7% of the patients. No periodic sharp-wave complexes were identified in EEG. Neuropathological investigations revealed uni- and multicentric prion protein (PrP) plaques in the cerebral cortex, putamen, thalamus, and cerebellum of one patient. Western blot analysis revealed 8-kDa proteinase-K-resistant PrP., Conclusions: This is the first report of the accumulation area of a PRNP P102L variant on the coast of Ariake Sea. The Ariake PRNP P102L variant can be characterized by a relatively long disease duration with sparse abnormalities in brain MRI and EEG relative to previous reports. Detailed interviews to obtain information on the birthplace and the family history of related symptoms are important to diagnosing a PRNP P102L variant., Competing Interests: The authors have no potential conflicts of interest to disclose., (Copyright © 2024 Korean Neurological Association.)

Background: Learning self-care for lymphedema is essential for patients to maintain their quality of life; however, it is sometimes difficult and stressful. There are only few studies about the psychological changes in patients hospitalized for conservative therapy. The purpose of this study was to evaluate the psychological changes in patients admitted for conservative therapy and training in self-care for lymphedema. Methods and Results: Nine patients who were hospitalized for conservative treatment of lymphedema of the lower limbs were administered the Profile of Moods States questionnaire twice: day of admission or the following day and after 5 days of hospital stay. Eight female patients and one male patient were included in this retrospective study. The mean age was 67.2 years. We provided standard compression therapy, manual lymph drainage, and exercise therapy to the patients. The Profile of Moods States 2nd edition, Japanese version of the Profile of Moods States, was used as an evaluation method of the psychological state. The results of the psychological tests were evaluated by a certified public psychologist. The scores for negative mood (anger or hostility, confusion or bewilderment, depression or rejection, fatigue or inertia, and tension and anxiety) were all lower on the fifth day of hospitalization compared with those at admission. In particular, the tension or anxiety scores decreased significantly ( p  = 0.019). However, the vigor or activity scores tended to increase. Conclusions: It was found that inpatient conservative therapy for lymphedema had a positive effect on the psychological state of the patients. Despite stressors such as a change in environment and introduction of new treatments (compression therapy and exercise therapy), the improvement in edema helped elevate the mood of the patients by the fifth day of hospitalization.

Laser-induced fluorescence (LIF) Doppler spectroscopy using an optical vortex beam with an asymmetric intensity distribution, referred to as aOVLIF, is proposed as a new method to measure plasma flow velocity. LIF spectra were calculated numerically using typical laboratory low-temperature plasma parameters, and it was revealed that an ion flow across the beam produces a frequency shift of the spectra. This method also has the capability of temperature measurements. The propagation effects of asymmetric optical vortex beams are discussed assuming an actual experiment, and it is found that the sensitivity to the transverse flow velocity is approximately unchanged. The aOVLIF method, which exploits the inhomogeneous phase structure of optical vortices, can be applied to the determination of three-dimensional velocity vectors and promises to enhance the usefulness of conventional LIF spectroscopy using plane waves., (© 2024. The Author(s).)

Nanobubble (NB) water has been shown to promote the growth of several types of plants and animals, but the mechanism underlying this promoting effect remains unclear. The present study evaluated the mechanism by which NBs maintain the freshness of cut flowers by keeping cut chrysanthemum ( Chrysanthemum morifolium Ramat.) flowers at the bud stage in vase water containing air NBs. The condition of petals and leaves was assessed to determine the vase life of these cut flowers. The NB treatment delayed bud opening and petal senescence of the inflorescences. Water absorption and transpiration by cut flower stems were lower in NB water than in distilled water (DW). Furthermore, when all the leaves were removed from the cut flower stems, no significant difference in vase life was observed between NB water and DW. These findings indicate that the inhibition of transpiration from leaves prolonged the vase life of NB-treated cut chrysanthemum flowers. In the early stage of the treatment, NB treatment significantly reduced transpiration without closing stomata, suggesting that the reduction in transpiration observed in the NB-treated plants might be due to the suppression of cuticular transpiration, defined as water loss through the epidermis. Surface tension, one of the important driving forces of water movement in plants, was not affected by the presence of NBs in water. To our knowledge, this is the first report to show that transpiration from leaves is inhibited by NB treatment., Competing Interests: None of the authors has any potential conflicts of interest to report., (© 2023 The Authors. Plant‐Environment Interactions published by New Phytologist Foundation and John Wiley & Sons Ltd.)

Tunable diode laser absorption spectroscopy (TDLAS) is a valuable method for measuring particle flow velocities in plasma. However, conventional TDLAS using a plane-wave beam is sensitive only to the laser propagation direction. This limitation is particularly unfavorable for the observation of the particle transportation perpendicularly incident on the material in the plasma-material interaction. In this paper, we show for the first time that flow measurements perpendicular to the beam direction are possible by replacing the probe beam with an optical vortex beam. Because an optical vortex has a helical wavefront, particles moving in its field experience an azimuthal Doppler shift in addition to the translational Doppler shift. Assuming a uniform gas flow across the optical vortex, the azimuthal Doppler shift of the absorption spectrum observed in the beam cross-section varies sinusoidally in the azimuthal direction. The transverse flow velocity is derived from the amplitude of this sinusoidal variation. At transverse velocities above 70 m/s, the measurement errors are found to be less than 15%, with a mean absolute percentage error of less than 8%., (© 2023. Springer Nature Limited.)

Background: Frozen shoulder (FS) is a pathological condition that involves a painful and stiff shoulder joint, most commonly in people aged 40-60 years. Most literature supports treatment with physical therapy (PT), although some studies have demonstrated years of continuing pain and functional deficits. Manipulation under anesthesia is effective at eliminating the contracture of intra-articular lesions for refractory FS. This study aimed to compare whether manipulation under anesthesia or PT is a more effective treatment in refractory FS., Methods: This study was a prospective observational study. A total of 102 patients with refractory FS were enrolled in this study in the medical records, all of whom had severe and multidirectional loss of motion and thickening of the joint capsule and coracohumeral ligament on magnetic resonance imaging. Fifty-one patients were in the manipulation under brachial plexus block (MUB) group (34 females, median age: 57 years), and 51 patients were in the PT group (34 females, median age: 59 years). The MUB procedure consisted of the conventional method with additional adduction manipulation, in which one examiner initially abducted the shoulder joint as much as possible. We recorded the visual analog scale, shoulder range of motion, and American Shoulder and Elbow Surgeons and Constant Scores at the initial baseline visit and at the 1-, 3-, 6-, and 12-month follow-ups. The total cost was calculated from the medical records, and cost-effectiveness was evaluated using quality-adjusted life year and incremental cost-effectiveness ratio., Results: Visual analog scale ( P  < .001), range of motion ( P  < .001), and American Shoulder and Elbow Surgeons and Constant Scores ( P  < .001) in the MUB group were significantly superior to those in the PT group at 1, 3, 6, and 12 months after treatment. The median cost and total quality-adjusted life year in the MUB and PT groups were $1375 versus $2751 and 2.95 versus 2.68, respectively, and the cost-effectiveness ratio between the MUB and PT groups was calculated as -$560., Conclusions: The new MUB procedure provides a shorter treatment period, better clinical outcomes, and higher cost-effectiveness in patients with refractory FS compared to PT., (© 2023 The Author(s).)

The surfaces of concrete structures are often coated with protective materials to minimize corrosion and weathering-based deterioration. Therefore, it is important to monitor the aging of the coating materials and their overall condition to extend the service lifetime of the structure effectively. Near-infrared spectroscopy (NIRS) is a contactless, nondestructive, rapid, and convenient method for material characterization; therefore, it is useful for onsite inspection of coating materials. Hence, in this study, we attempt to determine whether NIRS can be used for simple inspection for health monitoring of organic resin-based coating materials. In addition to identifying different severities of peeling damage, we characterize the ultraviolet-induced deterioration of coating materials with different thicknesses using diffuse reflection spectra acquired in the near-infrared wavelength region. For independent comparison with the NIR spectra, the state of the coating materials on the mortar specimens was analyzed using a combination of Fourier-transform infrared spectroscopy and scanning electron microscopy, while the state of the underlying mortar specimens was analyzed using permeability and salt-water immersion tests. The results confirm that the NIRS could detect the degradation of coating materials at early stages of deterioration before their permeability had been affected. NIRS offers the possibility of intermittent monitoring of coating deterioration. In addition, because the NIR spectrometer is portable, it can help in inspecting high-rise areas and areas that are difficult to reach. Therefore, we believe that NIRS is a simple, safe, and inexpensive method for inspection of surface coating materials., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Watanabe et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Cerebral small vessel disease (SVD) is one of the leading causes of stroke; each neuroimaging marker of SVD is correlated with vascular risk factors and associated with poor prognosis after stroke. However, longitudinal studies investigating the association between comprehensive SVD burden scoring system, "total SVD score" - which encompasses the established neuroimaging markers of lacunae, cerebral microbleeds (CMBs), white matter hyperintensities (WMH) including periventricular hyperintensities, and perivascular spaces in basal ganglia- and clinical outcomes are limited. The aim of this study is to determine the association between SVD burden and long-term prognosis in patients with ischemic stroke., Methods and Design: This prospective, single-center, observational study enrolled patients with acute ischemic stroke, including cerebral infarction and transient ischemic attack. Magnetic resonance imaging scans were performed, and then total SVD score (range, 0-4) was calculated. We recorded baseline characteristics and evaluated the relationships of long-term outcomes to SVD neuroimaging markers and total SVD score. Stroke recurrence was thought as primary outcome. Hazard ratios (HRs) of events during follow-up were calculated using Cox proportional hazards modeling with adjustments for age, sex, hypertension, dyslipidemia, diabetes mellitus, atrial fibrillation, and smoking. Cumulative event rates were estimated using the Kaplan-Meier method., Results: Consecutive 564 acute ischemic stroke patients were enrolled according to inclusion and exclusion criteria. A total of 467 participants with first-ever ischemic stroke were analyzed (median age 75.0 [interquartile range, 64.0-83.0] years, 59.3% male). Total SVD score was 0 point in 47 individuals (12.0%), 1 point in 83 (21.2%), 2 points in 103 (26.3%), 3 points in 85 (21.7%), and 4 points in 73 (18.7%). Twenty-eight recurrent stroke events were identified during follow-up. Total SVD score ≥ 2, presence of CMBs, and moderate-to-severe WMH were associated with increased risk of recurrent stroke events (HR 9.31, 95% confidence interval [CI] 2.33-64.23; HR 2.81, 95% CI 1.08-7.30; HR 2.90, 95% CI 1.22-6.88, respectively)., Conclusion: The accumulation of SVD biomarkers as determined by total SVD score offered a reliable predictor of stroke recurrence. This study established a firm understanding of SVD prognosis in clinical settings., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ikeda, Yakushiji, Tanaka, Nishihara, Ogata, Eriguchi, Ono, Kosugi, Suzuyama, Mizoguchi, Shichijo, Ide, Nagaishi, Mori, Ono, Yoshikawa, Ide, Minagawa, Iida, Kawamoto, Katsuki, Irie, Abe and Hara.)

Ammonium absorbed by roots is assimilated into amino acids. The glutamine synthetase/glutamate synthase (glutamine 2-oxoglutarate aminotransferase) (GS/GOGAT) cycle is essential to this biological process. In Arabidopsis thaliana , GLN1;2 and GLT1 are the GS and GOGAT isoenzymes induced in response to ammonium supply and playing key roles in ammonium utilization. Although recent studies suggest gene regulatory networks involved in transcriptional regulation of ammonium-responsive genes, direct regulatory mechanisms for ammonium-induced expression of GS/GOGAT remain unclear. In this study, we revealed that the expression of GLN1;2 and GLT1 in Arabidopsis is not directly induced by ammonium but is regulated by glutamine or post-glutamine metabolites produced by ammonium assimilation. Previously, we identified a promoter region required for ammonium-responsive expression of GLN1;2 . In this study, we further dissected the ammonium-responsive region of the GLN1;2 promoter and also performed a deletion analysis of the GLT1 promoter, which led to the identification of a conserved ammonium-responsive region. Yeast one-hybrid screening using the ammonium-responsive region of the GLN1;2 promoter as a decoy sequence revealed a trihelix family transcription factor DF1 that binds to this region. A putative DF1 binding site was also found in the ammonium-responsive region of the GLT1 promoter., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Kojima, Minagawa, Yoshida, Inoue, Takahashi and Ishiyama.)

Background: A Wilms' tumor 1 (WT1) oral vaccine, Bifidobacterium longum (B. longum) 420, in which the bacterium is used as a vector for WT1 protein, triggers immune responses through cellular immunity consisting of cytotoxic T lymphocytes (CTLs) and other immunocompetent cells (e.g., helper T cells). We developed a novel, oral, helper epitope-containing WT1 protein vaccine (B. longum 2656) to examine whether or not B. longum 420/2656 combination further accelerates the CD4 + T cell help-enhanced antitumor activity in a model of murine leukemia., Methods: C1498-murine WT1-a genetically-engineered, murine leukemia cell line to express murine WT1-was used as tumor cell. Female C57BL/6 J mice were allocated to the B. longum 420, 2656, and 420/2656 combination groups. The day of subcutaneous inoculation of tumor cells was considered as day 0, and successful engraftment was verified on day 7. The oral administration of the vaccine by gavage was initiated on day 8. Tumor volume, the frequency and phenotypes of WT1-specific CTLs in CD8 + T cells in peripheral blood (PB) and tumor-infiltrating lymphocytes (TILs), as well as the proportion of interferon-gamma (INF-γ)-producing CD3 + CD4 + T cells pulsed with WT1 35-52 peptide in splenocytes and TILs were determined., Results: Tumor volume was significantly smaller (p < 0.01) in the B. longum 420/2656 combination group than in the B. longum 420 group on day 24. WT1-specific CTL frequency in CD8 + T cells in PB was significantly greater in the B. longum 420/2656 combination group than in the B. longum 420 group at weeks 4 (p < 0.05) and 6 (p < 0.01). The proportion of WT1-specific, effector memory CTLs in PB increased significantly in the B. longum 420/2656 combination group than in the B. longum 420 group at weeks 4 and 6 (p < 0.05 each). WT1-specific CTL frequency in intratumoral CD8 + T cells and the proportion of IFN-γ-producing CD3 + CD4 + T cells in intratumoral CD4 + T cells increased significantly (p < 0.05 each) in the B. longum 420/2656 combination group than in the 420 group., Conclusions: B. longum 420/2656 combination further accelerated antitumor activity that relies on WT1-specific CTLs in the tumor compared with B. longum 420., (© 2023. The Author(s).)

Pleurotus ostreatus is an edible white-rot fungus with lignocellulosic biomass degrading enzymes that have been studied extensively. However, until now, lipolytic enzymes from P. ostreatus, which degrade extractives in lignocellulosic biomass, have not been purified and characterized. In this study, P. ostreatus was inoculated into the rapeseed oil containing culture to induce lipase. The lipase in the culture broth was successfully purified to homogeneity by chromatographic methods. The molecular weight of the purified lipase was 27 kDa, and its optimal pH and temperature were 5.0 and 30 °C, respectively. The purified lipase showed high activity with the substrates 4-methylumbelliferyl (4-MU) decanoate (C10:0) and 4-MU oleate (C18:1), and no activity with 4-MU acetate (C2:0) and 4-MU butyrate (C4:0). The amino acid sequences and substrate specificities of the purified lipase suggested that it belonged to class III. Kinetic parameters measurements (Km and Vmax) showed that 4-MU palmitate had a high affinity for the purified lipase, and it was the substrate most efficiently hydrolyzed by the purified lipase., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Wilms' tumor 1 (WT1) is a promising tumor-associated antigen for cancer immunotherapy. We developed an oral protein vaccine platform composed of WT1-anchored, genetically engineered Bifidobacterium longum (B. longum) and conducted an in vivo study in mice to examine its anticancer activity. Mice were orally treated with phosphate-buffered saline, wild-type B. longum105-A, B. longum 2012 displaying only galacto-N-biose/lacto-N-biose I-binding protein (GLBP), and WT1 protein- and GLBP-expressing B. longum 420. Tumor size reduced significantly in the B. longum 420 group than in the B. longum 105-A and 2012 groups (P < 0.00 l each), indicating B. longum 420's antitumor activity via WT1-specific immune responses. CD8 + T cells played a major role in the antitumor activity of B. longum 420. The proportion of CD103 + CD11b + CD11c + dendritic cells (DCs) increased in the Peyer's patches (PPs) from mice in the B. longum 420 group, indicating the definite activation of DCs. In the PPs, the number and proportion of CD8 + T cells capable of producing interferon-gamma were significantly greater in the B. longum 420 group than in the B. longum 2012 group (P < 0.05 or < 0.01). The production of WT1-specific IgG antibody was significantly higher in the B. longum 420 group than in the 2012 group (P < 0.05). The B. longum 420 group showed the most intense intratumoral infiltration of CD4 + and CD8 + T cells primed by activated DCs in the PPs of mice in the B. longum 420 group. Our findings provide insights into a novel, intestinal bacterium-based, cancer immunotherapy through intestinal immunity., (© 2022. The Author(s).)

In Japan, major mumps outbreaks still occur every 4-5 years because of low mumps vaccine coverage (30-40%) owing to the voluntary immunization program. Herein, to prepare for a regular immunization program, we aimed to reveal the nationwide and long-term molecular epidemiological trends of the mumps virus (MuV) in Japan. Additionally, we performed whole-genome sequencing (WGS) using next-generation sequencing to assess results from conventional genotyping using MuV sequences of the small-hydrophobic (SH) gene. We analyzed 1,064 SH gene sequences from mumps clinical samples and MuV isolates collected from 25 prefectures from 1986 to 2017. The results showed that six genotypes, namely B (110), F (1), G (900), H (3), J (41), and L (9) were identified, and the dominant genotypes changed every decade in Japan since the 1980s. Genotype G has been exclusively circulating since the early 2000s. Seven clades were identified for genotype G using SH sequence-based classification. To verify the results, we performed WGS on 77 representative isolates of genotype G using NGS and phylogenetically analyzed them. Five clades were identified with high bootstrap values and designated as Japanese clade (JPC)-1, -2, -3, -4, -5. JPC-1 and -3 accounted for over 80% of the total genotype G isolates (68.3 and 13.8%, respectively). Of these, JPC-2 and -5, were newly identified clades in Japan through this study. This is the first report describing the nationwide and long-term molecular epidemiology of MuV in Japan. The results provide information about Japanese domestic genotypes, which is essential for evaluating the mumps elimination progress in Japan after the forthcoming introduction of the mumps vaccine into Japan's regular immunization program. Furthermore, the study shows that WGS analysis using NGS is more accurate than results obtained from conventional SH sequence-based classification and is a powerful tool for accurate molecular epidemiology studies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kidokoro, Shiino, Yamaguchi, Nariai, Kodama, Nakata, Sano, Gotou, Kisu, Maruyama, Kuba, Sakata, Higashi, Kiyota, Sakai, Yahiro, Nagita, Watanabe, Hirokawa, Hamabata, Fujii, Yamamoto, Yokoi, Sakamoto, Saito, Shibata, Inada, Fujitani, Minagawa, Ito, Shima, Murano, Katoh, Kato, Takeda, Suga and The Surveillance Team for Mumps Virus in Japan.)

Simple, highly sensitive detection technologies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are crucial for the effective implementation of public health policies. We used the systematic evolution of ligands by exponential enrichment with a modified DNA library, including a base-appended base (uracil with a guanine base at its fifth position), to create an aptamer with a high affinity for the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein. The aptamer had a dissociation constant of 1.2 and < 1 nM for the RBD and spike trimer, respectively. Furthermore, enzyme-linked aptamer assays confirmed that the aptamer binds to isolated authentic SARS-CoV-2 wild-type and B.1.617.2 (delta variant). The binding signal was larger that of commercially available anti-SARS-CoV-2 RBD antibody. Thus, this aptamer as a sensing element will enable the highly sensitive detection of SARS-CoV-2., Competing Interests: Declaration of competing interest This work was partly supported by grants for the Japan Program for Infectious Diseases Research and Infrastructure (JP21wm0225003 and JP21fk0108104j) from Japan Agency for Medical Research and Development (AMED) and fund from NEC Solution Innovators, Ltd., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Wilms' tumor gene 1 (WT1) peptide vaccine and anti-programmed cell death-1 (anti-PD-1) antibody are expected as immunotherapies to improve the clinical outcome of glioblastoma. The aims of this study were to clarify how each immunotherapy affects tumor-infiltrating immune cells (TIIs) and to determine whether the combination of these two therapies could synergistically work., Methods: Mice were transplanted with WT1 and programmed cell death-ligand 1 doubly expressing glioblastoma cells into brain followed by treatment with WT1 peptide vaccine, anti-PD-1 antibody, or the combination of the two, and survival of each therapy was compared. CD45 + cells were positively selected as TIIs from the brains with tumors, and TIIs were compared between WT1 peptide vaccine and anti-PD-1 antibody therapies., Results: Most mice seemed to be cured by the combination therapy with WT1 peptide vaccine and anti-PD-1 antibody, which was much better survival than each monotherapy. A large number of CD4 + T cells, CD8 + T cells, and NK cells including WT1-specific CD8 + and CD4 + T cells infiltrated into the glioblastoma in WT1 peptide vaccine-treated mice. On the other hand, the number of TIIs did not increase, but instead PD-1 molecule expression was decreased on the majority of the tumor-infiltrating CD8 + T cells in the anti-PD-1 antibody-treated mice., Conclusion: Our results clearly demonstrated that WT1 peptide vaccine and anti-PD-1 antibody therapies worked in the different steps of cancer-immunity cycle and that the combination of the two therapies could work synergistically against glioblastoma., (© The Author(s) 2021. Published by Oxford University Press, the Society for Neuro-Oncology and the European Association of Neuro-Oncology.)

Background: Although it is known that malignancies can be associated with dermatomyositis, there are few reports on dermatomyositis associated with prostate cancer with neuroendocrine differentiation., Case Presentation: A 63-year-old man visited our hospital due to pollakiuria. High levels of PSA and NSE were observed, and prostate biopsy revealed an adenocarcinoma with neuroendocrine differentiation. Multiple metastases to the lymph nodes, bones, and liver were identified, and androgen deprivation therapy (ADT) was started immediately. Following 2 weeks of treatment, erythema on the skin, and muscle weakness with severe dysphagia appeared. The patient was diagnosed with dermatomyositis, and high-dose glucocorticoid therapy was initiated. ADT and subsequent chemotherapy with etoposide and cisplatin (EP) were performed for prostate cancer, which resulted in decreased PSA and NSE and reduction of all metastases. After the initiation of EP therapy, dermatomyositis improved, and the patient regained oral intake function. Although EP therapy was replaced by docetaxel, abiraterone, and enzalutamide because of adverse events, no cancer progression was consistently observed. Dermatomyositis worsened temporarily during the administration of abiraterone, but it improved upon switching from abiraterone to enzalutamide and dose escalation of glucocorticoid., Conclusions: We successfully treated a rare case of dermatomyositis associated with prostate adenocarcinoma with neuroendocrine differentiation.

Background: The combination of systematic evolution of ligands by exponential enrichment (SELEX) and deep sequencing is termed high-throughput (HT)-SELEX, which enables searching aptamer candidates from a massive amount of oligonucleotide sequences. A clustering method is an important procedure to identify sequence groups including aptamer candidates for evaluation with experimental analysis. In general, aptamer includes a specific target binding region, which is necessary for binding to the target molecules. The length of the target binding region varies depending on the target molecules and/or binding styles. Currently available clustering methods for HT-SELEX only estimate clusters based on the similarity of full-length sequences or limited length of motifs as target binding regions. Hence, a clustering method considering the target binding region with different lengths is required. Moreover, to handle such huge data and to save sequencing cost, a clustering method with fast calculation from a single round of HT-SELEX data, not multiple rounds, is also preferred., Results: We developed fast string-based clustering (FSBC) for HT-SELEX data. FSBC was designed to estimate clusters by searching various lengths of over-represented strings as target binding regions. FSBC was also designed for fast calculation with search space reduction from a single round, typically the final round, of HT-SELEX data considering imbalanced nucleobases of the aptamer selection process. The calculation time and clustering accuracy of FSBC were compared with those of four conventional clustering methods, FASTAptamer, AptaCluster, APTANI, and AptaTRACE, using HT-SELEX data (>15 million oligonucleotide sequences). FSBC, AptaCluster, and AptaTRACE could complete the clustering for all sequence data, and FSBC and AptaTRACE performed higher clustering accuracy. FSBC showed the highest clustering accuracy and had the second fastest calculation speed among all methods compared., Conclusion: FSBC is applicable to a large HT-SELEX dataset, which can facilitate the accurate identification of groups including aptamer candidates., Availability of Data and Materials: FSBC is available at http://www.aoki.ecei.tohoku.ac.jp/fsbc/.

Human C-reactive protein (CRP) and lactate dehydrogenase are important markers in clinical laboratory testing-the former is used to detect in vivo inflammation, and the latter is used to detect cell necrosis and tissue destruction. We developed aptamers that bind to human CRP and human lactate dehydrogenase-5 (LDH-5) with high affinities (dissociation constants of 6.2 pM and 235 pM, respectively), applying the systematic evolution of ligands by exponential enrichment (SELEX) method, and by using a modified DNA library containing the following base-appended base modifications: analog adenine derivative at the fifth position of uracil (U ad ), analog guanine derivative at the fifth position of uracil (U gu ), and analog adenine derivative at the seventh position of adenine (A ad ). A potential application of these aptamers as sensor elements includes high-sensitivity target detection in point-of-care testing.

We used base-appended base modification to develop a new adenine analog, which incorporates an adenine derivative at position 7 of adenine. Using the systematic evolution of ligands by exponential enrichment method with a modified DNA library including this analog, we obtained A ad1 , an aptamer that binds strongly to human β-defensin 2, a biomarker of physical stress found in saliva. The dissociation constant of A ad1 with respect to human β-defensin 2 was found to be low (6.8 nM), and was found to bind specifically to human β-defensin 2 in saliva spiked with the protein, as confirmed using pull-down with magnetic beads. To our knowledge, there are no prior reports of nucleic-acid aptamers that bind specifically to human β-defensin 2. However, our results indicated that such adenine analog-containing DNA libraries are extremely effective in the acquisition of high-affinity aptamers., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The field of care testing toward the analysis of blood and saliva lacks nowadays simple test techniques for biomarkers. In this study, we have developed a novel nucleobase analog, U gu , which is a uracil derivative bearing a guanine base at the 5-position. Moreover, we attempted the development of aptamers that can bind to secretory immunoglobulin A (SIgA), which has been examined as a stress marker in human saliva. It was observed that the acquired aptamer binds strongly and selectively to the SIgA dimer ( K d = 13.6 nM) without binding to the IgG and IgA monomers of human serum. Reduction of the aptamer length (41 mer) successfully improved 4-fold the binding affinity ( K d = 3.7 nM), compared to the original, longer aptamer (78 mer). Furthermore, the development of a simple detection system for human saliva samples by fluorescence polarization was investigated, using the reported human salivary α-amylase (sAA) and the SIgA-binding aptamer. Comparison of the present method with conventional enzyme-linked immunosorbent assay techniques highlighted a significant Pearson's correlation of 0.94 and 0.83 when targeting sAA and SIgA, respectively. It is thus strongly suggested that a new simple test of stress markers in human saliva can be quantified quickly without bound/free (B/F) separation.

Introduction: To carry out ultrasound-guided cervical nerve root block (CNRB) safely, we investigated the frequency of risky blood vessels around the target nerve root and within the imaginary needle pathway in the actual injecting position., Methods: 30 patients (20 men, 10 women) with cervical radiculopathy who received ultrasound-guided CNRB were included in this study. We defined a risky blood vessel as an artery existing within 4 mm from the center of the target nerve root or located in the range of 2 mm above or below the imaginary needle pathway., Results: Using the color Doppler method, the frequency of a risky blood vessel existing around 4 mm from the center of the C5 nerve root was 3.3% (1/30), whereas it was 3.3% (1/30) for the C6 nerve root and 23.3% (7/30) for the C7 nerve root. Hence, the C7 level had more blood vessels close to the target nerve root compared to the C5 and C6 levels, but there was no significant difference (p = 0.0523). On the other hand, the frequency of a risky blood vessel existing within 2 mm above and below the imaginary needle pathway was 3.3% (1/30) for the C5 nerve root, whereas it was 3.3% (1/30) for the C6 nerve root and 10.0% (3/30) for the C7 nerve root. The C7 level had more blood vessels within the needle pathway compared to the C5 and C6 levels, but there was no significant difference (p = 0.301)., Conclusions: To reduce the risk of unintended intravascular injections, more careful checking for the presence or absence of blood vessels at the C7 level using color Doppler is necessary., Competing Interests: Conflicts of Interest: The authors declare that there are no relevant conflicts of interest., (Copyright © 2020 by The Japanese Society for Spine Surgery and Related Research.)

Genotyping evidence that supports the interruption of endemic measles virus (MV) transmission is one of the essential criteria to be verified in achieving measles elimination. In Japan since 2014, MV genotype analyses have been performed for most of the measles cases in prefectural public health institutes nationwide. With this strong molecular epidemiological data, Japan was verified to have eliminated measles in March, 2015. However, even in the postelimination era, sporadic cases and small outbreaks of measles have been detected repeatedly in Japan. This study investigated the nationwide molecular epidemiology of MV between 2008 and 2017. The 891 strains in the total period between 2008 and 2017 belonged to seven genotypes (D5, D4, D9, H1, G3, B3, and D8) and 124 different MV sequence variants, based on the 450-nucleotide sequence region of the N gene (N450). The 311 MV strains in the postelimination era between 2015 and 2017 were classified into 1, 7, 8, and 32 different N450 sequence variants in D9, H1, B3, and D8 genotypes, respectively. Analysis of the detection period of the individual N450 sequence variants showed that the majority of MV strains were detected only for a short period. However, MV strains, MVs/Osaka.JPN/29.15/ [D8] and MVi/Hulu Langat.MYS/26.11/ [D8], which are named strains designated by World Health Organization (WHO), have been detected in many cases over 2 or 3 years between 2015 and 2017. The WHO-named strains have circulated worldwide, causing outbreaks in many countries. Epidemiological investigation revealed repeated importation of these WHO-named strains into Japan. To demonstrate the elimination status (interruption of endemic transmission) in situations with repeated importation of the same strains is challenging. Nevertheless, the detailed sequence analysis of individual MV strains and chronological analysis of these strains provided sufficient evidence to show that Japan has still maintained its measles elimination status in 2017.

This study aimed to obtain screening data on the maturity status of the tibial tuberosity in schoolchildren of higher elementary school grades for risk management of Osgood-Schlatter disease (OSD). The maturity stages and cartilage thicknesses at the tibial tuberosity were determined by ultrasonography on the occasion of a school-based musculoskeletal examination for 124 grade 5-6 elementary schoolchildren, and their associations with the students' demographic characteristics and OSD were examined. The time-dependent changes of the maturity status of the tibial tuberosity were also examined in grade 5 students ( n = 26) by a longitudinal survey. The cross-sectional survey showed that the epiphyseal stage was reached in 89% of girls and 35% of boys. The girls who had experienced menarche ( n = 28) were all in the epiphyseal stage and had a decreased cartilage thickness ( p = 0.004, after adjusting maturity stages). Students with OSD ( n = 5) were all girls in the epiphyseal stage, and only two of them had an increased cartilage thickness. During the longitudinal survey, a marked increase in cartilage thickness from the previous measurement was observed in three boys (without clinical symptoms) and a girl who newly developed OSD. Two students with OSD without chronic pain had thin cartilage. In conclusion, for schoolchildren of higher elementary school grades, the risk of OSD is higher among girls with the epiphyseal stage. Cartilage thickness may not contribute to the diagnosis of OSD, since thick cartilage is not very common in OSD. However, cartilage thickness may reflect the status of OSD.

Koplik spots are considered a disease-specific sign for measles, although comprehensive virological studies have not been conducted to date. In Japan, a national survey of 3023 measles and measles-suspected cases was conducted between 2009 and 2014 using polymerase chain reaction (PCR) or reverse transcription PCR (RT-PCR) to detect various rash/fever-associated viruses. Koplik spots were observed in 717 of 3023 cases (23.7%). Among these, the measles virus was detected in 202 cases (28.2%), while the rubella virus was detected in 125 cases (17.4%). Other viruses were detected in 51 cases having the spots (7.1%). In some of the cases with spots, two or three viruses, such as the rubella virus, parvovirus, and human herpesvirus type 6 were also detected. The sensitivity and specificity of Koplik spots as a diagnostic marker for measles were 48 and 80%, respectively. The results suggested that Koplik spots might appear not only in measles but also in other viral infections, such as rubella, as a clinical sign.

Background: Although the pronator teres muscle, a major dynamic stabilizer of elbow valgus stress during throwing, frequently presents stiffness, its relationship with elbow injuries in youth baseball players is unknown. This study investigated the relationship between the elasticity of the pronator teres muscle and elbow injuries in youth baseball players., Methods: The strain ratio (SR) of 15 individuals with osteochondritis dissecans of the humeral capitellum (OCD group), 67 individuals with medial epicondylar fragmentation (medial injury group), and 115 healthy individuals (control group) was measured as the index of the elasticity of the pronator teres muscle using ultrasound strain elastography. In addition, the forearm and glenohumeral joint rotation range of motion was measured., Results: The SR of the throwing arm was significantly higher in the OCD and medial injury groups than in the control group (both P <.001). In the OCD group, the SR was significantly higher in the throwing arm than in the nonthrowing arm (P <.001), whereas in the medial injury group, there was no significant difference between both arms. The glenohumeral joint external rotation range of motion of the throwing arm was moderately negatively correlated with the SR (r = -0.478, P <.001)., Conclusions: Stiffness of the pronator teres muscle was exhibited only in the throwing arm of individuals with OCD and in both arms in individuals with medial elbow injury. These findings may contribute to an accurate evaluation and prevention of elbow injuries in youth baseball players., (Copyright © 2018 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Background: Anhedonia is a core symptom of major depressive disorder (MDD). While recent evidence suggests that reduced motivation for reward may be a core feature of anhedonia, the abnormalities in modulatory neural responses to variable reward amounts in MDD patients remain unclear. We investigated whether MDD patients' ability to represent variable-sized monetary rewards in the striatum is disrupted., Methods: Twelve MDD patients and 12 healthy volunteers completed an assessment of psychometric status and participated in a functional magnetic resonance imaging (fMRI) task that involved the anticipation of financial reward (monetary incentive delay task). The size of the monetary reward was varied among trial conditions and was cued with geometric stimuli. Patients participated in additional fMRI sessions after a 6-week pharmacological treatment with escitalopram, an SSRI., Results: In healthy volunteers, striatal activity increased in proportion to the size of the monetary reward during reward anticipation. This pattern was altered in MDD patients, and significant group-by-reward size interaction effects were observed in the bilateral putamen and the left ventral striatum. Reward sensitivity in motor response and striatum activity at three regions were correlated in healthy controls. In MDD patients, this neurobehavioral coupling was not observed. In addition, changes in the neural reward sensitivity parameter at the left ventral striatum in response to treatment were positively correlated with a reduction of depressive symptoms., Conclusions: Patients with MDD exhibit reduced ability to modulate neural response when adjusting for variable amount of reward. This result suggests that reward size coding in the striatum may represent a neural correlate of motivational anhedonia in MDD patients., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

We have attained a chemically modified DNA aptamer against salivary α-amylase (sAA), which attracts researchers' attention as a useful biomarker for assessing human psychobiological and social behavioural processes, although high affinity aptamers have not been isolated from a random natural DNA library to date. For the selection, we used the base-appended base (BAB) modification, that is, a modified-base DNA library containing (E)-5-(2-(N-(2-(N6-adeninyl)ethyl))carbamylvinyl)-uracil in place of thymine. After eight rounds of selection, a 75 mer aptamer, AMYm1, which binds to sAA with extremely high affinity (K d  < 1 nM), was isolated. Furthermore, we have successfully determined the 36-mer minimum fragment, AMYm1-3, which retains target binding activity comparable to the full-length AMYm1, by surface plasmon resonance assays. Nuclear magnetic resonance spectral analysis indicated that the minimum fragment forms a specific stable conformation, whereas the predicted secondary structures were suggested to be disordered forms. Thus, DNA libraries with BAB-modifications can achieve more diverse conformations for fitness to various targets compared with natural DNA libraries, which is an important advantage for aptamer development. Furthermore, using AMYm1, a capillary gel electrophoresis assay and lateral flow assay with human saliva were conducted, and its feasibility was demonstrated.

We identified hypervirulent Streptococcus pyogenes in 27 and 420 isolates from patients with invasive and non-invasive diseases, respectively, in Aichi Prefecture, Japan, between 2003 and 2012, in an attempt to understand why the prevalence of streptococcal toxic shock syndrome (STSS) suddenly increased in this location during 2011. Hypervirulent strains belong to the emm1 genotype, with a mutation in the covR/S genes that regulate many other genes, encoding virulence determinants and resulting in the absence of the proteinase streptococcal exotoxin B and the production of virulence factors such as the superantigen streptococcal exotoxin A, the nuclease streptococcal DNase, the cytotoxin NAD-glycohydrolase, and the hemolysin streptolysin O. We found 1 strain from invasive disease and 1 from non-invasive disease with traits similar to those of hypervirulent strains, except that the sda1 gene was absent. We also found 1 non-emm1 strain with phenotypic and genetic traits identical to those of the emm1 hypervirulent strains except that it did not belong to emm1 genotype, from non-invasive diseases cases in 2011. These findings suggested that hypervirulent and hypervirulent-like strains from invasive and non-invasive disease cases could have at least partially contributed to the sudden increase in the number of patients with STSS in Aichi during 2011.

Acute gastroenteritis is a critical infectious disease that affects infants and young children throughout the world, including Japan. This retrospective study was conducted from September 2008 to August 2014 (six seasons: 2008/09-2013/14) to investigate the incidence of enteric viruses responsible for 1,871 cases of acute gastroenteritis in Aichi prefecture, Japan. Of the 1,871 cases, 1,100 enteric viruses were detected in 978 samples, of which strains from norovirus (NoV) genogroup II (60.9%) were the most commonly detected, followed by strains of rotavirus A (RVA) (23.2%), adenovirus (AdV) type 41 (8.2%), sapovirus (SaV) (3.6%), human astrovirus (HAstV) (2.8%), and NoV genogroup I (1.3%). Sequencing of the NoV genogroup II (GII) strains revealed that GII.4 was the most common genotype, although four different GII.4 variants were also identified. The most common G-genotype of RVA was G1 (63.9%), followed by G3 (27.1%), G2 (4.7%) and G9 (4.3%). Three genogroups of SaV strains were found: GI (80.0%), GII (15.0%), and GV (5.0%). HAstV strains were genotyped as HAstV-1 (80.6%), HAstV-8 (16.1%), and HAstV-3 (3.2%). These results show that NoV GII was the leading cause of sporadic acute viral gastroenteritis, although a variety of enteric viruses were detected during the six-season surveillance period., (© 2015 Wiley Periodicals, Inc.)

Background: An easy and reliable assay for detection of the rubella virus is required to strengthen rubella surveillance. Although a TaqMan RT-PCR assay for detection of the rubella virus has been established in Japan, its utility for diagnostic purposes has not been tested., Objectives: To allow introduction of the TaqMan RT-PCR into the rubella surveillance system in Japan, the sensitivity of the assay was determined using representative strains for all genotypes and clinical specimens., Study Design: The detection limits of the method for individual genotypes were examined using viral RNA extracted from 13 representative strains. The assay was also tested at 10 prefectural laboratories in Japan, designated as local reference laboratories for measles and rubella, to allow nationwide application of the assay., Results: The detection limits and amplification efficiencies of the assay were similar among all the representative strains of the 13 genotypes. The TaqMan RT-PCR could detect approximately 90% of throat swab and urine samples taken up to 5days of illness. These samples were determined positive by a highly sensitive nested RT-PCR., Conclusions: The TaqMan RT-PCR could detect at least 10 pfu of rubella virus. Although the sensitivity was somewhat lower than that of the conventional nested RT-PCR, the TaqMan RT-PCR could be more practical to routine tests for rubella laboratory diagnosis and detection in view of the rapid response and reducing risks of contamination., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Japan was verified as having achieved measles elimination by the Measles Regional Verification Commission in the Western Pacific Region in March 2015. Verification of measles elimination implies the absence of continuous endemic transmission. After the last epidemic in 2007 with an estimated 18,000 cases, Japan introduced nationwide case-based measles surveillance in January 2008. Laboratory diagnosis for all suspected measles cases is essentially required by law, and virus detection tests are mostly performed by municipal public health institutes. Despite relatively high vaccination coverage and vigorous response to every case by the local health center staff, outbreak of measles is repeatedly observed in Aichi Prefecture, Japan., Methods: Measles virus N and H gene detection by nested double RT-PCR was performed with all specimens collected from suspected cases and transferred to our institute. Genotyping and further molecular epidemiological analyses were performed with the direct nucleotide sequence data of appropriate PCR products., Results: Between 2010 and 2014, specimens from 389 patients suspected for measles were tested in our institute. Genotypes D9, D8, H1 and B3 were detected. Further molecular epidemiological analyses were helpful to establish links between patients, and sometimes useful to discriminate one outbreak from another. All virus-positive cases, including 49 cases involved in three outbreaks without any obvious epidemiological link with importation, were considered as import-related based on the nucleotide sequence information. Chain of transmission in the latest outbreak in 2014 terminated after the third generations, much earlier than the 2010-11 outbreak (6th generations)., Conclusion: Since 2010, almost all measles cases reported in Aichi Prefecture are either import or import-related, based primarily on genotypes and nucleotide sequences of measles virus detected. In addition, genotyping and molecular epidemiological analyses are indispensable to prove the interruption of endemic transmission when the importations of measles are repeatedly observed., (Copyright © 2015 Elsevier Ltd. All rights reserved.)