Fourth International Symposium on Atomic Technology

The Ministry of Education, Culture, Sports, Science and Technology of Japan started the Priority Assistance for the Formation of Worldwide Renowned Centers of Research – Global COE Program. This program is based on the competitive principle where a third party evaluation decides which program to support and to give priority support to the formation of world-class centers of research. Our program Center of Excellence for Advanced Structural and Functional Materials Design was selected as one of 13 programs in the field of Chemistry and Materials Science. This center is composed of two materials-related Departments in the Graduate School of Engineering: Materials and Manufacturing Science and Adaptive Machine Systems, and 4 Research Institutes: Center for Atomic and Molecular Technologies, Welding and Joining Research Institute, Institute of Scientific and Industrial Research and Research Center for Ultra-High Voltage Electron Microscopy. Recently, materials research, particularly that of metallic materials, has specialized only in individual elemental characteristics and narrow specialty fields, and there is a feeling that the original role of materials research has been forgotten. The 6 educational and research organizations which make up the COE program cooperatively try to develop new advanced structural and functional materials and achieve technological breakthrough for their fabrication processes from electronic, atomic, microstructural and morphological standpoints, focusing on their design and application: development of high performance structural materials such as space plane and turbine blades operating under a severe environment, new fabrication and assembling methods for electronic devices, development of evaluation technique for materials reliability, and development of new biomaterials for regeneration of biological hard tissues. The aim of this international conference was to report the scientific progress in our Global COE program and also to discuss related research topics. The organizing committee gratefully thanks participants for presenting their recent results and for discussions with our COE members and international attendees. November 2008 Professor Tomoyuki Kakeshita Chairman of the Conference Vice Dean, Graduate School of Engineering, Osaka University, Division of Materials and Manufacturing Science, Graduate School of Engineering Leader of Global COE Program, Osaka University, ‘‘Center of Excellence for Advanced Structural and Functional Materials Design’’ Organization Chairman: T Kakeshita (Osaka University) Advisory Board: H Mehrer (University Münster, Germany), E K H Salje (University of Cambridge, United Kingdom), H-E Schaefer (University of Stuttgart, Germany), P Veyssiere (CNRS-ONERA, France) Organizing Committee: T Kakeshita, H Araki, H Fujii, S Fujimoto, Y Fujiwara, A Hirose, S Kirihara, M Mochizuki, H Mori, T Nagase, H Nakajima, T Nakano, R Nakatani, K Nogi, Y Setsuhara, Y Shiratsuchi, T Tanaka, T Terai, H Tsuchiya, N Tsuji, H Utsunomiya, H Yasuda, H Yasuda (Osaka University) Executive Committee: T Kakeshita, S Fujimoto, Y Fujiwara, A Hirose, T Tanaka, H Yasuda (Osaka University) Conference Secretariat: Y Fujiwara (Osaka University) Proceedings Editors: T Kakeshita and Y Fujiwara (Osaka University)

This study examined the effects of amenorrhea on mucosal immune function and susceptibility to upper respiratory tract infection (URTI) in elite female distance runners. Based on their menstrual cycles during the prior year, 21 elite, collegiate, female distance runners were designated as eumenorrheic runners (ERs; n = 8; 19.9 ± 0.8 years) or amenorrheic runners (ARs; n n = 13; 20.0 ± 0.3 years). Resting saliva and blood samples were collected in the morning. The secretory immunoglobulin A (SIgA) concentration was measured using enzyme-linked immunosorbent assay. The SIgA secretion rate was calculated. Serum 17β-estradiol concentrations and serum progesterone concentrations were measured using radioimmunoassay. Subjects reported the appearance of URTI symptoms (sore throat, headache, runny nose, coughing, or fever), if any, during the prior month. The serum estradiol concentration and salivary SIgA secretion rate were significantly lower for ARs than for ERs (p < 0.05). Serum progesterone concentration was not significantly different between groups. Higher frequencies of headache, runny nose, coughing, and fever were observed in ARs than in ERs. Results show that athletic amenorrhea with low estrogen might accelerate downregulation of mucosal immune function in athletes and enhance susceptibility to infection.

Department of Ophthalmology, Osaka University Graduate School of Medicine, Osaka, Japan. Correspondence: Kohji Nishida, MD, PhD, Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan (e-mail: [email protected]). Supported in part by the Japan Agency for Medical Research and Development (AMED) under Grant Number JP19bm0504005 and JP19bk0104080, the Japan Science and Technology Agency (JST), The Ministry of Health, Labour and Welfare of Japan, and Grants-in-Aid for Scientific Research from The Ministry of Education, Culture, Sports, Science and Technology of Japan. The author has no conflicts of interest to disclose.

Artificial Intelligence Uncovered Clinical Factors for Cardiovascular Events in Myocardial Infarction Patients with Glucose Intolerance in the Cohort of Two Large-Scale ABC and PPAR Clinical Trials

The open reading frame of TTHA0613 from Thermus thermophilus HB8, consisting of 94 amino acid residues, was annotated as a hypothetical protein, and its function was not inferred. A PSI-BLAST search,1 using the amino acid sequence of TTHA0613 as the query, identified a total of 10 homologues with E-values below 0.005. In the Clusters of Orthologous Groups of proteins2 database, two of these homologues belong to COG0607: rhodanese-related sulfurtransferases. Rhodaneses catalyze the transfer of the sulfane sulfur from thiosulfate to cyanide, forming thiocyanate and sulfite in vitro. The 3-mercaptopyruvate sulfurtransferases (MST), which share strong sequence similarity with the rhodaneses, catalyze the same sulfane sulfur transfer reaction, using 3-mercaptopyruvate as a sulfur donor in vitro.3 Rhodanese homology domains are conserved in all three kingdoms. Their in vivo functions have been seriously debated. The ∼110-amino acid rhodanese homology domains exist as single domain proteins, as tandem repeats with a C-terminal domain hosting the properly structured active-site Cys residue, and as members of multidomain proteins. The rhodanese domain fold was first observed in the crystal structure of bovine mitochondrial rhodanese (Rhobov).4 Subsequently, that of Azotobacter vinelandii rhodanese (RhoA) was determined.5 Each structure consists of two rhodanese domain folds in the N- and C-terminal halves of the polypeptide. On the other hand, GlpE from Escherichia coli, which displays rhodanese activity in vitro,6 is composed of a single rhodanese domain fold.7 Rhodanese homology domains were also observed in the crystal structures of the human Cdc25a8 and Cdc25b9 phosphatases. Several genes encoding distinct rhodanese homology domains exist in the same genomes. This fact suggests that the various rhodanese homology domains might have diverse biological functions.10 Here we report the crystal structure of TTHA0613 from Thermus thermophilus HB8 at 2.0 Å resolution, and discuss our sequence and structural comparisons with its homologues. The open reading frame of TTHA0613 from T. thermophilus HB8 was cloned into the pET11a expression vector (Novagen). SeMet-substituted proteins were produced in E. coli Rosetta834 (DE3). The cell lysate was heated at 70°C for 15 min, and the proteins were purified by a series of HiTrap Q HP, Resource Q, and Superdex 75 HR 10/30 (Amersham Biosciences) column chromatography steps. The purified protein was concentrated to 12.7 mg/mL using an Amicon Ultra 4 filter (Millipore). The crystals were obtained by the microbatch method at 20°C. The precipitant solution was 100 mM MES-NaOH (pH 6.0–6.3) containing 35–40% polyethylene glycol 4000. The crystal was cryo-cooled to −173°C after soaking in a cryoprotectant solution [95% (v/v) Paraton-N and 5% (v/v) glycerol]. X-ray diffraction data of the SeMet-derivative crystal were obtained on beamline BL26B1 at the SPring-8 (Harima, Japan), using a Jupiter210 CCD detector (RIGAKU/MSC). Diffraction data were processed with the HKL200011 program suite. The crystal structure was solved by the multiwavelength anomalous dispersion (MAD)12 method, using the programs SOLVE13 and RESOLVE.14 The refinement was carried out using the program CNS.15 The final model comprises 173 amino acid residues and 132 water molecules in the asymmetric unit. Residues 1, 26–27, and 91–94 of molecule A, and residues 26–28 and 90–94 of molecule B were not visible, due to disorder, and were excluded from the final model. Data collection, MAD phasing, and refinement statistics are summarized in Table I. We have determined the crystal structure of TTHA0613 from T. thermophilus HB8 at 2.0 Å resolution. The crystal contains two molecules, A and B, per asymmetric unit [Fig. 1(A)]. The root mean square deviations of all of the atoms between molecule A and B are 0.65 Å. The monomeric structure consists of a five-stranded parallel β-sheet, surrounded by three α-helices [Fig. 1(B)]. As expected from the PSI-BLAST search results, the structure adopts a single rhodanese domain fold. A DALI16 search revealed that TTHA0613 shares high structural similarity to rhodanese homology domains. The Z-scores are 10.4 for GlpE from E. coli, 9.9 for the catalytic domain of RhoA, 9.6 for the catalytic domain of Rhobov, 8.7 for MST from Leishmania major,17 8.3 for Sud and 7.5 for Cdc25a. TTHA0613 tends to be structurally more similar to the sulfurtransferases than the phosphatase. A structure-based sequence alignment of TTHA0613 and the rhodanese homology domains is shown in Figure 1(C). The putative active-site loop motif of TTHA0613 (CEKGLL) lacks similarity to the other catalytic rhodanese homology domains with structures that have already been determined [Fig. 1(C)]. However, the conformation of the putative active-site loop closely matches those of the other rhodaneses. The Cα–Cα RMSDs calculated over the six residues of the active-site loop yielded the small values of 0.23 Å for GlpE, 0.12 Å for RhoA, 0.22 Å for Rhobov, 0.15 Å for MST from L. major, and 0.68 Å for Cdc25a. The conformation of the putative active site is also similar to those of the sulfurtransferases. A sequence comparison between TTHA0613 and its six closest homologues, each with a single rhodanese homology domain, is shown in Figure 1(D). The high sequence homologies of β4 and α3 are probably necessary to form the proper active-site loop conformation. Furthermore, high sequence homology exists in the N-terminal β1–α1–β2 region. This region might participate in interactions with other proteins that bind to the in vivo substrate. (A). Crystal packing of the molecules in the asymmetric unit. Molecules A and B are colored pink and blue, respectively. (B) Ribbon representation of the TTHA0613 monomer (stereo view). The α-helices and β-strands are colored red and green, respectively. The figure was generated with the graphics program CueMol (http://cuemol.sourceforge.jp/en/). (C) Structure-based sequence alignment of TTHA0613 and rhodanese homology domains. (D) Sequence comparison between TTHA0613 and its six closest homologs, which each have a single rhodanese homology domain. The figure was constructed by PSI-BLAST1 followed by CLUSTALW.18 Ll, Lactococcus lactis subsp; Aa, Arthrobacter aurescens; Tt, Thermus thermophilus; Lp, Legionella pneumophila subs; Ss, Streptococcus suis; Ps, Pseudomonas syringae. Strictly conserved and similar residues are represented within a red box and by a red letter, respectively. The figure was generated with ESpript 2.0.19 We thank M. Yamamoto for data collection at BL26B1 of SPring-8. This work was supported by the RIKEN Structural Genomics/Proteomics Initiative (RSGI), the National Project on Protein Structural and Functional Analysis, Ministry of Education, Culture, Sports, Science and Technology of Japan.

Since a fullerene was discovered in 1985, fullerene has attracted much attentions [1]. A unique feature of the fullerene that has the spherical structure with hollow interior allows a fullerene to accommodate any other atoms and molecules. Especially, the fullerenes containing other atoms which is called as endohedral fullerene have been expected to be applied in medical uses, electronics systems and so on [2]. In many cases, endohedral fullerenes have been synthesized by ion implantation, co-vaporization of carbon and metal atoms [3], or ion plasma irradiation technique for non-metal material [4]. However, these procedures have remained several issues. One of the critical issues is low production rate for endohedral fullerene synthesize due to difficulties in controlling a physical and a chemical property during synthesizing process. To increase the production efficiency of endohedral fullerene, it is required to optimize elementary process between fullerenes and injected atoms.Under these situations, we recently developed a novel procedure to synthesize endohedral fullerenes that induce the collision of vaporized fullerenes and ablated particles produced by the laser ablation of a solid material. Here, a Q-switched Nd:YAG Laser (Continuum, Powerlite DLS8000) with a wavelength of 1064 nm, a pulse-width of 9 ns, a laser energy of up to 1 J, and a pulse-frequency of up to 10Hz was used. Laser light was focused to be 50 m in diameter on the target surface. In this procedure, N@C60 was synthesized using the ablation of boron nitride material. From the previous studies [4], it is quite important for encapsulated particles to have about 100 eV. Optical emission spectrum of ablated particles by a polychromator (Bunkokeiki, CT-1000) with an intensified CCD detector (Princeton Instruments, PI-Max) was measured, and Doppler shift of the spectrum was estimated. The result showed that the kinetic energy of nitrogen ions was in the range of 20 to 65 eV by controlling the energy of the incident laser beam. The synthesis of N@C60 was conducted with a nitrogen kinetic energy of 65 eV, fullerenes vaporizing temperature of 800 degree Celsius, and a total reaction time of one hour. By optimizing the kinetic energy of ablated particles, synthesis of N@C60 was succeeded [5]. Existence of N@C60 in the toluene solution was confirmed by an electron spin resonance measurement (Elexsys E500, Bruker) without purification by high performance liquid chromatography [6]. The purity of produced N@C60 was two orders of magnitude higher than that achieved using alternative physical synthetic methods that use solid materials, such as arc discharge and co-evaporation methods. However, the purity must be increased by at least two orders of magnitude for the commercial use. Therefore, it is necessary to enhance the purity by optimizing experimental conditions such as laser energy, laser wavelength, amount of ion flux with appropriate energy, repetition rate of laser pulses, laser duration time, area of the laser ablation, and so on. In the meeting, novel synthesis procedure for endohedral-fullerenes using laser ablation plasma from solid material and vaporized fullerenes will be presented.AcknowledgmentsThis study was financially supported by JSPS KAKENHI Grant No. 25287157 from the Ministry of Education, Culture, Sports, Science and Technology of Japan.[1] H. W. Kroto, et.al., Nature 318, 162-163 (1985).[2] H. Shinohara, Rep. Prog. Phys. 63, 843-892 (2000).[3] Alexey A. Popov, et.al., Chem. Rev. 113, 5989-6113 (2013).[4] S.C. Cho, et.al., J. Appl. Phys. 117, 123301 (2015).[5] H. Itagaki, et al., AIP Advances 9-7 (075324), 1-7 (2019).[6] T.A. Murphy, et al., Phys. Rev. Lett. 77-6, 1075 (1996).

Pacemaker implantation with right ventricular pacing is widely used in clinical practice in the treatment of bradycardia, especially in patients with symptomatic AV block. However, right ventricular apical pacing (RVAP) sometimes induces electromechanical dyssynchrony, leading to adverse clinical impacts on clinical outcomes, including an increased risk of new-onset atrial arrhythmias. Physiological conduction system pacing (CSP), His bundle pacing (HBP), and left bundle area pacing (LBBAP) are recommended for patients with reduced left ventricular (LV) systolic function and substantial ventricular pacing (>20%) since CSP has been reported to improve clinical outcomes compared with RVAP. Although several studies suggest that CSP is associated with a lower incidence of new-onset atrial arrhythmias detected as atrial high-rate episodes (AHRE), the performance between HBP pacing and LBBAP on the risk of new-onset AHRE remains unclear. Pestrea et al. showed that HBP and LBBAP were associated with a similar incidence of device-detected new-onset AHRE during a medium-term follow-up period in patients with atrioventricular block.1 They compared the incidence of device-detected new-onset AHRE between the two groups of patients after HBP (n = 59) and those after LBBAP (n = 83) during a mean follow-up of 624 days. New-onset AHRE occurred in 8 (13.5%) in the HBP group and in 14 (16.8%) in the LBBAP group. Multivariable Cox regression analysis showed that HBP and LBBAP had similar predictive values for device-detected new-onset AHRE. Moreover, there was no significant difference between the two groups regarding the total burden of AHRE, which was less than 1% in almost all patients with new-onset AHRE, although there were several limitations such as using different criteria of the current 2020 ESC guideline, which indicated the device-programmed rate criterion for AHRE is greater than or equal to 175 bpm and the duration criterion is greater than or equal to 5 min.2 Cardiac electronic implantable devices such as pacemakers have the ability to monitor rhythm abnormalities, which allow us to recognize a new entity of AHRE easily. From a clinical point of view, AHRE has been associated with the development of clinical atrial fibrillation (AF) and an increase in stroke and death risk. There have been reports about the risk factors for AHRE including older age, left atrial volume, prior history of AF, white cell count, high levels of C reactive protein, and CHADS2 score.3 As for the issue related to new-onset AHRE after implantation, a high burden of RVAP is a risk for increased AHRE since RVAP induces paradoxical septal motion and ventricular dyssynchrony. As a result, increased filling pressure in each heart chamber leads to electric remodeling of the left atrium. CSP restores ventricular contraction synchrony by pacing the His-Purkinje conduction system directly, which allows for rapid and widespread dissemination of ventricular activation throughout the ventricle. CSP, including both HBP and LBBAP, has several advantages of LV function, subsequent events of heart failure hospitalization, and the incidence of new-onset AHRE during a follow-up compared to RVAP, especially in patients with LV dysfunction and a high burden of right ventricular pacing.4 There are some limitations of relatively low procedural success rates and the development of a high and unstable pacing threshold in terms of HBP, although it has demonstrated several clinical benefits. In addition, HBP is inefficient as physiological pacing if a patient has infra-Hisian distal conduction block. There are several strong points of LBBAP, including the wide target area of left bundle and Purkinje fibers on the LV septum and a stable low pacing threshold with no significant sensing issues. In fact, a previous meta-analysis revealed that LBBP was significantly associated with higher implant success rates (relative risk: 1.12), lower capture threshold at implantation (mean difference [MD]: 0.63 V at 0.5 ms) and lower capture threshold at follow-up (MD: 0.76 V at 0.5 ms) compared with HBP.5 These data suggest that pacing characteristics are better in LBBAP than in HBP; however, the incidence of new-onset AHRE during a follow-up may be similar between patients with LBBAP and those with HBP by taking the current issue into consideration. At this moment, it remains controversial which we should select: LBBAP or HBP. Large-scale, randomized control studies are warranted to reveal the true answer. Dr. Noda reports Grants-in-Aid for Scientific Research (22K08092) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and declares receiving fees for speakers from Medtronic Japan and Biotronik Japan.

The global mpox outbreak (2022-2024) highlights the need for effective and safe vaccines, particularly for vulnerable populations. The LC16m8 vaccine, an attenuated vaccinia virus strain for smallpox, shows promise in inducing immunity against the monkeypox virus (MPXV). We conducted a comprehensive immunological evaluation of LC16m8 in mice, non-human primates, and humans. LC16m8 induced strong humoural responses in BALB/c, C57BL/6J, and CAST/EiJ mice, targeting MPXV H3, A35, and M1R antigens, promoting germinal centre B cells and follicular helper T cells, essential for long-term immunity. Vaccinated CAST/EiJ mice showed reduced lung MPXV viral loads, demonstrating efficacy. In humans, LC16m8 enhanced neutralising antibodies against multiple MPXV clades, suggesting broad protection. In cynomolgus monkeys, systemic administration caused localised pox lesions without significantly affecting weight, temperature, or haematological parameters. This cross-species immunological analysis provides preclinical and early clinical insights into LC16m8's efficacy and safety against mpox. While LC16m8 enhanced antibody responses against MPXV clade Ia and Ib, further studies are required to evaluate its efficacy, particularly in naive and immunocompromised populations. This research was supported by AMED under Grant Numbers JP243fa727002, JP243fa727001s0703, and JP243fa627001h0003 (K.J.I), JP24jf0126002, JP24fk0108690, JP243fa627001h0003, and JP243fa727002 (K.S), JP243fa727002 (Y.T.), JP243fa727002 and JP243fa627007h0003 (Y.Y.), and by the Research Support Project for Life Science and Drug Discovery (BINDS) from AMED under Grant Number JP23ama121011 (J.T.), and JP23ama121010 (T.S.), and by the Ministry of Education, Culture, Sports, Science and Technology in Japan under Grant Number 23K06577 (E.S.). AMED under Grant Number JP233fa827017 and JP243fa827017 (F.Y.), JP22fk0108501 (M.K.).

Identifying clustering of cholera cases using geospatial analysis in Kolkata and surrounding districts: data from patients at tertiary care referral hospitals

The career educational program of Anan College of Technology was adopted in 2006 by the Ministry of Education, Culture, Sports, Science and Technology in Japan as the Support Program for Distinctive University Education (Good Practice in Modern Educational Needs) . The original program of career education is operated for students in lower grades and the support system is provided for the recruiting activity of students in the higher grades. Students can be instilled with a sense of purpose by receiving education regarding the structure of industrial society as an early step of the curriculum.

The Ministry of Education, Culture, Sports, Science and Technology in Japan (2011) started the ‘Foreign Language Activities’ in fifth grade based on the new Courses of Study in April of 2011. The ministry has considered beginning this course in third grade and the ‘English Course’ in 5th grade in order to improve students’ reading, writing, listening, and speaking skills. The ministry also developed supplemental teaching instruments, such as the CALL system. The purpose of this study is to model and analyse the effectiveness of a speaking-pen on English learning among elementary school children in consideration of individual differences using a Liner Mixed-Effect Model. The authors constructed models representing students’ overall abilities in four English skills, and analysed the effectiveness of the tools such as a speaking-pen and an audio CD on English learning in consideration of students’ backgrounds including their English learning experiences and individual differences.

Introduction Regular aerobic exercise reverses aging-induced deterioration of arterial stiffness via an increased arterial nitric oxide (NO) production. Concurrently, asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase, was decreased by aerobic exercise training. A recent study showed that the NOx/ADMA ratio reflects endothelial function and may be an index of the states of cardiovascular disease. However, whether changes in the NOx/ADMA ratio by aerobic exercise training are associated with a decrease in arterial stiffness in healthy middle-aged and older male and female adults remains unclear. Purpose This study aimed to clarify whether the relationship between plasma ADMA and NOx levels affected aerobic exercise training-induced reduction of arterial stiffness in middle-aged and older adults. Additionally, we examined whether the effect of AT on circulating ADMA levels differed according to sex. Methods Thirty-one healthy middle-aged and older male and female subjects (male = 13, female = 18, 66.4±1.3 years) participated in the study. The subjects were randomly divided into 2 groups: a training group (n=16 [male = 6 / female = 10], 64.8±2.0 years) and a control group (n=15 [male = 7 / female = 8], 68.1±1.6 years). Subjects in the training group performed the AT program, which consisted of cycling on a leg ergometer at 60–70% of peak oxygen uptake (V(·)O2peak)for 45 min/day, 3 days/week for 8 weeks. Before and after the 8-week aerobic exercise training intervention, V(·)O2peak, plasma ADMA levels and plasma NOx levels were measured in all subjects. Also, carotid β-stiffness as an index of arterial stiffness was determined with ultrasonography. Results Aerobic exercise training significantly increased V(·)O2peak (P&amp;lt;0.05) and decreased carotid b-stiffness (P&amp;lt;0.01). Moreover, plasma ADMA levels were significantly decreased, and plasma NOx levels and NOx/ADMA ratio were significantly increased by aerobic exercise training (P&amp;lt;0.01). Additionally, no sex differences in aerobic exercise training-induced changes in circulating ADMA and NOx levels, NOx/ADMA ratio, and carotid β-stiffness were observed. Furthermore, the aerobic exercise training-induced increase in circulating ADMA levels was negatively correlated with the increase in circulating NOx levels (r=−0.414, P&amp;lt;0.05), and aerobic exercise training induced increase in NOx/ADMA ratio was negatively correlated with the decrease in carotid β-stiffness (r=−0.514, P&amp;lt;0.01). Conclusion These results suggest that higher NOx/ADMA ratio affects aerobic exercise training-induced reduction of arterial stiffness, regardless of sex in middle-aged and older adults. Funding Acknowledgement Type of funding sources: Other. Main funding source(s): This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan

There is a high rate of comorbidity between autism spectrum disorder (ASD), obsessive-compulsive disorder (OCD) and eating disorders (EDs). Furthermore, neuropsychological studies and genetic studies support the presence of shared background among these disorders.1-4 All of these findings suggest shared pathophysiology of these disorders. NBEA encodes neurobeachin and has been strongly implicated in ASD risk by the identification of de novo loss-of-function variants of this gene in multiple unrelated cases.5-7 Neurobeachin regulates recycling of GluN2B-NMDA receptors, and is involved in neurotransmission, synaptic formation and plasticity by a haploinsufficient genetic mechanism.8 Although we also identified an ASD patient with rare deletion disrupting this gene in a study of copy number variation (CNV),2 only limited knowledge was available about the clinical implications of NBEA deletion. It is important to accumulate high-quality case reports to improve the clinical management of patients with such CNVs. Therefore, we report a 10-year clinical course of an ASD patient with NBEA deletion, who had comorbidities of OCD, ED, and intellectual disability (ID). The patient was a 41-year-old Japanese female. Her father had a broad autism phenotype. During infancy, language developmental delay, insistence on the same clothes and foods, tactile hypersensitivity, and impairments in communication and social interactions were noticed. She had very little social contact with her peers throughout her school career. After she graduated from junior college, she had several part-time jobs but soon quit because of interpersonal conflicts in the workplace. Her developmental history is summarized in Table S1. At the age of 31, she had recurrent episodes of overeating and rapidly gained weight. In response to this sudden weight gain, she began to restrict her food intake. Thereafter, her body mass index (BMI) decreased to 12.3 from 21.1 kg/m2 in 6 months. She was hospitalized with a diagnosis of unspecified feeding or eating disorder according to the Diagnostic and Statistical Manual of Mental Disorders-Fifth Edition (DSM-5) diagnostic criteria. Additionally, during the hospitalization, she was diagnosed with ASD. Psychoeducation about ED and ASD and supportive psychotherapy were provided. Although she did not meet the diagnostic criteria for major depressive disorders, pharmacotherapy was initiated to target her depressive symptoms. Gradually, her food intake increased, her BMI recovered to 15.4 kg/m2, and she was discharged in 3 months. After regaining weight, overeating was observed, but improved after discontinuing olanzapine medication. The patient continued to receive outpatient treatment, but obsessive-compulsive symptoms with obsessive thoughts appeared. She developed compulsive behaviors such as hand or body washing, and she spent a lot of time performing such behaviors. She was diagnosed with OCD and received medication (selective serotonin reuptake inhibitors and atypical antipsychotics) and cognitive behavioral therapy, but the symptoms were intractable. Since her last hospitalization, her symptoms have been under control. The clinical time course is summarized in Fig. 1a. Her intelligence quotient score was 62 (Table S2). Array comparative genomic hybridization using NimbleGen 720k Whole-Genome Tiling Array detected a 447-kb deletion spanning exons 11–41 of NBEA2 (Fig. 1b,c). Parental CNV analysis revealed that NBEA deletion was paternally inherited (Fig. 1d). Gene expression analysis revealed that the NBEA mRNA expression level was reduced below the 15th percentile of the control group (N = 29) (Fig. 1e, Appendix S1). We reported an ASD patient with OCD, ED, ID, and NBEA deletion. Neuropsychological studies support the presence of shared traits among ASD, OCD, and EDs.1 As shown in Table S2, ASD patients with NBEA deletion have been reported.6, 7 This is the first case report of an adult with NBEA deletion. Though previously reported cases did not mention OCD and EDs, these patients had not reached the peak age of onset of these disorders. Heterozygous Nbea knockout mice exhibit not only autistic behaviors but also impaired extinction of contextual fear memory, which is associated with OCD, and abnormal feeding behavior, episodically overconsuming tastants.9, 10 Thus, the clinical phenotypes of the patient were similar to those of heterozygous Nbea knockout mice, raising the possibility that exonic NBEA deletion was responsible for the symptoms in this patient. As Nbea knockout mice show hyperphagia, patients with NBEA deletion may be more prone to episodic overeating. The patient we reported showed rapid weight gain during olanzapine medication. Choosing medicines carefully and monitoring weight are important. In conclusion, NBEA deletion may be a shared genetic underpinning of ASD, OCD, and EDs. The accumulation of clinical data of patients with NBEA deletion including the present case may contribute to establishment of appropriate clinical management of these patients in the future. We thank the patient and her family members for participating in this study. We also thank Mami Yoshida, Kiyori Monta, and Yukari Mitsui for their technical assistance. This research was supported by research grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) and the Ministry of Health, Labour and Welfare of Japan; the Japan Agency for Medical Research and Development (AMED) under Grant Nos. JP20dm0107087, JP21wm0425007, JP21dm0207075, JP21ak0101113, JP21dk0307075, JP20dk0307081, JP21dk0307103, JP21ek0109488, JP21km0405216, and JP21ek0109411; the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Nos. 17H05090, 15K19720, 18H04040, 21K07543, 21H00194, 21H04815, and 20K20602; the Uehara Memorial Foundation; and the SENSHIN Medical Research Foundation. H.K., I.K., A.Y., K.I., K.I., and B.A. declare no conflict of interest. N.T. has received research support or speakers' honoraria from, or has served as a consultant to, Otsuka, Shionogi, Takeda, Janssen, and Pfizer, outside the submitted work. T.O. has received lecture fees from Shionogi and Mochida, outside the submitted work. N.O. has received research support or speakers' honoraria from, or has served as a consultant to, Sumitomo Dainippon, Eisai, Otsuka, KAITEKI, Mitsubishi Tanabe, Shionogi, Eli Lilly, Mochida, DAIICHI SANKYO, Nihon Medi-Physics, Takeda, Meiji Seika Pharma, EA Pharma, Pfizer, MSD, Lundbeck Japan, Taisho Pharma, outside the submitted work. This study was approved by the ethics committee of the Nagoya University Graduate School of Medicine and this study complied with all the provisions of the Declaration of Helsinki. The patient and her parents gave written informed consent for genetic testing and publishing of this report. Appendix S1. Supplementary Appendix. Methods of gene expression analysis Appendix S2. Author Contributions Table S1. Time course of the patient's developmental history. Table S2. Clinical summary of cases with NBEA deletion Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Brown and beige adipose tissue (BAT) promotes energy expenditure by thermogenesis mediated by uncoupling protein (UCP)-1. BAT have an endocrine system and secreted “batokines” play a crucial role in regulating metabolism. Ependymin-related protein 1 (EPDR1), which was recently identified as a novel batokine, is involved in development into a functional thermogenic adipocyte. However, the effects of habitual aerobic exercise on EPDR1 secretion of BAT remain unclear. PURPOSE: To clarify whether the secreted EPDR1 is associated with the fat burning and browning by habitual aerobic exercise in obese mice. METHODS: Eight-week-old ApoE-/- mice were used as an obese model and were randomly divided into the following two groups: sedentary control (ApoE-/- SED, n = 6) group and 16-wk voluntary wheel running aerobic exercise (ApoE-/- AT, n = 6) group. ApoE-/- mice of two groups were fed a high-fat diet during the experimental period. Age-matched C57BL/6 J wild-type mice were fed a normal chow and were used as a healthy sedentary control group (WT-SED, n = 6). RESULTS: Epididymal fat mass of the ApoE-/- SED group was significantly higher compared with WT-SED group (p < 0.05), and that of ApoE-/- AT group was significantly lower compared with ApoE-/- SED group (p < 0.05). Protein expression levels of PGC1-α, UCP-1, and EPDR1 in epididymal WAT (eWAT) were significantly higher in the ApoE-/- AT group as compared with ApoE-/- SED group (p < 0.05). Additionally, EPDR1 mRNA levels in interscapular BAT was higher in the ApoE-/- AT group as compared with ApoE-/-SED group. Plasma EPDR1 levels were higher in the ApoE-/- AT group as compared with ApoE-/-SED group (p < 0.05). Furthermore, EPDR1 protein expression levels in eWAT were positively correlated with PGC1-α (r = 0.431, p < 0.05) and UCP-1 (r = 0.325, p < 0.05) protein expression levels in eWAT. CONCLUSIONS: These findings suggest that the secreted EPDR1 is associated with the fat burning and browning via increase in UCP-1 protein expression by habitual aerobic exercise in obese mice. Supported by: Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (KAKENHI: 23 K24744 to M. Iemitsu)

Subchondral Fracture of the Femoral Head in a High School Badminton Player: A Case Report.