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In this study, we used the placement test scores, e-learning history, and posttest scores obtained in the pre-enrollment education at University A conducted in 2015, 2016, and 2017 to exploratorily classify seven types of students according to learning habits (types). It was revealed that more than 80% (with the exception of English in 2016) of the highest placement test scores (Japanese ≥ 90, English ≥ 74 in this study) were of learners with learning habits (types) of long-term completion (LTrf) or mid-term completion (MTrf). Although the percentage of learners with LTrf or MTrf learning habits (types) gradually decreased as placement test scores declined, there was no significant change in learning habit trends and learning habits (types) other than for LTrf or MTrf. In addition, learners with large increases in posttest scores (posttest score - placement score) after completing e-learning were classified with learning habits (types) of LTrf or MTrf. However, it is not only the learners who improved their test scores that were progressing without bias during the learning period (LTrf or MTrf). It was also found that more than 65% progressed in their learning (logged in) without being biased during the learning period. These findings suggest that there is no significant difference between learners' learning habits (types), placement test scores, and test score growth.

OBJECTIVE Various complications have been reported in the treatment of pediatric spinal deformities. Among these, instrument-related complications could be critical concerns and risks of reoperation. This study aimed to identify the incidence and causes of complications after primary definitive fusion for pediatric spine deformities. METHODS The authors retrospectively collected data from 14 institutions about patients who underwent primary definitive fusion between 2015 and 2017. There were 1490 eligible patients (1184 female and 306 male), with a mean age of 13.9 years. The incidence, causes, and reoperation rates were analyzed according to 4 etiologies of pediatric spine deformity (congenital, neuromuscular, syndromic, idiopathic). The complications were also categorized as screw-, hook-, or rod-related complications, implant loosening or backout, and junctional problems. RESULTS The incidence of overall instrument-related complications was 5.6% (84 cases). Regarding etiology, the incidence rates were 4.3% (idiopathic), 6.8% (syndromic), 7.9% (congenital), and 10.4% (neuromuscular) (p < 0.05). The most common causes were pedicle screw malposition (60.7%), followed by implant backout or loosening (15.4%), junctional problems (13.1%), rod breakage (4.8%), and other complications (6.0%). Univariate analysis showed that etiology, type of deformity (kyphosis), surgical procedure, operation time, and estimated blood loss were significant factors. Multivariate analysis revealed that etiology (neuromuscular), surgical procedure (combined approach), and operation time (> 5 hours) remained as significant risk factors. Among all patients with instrument-related complications, 45% (38/84) required revision surgery. Of these cases, > 50% were related to pedicle screw malposition. Medial breach was the most common complication regardless of location, from upper thoracic to lumbar spine. CONCLUSIONS Pedicle screw malposition was the primary cause of overall complications and subsequent reoperation. In addition to more precise screw insertion techniques, meticulous confirmation of pedicle screw placement, especially of medial breach, may reduce the overall instrument-related complications and revision rates.

Hydnum repando-orientale is an East Asian species closely related to H. boreorepandum and H. repandum; all three species produce edible mushrooms. We identified two ecological groups of H. repando-orientale in Japan: a temperate group occurring in Fagaceae-dominated forest at 1900 m a.s.l. (ROC). We re-examined the taxonomy of the two ecological groups of H. repando-orientale using integrative approaches. Phylogenies of the two ecological groups and other related species were inferred from the internal transcribed spacer (ITS) and gene portions encoding the large subunit of nc rRNA (LSU), translation elongation factor-1 alpha (TEF1), RNA polymerase II largest subunit (RPB1), and RNA polymerase II second-largest subunit (RPB2). The concatenated phylogenetic tree separated the two ecological groups into well-supported sister clades. Also, species delimitations based on the topological congruence (GCPSR) and multispecies coalescent model (GMYC and BP&P) supported to separate the two ecological groups. Morphological analysis showed that ROC specimens had significantly larger basidiospores, compared with ROF specimens. Mon-mon mating tests using six ROF, three ROC, and three H. boreorepandum strains each showed independent incompatible groups, whereas one ROC strain showed compatibility with both ROC and ROF populations. Based on these results, we defined the ROC group as a new species, H. subalpinum. Because H. repando-orientale and H. subalpinum have smaller genetic divergence in nc rDNA and maintain slight sexual compatibility, they may have recently speciated in East Asia.

Adolescent idiopathic scoliosis (AIS) is a serious health problem affecting 3% of live births all over the world. Many loci associated with AIS have been identified by previous genome wide association studies, but their biological implication remains mostly unclear. In this study, we evaluated the AIS-associated variants in the 7p22.3 locus by combining in silico, in vitro, and in vivo analyses. rs78148157 was located in an enhancer of UNCX, a homeobox gene and its risk allele upregulated the UNCX expression. A transcription factor, early growth response 1 (EGR1), transactivated the rs78148157-located enhancer and showed a higher binding affinity for the risk allele of rs78148157. Furthermore, zebrafish larvae with UNCX messenger RNA (mRNA) injection developed body curvature and defective neurogenesis in a dose-dependent manner. rs78148157 confers the genetic susceptibility to AIS by enhancing the EGR1-regulated UNCX expression. © 2022 American Society for Bone and Mineral Research (ASBMR).

Background Cast and brace treatment is a conservative treatment for early-onset scoliosis (EOS). We analyzed the clinical results of this treatment when extended into patients' schooldays. Methods Twenty-two children with EOS underwent alternatively repetitive cast and brace treatment (ARCBT). Cast was applied under general anesthesia and remained for some weeks, and the brace was continuously worn, which was repeatedly applied when scoliosis progressed. The change in the major curve angle, requirement for surgical intervention, and complications of ARCBT were analyzed. Results Six patients had idiopathic scoliosis (IS), 9 had syndromic scoliosis (SS), and 7 had neuromuscular scoliosis. Cast treatment started at 4.9±2.1 years of age, and the patients were followed up for 8.3 years. The average total number of cast applications was 5.5 per patient. The mean major scoliosis angle was 45.4±12.5 degrees at the first visit; this was corrected to 26.5±8.7 degrees at the first cast application and progressed to 75.4±22.1 degrees at the final visit. Nine patients whose scoliosis progressed finally underwent surgery. The mean age at surgery was 11.0 years (range, 8.8 to 13.1 y), and the mean period from first cast application to surgery was 6.1 years (range, 5.0 to 8.9 y). Four patients' IS rapidly progressed after 9 years of age, and all of them underwent final fusion surgery with a correction rate of 56.9%. Among patients with SS, 1 final fusion, and 2 growth-friendly surgeries were performed at a mean age of 9.2 years; the correction rate was 21.5%. Among patients with neuromuscular scoliosis, 2 final fusion surgeries were performed at a mean age of 11.3 years; the correction rate was 30.7%. The cast-related complication rate was 5.0%, and most complications were skin problems. Conclusions Long-term ARCBT extended into schooldays might be effective for IS. This should be considered as a time-saving treatment option for EOS. Level of evidence Level IV-case series.

Adolescent idiopathic scoliosis (AIS) is a common disease causing three-dimensional spinal deformity in as many as 3% of adolescents. Development of a method that can accurately predict the onset and progression of AIS is an immediate need for clinical practice. Because the heritability of AIS is estimated as high as 87.5% in twin studies, prediction of its onset and progression based on genetic data is a promising option. We show the usefulness of polygenic risk score (PRS) for the prediction of onset and progression of AIS. We used AIS genomewide association study (GWAS) data comprising 79,211 subjects in three cohorts and constructed a PRS based on association statistics in a discovery set including 31,999 female subjects. After calibration using a validation data set, we applied the PRS to a test data set. By integrating functional annotations showing heritability enrichment in the selection of variants, the PRS demonstrated an association with AIS susceptibility (p = 3.5 × 10-40 with area under the receiver-operating characteristic [AUROC] = 0.674, sensitivity = 0.644, and specificity = 0.622). The decile with the highest PRS showed an odds ratio of as high as 3.36 (p = 1.4 × 10-10 ) to develop AIS compared with the fifth in decile. The addition of a predictive model with only a single clinical parameter (body mass index) improved predictive ability for development of AIS (AUROC = 0.722, net reclassification improvement [NRI] 0.505 ± 0.054, p = 1.6 × 10-8 ), potentiating clinical use of the prediction model. Furthermore, we found the Cobb angle (CA), the severity measurement of AIS, to be a polygenic trait that showed a significant genetic correlation with AIS susceptibility (rg = 0.6, p = 3.0 × 10-4 ). The AIS PRS demonstrated a significant association with CA. These results indicate a shared polygenic architecture between onset and progression of AIS and the potential usefulness of PRS in clinical settings as a predictor to promote early intervention of AIS and avoid invasive surgery. © 2021 American Society for Bone and Mineral Research (ASBMR).

Despite the economic and ecological importance of Hydnum species, in vitro synthesis of ectomycorrhizae of this genus has not been reported due to difficulties in establishing pure cultures. We inoculated pure cultures of 12 monokaryotic and 3 dikaryotic mycelial strains of an undescribed Hydnum albidum-like species on roots of axenic Pinus densiflora seedlings to synthesize ectomycorrhizae and to evaluate their mycorrhiza-forming ability. Six months after inoculation, both monokaryotic and dikaryotic strains formed ectomycorrhizae with Hartig net hyphae at the root cortex. Monokaryotic and dikaryotic strains exhibited similar morpho-anatomical characteristics of ectomycorrhizae, with the exception for clamped septa of emanating and outer mantle hyphae in the latter. Between monokaryotic and descendant dikaryotic strains, there were no significant differences in number of mycorrhizae in pine seedlings, whereas monokaryotic strains showed a greater total number of root tips and lower colonization rates than the descendant dikaryotic strains. These results indicate that both monokaryotic and dikaryotic mycelia of the H. albidum-like species can form mycorrhizae under axenic condition, and that can be applied toward the cultivation of hedgehog mushrooms.

STUDY DESIGN A retrospective multicenter study. OBJECTIVE To determine the surgical site infection (SSI) rate, associated risk factors, and causative pathogens in pediatric patients with spinal deformity. SUMMARY OF BACKGROUND DATA There have been no extensive investigations of the risk factors for SSI in Japan. METHODS Demographic data, radiographic findings, and the incidence of SSI were retrospectively analyzed in 1449 pediatric patients who underwent primary definitive fusion surgery for spinal deformity at any of 15 institutions from 2015 to 2017. SSI was defined according to the US Centers for Disease Control and Prevention guideline. RESULTS The incidence of all SSIs was 1.4% and that of deep SSIs was 0.76%. The most common pathogenic microbes were methicillin-resistant staphylococci (n = 5) followed by gram-negative rods (n = 4), methicillin-sensitive staphylococci (n = 1), and others (n = 10). In univariate analysis, younger age, male sex, a diagnosis of kyphosis, type of scoliosis, American Society of Anesthesiologists (ASA) class ≥3, mental retardation urinary incontinence, combined anterior-posterior fusion, greater magnitude of kyphosis, three-column osteotomy, use of blood transfusion, and number of antibiotic administration were associated with the likelihood of SSI (all P

STUDY DESIGN A retrospective multicenter cohort study. OBJECTIVE The aim of this study was to identify the incidence and risk factors for UPROR within minimum 2-year follow-up in primary definitive fusion for pediatric spinal deformity. SUMMARY OF BACKGROUND DATA Several previous reports have elucidated the incidence of complications after pediatric scoliosis surgery; however, there has been no study that described the incidence and risk factors for unplanned return to the operating room (UPROR) with long-term follow-up in surgery for pediatric scoliosis with every etiology. METHODS We retrospectively extracted data of patients aged

We described the ectomycorrhizae (ECM) of the Japanese Lactarius section Deliciosi species L. akahatsu, L. hatsudake, L. laeticolor, and L. subindigo with the aim of promoting the use of mycorrhizae for taxonomy and expansion of available cultures. Lactarius tottoriensis (sect. Lactarius) was also included as a comparator. The ECM of the four species of sect. Deliciosi showed orange laticifers and plectenchymatous mantles, whereas those of L. tottoriensis showed white laticifers and pseudoparenchymatous mantles. The ECM of L. hatsudake and L. subindigo showed complex rhizomorphs with thicker walls and more developed vessel-like hyphae compared to the other three species. Cystidium-like cells were rarely found on the mantles of L. laeticolor and L. subindigo. Fungal cultures of the five Lactarius species from their ECM were morphologically characterized. Cultures produced laticifers on agar medium, but the five species showed different growth rates. Rhizomorphs of L. hatsudake and L. subindigo in cultures showed a similar morphology to their ECM. Molecular phylogenetic analyses of cultures from the ECM demonstrated that they were of the targeted Lactarius species and suggested the need for taxonomic studies of L. hatsudake, L. subindigo, and L. tottoriensis at the species and section levels.

Tricholoma foliicola was taxonomically reevaluated based on analyses of the holotype and newly collected materials. Basidiospores of T. foliicola were irregularly ellipsoid, showing a cyanophilic reaction with or without tubercles. Phylogenetic analyses of the internal transcribed spacer (ITS) and large subunit 28S regions of the fungal nuclear ribosomal RNA gene demonstrated that T. foliicola is a species of Gerhardtia characterized by irregularly shaped basidiospores. Thus, we transferred this species to Gerhardtia foliicola comb. nov. Cultured mycelia of G. foliicola on malt extract agar medium produced cystidia covered with granules and abundant thallic conidia (arthroconidia), with both schizolytic and rhexolytic secession.

Adolescent idiopathic scoliosis (AIS) is the most common pediatric spinal deformity. Several AIS susceptibility loci have been identified; however, they could explain only a small proportion of AIS heritability. To identify additional AIS susceptibility loci, we conduct a meta-analysis of the three genome-wide association studies consisting of 79,211 Japanese individuals. We identify 20 loci significantly associated with AIS, including 14 previously not reported loci. These loci explain 4.6% of the phenotypic variance of AIS. We find 21 cis-expression quantitative trait loci-associated genes in seven of the fourteen loci. By a female meta-analysis, we identify additional three significant loci. We also find significant genetic correlations of AIS with body mass index and uric acid. The cell-type specificity analyses show the significant heritability enrichment for AIS in multiple cell-type groups, suggesting the heterogeneity of etiology and pathogenesis of AIS. Our findings provide insights into etiology and pathogenesis of AIS., Adolescent idiopathic scoliosis (AIS) is a common pediatric disease leading to spinal deformities. Here, the authors report GWAS followed by genome-wide meta-analysis in up to 79,211 Japanese individuals, identifying 20 genetic loci for AIS, 14 of which were previously unreported, and perform in vitro validation for rs1978060.

The Entoloma clypeatum species complex, known as “Harushimeji” in Japan, associates with Rosaceae and Ulmaceae plant species. In this study, we successfully isolated cultures of this fungal group via basidiospore isolation from tentative four Harushimeji species using modified Norkrans's C (MNC) medium and MNC medium containing n-butyric acid. Colony formations were observed on 22 of 25 basidioma samples; however, most exhibited slow and unsteady growth. The isolated mycelia contained dikaryotic hyphae and were identified through molecular phylogenetic analyses of the internal transcribed spacer region of fungal ribosomal RNA. Six isolates showing steady growth were deposited in the fungal culture collection of the Fungus/Mushroom Resource and Research Center, Faculty of Agriculture, Tottori University, Japan. This result indicates that basidiospore isolation is a useful method for obtaining Harushimeji strains.

BackgroundCongenital scoliosis (CS) is a common vertebral malformation. Spondylocostal dysostosis (SCD) is a rare skeletal dysplasia characterised by multiple vertebral malformations and rib anomalies. In a previous study, a compound heterozygosity for a null mutation and a risk haplotype composed by three single-nucleotide polymorphisms inTBX6have been reported as a disease-causing model of CS. Another study identified bi-allelic missense variants in a SCD patient. The purpose of our study is to identifyTBX6variants in CS and SCD and examine their pathogenicity.MethodsWe recruited 200 patients with CS or SCD and investigatedTBX6variants. We evaluated the pathogenicity of the variants by in silico prediction and in vitro experiments.ResultsWe identified five 16p11.2 deletions, one splice-site variant and five missense variants in 10 patients. In vitro functional assays for missense variants identified in the previous and present studies demonstrated that most of the variants caused abnormal localisation of TBX6 proteins. We confirmed mislocalisation of TBX6 proteins in presomitic mesoderm cells induced from SCD patient-derived iPS cells. In induced cells, we found decreased mRNA expressions ofTBX6and its downstream genes were involved in somite formation. All CS patients with missense variants had the risk haplotype in the opposite allele, while a SCD patient with bi-allelic missense variants did not have the haplotype.ConclusionsOur study suggests that bi-allelic loss of function variants ofTBX6cause a spectrum of phenotypes including CS and SCD, depending on the severity of the loss ofTBX6function.

The ossification center of the femoral head reportedly appears at 7-8 months in 95% infants in the radiographs, but its appearance often delays in Japanese normal infants. The purpose of this study was to survey the age at appearance of the femoral head in the radiographs, evaluate the normal standards of its appearance in Japanese infants, and determine whether the acetabular state affects the time of its appearance.The patients comprised 436 infants consulted with our institution because of a limitation of abduction in flexion (LA), acetabular dysplasia, or suspected developmental dysplasia of the hip (DDH). Among these patients, 111 infants (222 hips) without radiographic appearance of the femoral head at the first visit were reviewed. The 222 hips were classified into three groups: α angle ≥30° (DDH + group),30° with LA (DDH-/LA + group), or 30° without LA (DDH-/LA-group). The age at appearance of the femoral head was investigated in the radiograph in each group.The 50th percentile of the age at appearance of the femoral head was 6 months in the DDH-/LA- and DDH-/LA + groups and 8 months in the DDH + group. The femoral head significantly appeared earlier in the DDH-/LA- and DDH-/LA + groups than in the DDH + group (P 0.01). The 95th percentile was 12 months in all groups.Radiographic appearance of the ossification center of the femoral head by 12 months of age is normal in Japanese infants. When the α angle is ≥ 30°, the age at appearance of the femoral head is sometimes delayed.

Background Patient-reported outcome measures are widely utilized to assess health-related quality of life (HRQOL) in patients with adolescent idiopathic scoliosis (AIS). However, the association between HRQOL and curve severity is mostly unknown. The aim of this study is to clarify the association between HRQOL and curve severity, and to determine the optimal cutoff values of patient-reported outcomes for major curve severity in female patients with AIS. Methods Female patients with AIS treated conservatively were recruited. The patients’ HRQOL outcomes were examined using the revised Scoliosis Research Society-22 (SRS-22r) and the Scoliosis Japanese Questionnaire-27 (SJ-27). The correlations of the SRS-22r and SJ-27 scores with the major Cobb angle were assessed using Spearman’s correlation coefficient analysis. The association between HRQOL issues in the SJ-27 and the major Cobb angle was evaluated by calculating Akaike’s Information Criterion (AIC). Furthermore, the optimal cutoff values of the SRS-22r and SJ-27 scores for the major Cobb angle were determined by AIC analysis. Results The study cohort comprised 306 female patients with AIS. The SRS-22r and SJ-27 scores were significantly correlated with the major Cobb angle. Questions in the SJ-27 regarding discomfort when wearing clothes showed a lower AIC value in patients with severe scoliosis. The optimal cutoff values were a SRS-22r score of 3.2 for the discrimination of severe scoliosis (Cobb angle ≥48°), and a SJ-27 score of 32 for the discrimination of moderate scoliosis (Cobb angle ≥33°). Conclusion Discomfort when wearing clothes was the most important HRQOL problem caused by severe scoliosis. The SRS-22r and SJ-27 scores are useful for the discrimination of clinical status in female patients with severe scoliosis or moderate scoliosis.

OBJECTIVE For instrumented correction surgery for adolescent idiopathic scoliosis (AIS), surgeons are increasingly switching from titanium (Ti) alloy rods to stiffer cobalt-chromium (CoCr) rods. The authors conducted the first multicenter randomized controlled clinical trial to investigate whether these materials affect the outcomes in terms of spine correction and quality of life (QOL). This trial was registered at UMIN Clinical Trials Registry on September 3, 2012, under the identifier UMIN000008838 (level of evidence 1). METHODS Female AIS patients (Lenke types 1–3, patient age 10–19 years) were recruited at 5 Japanese institutions and randomized into two cohorts: 6.0-mm-diameter Ti rods were placed in one group, and 6.0-mm-diameter CoCr rods were placed in the other. Patients were followed up at 2 weeks and 3, 6, and 12 months with radiographic examination to quantify the sagittal and coronal correction (Cobb angle, thoracic kyphosis, rib hump, and apical vertebral rotation). Patients completed questionnaires (Scoliosis Research Society–22r, 12-Item Short-Form Health Survey, and Scoliosis Japanese Questionnaire–27) at 6 and 12 months to assess QOL. RESULTS A total of 69 AIS patients were randomized to the demographically similar Ti (n = 37) or CoCr (n = 32) cohort. Four adverse events were recorded, two in each cohort, but these were not related to the rod material. At the final follow-up, both Ti and CoCr cohorts showed significant improvement in spinal correction, including the Cobb angle, thoracic kyphosis, and rib hump size. The correction rates were 68.4% and 67.1% for the Ti and CoCr cohorts, respectively. No parameters differed significantly between the cohorts at any time. Survey data showed improved but similar outcomes in both cohorts. CONCLUSIONS Both treatments (Ti and CoCr) produced similar results and were efficient in engendering clinically significant spine corrections. Clinical trial registration no.: UMIN000008838 (UMIN Clinical Trials Registry)