Generation of single-round infectious rotavirus with a mutation in the intermediate capsid protein VP6.

Rotavirus causes severe diarrhea in infants. Although live attenuated rotavirus vaccines are available, vaccine-derived infections have been reported, which warrants development of next-generation rotavirus vaccines. A single-round infectious virus is a promising vaccine platform; however, this platform has not been studied extensively in the context of rotavirus. Here, we aimed to develop a single-round infectious rotavirus by impairing the function of the viral intermediate capsid protein VP6. Recombinant rotaviruses harboring mutations in VP6 were rescued using a reverse genetics system. Mutations were targeted at VP6 residues involved in virion assembly. Although the VP6-mutated rotavirus expressed viral proteins, it did not produce progeny virions in wild-type cells; however, the virus did produce progeny virions in VP6-expressing cells. This indicates that the VP6-mutated rotavirus is a single-round infectious rotavirus. Insertion of a foreign gene, and replacement of the VP7 gene segment with that of human rotavirus clinical isolates, was successful. No infectious virions were detected in mice infected with the single-round infectious rotavirus. Immunizing mice with the single-round infectious rotavirus induced neutralizing antibody titers as high as those induced by wild-type rotavirus. Taken together, the data suggest that this single-round infectious rotavirus has potential as a safe and effective rotavirus vaccine. This system is also applicable for generation of safe and orally administrable viral vectors.IMPORTANCERotavirus, a leading cause of acute gastroenteritis in infants, causes an annual estimated 128,500 infant deaths worldwide. Although live attenuated rotavirus vaccines are available, they are replicable and may cause vaccine-derived infections. Thus, development of safe and effective rotavirus vaccine is important. In this study, we report the development of a single-round infectious rotavirus that can replicate only in cells expressing viral VP6 protein. We demonstrated that (1) the single-round infectious rotavirus did not replicate in wild-type cells or in mice; (2) insertion of foreign genes and replacement of the outer capsid gene were possible; and (3) it was as immunogenic as the wild-type virus. Thus, the mutated virus shows promise as a next-generation rotavirus vaccine. The system is also applicable to orally administrable viral vectors, facilitating development of vaccines against other enteric pathogens.

[1,2,4]Triazolo[1,5-a]pyrimidine derivatives: Structure-activity relationship study leading to highly selective ENPP1 inhibitors.

The STING (stimulator of interferon genes) pathway is one of the pathways that regulate innate immunity, and the extracellular hydrolytic enzyme ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) has been identified as its dominant negative regulator. Since activation of the innate immune system is a promising strategy for the treatment of various infectious diseases and cancers, ENPP1 inhibitors have attracted great attention as candidate drugs. We have previously identified small-molecule ENPP1 inhibitors having a [1,2,4]triazolo[1,5-a]pyrimidine scaffold by means of chemical screening using a fluorescence probe, TG-mAMP. In this study, we evaluated the structure-activity relationships of the hit and lead compounds in detail, and succeeded in developing compounds that strongly and selectively inhibit ENPP1 not only in vitro, but also in cellular systems.

Analysis of Rod Fracture at the Lumbosacral Junction Following Surgery for Adult Spinal Deformity.

STUDY DESIGN: Retrospective study. PURPOSE: This study aimed to investigate the differences in the characteristics of patients with rod fracture (RF) at the lumbosacral junction from those without RF following adult spinal deformity (ASD) surgery. OVERVIEW OF LITERATURE: RF is a major complication following ASD surgery and may require reoperation because of pain and correction loss. The lumbosacral junction is a common RF site. However, risk factors for RFs at the lumbosacral junction remain unknown. METHODS: The study included data from 100 patients who underwent ASD surgery between 2012 and 2020. Fifteen of these patients presented with RFs. Patient demographics, clinical outcomes, and radiographic parameters were evaluated in each group. RESULTS: RFs were significantly more frequent in patients with a medical history of total hip arthroplasty (THA; p=0.01) or severe obesity (p=0.04). However, no significant differences in clinical outcomes, preoperative or postoperative measurements, or changes were found between pre- and postoperative radiographic parameters within the groups. Both pre- (p=0.01) and postoperative (p=0.02) anterior disc heights were significantly greater in the RF group than in the non-RF group. In the RF group, the postoperative lordotic angles of the lumbosacral junction significantly decreased compared with preoperative angles (p=0.02). Multiple logistic regression analysis demonstrated that a THA history (odds ratio, 34.2), severe obesity (odds ratio, 14.0), and preoperative anterior disc height (odds ratio, 1.2) were significant risk factors for RFs. CONCLUSIONS: In this study, the greatest risk factors for postoperative lumbosacral RF after ASD surgery were THA history, severe obesity, and postoperative anterior disc height of ≥10. For patients at higher risk, the use of multirods is considered necessary.

Measurement of Postoperative Apical Vertebral Rotation Using Radiographic Images in Adolescent Idiopathic Scoliosis-Quantitative Evaluation Using Inverse Trigonometric Functions.

Introduction: Conventional methods for analyzing vertebral rotation are limited to postoperative patients who underwent posterior fusion. A previous methodology calculated vertebral rotation using inverse trigonometric functions based on the length of the pedicle screw (PS). Accordingly, this study evaluates rotational deformity in patients with postoperative adolescent idiopathic scoliosis (AIS) using inverse trigonometric functions. Methods: This retrospective single-center study includes patients with AIS who underwent posterior fusion surgery. Postoperative radiography and computed tomography (CT) scans were retrospectively evaluated. The magnification ratio was calculated using the rod diameter (δ=lateral/frontal rod diameter), and the visible screw lengths were measured using radiographs. The rotation angle was calculated using the apex of the main curve and the lower instrumented vertebra (LIV) (rotation angle=tan-1 (lateral PS length/(δ×frontal PS length))) immediately following the surgery as well as two and five years postoperatively. The correlation between the direct CT measurement and postoperative rotation angle progression was investigated. The crankshaft phenomenon (CSP) and distal adding-on (DAO) were evaluated as postoperative deformities. CSP was defined as a 5° increase in rotation angle. Results: Seventy-eight patients (age: 15.3±2.0 years, eight boys and seventy girls) were included. The rotation angle was strongly correlated with CT rotation measurements (r=0.87). The mean rotation angle at the apex and LIV did not change within five years postoperatively (mean: 0.5±3.6° and 0.4±3.4°, respectively). CSP and DAO were observed in 6.4% and 3.8% of patients, respectively. Conclusions: The inverse trigonometric method is useful to quantitatively evaluate the postoperative rotation angle and identify CSP.

Factors Influencing Optimal Bracing Compliance in Adolescent Idiopathic Scoliosis: A Single Center Prospective Cohort Study.

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: To identify factors contributing to optimal bracing compliance in adolescent idiopathic scoliosis (AIS). SUMMARY OF BACKGROUND DATA: Poor brace compliance is a key factor affecting brace treatment success in AIS. Predictive factors influencing optimal brace compliance to achieve brace treatment success remain unknown. MATERIALS AND METHODS: This study included AIS patients, aged 10-15, with a Cobb angle of 20-40 degrees. Demographics data, radiographic assessments, and patient-reported outcomes (including the SRS-22r patient questionnaire) were collected. Brace compliance was monitored using in-brace thermometers, defining optimal bracing time as more than 18 hours/day. Multivariable logistic regression analysis was employed to identify predictors of optimal bracing time from the demographic and patient- reported outcomes score before bracing. RESULTS: Among 122 patients, 59.0% achieved optimal bracing time by six months. The achieved group indicated higher scores in satisfaction domain before bracing (3.3±0.7 vs. 3.1±0.6; P=0.034). Multivariable logistic regression analysis demonstrated that Satisfaction domain before bracing was an independent factor associated with achievement of the optimal bracing time (OR 1.97 [95%CI 1.00 - 3.89], P=0.049). The model with bracing at 1-month follow-up also demonstrated the bracing at 1-month was a significant factor (OR 1.52 [95%CI 1.30 - 1.79], P<0.001). CONCLUSION: Optimal bracing compliance in AIS is significantly influenced by pre-bracing satisfaction and brace compliance at earlier time point. These findings highlight the need to address psychological factors and early compliance in AIS bracing treatment. SRS-22r can be useful to identify the need for psychological support.

The association between variations in the number of thoracic and lumbar vertebrae and rib morphology in adolescent idiopathic scoliosis.

PURPOSE: Preoperative counting of thoracic and lumbar vertebrae is crucial in adolescent idiopathic scoliosis (AIS) due to reported anatomical variations and potential surgical site misidentification. This study investigated characteristics associated with the vertebral number variations AIS, particularly focusing on rib morphology. METHODS: Based on three-dimensional computed tomography, patients were categorized into the non-variant number group, comprising individuals with 12 thoracic and 5 lumbar vertebrae, and the variant number group, comprising individuals with different numbers of vertebrae. Additionally, the most caudal rib morphology was classified as normal, unilateral, or hypoplastic. RESULTS: A total of 359 patients were included in our study (41 males, 318 females, age: 16.3 ± 3.1 years), with 44 patients (12.3%) assigned to the variant number group. Logistic regression analysis identified unilateral ribs (odds ratio [OR]: 10.50) and lumbosacral transitional vertebrae (LSTV) (OR 6.49) as significant risk factors associated with variations. Further analysis revealed hypoplastic ribs as a significant risk factor associated with LSTV (OR: 4.58). 8 CONCLUSION: Our study suggests that abnormal rib morphology may be associated with vertebral number variations. Close attention to rib morphology is, therefore, warranted in cases with atypical vertebral numbers. Accordingly, to ensure surgical safety and accuracy, spine surgeons must communicate these variations to the surgical team, standardize nomenclature for describing them, and intraoperatively verify fusion levels with them.

Risk factors for allogeneic red blood cell transfusion in adult spinal deformity surgery.

Study Design: Retrospective study. Purpose: To investigate the risk factors for allogeneic red blood cell (RBC) transfusion in adult spinal deformity (ASD) surgery. Overview of Literature: Studies have not thoroughly explored the roles of intraoperative hypothermia, autologous blood donation, and hemostatic agent administration, which would provide a better understanding of the risk for perioperative RBC transfusion in ASD surgery. Methods: The medical records of 151 patients with ASD who underwent correction surgery between 2012 and 2021 were retrospectively reviewed. Estimated blood loss and perioperative allogeneic transfusion were examined. Patients were categorized into two groups based on whether they received perioperative allogeneic blood transfusion. Logistic regression analysis was employed to investigate the effect of age, sex, blood type, body mass index, American Society of Anesthesiologists' physical status, preoperative hemoglobin level, autologous blood donation, global spine alignment parameters, preoperative use of anticoagulants or antiplatelet medicine and nonsteroidal anti-inflammatory drugs, number of instrumented fusion levels, total operative duration, three-column osteotomy, lateral interbody fusion, pelvic fixation, intraoperative hypothermia, use of gelatin-thrombin based hemostatic agents, and intraoperative tranexamic acid (TXA) with simultaneous exposure by two attending surgeons. Results: The estimated blood loss was 994.2±754.5 mL, and 71 patients (47.0%) received allogeneic blood transfusion. In the logistic regression analysis, the absence of intraoperative TXA use and simultaneous exposure (odds ratio [OR], 26.3; 95% confidence interval [CI], 7.6-90.9; p<0.001), lack of autologous blood donation (OR, 21.2; 95% CI, 4.4-100.0; p<0.001), and prolonged operative duration (OR, 1.6; 95% CI, 1.3-1.9; p<0.001) were significant independent factors for perioperative allogeneic blood transfusion in ASD surgery. Conclusions: Autologous blood storage, intraoperative TXA administration, and simultaneous exposure should be considered to minimize perioperative allogeneic blood transfusion in ASD surgery, particularly in patients with anticipated lengthy surgeries.

Facet Joint Bridging in Adolescent-onset Adult Idiopathic Scoliosis with Thoracolumbar/Lumbar Curves.

OBJECTIVE: This study aims to comprehend the natural history of Adolescent idiopathic scoliosis (AIS) patients and determine risk factors for facet joint bridging in adolescent-onset adult idiopathic scoliosis with thoracolumbar/lumbar (TL/L) curves. METHODS: We included 50 patients with residual AIS with TL/L curves (3 males, 47 females; age 41.5±17.3 years, TL/L Cobb angle 59.4±11.8°). They were >20 years old and diagnosed with AIS during their adolescence. Radiographic parameters were measured, and facet joint bridging was defined from axial computed tomography (CT) images. RESULTS: The sagittal vertical axis (SVA) significantly increased with age (r=0.71, p<0.01). Coronal Cobb angle of the TL/L curve, L4 tilt, C7 translation, lumbar lordosis (LL), Pelvic incidence (PI) -LL, pelvic tilt (PT), and thoracolumbar kyphosis (TLK) were also correlated to age (p<0.05). There were significant differences in age, SVA, PI-LL, vertebral bridging, facet tropism, and apical vertebral rotation (AVR) between the facet joint bridging group (n=10) and the non-facet joint bridging group (n=40). In the multivariate logistic regression analysis, SVA, vertebral bridging, and AVR emerged as notable risk determinants for facet joint bridging. The threshold for facet joint bridging based on SVA was 2.1 cm (area under the curve [AUC]: 0.801; sensitivity = 90%, specificity = 65%). CONCLUSION: This research revealed that large SVA, the presence of vertebral bridging, and large AVR are associated with facet joint bridging in adolescent-onset adult idiopathic scoliosis patients with TL/L curves. The cut-off value for facet joint bridging based on SVA was 2.1cm.

The Optimal Patient Position on the Surgical Table for Lateral Lumbar Interbody Fusion in Adult Spinal Deformity Using Three-Dimensional Computed Tomography: A Retrospective Study.

Introduction: Lateral lumbar interbody fusion (LLIF) techniques have been extensively used in adult spinal deformity surgery. Preoperative knowledge of the optimal position of the patient on the surgical table is essential for a safe procedure. Therefore, this study aims to determine the optimal angle for positioning the patient on the surgical table during LLIF using three-dimensional computed tomography (3DCT). Methods: Data from 59 patients (2 males, 57 females, mean age 66.3±8.6 years) with adult spinal deformities treated by performing corrective spinal surgery were included in this observational retrospective study. Simulated fluoroscopic images were obtained using 3DCT images rotated from the reference position with the spinous process of S1 as the midline to the position with the spinous process in the center of the bilateral pedicle of T12-L5. The rotation angle of each vertebra was measured and defined as the optimal rotation angle (ORA). The angle that bisected the angle between the maximum and minimum ORA was defined as the optimal mean angle of the maximum and minimum ORA (OMA) and considered the optimal angle for the patient's position on the surgical table, as this position could minimize the rotation angle of the surgical table during surgery. A multiple regression analysis was performed to predict OMA. Results: Multiple regression analysis revealed the following equation: OMA=1.959+(0.238×lumbar coronal Cobb angle)+(-0.208×sagittal vertical axis). Conclusions: When the patient is placed on the surgical table by rotating them at the OMA, the rotation of the surgical table can be reduced, ensuring a safe and efficient surgical procedure.

Association of Spinal Corrective Surgery With Abdominal Aorta Length in Patients With Adult Spinal Deformity.

Introduction This research aimed to explore the relationship between spinal characteristics and the length of the abdominal aorta in adult spinal deformity (ASD) patients who underwent corrective spinal surgery. We hypothesized that adjusting spinal alignment might affect the abdominal aorta's length. Methods This study included thirteen patients with ASD (average age: 63.0 ± 8.9 years; four males and nine females) who received spinal correction surgery. We measured both pre-operative and post-operative spinal parameters, including thoracolumbar kyphosis (TLK), and calculated their differences (Δ). The length of the aorta (AoL) was determined using an automated process that measures the central luminal line from the celiac artery's bifurcation to the inferior mesenteric artery. This measurement was made using contrast-enhanced computed tomography for three-dimensional aortic reconstruction. We compared the pre-operative and post-operative AoLs and their differences (Δ). The study examined the correlation between changes in spinal parameters and changes in AoL. Results Post-operatively, there was an increase in aortic length (ΔAoL: 4.2 ± 4.9 mm). There was a negative correlation between the change in TLK and the change in AoL (R2 = 0.45, p = 0.012, ß = -0.21). No significant correlations were found with other spinal parameters. Conclusions The abdominal aorta can elongate by 4.8% after spinal corrective surgery in patients with ASD. The degree of elongation of the abdominal aorta is associated with spinal alignment correction.

Vero cell-adapted SARS-CoV-2 strain shows increased viral growth through furin-mediated efficient spike cleavage.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) utilizes several host proteases to cleave the spike (S) protein to enter host cells. SARS-CoV-2 S protein is cleaved into S1 and S2 subunits by furin, which is closely involved in the pathogenicity of SARS-CoV-2. However, the effects of the modulated protease cleavage activity due to S protein mutations on viral replication and pathogenesis remain unclear. Herein, we serially passaged two SARS-CoV-2 strains in Vero cells and characterized the cell-adapted SARS-CoV-2 strains in vitro and in vivo. The adapted strains showed high viral growth, effective S1/S2 cleavage of the S protein, and low pathogenicity compared with the wild-type strain. Furthermore, the viral growth and S1/S2 cleavage were enhanced by the combination of the Δ68-76 and H655Y mutations using recombinant SARS-CoV-2 strains generated by the circular polymerase extension reaction. The recombinant SARS-CoV-2 strain, which contained the mutation of the adapted strain, showed increased susceptibility to the furin inhibitor, suggesting that the adapted SARS-CoV-2 strain utilized furin more effectively than the wild-type strain. Pathogenicity was attenuated by infection with effectively cleaved recombinant SARS-CoV-2 strains, suggesting that the excessive cleavage of the S proteins decreases virulence. Finally, the high-growth-adapted SARS-CoV-2 strain could be used as the seed for a low-cost inactivated vaccine; immunization with this vaccine can effectively protect the host from SARS-CoV-2 variants. Our findings provide novel insights into the growth and pathogenicity of SARS-CoV-2 in the evolution of cell-cell transmission. IMPORTANCE: The efficacy of the S protein cleavage generally differs among the SARS-CoV-2 variants, resulting in distinct viral characteristics. The relationship between a mutation and the entry of SARS-CoV-2 into host cells remains unclear. In this study, we analyzed the sequence of high-growth Vero cell-adapted SARS-CoV-2 and factors determining the enhancement of the growth of the adapted virus and confirmed the characteristics of the adapted strain by analyzing the recombinant SARS-CoV-2 strain. We successfully identified mutations Δ68-76 and H655Y, which enhance viral growth and the S protein cleavage by furin. Using recombinant viruses enabled us to conduct a virus challenge experiment in vivo. The pathogenicity of SARS-CoV-2 introduced with the mutations Δ68-76, H655Y, P812L, and Q853L was attenuated in hamsters, indicating the possibility of the attenuation of excessive cleaved SARS-CoV-2. These findings provide novel insights into the infectivity and pathogenesis of SARS-CoV-2 strains, thereby significantly contributing to the field of virology.

Machine learning algorithms for predicting Cobb angle beyond 25 degrees in female adolescent idiopathic scoliosis patients.

STUDY DESIGN: Retrospective cohort study. OBJECTIVE: To develop a machine learning (ML) model that predicts the progression of AIS using minimal radiographs and simple questionnaires during the first visit. SUMMARY OF BACKGROUND DATA: Several factors are associated with angle progression in patients with AIS. However, it is challenging to predict angular progression at the first visit. METHODS: Among female patients with AIS treated at a single institution from July 2011 to February 2023, 1119 cases were studied. Patient data, including demographic and radiographic data based on anterior-posterior and lateral whole-spine radiographs, were collected at the first and last visits. The last visit was defined differently based on treatment plans. For patients slated for surgery or bracing, the last visit occurred just before these interventions. For others, it was their final visit before turning 18 years. Angular progression was defined as a Cobb angle greater than 25 degrees for each of the proximal thoracic (PT), main thoracic (MT), and thoracolumbar/lumbar (TLL) curves at the last visit. ML algorithms were employed to develop individual binary classification models for each type of curve (PT, MT, and TLL) using PyCaret in Python. Multiple models were explored and analyzed, with the selection of optimal models based on the area under the curve (AUC) and Recall scores. Feature importance was evaluated to understand the contribution of each feature to the model predictions. RESULTS: For PT, MT, and TLL progression, the top-performing models exhibit AUC values of 0.94, 0.89, and 0.84, and achieve recall rates of 0.90, 0.85, and 0.81. The most significant factors predicting progression varied for each curve: initial Cobb angle for PT, presence of menarche for MT, and Risser grade for TLL. CONCLUSIONS: This study introduces an ML-based model using simple data at the first visit to precisely predict angle progression in female patients with AIS.