Boric acid intercepts 80S ribosome migration from AUG-stop by stabilizing eRF1.

In response to environmental changes, cells flexibly and rapidly alter gene expression through translational controls. In plants, the translation of NIP5;1, a boric acid diffusion facilitator, is downregulated in response to an excess amount of boric acid in the environment through upstream open reading frames (uORFs) that consist of only AUG and stop codons. However, the molecular details of how this minimum uORF controls translation of the downstream main ORF in a boric acid-dependent manner have remained unclear. Here, by combining ribosome profiling, translation complex profile sequencing, structural analysis with cryo-electron microscopy and biochemical assays, we show that the 80S ribosome assembled at AUG-stop migrates into the subsequent RNA segment, followed by downstream translation initiation, and that boric acid impedes this process by the stable confinement of eukaryotic release factor 1 on the 80S ribosome on AUG-stop. Our results provide molecular insight into translation regulation by a minimum and environment-responsive uORF.

Epitopes of an antibody that neutralizes a wide range of SARS-CoV-2 variants in a conserved subdomain 1 of the spike protein.

The evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has continued, enabling the virus to escape from host immunity by changing its spike antigen, while biased toward the receptor-binding domain and N-terminal domain. Here, we isolated a novel pan-SARS-CoV-2 neutralizing antibody (which we named MO11) for even the recent dominators XBB.1.16 and EG.5.1, from a convalescent patient who had received three doses of an original mRNA COVID-19 vaccination. A cryo-electron microscopy analysis of the spike-MO11 complex at 2.3 Å atomic resolution revealed that it recognizes a conserved epitope hidden behind a glycan shield at N331 on subdomain 1 (SD1), holding both the N- and C-terminal segments comprising SD1. Our identification of MO11 unveiled the functional importance of SD1 for the spike's function, and we discuss the potential availability of a novel common epitope among the SARS-CoV-2 variants.IMPORTANCENovel severe acute respiratory syndrome coronavirus 2 variants with immune evasion ability are still repeatedly emerging, nonetheless, a part of immunity developed in responding to the antigen of earlier variants retains efficacy against recent variants irrespective of the numerous mutations. In exploration for the broadly effective antibodies, we identified a cross-neutralizing antibody, named MO11, from the B cells of the convalescent patient. MO11 targets a novel epitope in subdomain 1 (SD1) and was effective against all emerging variants including XBB.1.16 and EG.5.1. The neutralizing activity covering from D614G to EG.5.1 variants was explained by the conservation of the epitope, and it revealed the importance of the subdomain on regulating the function of the antigen for viral infection. Demonstrated identification of the neutralizing antibody that recognizes a conserved epitope implies basal contribution of such group of antibodies for prophylaxis against COVID-19.

Reliability of the Risser+ grade for assessment of bone maturity in pediatric scoliosis cases: Investigation using standing and supine whole-spine radiograph.

BACKGROUND: Although several radiography-based systems for assessing skeletal maturity are available to clinicians, the classical Risser grading system remains a clinical gold standard. For scoliosis follow-up, a standing whole-spine radiograph is usually used. However, in our clinical practice, we have occasionally encountered cases in which ossification of the iliac crest is seen differently in the standing and supine whole-spine radiography. Here, we aimed to clarify the reliability of the Risser+ grading system for supine versus standing position radiographs. METHODS: This study recruited patients with all types of scoliosis who had been radiographed in both the standing and supine positions. We retrospectively evaluated the Risser+ grade of standing and supine whole-spine radiographs taken consecutively. Kappa statistics were computed to investigate the agreement between standing and supine Risser+ grades for this study. RESULTS: We evaluated 111 patients (age: 12.6 ± 2.0; male-to-female = 23:88). The Kappa value for the standing and supine Risser+ grade systems was 0.74. The degree of agreement between the two positions for each Risser+ grade revealed high agreement for grades 0 and 5 in all cases, whereas grades 2 and 3 had low agreement. CONCLUSIONS: Overall, there was substantial agreement between the Risser+ grades assigned to standing and supine position radiographs. However, disagreement was observed between standing and supine position radiographs assigned Risser+ grades of 2 or 3. Therefore, we have found a wide range in the visibility of iliac apophysis ossification of the iliac depending on the posture, and there are limitations in assessing bone maturity using the Risser+ grade alone. Clinicians should use other evaluation systems, in addition to the Risser+ system, to achieve a more accurate bone maturity assessment, especially for cases with standing position radiographs assigned Risser grades of 2 or 3.

Structural insights into thermophilic chaperonin complexes.

Group I chaperonins are dual heptamer protein complexes that play significant roles in protein homeostasis. The structure and function of the Escherichia coli chaperonin are well characterized. However, the dynamic properties of chaperonins, such as large ATPase-dependent conformational changes by binding of lid-like co-chaperonin GroES, have made structural analyses challenging, and our understanding of these changes during the turnover of chaperonin complex formation is limited. In this study, we used single-particle cryogenic electron microscopy to investigate the structures of GroES-bound chaperonin complexes from the thermophilic hydrogen-oxidizing bacteria Hydrogenophilus thermoluteolus and Hydrogenobacter thermophilus in the presence of ATP and AMP-PNP. We captured the structure of an intermediate state chaperonin complex, designated as an asymmetric football-shaped complex, and performed analyses to decipher the dynamic structural variations. Our structural analyses of inter- and intra-subunit communications revealed a unique mechanism of complex formation through the binding of a second GroES to a bullet-shaped complex.

Which patients do we need to consider augmentation of muscle active potentials regarding transcranial electrical stimulation motor-evoked potentials monitoring before spine surgery?

BACKGROUND CONTEXT: Transcranial electrical stimulation motor-evoked potentials (Tc-MEPs) are the current trend and are important in preventing intraoperative neurological deficits. Posttetanic Tc-MEPs (p-MEP) can augment the amplitudes of compound muscle active potentials (CMAPs), especially in the case of insufficient conventional Tc-MEPs (c-MEP). PURPOSE: To retrospectively investigate pre- and intraoperative factors necessitating p-MEP monitoring and to examine changes in the success rates of baseline Tc-MEP monitoring before and after tetanic stimulation in patients with such factors. STUDY DESIGN: Retrospective observational study. PATIENT SAMPLE: Patients (n=184) who underwent spinal surgery with Tc-MEP monitoring in our department between August 2020 and July 2022. OUTCOME MEASURES: Manual muscle testing (MMT) scores were calculated to identify patients with preoperative motor deficits. c-MEP and p-MEP amplitudes were recorded from the defined muscles. METHODS: We compared preoperative and intraoperative factors between the c-MEP and p-MEP groups (study 1). In cases where the factors were identified, we investigated the success rate of the baseline MEP measurement of each muscle before and after tetanic stimulation (study 2). RESULTS: One hundred fifty-seven patients were included. Of those, 87 showed sufficient CMAPs with c-MEP. Meanwhile, 70 needed p-MEP because of insufficient CMAPs. In univariate analysis, cervical/thoracic surgery (p<.001), preoperative MMT 3 or below (p=.009), shorter duration of illness (p=.037), previous cerebrovascular disease (p=.014), and dialysis (p=.031) were significantly associated with p-MEP group. Preoperative MMT 3 or below was the only factor requiring p-MEP (odds ratio, 3.34; 95% confidence interval, 1.28-8.73, p=.014) in multivariate analysis. In the p-MEP group, 24 patients had preoperative motor deficits; 16 patients with complete data were included in the analysis (study 2). The success rates of MEP monitoring before and after tetanic stimulation of the entire lower-extremity muscles were 42.7 and 57.3%, respectively (p<.001). The success rates for each muscle before and after tetanic stimulation were abductor pollicis brevis: 81.3% and 96.9%, tibialis anterior: 34.4% and 50.0%, gastrocnemius: 25% and 40.6%, and abductor hallucis: 68.8% and 81.3%, respectively. No significant differences were observed in success rates for any of the muscles. CONCLUSIONS: Patients with preoperative MMT 3 or below highly needed p-MEP. The success rate of baseline MEP monitoring increased with tetanic stimulation, even in patients with preoperative motor deficits. We believe that p-MEP monitoring can result in reliable CMAP recording, especially in cases of preoperative motor deficits with MMT scores of 3 or below.

A novel technique for posterior lumbar interbody fusion to obtain a good local lordosis angle: anterior-release posterior lumbar interbody fusion.

Herein, we describe a novel posterior lumbar interbody fusion (PLIF) technique with annulus fibrosus (AF) release and the use of expandable cages (called "anterior-release PLIF" [ARPLIF]). In this technique, posterior column osteotomy (PCO) and AF release provide excellent intervertebral mobility. AF release involves circumferentially peeling off the AF above or below the endplate between the fixed vertebrae under radiographic guidance without cutting the AF and anterior longitudinal ligament. Subsequently, high-angle variable-angle expandable cages are used to simultaneously expand both sides before inserting the percutaneous pedicle screws and correcting to achieve good local lumbar lordosis. PCO and AF release achieve excellent intervertebral mobility. Intervertebral mobility and simultaneous expansion of both cages disperse the force on the endplates, reducing cage subsidence, and the high-angle cages facilitate high intervertebral angle creation. The novel ARPLIF intervertebral manipulation technique can promote good local lumbar lordosis formation.

Anterior Placement of Cages in Posterior Lumbar Interbody Fusion for Obtaining Good Lumbar Lordosis Formation.

Introduction: Posterior lumbar interbody fusion (PLIF) is a common treatment for nerve root disease associated with lumbar foraminal stenosis or lumbar spondylolisthesis. At our institution, PLIF is usually performed with high-angle cages and posterior column osteotomy (PLIF with HAP). However, not all patients achieve sufficient segmental lumbar lordosis (SLL). This study determined whether the location of PLIF cages affect local lumbar lordosis formation. Methods: A total of 59 patients who underwent L4/5 PLIF with HAP at our hospital, using the same titanium control cage model, were enrolled in this cohort study. The mean ratio of the distance from the posterior edge of the cage to the posterior wall of the vertebral body/vertebral length (RDCV) immediately after surgery was 16.5%. The patients were divided into two groups according to RDCV <16.5% (group P) and ≥16.5% (group G). The preoperative and 6-month postoperative slip rate (%slip), SLL, local disk angle (LDA), ratio of disk height/vertebral height (RDV), 6-month postoperative RDCV, ratio of cage length/vertebral length (RCVL), and ratio of posterior disk height/anterior disk height at the fixed level (RPA) were evaluated via simple lumbar spine X-ray. The preoperative and 6-month postoperative Japanese Orthopedic Association (JOA) and low back pain visual analog scale (VAS) scores were also evaluated. Results: Groups G and P included 31 and 28 patients, respectively. The preoperative %slip, SLL, LDA, RDV, JOA score, and low back pain VAS score were not significantly different between the groups. In groups G and P, 6-month postoperative %slip, SLL, LDA, RDV, RDCV, RCVL, and RPA were 3.3% and 7.9%, 18.6° and 15.4°, 9.7° and 8.0°, 36.6% and 40.3%, 21.1% and 10.1%, 71.4% and 77.0%, and 56.1% and 67.7%, respectively. The 6-month postoperative SLL, LDA, RDV, RDCV, RCVL, and RPA significantly differed (p=0.03, 0.02, 0.02, <0.001, <0.001, and <0.001, respectively). Conclusions: Anterior PLIF cage placement relative to the vertebral body is necessary for good SLL in PLIF.

NDT-C11 as a Viable Novel Detergent for Single Particle Cryo-EM.

Single particle cryo electron microscopy (cryo-EM) is now the major method for the determination of integral membrane protein structure. For the success of a given project the type of membrane mimetic used for extraction from the native cell membrane, purification to homogeneity and finally cryo-grid vitrification is crucial. Although small molecule amphiphiles - detergents - are the most widely used membrane mimetic, specific tailoring of detergent structure for single particle cryo-EM is rare and the demand for effective detergents not satisfied. Here, we compare the popular detergent lauryl maltose-neopentyl glycol (LMNG) with the novel detergent neopentyl glycol-derived triglucoside-C11 (NDT-C11) in its behavior as free detergent and when bound to two types of multisubunit membrane protein complexes - cyanobacterial photosystem I (PSI) and mammalian F-ATP synthase. We conclude that NDT-C11 has high potential to become a very useful detergent for single particle cryo-EM of integral membrane proteins.

Structural transitions in kinesin minus-end directed microtubule motility.

Kinesin motor proteins hydrolyze ATP to produce force for spindle assembly and vesicle transport, performing essential functions in cell division and motility, but the structural changes required for force generation are uncertain. We now report high-resolution structures showing new transitions in the kinesin mechanochemical cycle, including power stroke fluctuations upon ATP binding and a post-hydrolysis state with bound ADP + free phosphate. We find that rate-limiting ADP release occurs upon microtubule binding, accompanied by central ß-sheet twisting, which triggers the power stroke - stalk rotation and neck mimic docking - upon ATP binding. Microtubule release occurs with ß-strand-to-loop transitions, implying that ß-strand refolding induces Pi release and the recovery stroke. The strained ß-sheet during the power stroke and strand-to-loop transitions identify the ß-sheet as the long-sought motor spring.