Development of Orally Bioavailable Peptides Targeting an Intracellular Protein: From a Hit to a Clinical KRAS Inhibitor.

Cyclic peptides as a therapeutic modality are attracting a lot of attention due to their potential for oral absorption and accessibility to intracellular tough targets. Here, starting with a drug-like hit discovered using an mRNA display library, we describe a chemical optimization that led to the orally available clinical compound known as LUNA18, an 11-mer cyclic peptide inhibitor for the intracellular tough target RAS. The key findings are as follows: (i) two peptide side chains were identified that each increase RAS affinity over 10-fold; (ii) physico-chemical properties (PCP) including Clog P can be adjusted by side-chain modification to increase membrane permeability; (iii) restriction of cyclic peptide conformation works effectively to adjust PCP and improve bio-activity; (iv) cellular efficacy was observed in peptides with a permeability of around 0.4 × 10-6 cm/s or more in a Caco-2 permeability assay; and (v) while keeping the cyclic peptide's main-chain conformation, we found one example where the RAS protein structure was changed dramatically through induced-fit to our peptide side chain. This study demonstrates how the chemical optimization of bio-active peptides can be achieved without scaffold hopping, much like the processes for small molecule drug discovery that are guided by Lipinski's rule of five. Our approach provides a versatile new strategy for generating peptide drugs starting from drug-like hits.

Validation of a New Methodology to Create Oral Drugs beyond the Rule of 5 for Intracellular Tough Targets.

Establishing a technological platform for creating clinical compounds inhibiting intracellular protein-protein interactions (PPIs) can open the door to many valuable drugs. Although small molecules and antibodies are mainstream modalities, they are not suitable for a target protein that lacks a deep cavity for a small molecule to bind or a protein found in intracellular space out of an antibody's reach. One possible approach to access these targets is to utilize so-called middle-size cyclic peptides (defined here as those with a molecular weight of 1000-2000 g/mol). In this study, we validated a new methodology to create oral drugs beyond the rule of 5 for intracellular tough targets by elucidating structural features and physicochemical properties for drug-like cyclic peptides and developing library technologies to afford highly N-alkylated cyclic peptide hits. We discovered a KRAS inhibitory clinical compound (LUNA18) as the first example of our platform technology.

Electron-Rich Gold Clusters Stabilized by Poly(vinylpyridines) as Robust and Active Oxidation Catalysts.

In this report, we introduced poly(n-vinylpyridine) (PnVP, n = 2, 4) as an electron-donating stabilizer for small (<2 nm) Au clusters and elucidated how coordinating pyridines affect the physical, optical, chemical, and catalytic properties of Au clusters. Spectroscopic measurements and theoretical calculation suggested the high electron-donating ability of PnVP. PnVP-stabilized Au clusters improved robustness in aerobic oxidation of alcohols compared to poly(N-vinyl-2-pyrrolidone)-stabilized ones, while retaining catalytic activities.

Identification of a selective inhibitor of transforming growth factor ß-activated kinase 1 by biosensor-based screening of focused libraries.

Transforming growth factor-ß activated kinase 1 (TAK1), a member of the mitogen-activated protein kinase kinase kinase family, plays an essential role in mediating signals from various pro-inflammatory cytokines and therefore may be a good target for developing anti-inflammation agents. Herein, we report our efforts to identify TAK1 inhibitors with a good selectivity profile with which to initiate medicinal chemistry. Instead of resorting to a high-throughput screening campaign, we performed biosensor-based biophysical screening for a limited number of compounds by taking advantage of existing knowledge on kinase inhibitors. Rather than focusing on one specific inhibition mode, we searched for three different types, Type I (ATP-competitive, DFG-in), Type II (DFG-out), and Type III binders (non-ATP competitive) in parallel, and succeeded in identifying candidates in all three categories efficiently and rapidly. Finally, the biosensor-based binding kinetics for the active and inactive forms of TAK1 were measured to prioritize the Type I and Type II inhibitors. The effort resulted in the identification of a new TAK1-selective Type I compound with a thienopyrimidine scaffold that served as a good starting point for medicinal chemistry.

Intraoperative and Postoperative Ultrasonographic Spinal Cord Evaluation of Cervical Double-Door Laminoplasty.

BACKGROUND: Ultrasonography is a useful tool for the localization, morphology, and characterization of lesions and is increasingly being applied to spinal cord evaluation in cervical spine diseases. However, in conventional cervical laminoplasty, detailed evaluation is difficult because of ultrasound attenuation. Therefore, the purpose of this study was to perform a cervical laminoplasty using a modified surgical technique and evaluate the spinal cord. METHODS: The spinal cord was evaluated intraoperatively and one week postoperatively in 11 patients with cervical compressive myelopathy. Double-door laminoplasty was selected as the surgical method, and the shape and placement of the bone graft between the expanded laminas were devised to reduce ultrasonic attenuation, such that there was a large space in which the dura mater was visible. RESULTS: Intraoperative and postoperative spinal cord decompression, claudication, and pulsation were confirmed in all cases. A more precise diagnosis was possible using ultrasound attenuation using the grafted bone between the laminas as an indicator. CONCLUSION: Intraoperative and postoperative spinal cord decompression status and wave patterns of modified cervical double-door laminoplasty can be evaluated using ultrasonography. Ultrasound-based evaluations of the spinal cord may provide new insights.

A Modified Sliding-Lengthening Approach to Tendon Lengthening with a Locking Mechanism Suture: A Technical Note.

There are various techniques used for tendon lengthening, of which Z-lengthening and sliding-lengthening are the most frequently performed. In patients with cerebral palsy, tendon lengthening may often be necessary at multiple sites. However, they can cause various complications, such as inaccurate extension, overextension, and a lack of tendon continuity. We modified the sliding-lengthening technique with a locking mechanism to address these issues. This technical note aims to describe the surgical technique and pitfalls associated with the modified sliding-lengthening approach and suture locking mechanism. The tendon was exposed and stabilized using sterilized spitz tubes and was then threaded so that each loop length was equivalent to the amount of tendon extension. Symmetrical hemisection of both ends of the tendon was performed, and the tendon was carefully extended to create a tense loop. The modified sliding-lengthening technique with the locking suture mechanism may be an advantageous method that accurately addresses extension volume, prevents hyperextension, and maintains tendon continuity, even when smaller incisions are used.

Preliminary Clinical Experience with a High-Definition Three Dimensional Exoscope for Spinal Surgery.

We retrospectively evaluated spinal surgeries performed using the high-definition three-dimensional exoscopic system, which became available at our institution in August 2020. Eleven patients (4 with cervical disease and 7 with lumbar disease) underwent surgery with the system. There were no surgical complications related to the system, and the results were satisfactory. The small, flexible camera of the exoscope allows the surgeon to view the surgical field from various angles, facilitating both the approach and technique. In addition, it allows the surgeon to operate in an upright position without strain on the head and neck. Although further surgical experience is needed, this system has the potential to improve the visualization of the surgical field in spinal surgery.

Effects of peripheral sensory nerve stimulation plus task-oriented training on upper extremity function in patients with subacute stroke: a pilot randomized crossover trial.

OBJECTIVE: To investigate the feasibility of peripheral sensory nerve stimulation combined with task-oriented training in patients with stroke during inpatient rehabilitation. DESIGN: A pilot randomized crossover trial. SETTING: Two rehabilitation hospitals. SUBJECTS: Twenty-two patients with subacute stroke. INTERVENTIONS: Participants were randomly assigned to two groups and underwent two weeks of training in addition to conventional inpatient rehabilitation. The immediate group underwent peripheral sensory nerve stimulation combined with task-oriented training in the first week, followed by another week with task-oriented training alone. The delayed group underwent the same training in reverse order. MAIN MEASURES: Outcome measures were the level of fatigue and Wolf Motor Function Test. Patients were assessed at baseline, one and two weeks. RESULTS: All participants completed the study with no adverse events. There was no significant difference in level of fatigue between each treatment. From baseline to one week, the immediate group showed larger improvements than the delayed groups in the Wolf Motor Function Test (decrease in mean time (± SD) from 41.9 ± 16.2 seconds to 30.6 ± 11.4 seconds versus from 46.8 ± 19.4 seconds to 42.9 ± 14.7 seconds, respectively) but the difference did not reach significance after Bonferroni correction (P = 0.041). Within-group comparison showed significant improvements in the Wolf Motor Function Test mean time after the peripheral sensory nerve stimulation combined with task-oriented training periods in each group (P < 0.01). CONCLUSION: Peripheral sensory nerve stimulation is feasible in clinical settings and may enhance the effects of task-oriented training in patients with subacute stroke.

Angulated Salter osteotomy in the treatment of developmental dysplasia of the hip.

Salter osteotomy is widely used to improve acetabular coverage in the treatment of developmental dysplasia of the hip. Herein we describe angulated Salter osteotomy (ASO) as the modified Salter osteotomy, which creates a two-point contact between the proximal and distal fragments and better stabilizes the fixation of the fragments. We reported our results of ASO and compared it with that of Salter osteotomy performed previously by us. We retrospectively reviewed 41 unilateral hips that underwent ASO, with no other accompanying procedures, between 2012 and 2018. We investigated the radiographic measurements included the preoperative values of the acetabular index and center-edge angle (CEA), immediate postoperative values of distance d (lateral displacement of the distal fragment), lateral rotation angle (LRA), the ratio of the obturator height (ROH), pelvic height increase percentage (PHIP) and the values of acetabular index and CEA during the last follow-up. Measurements were compared with 20 unilateral hips that underwent Salter osteotomy. The mean age at the time of surgery was 5.4 years, and the mean follow-up duration was 3.3 years. Immediately after surgery, the mean distance d, LRA, ROH and PHIP were 8 mm, 19°, 70 and 1%, respectively. The last follow-up values of acetabular index and CEA significantly improved from the preoperative values by 18° and 21°, respectively. Patients treated with ASO showed significantly larger distance d, more improvement in CEA, and lesser PHIP than those treated with Salter osteotomy. The short-term outcomes of ASO are favorable. ASO was as effective as or better than Salter osteotomy in pulling out and stabilizing the distal fragment anterolaterally. ASO prevents elongation of the ilium, which causes pelvic obliquity.

Computed Tomographic Epidurography in Patients with Low Back Pain and Leg Pain: A Single-Center Observational Study.

This study was conducted to analyze the findings and benefits of computed tomography (CT) epidurography in patients with low back and leg pain and compare these findings with those of magnetic resonance imaging (MRI) images. In total, 495 intervertebral discs from 99 patients with low back and leg pain who underwent percutaneous epidural adhesiolysis (epidural neuroplasty or percutaneous adhesiolysis) were examined. The axial views of CT epidurography were classified into six types to examine each intervertebral disc: round type, ellipse type, spike type, Benz mark, incomplete block, complete block, and non-contrast. MRI images were graded from A to D using the Schizas classification. Notably, 176 images were round-type and ellipse-type axial views, and 138 were spike-type and Benz-mark views; Schizas classification Grades A and B were observed in 272 and 47 MRI images, respectively. The incomplete block and complete block axial images did not significantly differ in CT epidurography and Schizas classification Grades C and D. The images showing Benz marks existed only at the L4/5 and L5/S intervertebral levels and only in 14.7% of patients. The ratio of normal shadows differed between MRI images and CT epidurography. Therefore, CT epidurography may enable a detailed evaluation of the epidural space.

Femoral varus derotational osteotomy without pelvic osteotomy in nonambulatory children with cerebral palsy: Minimum 5 years follow-up.

ABSTRACT: Whether femoral varus derotational osteotomy (VDRO) alone or a combination of femoral and pelvic osteotomies should be performed for hip dislocation in nonambulatory children with cerebral palsy (CP) remains controversial. Few studies have reported radiographical results after the surgical treatment in nonambulatory children with CP. This study aimed to assess the results and determine predictors indicating progressive hip subluxation and redislocation after VDRO without pelvic osteotomy. We retrospectively analyzed 22 hips in 15 nonambulatory children with CP. All patients underwent VDRO without pelvic osteotomy and were followed up for at least 5 years. The mean follow-up period was 7.3 ±â€Š1.9 years. In radiological assessments, we investigated migration percentage (MP), center-edge angle, neck-shaft angle, teardrop distance, break in Shenton's line (SL), sharp's angle, acetabular ridge angle (ARA), and the change ratio of MP (Change MP). We classified patients with an MP of <40% at final follow-up in the Good group and those with an MP of ≥40% in the Poor group. The Good group included 10 children (14 hips), and the Poor group included 8 children (8 hips). No preoperative differences were found in the means of all the radiographical parameters. However, MP was significantly different between the groups from 1 year postoperatively. ARA showed improvement 5 years after surgery in the Good group. Change MP in the Good group was maintained from immediately after surgery to the final follow-up. Multivariate logistic regression analyses revealed that preoperative break in SL and Change MP immediately after surgery were parameters to predict MP at the final follow-up. In the receiver operating characteristic analysis, the cut-off values were estimated to be 19.2 mm for preoperative SL and 79.0% for Change MP immediately after surgery. Within 7.3 years of follow-up, 63.6% of the patients who underwent VDRO without pelvic osteotomy had good results. Preoperative SL and postoperative Change MP can be considered as predictors of postoperative subluxation and/or dislocation.

Effects of plantar perception training on standing posture balance in the old old and the very old living in nursing facilities: a randomized controlled trial.

OBJECTIVE: To determine whether plantar perception training using a hardness discrimination task efficiently improves stabilization of standing posture balance in the old old as well as the very old. DESIGN: A randomized two-group parallel controlled trial. SETTING: Nursing homes. PARTICIPANTS: Forty-six elderly persons 75 years of age or older living in nursing facilities were randomly assigned evenly to either an intervention or a control group. INTERVENTION: The intervention group was given a task to discriminate hardness differences while standing on foam rubber of different levels of hardness, while the control group was given the task to simply remain standing on foam rubber. The tasks were imposed for 10 successive days. OUTCOME MEASURES: Outcome assessment was made by determinations of centre-of-gravity sway in the standing position and the Functional Reach Test. RESULTS: Planter perception was significantly improved and centre-of-gravity sway in the standing position was also significantly reduced in the intervention group after the intervention. In the control group, however, there were no significant changes in perception or in sway (P < 0.01) There was a significant difference in the Functional Reach Test values between the two groups: an increment of 12.3 ± 10.1 cm in the intervention group vs. 2.3 ± 5.8 cm in the control group (P < 0.001). CONCLUSION: These results suggested that plantar perception exercises might efficiently stabilize standing postural balance in the old old as well as the very old.

[Survey of ocular injury by solar eclipse 2009].

PURPOSE: Incidence of ocular injury caused by the solar eclipse of 2009 has been investigated using the questionnaire of the Japan Committee of the International Year of Astronomy 2009. METHODS: The questionnaire was available to the public on the website. RESULTS: Answers were obtained from 14 people (12 men, 2 women), aged from sixteen to fifty-four. All had looked at the sun either with the naked eye or through some shading device (plastic sheet, plastic bag, compact disc, etc.) the safty of which is unknown. Reported symptoms included a sense of incongruity, heat, pain, central scotoma or visual disturbance, classified into four periods of duration. Three people with transient symptoms watched the eclipse with the naked eye for one minute in cloudy weather. Three people with symptoms lasting for one day and three people with symptoms lasting for one week had watched it with the naked eye and some shading device for 10 minutes in cloudy weather. Five people with symptoms lasting for more than one week watched the eclipse with the naked eye and through some sort of device in fine or slightly cloudy weather. Three people consulted their ophthalmologist. CONCLUSIONS: Those who reported adverse symptoms did not comply with the instructions for safe watching of the eclipse and the duration of the symptoms depended on the weather and the type of device used.

Influence of Stochastic Resonance on Manual Dexterity in Children With Developmental Coordination Disorder: A Double-Blind Interventional Study.

Background: There is increasing evidence that the stochastic resonance (SR) phenomenon provided by subthreshold mechanical noise stimulation improves the sensory-motor system. However, the effect of SR on children with developmental coordination disorder (DCD) is unclear. The purpose of this study was to assess whether SR activated by subthreshold vibrotactile noise stimulation of the wrist influences manual dexterity in children with DCD. Methods: A double-blind interventional study was conducted. Participants were 30 children (age: 9.3 ± 1.44 years, range 6-11 years; 27 male, three female; 25 right-handed, five left-handed) meeting DCD diagnostic criteria in DSM-5. The manual dexterity test was administered the day before SR intervention (baseline-data). SR was elicited using subthreshold vibrotactile noise stimulation at 60% of the vibrotactile threshold measured at the wrist. SR was delivered two times and the manual dexterity test was administered during each SR stimulation block (SR-on condition) and after each SR stimulation block (SR-off), for a total of four measurements. Target outcomes were the component score, the standard score, and the percentile score of the manual dexterity test. Results: The manual dexterity test scores in the SR-on condition were significantly improved compared to scores at the baseline and in the SR-off condition (p < 0.001). Conclusions: The present study showed that subthreshold noise stimulation eliciting SR significantly improved manual dexterity outcomes in children with DCD during stimulation but not after stimulation. Future studies will need to investigate the carry-over effects of SR stimulation.

In Vivo Evaluation of the Mechanical Strength of a Slide Lengthening Technique With a Locking Mechanism Using a Rabbit Model.

Background There are many reports of Achilles tendon lengthening procedures for equinus deformity of the ankle. We previously modified an Achilles tendon lengthening to prevent overextension with a locking mechanism suture before performing a sliding lengthening. The purpose of this study was to compare the biomechanical properties of the locking mechanism suture with sliding lengthening (L-SL) and Z-lengthening (ZL) using a rabbit model. Methods Thirty-six male Japanese white rabbits were assigned to two groups - half undergoing the L-SL technique and half undergoing the ZL technique on the flexor hallucis longus (FHL) tendon. Six rabbits in each group were sacrificed at one week, three weeks, and six weeks postoperatively and evaluated, while five rabbits underwent radiographical and biomechanical evaluation and one underwent histological evaluation. Results In extension length, L-SL was significantly lower than ZL one week postoperatively. In the L-SL group, elongation one week postoperatively was significantly lower than that three and six weeks postoperatively. In the ultimate failure load, L-SL was significantly higher than ZL one and three weeks after lengthening. In the L-SL group, the ultimate failure load one week postoperatively was significantly lower than that three and six weeks postoperatively. In the ZL group, there were significant differences at all time points. Conclusion  L-SL had higher mechanical property in vivo.

Transposition osteotomy of the acetabulum for advanced-stage osteoarthritis of the hips.

BACKGROUND: Transposition osteotomy of the acetabulum (TOA) was the first periacetabular osteotomy in which the acetabulum was transposed with articular cartilage. TOA improves coverage of the femoral head and joint congruity. The purpose of this study was to investigate whether TOA is an appropriate option for treating osteoarthritis of the hips at the advanced stage by comparing it with matched control hips at the early stage. METHODS: Between 1998 and 2001, TOA was performed in 104 hips of 98 patients. Altogether, 16 of 17 hips (94%) with osteoarthritis at the advanced stage were examined and compared with 37 matched control hips at the early stage. The mean age at the operation was 48 years (38-56 years), and the mean follow-up period was 88 months (65-107 months). RESULTS: TOA corrected the acetabular dysplasia and significantly improved containment of the femoral head. Clinical scores were also significantly improved in both groups. In the advanced osteoarthritis cases, there was a tendency for abduction congruity before transposition osteotomy of the acetabulum to reflect the clinical outcome. CONCLUSIONS: TOA is a promising treatment option for advanced osteoarthritis of the hips as well as for patients at an early stage when preoperative radiographs show good congruity or containment of the joint.

Subthreshold Vibrotactile Noise Stimulation Immediately Improves Manual Dexterity in a Child With Developmental Coordination Disorder: A Single-Case Study.

Developmental coordination disorder (DCD) is the most common childhood movement disorder. It is characterized by clumsiness of fine and gross motor skills in developing children. Children with DCD have low ability to effectively use tactile information for movements, instead relying on visual information. In addition, children with DCD have deficits in visuo-motor temporal integration, which is important in motor control. These traits subsequently lead to clumsiness of movements. Conversely, however, imperceptible vibrotactile noise stimulation (at 60%-intensity of the sensory threshold) to the wrist provides stochastic resonance (SR) phenomenon to the body, improving the sensory and motor systems. However, the effects of SR have not yet been validated in children with DCD. Thus, we conducted a single case study of a 10-year-old boy with a diagnosis of DCD to investigate the effect of SR on visual dependence, visuo-motor temporal integration, and manual dexterity. SR was provided by vibrotactile noise stimulation (at an intensity of 60% of the sensory threshold) to the wrist. Changes in manual dexterity (during the SR on- and off-conditions) were measured using the manual dexterity test of the Movement Assessment Battery for Children-2nd edition. The point of subjective equality measured by visual or tactile temporal order judgment task served as a quantitative indicator reflecting specific sensory dependence. The delay detection threshold and steepness of delay detection probability curve, which were measured using the delayed visual feedback detection task, were used as quantitative indicators of visuo-motor temporal integration. The results demonstrated alleviated visual dependence and improved visuo-motor temporal integration during the SR on-conditions rather than the SR off-conditions. Most importantly, manual dexterity during the SR on-conditions was significantly improved compared to that during the SR off-conditions. Thus, the present results highlighted that SR could contribute to improving poor movement in children with DCD. However, since this was a single case study, a future interventional study with a large sample size is needed to determine the effectiveness of SR for children with DCD.

Stochastic resonance improves visuomotor temporal integration in healthy young adults.

Mechanical and electrical noise stimulation to the body is known to improve the sensorimotor system. This improvement is related to stochastic resonance (SR), a phenomenon described as a "noise benefit" to various sensory and motor systems. The current study investigated the influence of SR on visuomotor temporal integration and hand motor function under delayed visual feedback in healthy young adults. The purpose of this study was to measure the usefulness of SR as a neurorehabilitation device for disorders of visuomotor temporal integration. Thirty healthy volunteers underwent detection tasks and hand motor function tests under delayed visual feedback, with or without SR. Of the 30 participants, 15 carried out the tasks under delayed visual feedback in the order of SR on-condition, off-condition, off-condition, and on-condition. The remaining 15 participants conducted the experimental tasks in the order of SR off-condition, on-condition, on-condition, and off-condition. Comparisons of the delay detection threshold (DDT), steepness of the delay detection probability curves, box and block test (BBT) scores, and nine-hole peg test (NHPT) scores between the SR on- and off-conditions were performed. The DDT under the SR on-condition was significantly shortened compared with the SR off-condition. There was no significant difference between the SR on- and off-conditions for the steepness of the delay detection probability curves, BBT scores, and NHPT scores. SR improved visuomotor temporal integration in healthy young adults, and may therefore improve movement disorders in patients with impaired visuomotor temporal integration. However, because the current results showed that SR did not improve hand motor function under delayed visual feedback, it may not improve motor function when a large distortion of visuomotor temporal integration is present. Further studies are required considering several limitations of the current study, and future clinical trials are necessary to verify the effects of motor training using SR for the treatment of visuomotor temporal integration disorders.

Factors Contributing to Satisfaction with Changes in Physical Function after Orthopedic Surgery for Musculoskeletal Dysfunction in Patients with Cerebral Palsy.

BACKGROUND: The recognition of required treatments for cerebral palsy (CP) patients, including orthopedic surgery, differs according to region. This study was performed to identify factors associated with satisfactory changes in physical function after orthopedic surgery. METHODS: 358 patients were selected for the questionnaire survey. The following information was collected: gender, primary disease, age of initial surgery, total procedural count, operated sites, satisfaction of postoperative rehabilitation frequency, ideal amount of postoperative rehabilitation sessions per week, frequency of voluntary home training per week, satisfaction of the timing of surgery and the current satisfaction with the changes in physical function after the orthopedic surgery. We classified the patients into the satisfied and dissatisfied group according to satisfactory changes in physical function after the surgery. We performed unpaired t-tests and chi-square tests to determine the variables that differed significantly between the groups. Variables with a p value of <0.2 were included in the multivariate logistic regression analysis. RESULTS: The logistic model was revised and summed up to two potential predictors of postsurgical satisfaction with physical function: satisfaction with the frequency of postoperative rehabilitation sessions and the orthopedic surgery of the hip (distinction hit ratio, 75.4%). CONCLUSIONS: This study demonstrated that the frequency of postoperative rehabilitation and history of hip surgery seemed to be related to the satisfaction with the changes in physical function after orthopedic surgery.