BRD2-specific inhibitor, BBC0403, inhibits the progression of osteoarthritis pathogenesis in osteoarthritis-induced C57BL/6 male mice.

BACKGROUND AND PURPOSE: The discovery of new bromo- and extra-terminal inhibitors presents new drugs to treat osteoarthritis (OA). EXPERIMENTAL APPROACH: The new drug, BBC0403, was identified in the DNA-encoded library screening system by searching for compounds that target BRD (bromodomain-containing) proteins. The binding force with BRD proteins was evaluated using time-resolved fluorescence energy transfer (TR-FRET) and binding kinetics assays. Subsequently, in vitro and ex vivo analyses demonstrated the effects of the BRD2 inhibitor, BBC0403, on OA. For animal experiments, medial meniscus destabilization was performed to create a 12-week-old male C57BL/6 mouse model, and intra-articular (i.a.) injections were administered. Histological and immunohistochemical analyses were then performed. The underlying mechanism was confirmed by gene set enrichment analysis (GSEA) using RNA-seq. KEY RESULTS: TR-FRET and binding kinetics assays revealed that BBC0403 exhibited higher binding specificity for BRD2 compared to BRD3 and BRD4. The anti-OA effects of BBC0403 were tested at concentrations of 5, 10 and 20 µM (no cell toxicity in the range tested). The expression of catabolic factors, prostaglandin E2 (PGE2) production and extracellular matrix (ECM) degradation was reduced. Additionally, the i.a. injection of BBC0403 prevented OA cartilage degradation in mice. Finally, BBC0403 was demonstrated to suppress NF-κB and MAPK signalling pathways. CONCLUSION AND IMPLICATIONS: This study demonstrated that BBC0403 is a novel BRD2-specific inhibitor and a potential i.a.-injectable therapeutic agent to treat OA.

Novel molecule BBC0901 inhibits BRD4 and acts as a catabolic regulator in the pathogenesis of osteoarthritis.

Osteoarthritis (OA) is induced by matrix degradation and inflammation mediated by bromo-domain-containing protein 4 (BRD4)-dependent catabolic factors. BRD4 acts as both a transcriptional regulator and an epigenetic reader. BBC0901 was identified as an inhibitor of BRD4 using a DNA-encoded library screening system. We aimed to demonstrate the effects of BBC0901 on OA pathogenesis by in vitro, ex vivo, and in vivo analyses. BBC0901 inhibited the expression of catabolic factors that degrade cartilage without significantly affecting the viability of mouse articular chondrocytes. Additionally, ex vivo experiments under conditions mimicking OA showed that BBC0901 suppressed extracellular matrix degradation. RNA sequencing analysis of gene expression patterns showed that BBC0901 inhibited the expression of catabolic factors, such as matrix metalloproteinases (MMPs) and cyclooxygenase (COX)2, along with reactive oxygen species (ROS) production. Furthermore, intra-articular (IA) injection of BBC0901 into the knee joint blocked osteoarthritic cartilage destruction by inhibition of MMP3, MMP13, COX2, interleukin (IL)6, and ROS production, thereby obstructing the nuclear factor kappa-light-chain-enhancer of activated B cell and mitogen activated protein kinase signaling. In conclusion, BBC0901-mediated BRD4 inhibition prevented OA development by attenuating catabolic signaling and hence, can be considered a promising IA therapeutic for OA.

Red Ginseng Dietary Fiber Shows Prebiotic Potential by Modulating Gut Microbiota in Dogs.

Red ginseng, widely used in traditional medicine for various conditions, imparts health benefits mainly by modulating the gut microbiota in humans. Given the similarities in gut microbiota between humans and dogs, red ginseng-derived dietary fiber may have prebiotic potential in dogs; however, its effects on the gut microbiota in dogs remain elusive. This double-blinded, longitudinal study investigated the impact of red ginseng dietary fiber on the gut microbiota and host response in dogs. A total of 40 healthy household dogs were randomly assigned to low-dose (n = 12), high-dose (n = 16), or control (n = 12) groups and fed a normal diet supplemented with red ginseng dietary fiber (3 g/5 kg body weight per day, 8 g/5 kg per day, or no supplement, respectively) for 8 weeks. The gut microbiota of the dogs was analyzed at 4 weeks and 8 weeks using 16S rRNA gene sequencing of fecal samples. Alpha diversity was significantly increased at 8 and 4 weeks in the low-dose and high-dose groups, respectively. Moreover, biomarker analysis showed that short-chain fatty acid producers such as Sarcina and Proteiniclasticum were significantly enriched, while potential pathogens such as Helicobacter were significantly decreased, indicating the increased gut health and pathogen resistance by red ginseng dietary fiber. Microbial network analysis showed that the complexity of microbial interactions was increased by both doses, indicating the increased stability of the gut microbiota. These findings suggest that red ginseng-derived dietary fiber could be used as a prebiotic to modulate gut microbiota and improve gut health in dogs. IMPORTANCE The canine gut microbiota is an attractive model for translational studies, as it responds to dietary interventions similarly to those in humans. Investigating the gut microbiota of household dogs that share the environment with humans can produce highly generalizable and reproducible results owing to their representativeness of the general canine population. This double-blind and longitudinal study investigated the impact of dietary fiber derived from red ginseng on the gut microbiota of household dogs. Red ginseng dietary fiber altered the canine gut microbiota by increasing diversity, enriching short-chain fatty acid-producing microbes, decreasing potential pathogens, and increasing the complexity of microbial interactions. These findings indicate that red ginseng-derived dietary fiber may promote canine gut health by modulating gut microbiota, suggesting the possibility of its use as a potential prebiotic.

Accelerated degeneration of the discoid lateral meniscus after medial opening wedge high tibial osteotomy.

PURPOSE: This study was undertaken to report clinical outcomes after high tibial osteotomy (HTO) in patients with a discoid lateral meniscus and to determine (1) whether discoid lateral meniscus degeneration by magnetic resonance imaging (MRI) progresses after HTO and (2) whether this progression adversely affects clinical results. METHODS: The records of 292 patients (292 knees) who underwent medial opening HTO were retrospectively reviewed, and discoid types and grades of lateral meniscus degeneration as determined by MRI were recorded preoperatively. Of the 292 patients, 17 (5.8%) had a discoid lateral meniscus, and postoperative MR images were obtained at least 2 years after HTO for 15 of these 17 patients. RESULTS: American Knee Society (AKS) pain, knee and function scores significantly improved in the 15 patients after surgery (p < 0.001). Eight (53%) had an incomplete and 7 (47%) had a complete discoid lateral meniscus. By preoperative MRI, the distribution of meniscal degeneration was as follows: grade 1, 4 patients; grade 2, 7 patients; and grade 3, 4 patients. At the final follow-up, the distribution of degeneration was as follows: grade 1, 2 patients; grade 2, 5 patients; and grade 3, 8 patients. Two patients with grade 3 degeneration who did not undergo partial meniscectomy showed tear progression. Thus, 8 of the 15 patients (53%) experienced progressive discoid meniscal degeneration after HTO. Median AKS pain score was significantly lower in the progression group than in the non-progression group (40 vs 45, respectively). CONCLUSION: The results of this study suggest that increased load on the lateral compartment after HTO can accelerate discoid lateral meniscus degeneration by MRI and caution that when a discoid lateral meniscus is found by preoperative MRI, progressive degeneration may occur after HTO and clinical outcome may be adversely affected. LEVEL OF EVIDENCE: Therapeutic study, Level IV.

Agent-based modeling of physical activity behavior and environmental correlations: an introduction and illustration.

PURPOSE: To introduce Agent-Based Model (ABM) to physical activity (PA) research and, using data from a study of neighborhood walkability and walking behavior, to illustrate parameters for an ABM of walking behavior. METHOD: The concept, brief history, mechanism, major components, key steps, advantages, and limitations of ABM were first introduced. For illustration, 10 participants (age in years: mean = 68, SD = 8) were recruited from a walkable and a nonwalkable neighborhood. They wore AMP 331 triaxial accelerometers and GeoLogger GPA tracking devices for 21 days. Data were analyzed using conventional statistics and highresolution geographic image analysis, which focused on a) path length, b) path duration, c) number of GPS reporting points, and d) interaction between distances and time. RESULTS: Average steps by subjects ranged from 1810-10,453 steps per day (mean = 6899, SD = 3823). No statistical difference in walking behavior was found between neighborhoods (Walkable = 6710 ± 2781, Nonwalkable = 7096 ± 4674). Three environment parameters (ie, sidewalk, crosswalk, and path) were identified for future ABM simulation. CONCLUSION: ABM should provide a better understanding of PA behavior's interaction with the environment, as illustrated using a real-life example. PA field should take advantage of ABM in future research.

Crosstalk reduction in auto-stereoscopic projection 3D display system.

In auto-stereoscopic multi-views 3D display systems, the crosstalk and low resolution become problems for taking a clear depth image with the sufficient motion parallax. To solve these problems, we propose the projection-type auto-stereoscopic multi-view 3D display system, in which the hybrid optical system with the lenticular-parallax barrier and multi projectors. Condensing width of the projected unit-pixel image within the lenslet by hybrid optics is the core concept in this proposal. As the result, the point crosstalk is improved 53% and resolution is increased up to 5 times.

Effect of Ultra-marathon (308 km) Race on Bone Metabolism and Cartilage Damage Biomarkers.

OBJECTIVE: To evaluate the potential effects of a 308-km ultra-marathon on bone and cartilage biomarkers. METHOD: Venous blood samples were collected at pre-race, 100 km, 200 km, and 308 km checkpoints. The following markers of cartilage damage and bone metabolism were studied: osteocalcin (OC), osteoprotegerin (OPG), and calcium, phosphorous, and cartilage oligomeric matrix protein (COMP). RESULTS: Blood samples were taken from 20 male runners at four different checkpoints. Serum COMP was increased by 194.1% (130.7% at 100 km and 160.4% at 200 km). Serum OPG was significantly increased by 158.57% at 100 km and 114.1% at 200 km compared to the pre-race measures. OC was transiently suppressed at 200 km. Serum calcium and phosphorous concentrations decreased compared to the pre-race measures. CONCLUSION: This study showed that the 308-km ultra-marathon induced several changes, including transient uncoupling of bone metabolism, increased bone resorption, suppressed bone formation, and bone turnover and had a major impact on cartilage structure.

Prognostic factors of arthroscopic pull-out repair for a posterior root tear of the medial meniscus.

BACKGROUND: Repair of a posterior root tear of the medial meniscus (MRT) decreases peak contact pressure by restoring hoop tension and is expected to prevent progression to osteoarthritis. PURPOSE: The purposes of this study were (1) to report the clinical and magnetic resonance imaging (MRI) results of arthroscopic pull-out repair of the MRT and (2) to identify prognostic factors of poor outcome. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Fifty-one patients (47 women, 4 men) who underwent arthroscopic pull-out repair of the MRT by a single surgeon were enrolled. Mean follow-up after surgery was 33 months (range, 24-44 months). To identify factors affecting final outcome, patient-specific factors, such as gender, age, body mass index, meniscus extrusion, extrusion increase, subchondral edema, degree of varus alignment (<5° or >5°), and cartilage status in the medial compartment (Outerbridge grade 1 or 2 lesion vs grade 3 or 4 lesion), were investigated. Final clinical outcomes were determined using a visual analog scale (VAS) for pain and patient satisfaction scores, American Knee Society (AKS) scores, and Lysholm scores, and MRI outcomes were determined by evaluating meniscus extrusion and articular cartilage status. Multiple regression analysis was performed to identify variables that independently affected clinical and MRI-determined outcomes. RESULTS: All clinical outcome measures significantly improved after surgery. Patients with Outerbridge grade 3 or 4 chondral lesions had poorer results than those with grade 1 or 2 lesions in terms of AKS function and Lysholm scores. Patients with varus alignment of >5° had poorer results than those with varus alignment of <5° in terms of VAS satisfaction, AKS function, and Lysholm scores. Mean meniscus extrusion increased from 3.6 mm preoperatively to 5.0 mm postoperatively. Chondral lesions progressed in 3 (9.7%) of 31 patients. Preoperative meniscus extrusion was found to be positively correlated with final extrusion. CONCLUSION: At a mean follow-up of 33 months after pull-out repair, extrusion of the meniscus was found to have progressed. Nevertheless, this technique provided patients with a clinical benefit. Outerbridge grade 3 or 4 chondral lesions and varus alignment of >5° were found to independently predict an inferior clinical outcome.

Arthroscopic pullout repair of posterior root tear of the medial meniscus: the anterior approach using medial collateral ligament pie-crusting release.

Posterior root tears of the medial meniscus are frequently encountered and should be repaired if possible to prevent osteoarthritis of the medial compartment. Various surgical techniques have been proposed to repair posterior root tears. The anterior arthroscopic approach can cause an iatrogenic chondral injury due to the narrow medial joint space. The posterior approaches might be technically unfamiliar to many surgeons because they require the establishment of a posteromedial or trans-septal portal. This paper describes the medial collateral ligament pie-crusting release technique for arthroscopic double transosseous pullout repair of posterior root tears of the medial meniscus through the anterior approach to provide the good visualization of the footprint and sufficient working space.

Iatrogenic suprascapular nerve injury after repair of type II SLAP lesion.

Suprascapular neuropathy after an arthroscopic repair of a SLAP lesion is theoretically possible, but it has been rarely reported. We present a case of suprascapular nerve injury at the spinoglenoid notch as a complication of an improperly inserted suture anchor after repair of a type II SLAP lesion. The diagnosis was confirmed by the magnetic resonance imaging findings and an electrodiagnostic study, and direct compression of the nerve was visualized under repeat arthroscopy. An anatomic study of the superior glenoid shows that the available bone stock of the superior glenoid rim for the anchor insertion is found to decrease posteriorly. During the repair of a SLAP lesion, surgeons should consider the possibility of an iatrogenic injury to the suprascapular nerve by an improperly inserted suture anchor.

A primer-test centered equating method for setting cut-off scores.

This study evaluated the use of a new primary field test method based on test equating to address inconsistent classification among field tests. We analyzed students' information on the Progressive Aerobic Cardiovascular Endurance Run (PACER), mile run (MR), and VO2max from three data sets (college: n = 94; middle school: n = 39; elementary school: n = 96). Using the college and elementary school data, the equivalent relationship between PACER and MR scores was first established by the Kernel equating method. This yielded MR scores derived from PACER (mile run PACER equated [MR PEQ]), which were used to predict maximal oxygen uptake (VO2max) and classify students according to the FITNESSGRAM Healthy Fitness Zones. We compared the results to the predictions and classifications based on measured VO2max, MR and PACER-predicted VO2max and cross-validated the relationships using the middle school data. We found the test conversion to be accurate and that the MR PEQ scores functioned similarly to the original MR scores. Both performed better than the original PACER scores in predicting VO2max and classifying students. The middle school data generally supported these results. The proposed method is accurate and effective in setting a new field test onto the same scale of a primary field test and determining its cut-off scores.

Generalizability theory analysis of balance error scoring system reliability in healthy young adults.

CONTEXT: An assessment of postural control is commonly included in the clinical concussion evaluation. Previous investigators have demonstrated learning effects that may mask concussion-induced balance decrements. OBJECTIVE: To establish the test-retest reliability of the Balance Error Scoring System (BESS) and to provide recommendations that account for known learning effects. DESIGN: Test-retest generalizability study. SETTING: Balance research laboratory. PATIENTS OR OTHER PARTICIPANTS: Young adults (n = 48) free from injuries and illnesses known to affect balance. INTERVENTION(S): Each participant completed 5 BESS trials on each of the assessment dates, which were separated by 50 days. MAIN OUTCOME MEASURE(S): Total score of the BESS was used in a generalizability theory analysis to estimate the overall reliability of the BESS and that of each facet. A decision study was completed to estimate the number of days and trials needed to establish clinical reliability. RESULTS: The overall reliability of the BESS was G = 0.64. The test-retest reliability was improved when male (0.92) and female (0.91) participants were examined independently. Clinically acceptable reliability (greater than 0.80) was established when 3 BESS trials were administered in a single day or 2 trials were administered at different time points. CONCLUSIONS: Learning effects have been noted in individuals with no previous exposure to the BESS. Our findings indicate that clinicians should consider interpreting the mean score from 3 BESS administrations on a given occasion for both normative data comparison and pretest and posttest design. The multiple assessment technique yields clinically reliable scores and provides the sports medicine practitioner with accurate data for clinical decision making.

Arthroscopic suprascapular nerve decompression at the suprascapular notch.

Release of a transverse scapular ligament (TSL) is indicated for the entrapment of the suprascapular nerve (SSN). Previous arthroscopic techniques use step-wise reference landmarks leading to the notch to identify a TSL, and the key landmarks are the conoid ligament of the coracoclavicular ligament and the coracoid. This technique needs considerable amount of fibro-fatty tissue removal, which is time-consuming procedure. The technique described herein uses the superior border of scapula as a key landmark. A lateral portal is used as a viewing portal, and an anterolateral portal, SSN portal, and accessory portal are required for the working portals. To identify the superior border of the scapula, dissection proceeds along the anterior border of the supraspinatus and advances medially into the supraspinatus fossa. Then, the TSL could be identified by palpating laterally along the superior border of scapula as a dimpling portion.

Medial opening wedge high-tibial osteotomy using a kinematic navigation system versus a conventional method: a 1-year retrospective, comparative study.

High tibial osteotomy is a realignment procedure to transfer weight-bearing load to the intact compartment of the knee to alleviate symptoms, slow disease progression, and defer subsequent total knee arthroplasty. To prevent overcorrection or undercorrection, it is not only important to have an exact preoperative calculation of the desired correction angle, but it is also critical to have an accurate intraoperative technique. 85 consecutive patients (90 knees) were enrolled, who were available at 1-year follow-up after a medial opening wedge high tibial osteotomy using a kinematic navigation system or a conventional method, for medial unicompartmental osteoarthritis. On radiographic assessment, the navigation group showed better results than the conventional group in both the mechanical axis and the coordinate of the weight-bearing line on a full-length standing anteroposterior radiograph (3.9 degrees +/- 1.0 degrees vs. 2.7 degrees +/- 2.2 degrees of valgus, P < 0.01), (62.3 +/- 2.9% vs. 58.7 +/- 6.6% coordinate at the tibial plateau, P < 0.01). There was no significant difference in the alteration of tibial slope between the two groups. On clinical assessment, the navigation group showed better results in both the mean Hospital for Special Surgery knee score (84 +/- 8 vs. 79 +/- 7, P < 0.01) and the mean Lysholm knee score (85 +/- 6 vs. 83 +/- 5, P < 0.05). There was no significant difference in operation times between the two groups. Kinematic navigation-guided high tibial osteotomy is a reproducible and reliable procedure compared to conventional high tibial osteotomy.

Arthroscopic treatment of patellofemoral soft tissue impingement after posterior stabilized total knee arthroplasty.

Patellar clunk syndrome is due to a suprapatellar fibrous nodule which develops at the junction between the proximal patella and the quadriceps tendon after total knee arthroplasty. Twelve patients (12 knees) who underwent arthroscopic treatment for the painful patellar crepitus or clunk after total knee arthroplasty had been followed at least for 1 year were enrolled. The average onset of symptom was 10.2 months (from 3 to 21 months) after total knee arthroplasty. All of the patients presented a painful crepitus, catching or clunk at the proximal patella, especially climbing stairs or rising from sitting. Both radiographic and clinical evaluations were done and for the clinical assessment, Knee Society Scores were recorded preoperatively and postoperatively. On their radiographic assessment, there was no evidence of component failure and in only one patient, joint line was elevated more than 8 mm. On arthroscopic examination, hypertrophic synovial tissue was identified at the junction of patella and quadriceps tendon in all patients. The hypertrophic synovial tissue did not engage completely in the intercondylar notch during flexion, but on extension, the lesion impinged on the distal part of anterior flange of femoral component at about 30 degrees to 40 degrees of knee flexion. On clinical assessment at the latest follow-up, the average knee score and function score increased from 63.8+/-4.6 to 90.9+/-3.2, 65.4+/-3.2 to 90.4+/-4.3 respectively. Arthroscopic debridement for soft tissue impingement at the patellofemoral joint following total knee arthroplasty showed satisfactory results.

Effect of extracts from safflower seeds on osteoblast differentiation and intracellular calcium ion concentration in MC3T3-E1 cells.

Although safflower seeds have long been used in Korea as herbal medicines, very little research has been published on the effects of safflower seed on bone formation or bone density. The study reported here therefore examined bone nodule formation, calcium uptake, alkaline phosphatase activity, and intracellular concentration of calcium ion [Ca(2+)](i) in murine osteoblastic cells of the MC3T3-E1 line that were cultured on modified Eagle's minimal essential medium alone (controls) or with addition of 0.1% crude extract of safflower seed (experimental group I) or 0.1% aqueous fraction of safflower seed (experimental group II). Fluorescence spectrometry measurement of ([Ca(2+)](i)) showed significantly accelerated rates of osteoblast differentiation in experimental group I (3 microL of crude extract in 8 x 10(4) cells) and experimental group II (2 microL of aqueous fraction in 8 x 10(4) cells) compared to the control group.

Pulmonary arterial reactivity during induced infection of single lung allografts.

OBJECTIVE: Infection is a major cause of mortality in the first year following single lung transplantation and is a distinct risk factor for the development of obliterative bronchiolitis. However, little is known about changes in pulmonary vascular activity in the setting of infection, which might affect and limit function of the graft. Therefore, the aim of this study was to determine how acute infection altered pulmonary arterial reactivity in single lung allografts. Such information could help to develop better diagnostic and therapeutic targets to improve outcome when grafts become infected. METHODS: Following single lung transplantation, dogs were immunosuppressed with methylprednisolone acetate, cyclosporine and azathioprine. On postoperative day 5, infection was induced in one group of dogs by endobronchial inoculation of antibiotic resistant Eschericia coli (infection group, n=5); in the second group, the same amount of culture media without bacteria was flushed into the bronchus (control group, n=4). All animals were medicated under the same drug protocol. On postoperative day 8, lungs were removed, reviewed for histological assessment, pulmonary arteries were isolated, cut into rings and suspended for pharmacological characterization in organ chambers. RESULTS: With acute infections, contractions to phenylephrine and angiotensin-1, but not endothelin-1, were reduced in pulmonary arteries with but not without endothelium. Inhibition of nitric oxide synthase with N(G)-monomethyl-L-arginine, monoacetate salt (L-NMMA) restored these contractions. Endothelium-dependent relaxations to adenosine diphosphate and calcium ionophore, which stimulate release of endothelium-derived nitric oxide by a receptor and non-receptor mediated process, respectively, were not different between groups. Relaxations to nitric oxide also were not different between groups. CONCLUSION: These results suggest that acute infection selectively reduces contractions of pulmonary arteries in part through receptor-mediated release of nitric oxide from the endothelium. Inhibiting nitric oxide during episodes of acute infection may help to improve graft perfusion during episodes of acute infection.