Withaferin A Effectively Targets Soluble Vimentin in the Glaucoma Filtration Surgical Model of Fibrosis

Withaferin A (WFA) is a natural product that binds to soluble forms of the type III intermediate filament (IF) vimentin. Currently, it is unknown under what pathophysiological contexts vimentin is druggable, as cytoskeltal vimentin-IFs are abundantly expressed. To investigate druggability of vimentin, we exploited rabbit Tenon\u27s capsule fibroblast (RbTCF) cell cultures and the rabbit glaucoma filtration surgical (GFS) model of fibrosis. WFA potently caused G₀/G₁ cell cycle inhibition (IC₅₀ 25 nM) in RbTCFs, downregulating ubiquitin E3 ligase skp2 and inducing p27(Kip1) expression. Transforming growth factor (TGF)-ß-induced myofibroblast transformation caused development of cell spheroids with numerous elongated invadopodia, which WFA blocked potently by downregulating soluble vimentin and α-smooth muscle actin (SMA) expression. In the pilot proof-of-concept study using the GFS model, subconjunctival injections of a low WFA dose reduced skp2 expression in Tenon\u27s capsule and increased p27(Kip1) expression without significant alteration to vimentin-IFs. This treatment maintains significant nanomolar WFA concentrations in anterior segment tissues that correspond to WFA\u27s cell cycle targeting activity. A ten-fold higher WFA dose caused potent downregulation of soluble vimentin and skp2 expression, but as found in cell cultures, no further increase in p27(Kip1) expression was observed. Instead, this high WFA dose potently induced vimentin-IF disruption and downregulated α-SMA expression that mimicked WFA activity in TGF-ß-treated RbTCFs that blocked cell contractile activity at submicromolar concentrations. These findings illuminate that localized WFA injection to ocular tissues exerts pharmacological control over the skp2-p27(Kip1) pathway by targeting of soluble vimentin in a model of surgical fibrosis

Functional Multijoint Position Reproduction Acuity in Overhead-Throwing Athletes

CONTEXT: Baseball players rely on the sensorimotor system to uphold the balance between upper extremity stability and mobility while maintaining athletic performance. However, few researchers have studied functional multijoint measures of sensorimotor acuity in overhead-throwing athletes. OBJECTIVE: To compare sensorimotor acuity between 2 high-demand functional positions and among planes of motion within individual joints and to describe a novel method of measuring sensorimotor function. DESIGN: Single-session, repeated-measures design. SETTING: University musculoskeletal research laboratory. PATIENTS OR OTHER PARTICIPANTS: Twenty-one National Collegiate Athletic Association Division I baseball players (age = 20.8 +/- 1.5 years, height = 181.3 +/- 5.1 cm, mass = 87.8 +/- 9.1 kg) with no history of upper extremity injury or central nervous system disorder. MAIN OUTCOME MEASURE(S): We measured active multijoint position reproduction acuity in multiple planes using an electromagnetic tracking device. Subjects reproduced 2 positions: arm cock and ball release. We calculated absolute and variable error for individual motions at the scapulothoracic, glenohumeral, elbow, and wrist joints and calculated overall joint acuity with 3-dimensional variable error. RESULTS: Acuity was significantly better in the arm-cock position compared with ball release at the scapulothoracic and glenohumeral joints. We observed significant differences among planes of motion within the scapulothoracic and glenohumeral joints at ball release. Scapulothoracic internal rotation and glenohumeral horizontal abduction and rotation displayed less acuity than other motions. CONCLUSIONS: We established the reliability of a functional measure of upper extremity sensorimotor system acuity in baseball players. Using this technique, we observed differences in acuity between 2 test positions and among planes of motion within the glenohumeral and scapulothoracic joints. Clinicians may consider these differences when designing and implementing sensorimotor system training. Our error scores are similar in magnitude to those reported using single-joint and single-plane measures. However, 3-dimensional, multijoint measures allow practical, unconstrained test positions and offer additional insight into the upper extremity as a functional unit

Functional Multijoint Position Reproduction Acuity in Overhead-Throwing Athletes

Context: Baseball players rely on the sensorimotor system to uphold the balance between upper extremity stability and mobility while maintaining athletic performance. However, few researchers have studied functional multijoint measures of sensorimotor acuity in overhead-throwing athletes. Objective: To compare sensorimotor acuity between 2 high-demand functional positions and among planes of motion within individual joints and to describe a novel method of measuring sensorimotor function. Design: Single-session, repeated-measures design. Setting: University musculoskeletal research laboratory. Patients or Other Participants: Twenty-one National Collegiate Athletic Association Division I baseball players (age = 20.8 ± 1.5 years, height = 181.3 ± 5.1 cm, mass = 87.8 ± 9.1 kg) with no history of upper extremity injury or central nervous system disorder. Main Outcome Measure(s): We measured active multijoint position reproduction acuity in multiple planes using an electromagnetic tracking device. Subjects reproduced 2 positions: arm cock and ball release. We calculated absolute and variable error for individual motions at the scapulothoracic, glenohumeral, elbow, and wrist joints and calculated overall joint acuity with 3-dimensional variable error. Results: Acuity was significantly better in the arm-cock position compared with ball release at the scapulothoracic and glenohumeral joints. We observed significant differences among planes of motion within the scapulothoracic and glenohumeral joints at ball release. Scapulothoracic internal rotation and glenohumeral horizontal abduction and rotation displayed less acuity than other motions. Conclusions: We established the reliability of a functional measure of upper extremity sensorimotor system acuity in baseball players. Using this technique, we observed differences in acuity between 2 test positions and among planes of motion within the glenohumeral and scapulothoracic joints. Clinicians may consider these differences when designing and implementing sensorimotor system training. Our error scores are similar in magnitude to those reported using single-joint and single-plane measures. However, 3-dimensional, multijoint measures allow practical, unconstrained test positions and offer additional insight into the upper extremity as a functional unit