Reliability, validity, and responsiveness of a novel subscapularis outcome score: the Baltimore Orthopedic Subscapularis Score.

Background: Available surveys that evaluate shoulder strength and pain often combine rotator cuff muscles making the test unable to differentiate subscapularis tears from other pathology including concomitant supraspinatus, infraspinatus tears. The purpose of this study was to validate a subscapularis-specific shoulder survey (Baltimore Orthopedic Subscapularis Score) as a viable clinical outcome assessment through analysis of psychometric properties. Methods: A 5-question survey was given to a study population of 390 patients, 136 of whom had full thickness rotator cuff tears with a minimum score of 5 (better) and a maximum score of 25 (worse). Surveys were given during the initial consultation, preoperative visit, and postoperative visit. Content validity, construct validity, test-retest reliability, responsiveness to change, internal consistency, and minimal clinically important difference using distribution and anchor-based methods were determined for our subscapularis function survey. Results: A high correlation was reported on test-retest reliability (intraclass correlation coefficient = 0.89). An acceptable internal consistency was reported for all patients surveyed (Cronbach alpha = 0.91). Floor and ceiling effects for patients with rotator cuff pathology were minimized (1% for both). Patients with an isolated subscapularis tear scored worse than supraspinatus/infraspinatus tears and exhibited similar dysfunction as patients with a supraspinatus/infraspinatus/subscapularis tear. An acceptable construct validity was reported with subscapularis-involved tears demonstrating higher scores with significance (P < .05). There was excellent responsiveness to change with a standardized response mean of 1.51 and effect size of 1.27 (large > 0.8). The minimal clinically important difference using a distribution and anchor-based method was 4.1 and 4.6, respectively. Among patients with rotator cuff tears in this population, a score of 22 or higher predicts a subscapularis tear 75% of the time, in spite of its low overall prevalence. Conclusion: The subscapularis shoulder score demonstrated acceptable psychometric performance for outcomes assessment in patients with rotator cuff disease. This survey can be used as an effective clinical tool to assess subscapularis function.

Glenoid bony morphology of osteoarthritis prior to shoulder arthroplasty: what the surgeon wants to know and why.

Shoulder arthroplasty is performed with increasing frequency, and osteoarthritis is the most common indication for this procedure. However, the glenoid side of the joint is widely recognized as a limiting factor in the long-term durability of shoulder replacement, and osteoarthritis leads to characteristic bony changes at the glenoid which can exacerbate this challenge by reducing the already limited glenoid bone stock, by altering biomechanics, and by interfering with operative exposure. This article reviews the Walch classification system for glenoid morphology. Several typical findings of osteoarthritis at the glenoid are discussed including central bone loss, posterior bone loss, retroversion, biconcavity, inclination, osteophyte formation, subchondral bone quality, and bone density. The three primary types of shoulder arthroplasty are reviewed, along with several techniques for addressing glenoid deformity, including eccentric reaming, bone grafting, and the use of augmented glenoid components. Ultimately, a primary objective at shoulder arthroplasty is to correct glenoid deformity while preserving bone stock, which depends critically on characterizing the glenoid at pre-operative imaging. Understanding the surgical techniques and the implications of glenoid morphology on surgical decision-making enables the radiologist to provide the morphologic information needed by the surgeon.

Therapeutic postoperative anticoagulation is a risk factor for wound complications, infection, and revision after shoulder arthroplasty.

BACKGROUND: The purpose of the present study was to examine the relationship between postoperative therapeutic anticoagulation, wound complications, infection, and revision. METHODS: Using a national insurance database from 2007 to 2016, patients who underwent shoulder arthroplasty with an indication for postoperative therapeutic anticoagulation in the case of atrial fibrillation or acute postoperative venous thromboembolism were identified. Those with a prescription for a therapeutic anticoagulant within 2 weeks of surgery were identified and compared with controls without postoperative therapeutic anticoagulant prescriptions. Wound complications and postoperative infection at 3 and 6 months, and revision shoulder arthroplasty at 6 months and all time points were then compared in the database using a multivariable logistic regression analysis. RESULTS: A total of 17,272 patients were included, including 684 patients who received therapeutic anticoagulation and 16,588 controls. Patients receiving therapeutic anticoagulation experienced increased wound complications at 3 months (odds ratio [OR] 3.0, 95% confidence interval [CI] 2.0-4.6, P < .0001) and 6 months (OR 2.5, 95% CI 1.7-3.8, P < .0001). Patients receiving therapeutic anticoagulation also experienced increased rates of wound infection at 3 months (OR 1.5, 95% CI 1.1-2.0, P = .007) and 6 months (OR 1.8, 95% CI 1.4-2.3, P < .0001). Finally, patients receiving therapeutic anticoagulation experienced increased rates of revision surgery at 6 months (OR 1.8, 95% CI 1.3-2.5, P = .0003) and within 9 years (OR 1.5, 95% CI 1.1-2.0, P = .007). CONCLUSIONS: Wound complications and revision rates in patients undergoing shoulder arthroplasty who require postoperative therapeutic anticoagulation are significantly elevated compared with controls.

Increasing Burden of Youth Baseball Elbow Injuries in US Emergency Departments.

BACKGROUND: Youth athletes are starting sports earlier and training harder. Intense, year-round demands are encouraging early sports specialization under the perception that it will improve the odds of future elite performance. Unfortunately, there is growing evidence that early specialization is associated with increased risk of injury and burnout. This is especially true of pediatric and adolescent baseball players. PURPOSE/HYPOTHESIS: The purpose of this investigation was to analyze national injury trends of youth baseball players. We hypothesized that while the total number of baseball injuries diagnosed over the past decade would decrease, there would be an increase in adolescent elbow injuries seen nationally. A further hypothesis was that this trend would be significantly greater than other injuries to the upper extremity and major joints. STUDY DESIGN: Descriptive epidemiology study. METHODS: Injury data from the National Electronic Injury Surveillance System, a United States Consumer Product Safety Commission database, were analyzed between January 1, 2006, and December 31, 2016, for baseball players aged ≤18 years. Data were collected on the location of injury, diagnosis, and mechanism of injury. RESULTS: Between 2006 and 2016, an estimated 665,133 baseball injuries occurred nationally. The mean age of the injured players was 11.5 years. The most common injuries diagnosed included contusions (26.8%), fractures (23.6%), and strains and sprains (18.7%). Among major joints, the ankle (25.6%) was most commonly injured, followed by the knee (21.3%), wrist (19.2%), elbow (17.7%), and shoulder (16.2%). The incidence of the ankle, knee, wrist, and shoulder injuries decreased over time, while only the incidence of elbow injuries increased. A linear regression analysis demonstrated that the increasing incidence of elbow injuries was statistically significant against the decreasing trend for all baseball injury diagnoses, as well as ankle, knee, wrist, hand, and finger injuries (P < .05). Additionally, the only elbow injury mechanism that increased substantially over time was throwing. CONCLUSION: The current investigation found that while the incidence of baseball injuries sustained by youth players is decreasing, elbow pathology is becoming more prevalent and is more commonly being caused by throwing. Given that the majority of elbow injuries among adolescent baseball players are overuse injuries, these findings underscore the importance of developing strategies to continue to ensure the safety of these youth athletes.

CT-Based 3D Printing of the Glenoid Prior to Shoulder Arthroplasty: Bony Morphology and Model Evaluation.

To demonstrate the 3D printed appearance of glenoid morphologies relevant to shoulder replacement surgery and to evaluate the benefits of printed models of the glenoid with regard to surgical planning. A retrospective review of patients referred for shoulder CT was performed, leading to a cohort of nine patients without arthroplasty hardware and exhibiting glenoid changes relevant to shoulder arthroplasty planning. Thin slice CT images were used to create both humerus-subtracted volume renderings of the glenoid, as well as 3D surface models of the glenoid, and 11 printed models were created. Volume renderings, surface models, and printed models were reviewed by a musculoskeletal radiologist for accuracy. Four fellowship-trained orthopaedic surgeons specializing in shoulder surgery reviewed each case individually as follows: First, the source CT images were reviewed, and a score for the clarity of the bony morphologies relevant to shoulder arthroplasty surgery was given. The volume rendering was reviewed, and the clarity was again scored. Finally, the printed model was reviewed, and the clarity again scored. Each printed model was also scored for morphologic complexity, expected usefulness of the printed model, and physical properties of the model. Mann-Whitney-Wilcoxon signed rank tests of the clarity scores were calculated, and the Spearman's ρ correlation coefficient between complexity and usefulness scores was computed. Printed models demonstrated a range of glenoid bony changes including osteophytes, glenoid bone loss, retroversion, and biconcavity. Surgeons rated the glenoid morphology as more clear after review of humerus-subtracted volume rendering, compared with review of the source CT images (p = 0.00903). Clarity was also better with 3D printed models compared to CT (p = 0.00903) and better with 3D printed models compared to humerus-subtracted volume rendering (p = 0. 00879). The expected usefulness of printed models demonstrated a positive correlation with morphologic complexity, with Spearman's ρ 0.73 (p = 0.0108). 3D printing of the glenoid based on pre-operative CT provides a physical representation of patient anatomy. Printed models enabled shoulder surgeons to appreciate glenoid bony morphology more clearly compared to review of CT images or humerus-subtracted volume renderings. These models were more useful as glenoid complexity increased.

Symptomatic, Unstable Os Acromiale.

Management of a persistently symptomatic, unstable os acromiale remains controversial. An unstable os acromiale is an easy diagnosis to miss and should be specifically evaluated for in patients with shoulder pain and a high degree of clinical suspicion. Surgical options include open or arthroscopic excision and open reduction and internal fixation. Open excision of large fragments has had poor results. Arthroscopic treatment is commonly used for small fragments (preacromion), and the technique has also been used in larger fragments (meso-os), but concerns persist over postoperative weakness secondary to shortening of the deltoid lever arm. Open reduction and internal fixation through a transacromial approach has been shown to have predictable union rates but can be complicated by symptomatic implant. Recent biomechanical studies have expanded our understanding of optimal fixation constructs, which may also decrease implant-related issues. Ultimately, the choice of which procedure to use will be dictated by patient factors such as age, activity level, and the nature of rotator cuff pathology. LEVEL OF EVIDENCE:: Level V.

Determining glenoid component version after total shoulder arthroplasty.

BACKGROUND: Glenoid component loosening after total shoulder arthroplasty (TSA) may occur if retroversion is not corrected to <10°. However, accurately measuring postoperative glenoid component version has been difficult without postoperative computed tomography (CT), adding cost and radiation exposure outside of the standard radiographic follow-up. We present a new method to assess glenoid component version after TSA using only routine preoperative CT and postoperative radiographs (x-rays). METHODS: Preoperative glenoid version was measured using established methods with an axillary x-ray, 2-dimensional CT, and Glenosys software (Imascap, Plouzané, France). Postoperative glenoid component version and inclination were measured for 61 TSA patients using Mimics software (Materialise, Leuven, Belgium) with preoperative CT and postoperative x-rays. Four patients also had postoperative CTs. Glenoid implantation and imaging were performed on 14 cadavers, allowing validation of results against the gold standard postoperative CT glenoid retroversion measurement. RESULTS: Compared with the gold standard, retroversion and inclination measurement error was 2° ± 1° and 2° ± 1°, respectively. Average postoperative version correction was 6° ± 7°, with 35 of 61 patients (57%) corrected to <10° of retroversion. Correlation between preoperative version measurement methods was good to very good, except on the axillary x-ray. Patients not corrected to <10° of retroversion had significantly higher preoperative retroversion (14° ± 6°) than those corrected to <10° (6° ± 7°; P < .00001). CONCLUSIONS: Glenoid component retroversion after TSA can be accurately measured with a method using only routine preoperative CT and postoperative x-rays, validated to within 1.9° of the gold standard postoperative CT measurement. Future studies using this method may correlate glenoid retroversion correction with glenoid component longevity to help optimize shoulder arthroplasty outcomes.

Shoulder Instability: Common Problems and Solutions.

Operative treatment of the unstable shoulder historically has a high success rate. However, the complication rate has risen. This article reviews the pearls and pitfalls to attempt to elucidate the etiology for these complications and failures. Preoperative assessment of the unstable shoulder ultimately is critical to avoid complications, including history, physical examination, and key radiographic features. Intraoperative techniques include appropriate soft tissue mobilization, multiple points of fixation, avoidance of hardware-related problems, and appropriate management of the capsule and bone defects. Finally, postoperative rehabilitation is equally important to regain physiologic range of motion in a safe, supervised fashion.

Outcomes of lesser tuberosity osteotomy in revision anatomic shoulder arthroplasty.

BACKGROUND: Lesser tuberosity osteotomy (LTO) is a common surgical approach during anatomic shoulder arthroplasty. Outcomes of LTO have been shown to be similar to subscapularis tenotomy and peel techniques, but little is known about the outcomes of LTO during revision arthroplasty. METHODS: This retrospective case series included 10 consecutive patients who underwent LTO during revision shoulder arthroplasty at a single institution from 2012 to 2016. Patients underwent a preoperative computed tomography scan to evaluate the lesser tuberosity bone stock. Demographic information, radiographic evidence of LTO healing, outcomes of range of motion, subscapularis strength, and visual analog scale pain scores were analyzed. RESULTS: Revision total shoulder arthroplasty with LTO was performed for glenoid arthritis after hemiarthroplasty in 10 patients. Average age at surgery was 59.8 years, and no humeral stems were revised. Eight of 10 patients had prior subscapularis tenotomy. Average follow-up after revision surgery was 9.2 months. LTO union was documented in 80% and nondisplaced nonunion in 20%. At follow-up, 50% reported mild pain. Subscapularis strength testing was graded normal in 80% and weak in 20%. Average visual analog scale pain improved from 9.4 prerevision to 4.8 postrevision (P < .05). On average, range of motion improved in active forward elevation from 123° to 141° and remained unchanged in active external rotation from 42° to 42°. CONCLUSION: Patients undergoing LTO during revision anatomic shoulder arthroplasty demonstrate successful LTO bony healing, improvement in pain, and improved forward elevation. In select patients not requiring humeral stem revision, LTO is a safe and effective surgical approach to subscapularis management during revision anatomic shoulder arthroplasty.

Management of Failed Shoulder Surgery.

The popularity of shoulder surgery has increased in the past several decades, which has resulted in a concomitant increase in the number of complications that occur in patients who undergo shoulder surgery. Surgeons should understand the complications that may occur in patients who undergo common shoulder procedures. A plan for the management of shoulder surgery complications is necessary regardless whether a surgeon is an expert shoulder surgeon or a novice shoulder surgeon.

Postoperative Deep Vein Thrombosis, Pulmonary Embolism, and Myocardial Infarction: Complications After Therapeutic Anticoagulation in the Patient With Spine Trauma.

STUDY DESIGN: A retrospective review (2001-2014) was conducted using prospectively collected data at a level I trauma center. OBJECTIVE: We sought to determine the incidence and characteristics of complications occurring secondary to therapeutic anticoagulation in adult spine trauma patients. SUMMARY OF BACKGROUND DATA: Numerous studies have assessed prophylactic anticoagulation after spine surgery, but none has investigated the risks of therapeutic doses of anticoagulation for treatment of postoperative thromboembolic events. METHODS: Patients were included if they sustained a postoperative thromboembolic event (deep venous thrombosis, pulmonary embolism, or myocardial infarction). Patients were excluded if anticoagulation was subtherapeutic. Of 1712 patients, 62 who received therapeutic anticoagulation and 174 propensity-matched control patients who did not receive therapeutic anticoagulation were included in the study. RESULTS: Initial anticoagulation was obtained by heparin infusion (51%), low-molecular-weight heparin (LMWH, 46%), and warfarin (3%). Complications requiring unplanned reoperation occurred in 18% of anticoagulated patients and 10% of nonanticoagulated patients (P = 0.17). The reoperation rate after heparin infusion was 31% and after LMWH was 6.5% (P = 0.02). Epidural hematoma occurred in 3% and 1% of anticoagulated and nonanticoagulated patients, respectively. Multivariate logistic regression analysis of the two groups showed a trend toward increased risk of reoperation in the anticoagulation group. Analysis of the heparin infusion subgroup separate from the LMWH subgroup compared with the control group showed an increased risk of reoperation for any complication (odds ratio, 3.57; P = 0.01) and for bleeding complications (odds ratio, 43.1; P = 0.01). CONCLUSION: This is the first study to quantify complications secondary to postoperative therapeutic anticoagulation in spine patients. Postoperative spine trauma patients who underwent therapeutic anticoagulation experienced an unplanned reoperation rate of 18%, including a 3% incidence of spinal epidural hematoma. Therapeutic anticoagulation using heparin infusion seems to drive the overall rate of reoperation (31%) compared with LMWH. LEVEL OF EVIDENCE: 3.

Maximizing Surgical Skills During Fellowship Training.

Orthopaedic surgery fellowship provides an opportunity to further develop skills in a particular subspecialty. However, the condensed time frame, complex skill acquisition, and clinical demands require efficient and effective learning techniques to achieve mastery. As with any advanced task, success during fellowship training can be achieved with active participation and a goal-directed approach. Skill acquisition can be successfully achieved by following a framework that includes preparation, execution, and reflection for every surgical case.

What is the Optimal Reconstruction Option after the Resection of Proximal Humeral Tumors? A Systematic Review.

PURPOSE: The proximal humerus is a common location for both primary and metastatic bone tumors. There are numerous reconstruction options after surgical resection. There is no consensus on the ideal method of reconstruction. METHODS: A systematic review was performed with a focus on the surgical reconstructive options for lesions involving the proximal humerus. RESULTS: A total of 50 articles and 1227 patients were included for analysis. Reoperation rates were autograft arthrodesis (11%), megaprosthesis (10%), RSA (17%), hemiarthroplasty (26%), and osteoarticular allograft (34%). Mechanical failure rates, including prosthetic loosening, fracture, and dislocation, were highest in allograft-containing constructs (APC, osteoarticular allograft, arthrodesis) followed by arthroplasty (hemiarthroplasty, RSA, megaprosthesis) and lowest for autografts (vascularized fibula, autograft arthrodesis). Infections involving RSA (9%) were higher than hemiarthroplasty (0%) and megaprosthesis (4%). Postoperative function as measured by MSTS score were similar amongst all prosthetic options, ranging from 66% to 74%, and claviculo pro humeri (CPH) was slightly better (83%). Patients were generally limited to active abduction of approximately 45° and no greater than 90°. With resection of the rotator cuff, deltoid muscle or axillary nerve, function and stability were compromised even further. If the rotator cuff was sacrificed but the deltoid and axillary nerve preserved, active forward flexion and abduction were superior with RSA. DISCUSSION: Various reconstruction techniques for the proximal humerus lead to relatively similar functional results. Surgical choice should be tailored to anatomic defect and functional requirements.

Anatomical Relationship of the Axillary Nerve to the Pectoralis Major Tendon Insertion.

Axillary nerve injury is a risk of the deltopectoral approach to the proximal humerus. The anterior motor branch is potentially vulnerable during subdeltoid dissection. Insertion of the pectoralis major tendon is an easily identifiable landmark on the humerus. This anatomical study explored whether the superior aspect of the pectoralis major tendon is a useful landmark for localizing the anterior motor branch of the axillary nerve as it travels under the lateral and anterior deltoid muscle. A total of 30 fresh-frozen human bilateral cadaveric upper extremities were examined. A deltopectoral approach was used to expose the pectoralis major tendon insertion and the anterior motor branch of the nerve under the deltoid muscle. The distance between the nerve as it crossed the posterolateral humerus and superior border of the pectoralis major tendon was measured. The axillary nerve was a mean 3.2 mm (range, 0-8 mm) distal to the superior border of the pectoralis major tendon insertion. No significant differences were observed in this anatomical relationship with the shoulder in abduction or external rotation. The nerve was not proximal to the superior border of the pectoralis major tendon in any specimen. The superior border of the pectoralis major tendon insertion represents a reliable landmark for the anterior motor branch of the axillary nerve as it travels under the deltoid muscle. The nerve is located at the level of the proximal centimeter of the pectoralis major tendon. Appreciation of this relationship may decrease risk of injury to the nerve when using a deltopectoral approach. [Orthopedics. 2017; 40(3):e460-e464.].

Os acromiale fixation: a biomechanical comparison of polyethylene suture versus stainless steel wire tension band.

BACKGROUND: Symptomatic hardware is a commonly reported complication after surgical fixation of an unstable meso-type os acromiale. This study compared the biomechanical properties of a cannulated screw tension band construct using a metal wire tension band vs. a suture tension band, considering that the suture construct could allow for decreased hardware burden in the clinical setting. METHODS: A meso-type os acromiale was created in 16 cadaveric shoulders. Two cannulated 4-mm screws were placed in each specimen. Tension band augmentation was accomplished with a 1-mm stainless steel wire (wire group) or a #5 braided polyethylene suture (suture group), with 8 specimens in each group. An inferiorly directed force was applied to the anterior acromion at 1 mm/s on a materials testing machine. Stiffness and ultimate failure load were recorded and analyzed. RESULTS: No significant difference (P = .22) was observed in the ultimate failure load between the wire (228 ± 85 N; range, 114-397 N) and the suture (275 ± 139 N; range, 112-530 N). No significant difference (P = .17) was observed in the stiffness between the wire (28 ± 12 N/mm; range, 18-53 N/mm) and the suture (38 ± 25 N/mm; range, 10-83 N/mm). CONCLUSIONS: Stainless steel wire and polyethylene suture have similar biomechanical strength in the cannulated screw tension band fixation of meso-type os acromiale at time zero.

A Biomechanical Analysis of Anchor Placement for Bankart Repair: Effect of Portal Placement.

During arthroscopic Bankart repair, penetration of suture anchors through the far cortex can compromise the initial biomechanical characteristics of anchor stability and repair integrity. This study compared the placement of suture anchors through a low anterior-inferior rotator interval portal (AI) vs a trans-subscapularis portal to evaluate the rate of anchor perforation as well as biomechanical strength. Ten matched pairs of cadaveric shoulders were randomized to an AI or a trans-subscapularis portal for placement of suture anchors at the 3 o'clock and 5:30 positions. The following measurements were obtained: (1) distance from the portal to the cephalic vein; (2) presence and length of anchor penetration through the inferior glenoid; and (3) ultimate failure strength of the anchors. The distance from the portal to the cephalic vein was significantly greater with the AI vs the trans-subscapularis portal across all specimens (29.9 vs 11.2 mm, P<.05). The rate of anchor penetration was significantly increased in the AI group vs the trans-subscapularis group at the 5:30 position (60% vs 10%, P=.014) but not at the 3 o'clock position (P=.33). Mean pullout strength of the anchors at the 5:30 position trended higher in the trans-subscapularis group, but the difference was not significant (132.8 vs 112.6 N, P=.18). The cephalic vein is closer to the trans-subscapularis portal than to the AI, but is at a safe distance. Both the rate and the degree of glenoid suture anchor penetration were lower with the trans-subscapularis portal compared with the AI at the 5:30 position. Placing anchors through the trans-subscapularis portal provides a safe alternative method, with improved positioning of the inferiormost anchor compared with the traditional AI.

Muscarinic receptor subtype-3 gene ablation and scopolamine butylbromide treatment attenuate small intestinal neoplasia in Apcmin/+ mice.

M3 subtype muscarinic receptors (CHRM3) are over-expressed in colon cancer. In this study, we used Apc(min/+) mice to identify the role of Chrm3 expression in a genetic model of intestinal neoplasia, explored the role of Chrm3 in intestinal mucosal development and determined the translational potential of inhibiting muscarinic receptor activation. We generated Chrm3-deficient Apc(min/+) mice and compared intestinal morphology and tumor number in 12-week-old Apc(min/+)Chrm3(-/-) and Apc(min/+)Chrm3(+/+) control mice. Compared with Apc(min/+)Chrm3(+/+) mice, Apc(min/+)Chrm3(-/-) mice showed a 70 and 81% reduction in tumor number and volume, respectively (P < 0.01). In adenomas, ß-catenin nuclear staining was reduced in Apc(min/+)Chrm3(-/-) compared with Apc(min/+)Chrm3(+/+) mice (P < 0.02). Whereas Apc gene mutation increased the number of crypt and Paneth cells and decreased villus goblet cells, these changes were absent in Apc(min/+)Chrm3(-/-) mice. To determine whether pharmacological inhibition of muscarinic receptor activation attenuates intestinal neoplasia, we treated 6-week-old Apc(min/+) mice with scopolamine butylbromide, a non-subtype-selective muscarinic receptor antagonist. After 8 weeks of continuous treatment, scopolamine butylbromide-treated mice showed a 22% reduction in tumor number (P = 0.027) and a 36% reduction in tumor volume (P = 0.004) as compared with control mice. Compared with Chrm3 gene ablation, the muscarinic antagonist was less efficacious, most probably due to shorter duration of treatment and incomplete blockade of muscarinic receptors. Overall, these findings indicate that interplay of Chrm3 and ß-catenin signaling is important for intestinal mucosal differentiation and neoplasia and provide a proof-of-concept that pharmacological inhibition of muscarinic receptor activation can attenuate intestinal neoplasia in vivo.

Scopolamine treatment and muscarinic receptor subtype-3 gene ablation augment azoxymethane-induced murine liver injury.

Previous work suggests that vagus nerve disruption reduces hepatocyte and oval cell expansion after liver injury. The role of postneuronal receptor activation in response to liver injury has not been ascertained. We investigated the actions of scopolamine, a nonselective muscarinic receptor antagonist, and specific genetic ablation of a key cholinergic receptor, muscarinic subtype-3 (Chrm3), on azoxymethane (AOM)-induced liver injury in mice. Animal weights and survival were measured as was liver injury using both gross and microscopic examination. To assess hepatocyte proliferation and apoptosis, ductular hyperplasia, and oval cell expansion, we used morphometric analysis of 5-bromo-2'-deoxyuridine-, activated caspase-3-, hematoxylin and eosin-, cytokeratin-19-, and epithelial cell adhesion molecule-stained liver sections. Sirius red staining was used as a measure of collagen deposition and its association with oval cell reaction. In AOM-treated mice, both muscarinic receptor blockade with scopolamine and Chrm3 ablation attenuated hepatocyte proliferation and augmented gross liver nodularity, apoptosis, and fibrosis. Compared with control, scopolamine-treated and Chrm3(-/-) AOM-treated mice had augmented oval cell reaction with increased ductular hyperplasia and oval cell expansion. Oval cell reaction correlated robustly with liver fibrosis. No liver injury was observed in scopolamine-treated and Chrm3(-/-) mice that were not treated with AOM. Only AOM-treated Chrm3(-/-) mice developed ascites and had reduced survival compared with AOM-treated wild-type controls. In AOM-induced liver injury, inhibiting postneuronal cholinergic muscarinic receptor activation with either scopolamine treatment or Chrm3 gene ablation results in prominent oval cell reaction. We conclude that Chrm3 plays a critical role in the liver injury response by modulating hepatocyte proliferation and apoptosis.

Bax activates endophilin B1 oligomerization and lipid membrane vesiculation.

Endophilins participate in membrane scission events that occur during endocytosis and intracellular organelle biogenesis through the combined activity of an N-terminal BAR domain that interacts with membranes and a C-terminal SH3 domain that mediates protein binding. Endophilin B1 (Endo B1) was identified to bind Bax, a Bcl-2 family member that promotes apoptosis, through yeast two-hybrid protein screens. Although Endo B1 does not bind Bax in healthy cells, during apoptosis, Endo B1 interacts transiently with Bax and promotes cytochrome c release from mitochondria. To explore the molecular mechanism of action of Endo B1, we have analyzed its interaction with Bax in cell-free systems. Purified recombinant Endo B1 in solution displays a Stokes radius indicating a tetrameric quarternary structure. However, when incubated with purified Bax, it assembles into oligomers more than 4-fold greater in molecular weight. Although Endo B1 oligomerization is induced by Bax, Bax does not stably associate with the high molecular weight Endo B1 complex. Endo B1 oligomerization requires its C-terminal Src homology 3 domain and is not induced by Bcl-xL. Endo B1 combined with Bax reduces the size and changes the morphology of giant unilamellar vesicles by inducing massive vesiculation of liposomes. This activity of purified Bax protein to induce cell-free assembly of Endo B1 may reflect its activity in cells that regulates apoptosis and/or mitochondrial fusion.