Complement contributes to antibody-mediated protection against repeated SHIV challenge.

The first clinical efficacy trials of a broadly neutralizing antibody (bNAb) resulted in less benefit than expected and suggested that improvements are needed to prevent HIV infection. While considerable effort has focused on optimizing neutralization breadth and potency, it remains unclear whether augmenting the effector functions elicited by broadly neutralizing antibodies (bNAbs) may also improve their clinical potential. Among these effector functions, complement-mediated activities, which can culminate in the lysis of virions or infected cells, have been the least well studied. Here, functionally modified variants of the second-generation bNAb 10-1074 with ablated and enhanced complement activation profiles were used to examine the role of complement-associated effector functions. When administered prophylactically against simian-HIV challenge in rhesus macaques, more bNAb was required to prevent plasma viremia when complement activity was eliminated. Conversely, less bNAb was required to protect animals from plasma viremia when complement activity was enhanced. These results suggest that complement-mediated effector functions contribute to in vivo antiviral activity, and that their engineering may contribute to the further improvements in the efficacy of antibody-mediated prevention strategies.

Polyfunctional Tier 2-Neutralizing Antibodies Cloned following HIV-1 Env Macaque Immunization Mirror Native Antibodies in a Human Donor.

Vaccine efforts to combat HIV are challenged by the global diversity of viral strains and shielding of neutralization epitopes on the viral envelope glycoprotein trimer. Even so, the isolation of broadly neutralizing Abs from infected individuals suggests the potential for eliciting protective Abs through vaccination. This study reports a panel of 58 mAbs cloned from a rhesus macaque (Macaca mulatta) immunized with envelope glycoprotein immunogens curated from an HIV-1 clade C-infected volunteer. Twenty mAbs showed neutralizing activity, and the strongest neutralizer displayed 92% breadth with a median IC50 of 1.35 µg/ml against a 13-virus panel. Neutralizing mAbs predominantly targeted linear epitopes in the V3 region in the cradle orientation (V3C) with others targeting the V3 ladle orientation (V3L), the CD4 binding site (CD4bs), C1, C4, or gp41. Nonneutralizing mAbs bound C1, C5, or undetermined conformational epitopes. Neutralization potency strongly correlated with the magnitude of binding to infected primary macaque splenocytes and to the level of Ab-dependent cellular cytotoxicity, but did not predict the degree of Ab-dependent cellular phagocytosis. Using an individualized germline gene database, mAbs were traced to 23 of 72 functional IgHV alleles. Neutralizing V3C Abs displayed minimal nucleotide somatic hypermutation in the H chain V region (3.77%), indicating that relatively little affinity maturation was needed to achieve in-clade neutralization breadth. Overall, this study underscores the polyfunctional nature of vaccine-elicited tier 2-neutralizing V3 Abs and demonstrates partial reproduction of the human donor's humoral immune response through nonhuman primate vaccination.

Effects of implant rotational malposition on contact surface area after implantation of the augmented glenoid baseplate in the setting of glenoid bone loss.

AIM OF THE STUDY: Augmented glenoid baseplates are utilized in reverse total shoulder arthroplasty in the setting of glenoid bone loss. These implants permit lateralization of the joint line and correction of bony version abnormalities. To allow bone preservation in the setting of abnormal bony version or deficiency, the backside of the augmented glenoid baseplate is not perpendicular to the axis of the central post/screw. Thus, if the baseplate is implanted with any rotational malposition, this could affect the backside contact area available for ingrowth. The purpose of this study was to assess if rotational malpositioning of a full-wedge augmented baseplate alongside the axis of the central screw significantly affects the glenoid implant backside contact area. METHODS: Seven synthetic scapulas (Sawbones, Vashon, WA) were used to implant a 15° full-wedge glenoid baseplate (Wright Medical, Memphis, TN) according to the manufacturer's technique. The contact pressure between the baseplate and the glenoid surface at rotational positions 5°, 10°, and 15° clockwise (CW) and counterclockwise (CCW) from the central axis was measured with Extreme Low Fujifilm Prescale (Tekscan, Boston, MA). The data was analyzed digitally to obtain a percentage of contact surface area. To evaluate gross contact, a computed tomography (CT) scan was performed and manual measurements of contact between the glenoid and the baseplate were conducted using a standardized axial CT slice. RESULTS: The average contact area at zero degrees of malrotation was 37.26 ± 3.27%. Average contact areas for the simulated malposition cases were 13.99 ± 9.39% at 15° CCW, 24.89 ± 5.11% at 10° CW, and 19.32 ± 3.13% at 15° CW. Each of these results was significant (p < 0.003). On computed tomography, at 15° CCW, the contact area decreased by 39%; at 15° CW, the contact area decreased by 38%. DISCUSSION: The use of augmented glenoid baseplates presents a technical challenge. It is difficult to avoid implant malrotation along the axis of the central peg/screw, because the final rotation of the baseplate must be chosen while the implant is several centimeters away from the bone. This study found that 10° and 15° malrotation about the glenoid baseplate's central axis leads to significant decreases in the implant-bone contact area. CONCLUSIONS: When implanting an augmented baseplate for total shoulder arthroplasty, it is important to minimize baseplate malrotation to decrease the risk of baseplate loosening.

Evaluation of rotator cuff abduction moment arms for superior capsular reconstruction and reverse total shoulder arthroplasty.

PURPOSE: The rotator cuff (RC) muscles contribute to dynamic stability and rotational actions of the glenohumeral joint. Moment arm can be used to demonstrate the potential work a muscle contributes to a musculoskeletal joint rotation. This study aimed to understand the moment arm contributions of the RC muscles and explore changes following a complete supraspinatus tear treated with either superior capsular reconstruction (SCR) or reverse total shoulder arthroplasty (rTSA). METHODS: Five fresh-frozen cadaveric specimens were prepared and mounted in an apparatus where each intact RC muscle was held in tension with a line of action toward its origin on the scapula. Mean moment arms for each muscle were determined experimentally based on Optotrak data collected during cadaveric shoulder arm abduction. RESULTS: Using ANOVA testing, our analysis demonstrated significant differences (p < 0.001) in infraspinatus and teres minor moment arms after rTSA compared to the intact shoulder model. After SCR, significant differences (p < 0.001) were seen in teres minor, with these differences being statistically similar to the changes seen in teres minor after rTSA. Subscapularis showed no significant difference in moment arm values between the models (p = 0.148). CONCLUSION: Our results illustrate that mean moment arms were preserved in the RC muscles after complete supraspinatus tear. This study also shows evidence that subscapularis function may be maintained after SCR or rTSA. After SCR, infraspinatus may maintain similar abduction ability compared to the anatomical shoulder, while teres minor ability may increase. Infraspinatus may have decreased abduction ability after rTSA while teres minor may have increased ability.

Antibody-mediated complement activation in pathology and protection.

Antibody-dependent complement activity is associated not only with autoimmune morbidity, but also with antitumor efficacy. In infectious disease, both recombinant monoclonal antibodies and polyclonal antibodies generated in natural adaptive responses can mediate complement activity to protective, therapeutic or disease-enhancing effect. Recent advances have contributed to the structural resolution of molecular complexes involved in antibody-mediated complement activation, defining the avid nature of participating interactions and pointing to how antibody isotype, subclass, hinge flexibility, glycosylation state, amino acid sequence and the contextual nature of the cognate antigen/epitope are all factors that can determine complement activity through impact on antibody multimerization and subsequent recruitment of complement component 1q. Beyond the efficiency of activation, complement activation products interact with various cell types that mediate immune adherence, trafficking, immune education and innate functions. Similarly, depending on the anatomical location and extent of activation, complement can support homeostatic restoration or be leveraged by pathogens or neoplasms to enhance infection or promote tumorigenic microenvironments, respectively. Advances in means to suppress complement activation by intravenous immunoglobulin (IVIG), IVIG mimetics and complement-intervening antibodies represent proven and promising exploratory therapeutic strategies, while antibody engineering has likewise offered frameworks to enhance, eliminate or isolate complement activation to interrogate in vivo mechanisms of action. Such strategies promise to support the optimization of antibody-based drugs that are able to tackle emerging and difficult-to-treat diseases by improving our understanding of the synergistic and antagonistic relationships between antibody mechanisms mediated by Fc receptors, direct binding and the products of complement activation.

1-D imaging of rotation-vibration non-equilibrium from pure rotational ultrafast coherent anti-Stokes Raman scattering.

We present one-dimensional (1-D) imaging of rotation-vibration non-equilibrium measured by two-beam pure rotational hybrid femtosecond/picosecond coherent anti-Stokes Raman scattering (fs/ps CARS). Simultaneous measurements of the spatial distribution of molecular rotation-vibration non-equilibrium are critical for understanding molecular energy transfer in low temperature plasmas and hypersonic flows. However, non-equilibrium CARS thermometry until now was limited to point measurements. The red shift of rotational energy levels by vibrational excitation was used to determine the rotational and vibrational temperatures from 1-D images of the pure rotational spectrum. Vibrational temperatures up to 5500 K were detected in a CH4/N2 nanosecond-pulsed pin-to-pin plasma within 2 mm near the cathode. This approach enables study of non-equilibrium systems with 40 µm spatial resolution.

Electric field vector measurements via nanosecond electric-field-induced second-harmonic generation.

Electric-field-induced second-harmonic generation, or E-FISH, has received renewed interest as a nonintrusive tool for probing electric fields in gas discharges and plasmas using ultrashort laser pulses. An important contribution of this work lies in establishing that the E-FISH method works effectively in the nanosecond regime, yielding field sensitivities of about a kV/cm at atmospheric pressure from a 16 ns pulse. This is expected to broaden its applicability within the plasma community, given the wider access to conventional nanosecond laser sources. A Pockels-cell-based pulse-slicing scheme, which may be readily integrated with such nanosecond laser systems, is shown to be a complementary and cost-effective option for improving the time resolution of the electric field measurement. Using this scheme, a time resolution of ∼3 ns is achieved, without any detriment to the signal sensitivity. This could prove invaluable for nonequilibrium plasma applications, where time resolution of a few nanoseconds or less is often critical. Finally, we take advantage of the field vector sensitivity of the E-FISH signal to demonstrate simultaneous measurements of both the horizontal and vertical components of the electric field.

Biomechanical effects of superior capsular reconstruction in a rotator cuff-deficient shoulder: a cadaveric study.

BACKGROUND: Superior capsular reconstruction (SCR) has been gaining popularity as a treatment for irreparable rotator cuff tears (RCTs), especially in younger patients. This biomechanical study aimed to investigate how SCR affects functional abduction force, humeral head migration, and passive range of motion following an irreparable RCT. We hypothesized that SCR will restore these parameters to nearly intact shoulder levels. METHODS: Six fresh-frozen cadaveric shoulders were evaluated using a custom biomechanical testing apparatus. Each shoulder was taken through 3 conditions: (1) intact (control); (2) irreparable, complete supraspinatus (SS) tear; and (3) SCR. Functional abduction force, superior humeral head migration, and passive range of motion, including axial shoulder rotation, were measured in static condition at 0°, 30°, and 60° of glenohumeral abduction. Data were analyzed using the paired Student t test or Wilcoxon signed rank test, depending on the results of normality testing. RESULTS: The irreparable SS tear resulted in significantly lower functional abduction force at 30° of abduction (P = .01) and a trend toward a decrease (P = .17) at 60° compared with the intact configuration. SCR shoulders produced greater functional force at 0° compared with the tear configuration (P = .046). Humeral head migration was significantly increased by 4.4 and 3.0 mm at 0° and 30° of abduction, respectively, when comparing the intact vs. SS tear configurations (P = .001). SCR decreased superior migration down to levels of intact shoulders at 0° and 30° of abduction (P = .008 and P = .013, respectively) and was not significantly different from the intact configuration at any angle. SCR decreased passive shoulder extension compared with the tear configuration and increased abduction compared with the intact configuration (P = .007 and P = .03, respectively). The overall arc of axial rotation was not significantly different between SCR and the intact configuration at any angle. CONCLUSIONS: In the setting of an irreparable SS tear, SCR restores key biomechanical parameters of the shoulder to intact levels. SCR should be considered for qualifying patients with irreparable RCTs.

Do Demographic or Anthropometric Factors Affect Lateralization of the Tibial Tubercle in a Diverse Population? A Magnetic Resonance Imaging Analysis.

INTRODUCTION: The tibial tubercle-trochlear groove (TT-TG) distance is commonly used to evaluate and guide treatment for patellar instability. There is limited data available regarding TT-TG variability based on patient demographics and anthropometric factors. MATERIALS AND METHODS: TT-TG was measured on magnetic resonance imaging (MRI) for 384 consecutive adult patients. Demographic information for the corresponding was then gathered from the medical record and analyzed. Demographic variables analyzed included age, sex, race, height, weight, and body mass index (BMI). RESULTS: Mean TT-TG among the 384 patients was 12.68mm (standard deviation [SD]: 4.13mm, 95% confidence interval [CI] 12.26-13.10mm, range, 3.2-27.0mm), and there was a significant correlation with height (p=0.009), weight (p=0.017), and race (p<0.001). However, there was no significant correlation seen with sex (p=0.854), BMI (p=0.253), or age (p=0.096). Height and African American race were identified as independent predictors of increased TT-TG (p=0.007 and p<0.001, respectively); and females were found to have an increased TT-TG relative to height (p=0.015). CONCLUSION: Tibial tubercle-trochlear groove distance was significantly correlated with race and height in the 384 patients examined. These findings may help explain clinical differences in these patients and help establish "norms" for patients of various ethnic and anthropometric variability.

The biomechanics of the supraspinatus-deficient shoulder treated with superior capsular reconstruction vs. reverse total shoulder arthroplasty-experimental study.

AIM OF THE STUDY: Our objective was to compare biomechanical effects of superior capsular reconstruction (SCR) and reverse total shoulder arthroplasty (rTSA) on shoulder motion, in the setting of an irreparable supraspinatus (SS) tear. We hypothesized that rTSA would produce greater improvement in abduction force and shift the humerus inferiorly, while SCR would produce greater range of motion (ROM) and prevent superior migration of the humerus during abduction. METHODS: Six cadaveric shoulders were evaluated using a custom biomechanical apparatus. Each shoulder underwent four experimental conditions: (1) intact/control, (2) irreparable SS tear, (3) SCR using dermal allograft, and (4) rTSA without SCR. Deltoid abduction force, superior humeral head translation, and passive range of motion were measured in static tendon loading condition at 0, 30, and 60° of glenohumeral abduction. RESULTS: Both rTSA and SCR restored abduction force to intact levels at all abduction angles. rTSA significantly increased abduction force compared with the SS-deficient shoulder at 0, 30, 60° (p = 0.04), while SCR produced a significant increase at 0° (p = 0.05) abduction. rTSA inferiorly shifted the humeral head by 27 mm (p = 0.002). SCR restored superior humeral head translation to intact SS levels. Compared with SCR, rTSA resulted in 25° less passive abduction (p = 0.001) without significant differences in forward flexion/extension. Compared with SCR, rTSA achieved 10° less passive internal rotation at 0° abduction (p = 0.03) and 26° and 17° greater external rotation at 30° and 60° abduction, respectively (p = 0.03, p = 0.04). DISCUSSION: Our investigation found that abduction force was restored to intact cuff levels by both procedures, without significant differences between the two techniques. SCR restored superior humeral head migration and rTSA translated the humerus inferiorly. rTSA resulted in decreased passive abduction ROM and increased external rotation, compared with SCR. CONCLUSION: Both SCR and rTSA restore key biomechanical parameters following an irreparable SS tear, although SCR offers superior passive abduction ROM.

1D time evolving electric field profile measurements with sub-ns resolution using the E-FISH method.

We present an approach for the measurement of time evolving electric field profiles in atmospheric pressure plasma discharges using electric field induced second harmonic generation (E-FISH). While the E-FISH effect has been known of for some time, recent advances in laser and detection technology have allowed the method to be utilized for spatial measurements of an arbitrarily applied electric field. A cylindrical lens is used to focus the femtosecond laser light to a line and an intensified charge coupled device is used for detection, allowing for one-dimensional (1D) spatial resolution on the order of ∼50 µm. Measurements have been carried out verifying the spatial resolution using a spatially periodic, localized electric field. Calibrated 1D electric field measurements have been completed with a time resolution of 500 ps in a laminar cold atmospheric pressure plasma jet with argon core flow and N2 co-flow powered by a nanosecond (ns) pulse dielectric barrier discharge. The field was shown to propagate as an ionization wave, with a velocity of ∼0.3 mm/ns.

Posterior dislocation of the long head biceps tendon: a case report, specific radiographic finding, and review of the literature.

Posterior dislocation of the long head of the biceps tendon uncommonly occurs with traumatic shoulder injury. The diagnosis is almost always associated with anterior shoulder dislocation which often cannot be reduced. We present a case of traumatic posterior dislocation of the long head of the biceps tendon with no reported history of shoulder dislocation, but instead a rare and specific radiographic finding. The imaging features are described, the relevant literature reviewed, and salient features discussed.

In Hospital Outcomes of Total Hip Arthroplasty in the Medicaid Population.

Total hip arthroplasty (THA) is one of the most common orthopaedic procedures. This study's purpose was to evaluate national trends, patient demographics and hospital outcomes for Medicaid patients who underwent a primary THA. The National Hospital Discharge Survey (NHDS) database was queried for patients undergoing THA from 2001-2010. Patients were stratified into two groups based on insurance. We found from 2001-2005, Medicaid accounted for 2.38% of all THA performed, increasing insignificantly to 2.61% between 2006-2010. The Medicaid group was younger (50.3 vs. 65.6 years, p < 0.01). Length of stay was longer for the Medicaid group (4.6 vs. 4.0 days, p < 0.01). Medicaid patients were more likely to be discharged home (53.7% vs. 47.2%, p < 0.01) and less likely to be discharged to rehabilitation facilities (24.4% vs. 29.0%, p < 0.05). In conclusion, we discovered that the rate of Medicaid insurance in patients undergoing primary THA was stable through 2010, prior to the Affordable Care Act. We found Medicaid THA patients had longer length of stay, despite being a mean 15 years younger than the non-Medicaid cohort. Medicaid insurance status should be factored into risk adjustment models to avoid creating additional disincentive to treat the Medicaid population. (Journal of Surgical Orthopaedic Advances 28(4):281-284, 2019).

Biomechanical evaluation of distal biceps tendon repair using tension slide technique and knotless fixation technique.

BACKGROUND: Acute distal biceps tendon ruptures are uncommon injuries that often affect young active males, typically resulting from an eccentric load on their dominant extremity. The purpose of this study was to compare pullout strength and tendon gapping in the tension slide technique (TST) versus a knotless fixation technique (KFT). METHODS: Two sets of experiments were performed using cadaveric elbow specimens. In the first experiment, eight elbows from different cadavers were tested to compare TST with a standard locking whipstitch with KFT, four elbows in each group, using a standard locking whipstitch. In the second experiment, 12 elbows were used to study the differences between TST with a standard locking whipstitch with KFT using suture tape reinforced whipstitch (RKFT), using the TST data from the first and second experiment. Each experiment evaluated gapping after cyclic loading and the second experiment also tested the construct to load to failure. RESULTS: Gapping for KFT with a standard locking whipstitch was 10.64 mm versus 2.69 mm for the TST after 1000 cycles (P = 0.016). A reinforced whipstitch significantly improved the failure to gap on the KFT with no significant difference in gapping when compared to TST after 3000 cycles (P = 0.36). The resultant gapping for TST and KST was 2.08 mm and 2.99 mm (P = 0.91), respectively. Load to failure for TST and KFT were 282 Nm and 328 Nm (P = 0.20), respectively. CONCLUSION: Bone-tendon gap resistance of a KFT repair of a torn distal biceps tendon is limited by suture technique. Using a tape reinforced locking whipstitch, the repair is as strong as TST repair. LEVELS OF EVIDENCE: Basic Science.

Survey study suggests that reverse total shoulder arthroplasty is becoming the treatment of choice for four-part fractures of the humeral head in the elderly.

PURPOSE: The role of reverse total shoulder arthroplasty (RTSA) for three and four-part proximal humerus fractures is evolving. However, there does not appear to be a clear consensus amongst surgeons. The purpose of this study is to further define the standard of care, assessing surgeon preference and treatment considerations for management of such fractures. METHODS: Orthopaedic surgeons were surveyed on their training, practice setting, and experience regarding management of four-part proximal humerus fractures. The survey also presented five representative cases to assess treatment preferences. RESULTS: Two hundred five surgeons responded to the survey with fellowship training in shoulder and elbow surgery (114), orthopaedic trauma (35) or sports medicine/other training (56). There was no difference between respondents with years in practice and confidence with performing RTSA, however, surgeons in the academic setting were more confident in performing the surgery. Surgeons preferred RTSA for management of four-part fractures in patients over age 65. However, they also trended to favour hemiarthroplasty with higher co-morbidities. Physicians with more than 11 years of experience were more likely to choose hemiarthroplasty for older and high comorbidity patients. RTSA was not the preferred treatment method for younger, active patients. Patient age and fracture pattern had a greater influence on the surgeon's decision. CONCLUSIONS: There is a consensus in our study population that RTSA is the preferred treatment for four-part proximal humerus fractures for elderly patients with patient age and fracture pattern being the most important factors in making management decisions. LEVEL OF EVIDENCE: Level III - Case controlled study.

Glenoid version and size: does gender, ethnicity, or body size play a role?

INTRODUCTION: Variations in glenoid morphology among patients of different gender, body habitus, and ethnicity have been of interest for surgeons. Understanding these anatomical variations is a critical step in restoring normal glenohumeral structure during shoulder reconstruction surgery. METHODS: Retrospective review of 108 patient shoulder CT scans was performed and glenoid version, AP diameter and height were measured. Statistical multiple regression models were used to investigate the ability of gender and ethnicity to predict glenoid AP diameter, height, and version independently of patient weight and height. RESULTS: The mean glenoid AP diameter was 24.7 ± 3.5, the mean glenoid height was 31.7 ± 3.7, and the mean glenoid version was 0.05 ± 9.05. According to our regression models, males would be expected to exhibit 8.4° more glenoid retroversion than females (p = 0.003) and have 2.9 mm larger glenoid height compared to females (p = 0.002). The predicted male glenoid AP diameter was 3.4 mm higher than that in females (p < 0.001). Hispanics demonstrated 6.4° more glenoid anteversion compared to African-Americans (p = 0.04). Asians exhibited 4.1 mm smaller glenoid AP diameters than African-Americans (p = 0.002). An increase of 25 kg in patient weight resulted in 1 mm increase in AP diameter (p = 0.01). CONCLUSIONS: Gender is the strongest independent predictor of glenoid size and version. Males exhibited a larger size and more retroverted glenoid. Patient height was found to be predictive of glenoid size only in patients of the same gender. Although variations in glenoid size and version are observed among ethnicities, larger sample size ethnic groups will be necessary to explore the precise relations. Surgeons should consider gender and ethnic variations in the pre-operative planning and surgical restoration of the native glenohumeral relationship. LEVEL OF EVIDENCE: Anatomic Study.

Can high-friction intraannular material increase screw pullout strength in osteoporotic bone?

BACKGROUND: Osteoporotic bone brings unique challenges to orthopaedic surgery, including a higher likelihood of problematic screw stripping in cancellous bone. Currently, there are limited options to satisfactorily repair stripped screws. Additionally, nonstripped screws hold with less purchase in osteoporotic bone. QUESTIONS/PURPOSES: This study attempts to answer the following questions: (1) Does high-friction intraannular (HFIA) augmentation increase pullout strength in osteoporotic and in severely osteoporotic bone; and (2) can HFIA repair stripped bone thread in osteoporotic and severely osteoporotic bone? METHODS: We measured screw pullout strength using a synthetic bone model in three groups: (1) predrilled nonstripped control holes as controls; (2) predrilled nonstripped augmented with HFIA; and (3) predrilled stripped holes repaired with HFIA. We tested this in osteoporotic and severely osteoporotic synthetic bone for a total of six test groups. We measured screw pullout force using an electromechanical tensile-testing machine comparing pullout force between the test groups and controls. RESULTS: HFIA augmentation did not increase pullout force compared with the control group in the osteoporotic bone model (489 ± 175 versus 607 ± 76, respectively; effect size = 0.94 [95% confidence interval {CI}, -1.75 to 0.08], p = 0.06). However, in severely osteoporotic cancellous bone that was augmented, the HFIA material generated more pullout force than the control (51 ± 18 versus 35 ± 16, respectively; effect size = 0.94 [95% CI, -0.02 to 1.82], p = 0.05). In stripped holes, HFIA partially restored pullout strength but remained weaker than controls in both osteoporotic and severely osteoporotic bone models (osteoporotic: 320 ± 59 versus 607 ± 76, respectively; effect size = -4.28 [95% CI, -5.57 to -2.51], p < 0.001; severely osteoporotic: 21 ± 8 versus 35 ± 16, respectively; effect size = -1.13 [95% CI, -2.0 to 0.12], p = 0.027). CONCLUSIONS: HFIA effectively augmented severely osteoporotic bone for screw purchase, but this effect was not seen for osteoporotic bone. In a model simulating both osteoporotic and severely osteoporotic bone, we found that HFIA can be used to repair stripped screw holes, but the resulting construct remains weaker than nonstripped controls. CLINICAL RELEVANCE: The HFIA material looks promising as a potential solution to stripped screws in osteoporotic bone. However, this material has yet to be tested in human bone. Furthermore, the fine mesh material could be damaged by autoclaving and could break off in vivo causing unknown tissue reactions. We recommend additional testing in a living animal model to better understand how living bone will react to the HFIA material.

National trends and perioperative outcomes in primary and revision total shoulder arthroplasty: Trends in total shoulder arthroplasty.

INTRODUCTION: Total shoulder arthroplasty (TSA) is a highly successful procedure for management of glenohumeral arthritis, fractures and rotator cuff tears. The purpose of this study was to evaluate patient demographics, perioperative outcomes and assess recent national trends in both primary and revision TSA. METHODS: The National Hospital Discharge Survey database was searched for patients admitted to US hospitals for primary and revision TSA from 2001 to 2010. RESULTS: A total of 1,297 patients who underwent primary TSA and 184 patients who underwent revision TSA were identified. The rates of primary TSA (r = 0.88) and revision TSA (r = 0.85) both demonstrated a strong positive correlation with time. The mean patient age of the primary group was significantly higher than the revision group. Gender was not significantly different between the groups. There was no significant difference in the racial make-up between the revision and primary groups. African Americans accounted for 3.3 % of primaries versus 4.3 % of revisions (p = 0.615). Revision TSA patients had a significantly longer average LOS (3.06 days vs 2.46 days, p < 0.01), more medical comorbidities (6.0 vs 5.1 comorbidities, p < 0.01) and a higher rate of developing a myocardial infarction (2.2 % versus 0 %, p < 0.01) than the primary TSA group. CONCLUSIONS: This study demonstrates that the rate of TSA is rapidly increasing in the US, with over a four-fold increase in revisions and five-fold increase in primaries over the ten years studied.

BenzoHTag, a fluorogenic self-labeling protein developed using molecular evolution.

Self-labeling proteins are powerful tools in chemical biology as they enable the precise cellular localization of a synthetic molecule, often a fluorescent dye, with the genetic specificity of a protein fusion. HaloTag7 is the most popular self-labeling protein due to its fast labeling kinetics and the simplicity of its chloroalkane ligand. Reaction rates of HaloTag7 with different chloroalkane-containing substrates is highly variable and rates are only very fast for rhodamine-based dyes. This is a major limitation for the HaloTag system because fast labeling rates are critical for live-cell assays. Here, we report a molecular evolution system for HaloTag using yeast surface display that enables the screening of libraries up to 108 variants to improve reaction rates with any substrate of interest. We applied this method to produce a HaloTag variant, BenzoHTag, which has improved performance with a fluorogenic benzothiadiazole dye. The resulting system has improved brightness and conjugation kinetics, allowing for robust, no-wash fluorescent labeling in live cells. The new BenzoHTag-benzothiadiazole system has improved performance in live-cell assays compared to the existing HaloTag7-silicon rhodamine system, including saturation of intracellular enzyme in under 100 seconds and robust labeling at dye concentrations as low as 7 nM. It was also found to be orthogonal to the silicon HaloTag7-rhodamine system, enabling multiplexed no-wash labeling in live cells. The BenzoHTag system, and the ability to optimize HaloTag for a broader collection of substrates using molecular evolution, will be very useful for the development of cell-based assays for chemical biology and drug development.

Multiplexed no-wash cellular imaging using BenzoTag, an evolved self-labeling protein.

Self-labeling proteins are powerful tools for exploring biology as they enable the precise cellular localization of a synthetic molecule, often a fluorescent dye. HaloTag7 is the most popular self-labeling protein due to its broad utility, its bio-orthogonality, and the simplicity of its chloroalkane ligand. However, reaction rates of HaloTag7 with different chloroalkane-containing substrates are highly variable and rates are only very fast for rhodamine-based dyes. This is a major limitation for the HaloTag system because fast labeling rates are critical for live-cell assays. Here, we use yeast surface display to produce a HaloTag variant, BenzoTag, with improved performance with a fluorogenic benzothiadiazole dye. Molecular evolution improved conjugation kinetics and increased the signal from the dye-protein complex, allowing for robust, no-wash fluorescence labeling in live cells. The new BenzoTag-benzothiadiazole system has improved performance compared to the best existing HaloTag7-silicon rhodamine system, including saturation of intracellular enzyme in under 100 seconds and robust labeling at dye concentrations as low as 7 nM. The BenzoTag system was also found to be sufficiently orthogonal to the HaloTag7-silicon rhodamine system to enable multiplexed no-wash labeling in live cells. The BenzoTag system will be immediately useful for a large variety of cell-based assays monitoring biological processes and drug action in real time.