MPAPASS software enables stitched multiplex, multidimensional EV repertoire analysis and a standard framework for reporting bead-based assays.

Extracellular vesicles (EVs) of various types are released or shed from all cells. EVs carry proteins and contain additional protein and nucleic acid cargo that relates to their biogenesis and cell of origin. EV cargo in liquid biopsies is of widespread interest owing to its ability to provide a retrospective snapshot of cell state at the time of EV release. For the purposes of EV cargo analysis and repertoire profiling, multiplex assays are an essential tool in multiparametric analyte studies but are still being developed for high-parameter EV protein detection. Although bead-based EV multiplex analyses offer EV profiling capabilities with conventional flow cytometers, the utilization of EV multiplex assays has been limited by the lack of software analysis tools for such assays. To facilitate robust EV repertoire studies, we developed multiplex analysis post-acquisition analysis (MPAPASS) open-source software for stitched multiplex analysis, EV database-compatible reporting, and visualization of EV repertoires.

Glioma-Derived miRNA-Containing Extracellular Vesicles Induce Angiogenesis by Reprogramming Brain Endothelial Cells.

Glioblastoma (GBM) is characterized by aberrant vascularization and a complex tumor microenvironment. The failure of anti-angiogenic therapies suggests pathways of GBM neovascularization, possibly attributable to glioblastoma stem cells (GSCs) and their interplay with the tumor microenvironment. It has been established that GSC-derived extracellular vesicles (GSC-EVs) and their cargoes are proangiogenic in vitro. To further elucidate EV-mediated mechanisms of neovascularization in vitro, we perform RNA-seq and DNA methylation profiling of human brain endothelial cells exposed to GSC-EVs. To correlate these results to tumors in vivo, we perform histoepigenetic analysis of GBM molecular profiles in the TCGA collection. Remarkably, GSC-EVs and normal vascular growth factors stimulate highly distinct gene regulatory responses that converge on angiogenesis. The response to GSC-EVs shows a footprint of post-transcriptional gene silencing by EV-derived miRNAs. Our results provide insights into targetable angiogenesis pathways in GBM and miRNA candidates for liquid biopsy biomarkers.

INFIX versus plating for pelvic fractures with disruption of the symphysis pubis.

PURPOSE: The purpose of this study is to compare INFIX to plating in the treatment of unstable pelvic ring injuries with disruption of the symphysis. METHODS: Twenty-four patients treated with INFIX were compared to 28 patients fixed by plating. All patients had anterior and posterior fixation. Injuries were classified using the Young and Burgess and AO/OTA classification systems. Reductions of the pelvic ring were assessed using the pelvic deformity index (PDI) and symphyseal widening. Patients were contacted to get functional outcomes using the Majeed scoring system and complications were tabulated . RESULTS: INFIX was inferior to plating at reducing symphyseal widening (INFIX 10.72± 5.0 Plates 6.97 ± 3.39 P = 0.012) but similar in reducing the pelvic deformity index. (INFIX 0.0221± .015 Plates 0.0190 ± .0105 P = 0 .38). Majeed scores were similar 83.95 ± 15.2 (median 89, range 51-100) for INFIX and 77.67± 16.7 (median 79, range 54-100) for plating. Complications included infection (1 (4%) INFIX , 4 (14%) plates), improper hardware placement or failure (2 (8%) INFIX, 3 (11%) plates), and heterotopic ossification (11 (46%) INFIX, 16 (57.1%) plates). Infection in the plated patients was related to urological injury in 3/4 cases. CONCLUSIONS: Plating provides better reduction of the pubic symphysis and requires only one surgery. Outcomes scores were similar. INFIX may be preferable in obese patients, young women of childbearing age or those with urological injury.

Midterm Radiographic and Functional Outcomes of the Anterior Subcutaneous Internal Pelvic Fixator (INFIX) for Pelvic Ring Injuries.

OBJECTIVE: To describe our experience using the anterior internal pelvic fixator (INFIX) for treating pelvic ring injuries. DESIGN: Case Series. SETTING: Level 1 Trauma Center. PATIENTS: Eighty-three patients with pelvic ring injuries were treated with INFIX. Follow-up average was 35 months (range 12-80.33). INTERVENTION: Surgical treatment of pelvic ring injuries included reduction, appropriate posterior fixation, and INFIX placement. OUTCOME MEASUREMENTS: Reduction using the pelvic deformity index and pubic symphysis widening, Majeed functional scores, complications; infection, implant failure, heterotopic ossification (HO), nerve injury, and pain. RESULTS: All patients healed in an appropriate time frame (full weight bearing 12 weeks postoperation). The average pelvic deformity index reduction (injury = 0.0420 ± 0.0412, latest FU = 0.0254 ± 0.0243) was 39.58%. The average reduction of pubic symphysis injuries was 56.92%. The average Majeed score of patients at latest follow-up was 78.77 (range 47-100). Complications were 3 infections, 1 case of implant failure, 2 cases implantation too deep, 7 cases of lateral femoral cutaneous nerve irritation, and 3 cases of pain associated with the device. HO was seen in >50% of the patients, correlated with increased age (P < 0.007), injury severity score (P < 0.05) but only 1 case was symptomatic. CONCLUSIONS: The pelvic injuries had good functional and radiological outcomes with INFIX and the appropriate posterior fixation. The downside is removal requiring a second anesthetic, there is a learning curve, HO often occurs, the lateral femoral cutaneous nerve may get irritated which often resolves once the implants are removed. Surgery-specific implants need to be developed. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Investigating the Universality of Preoperative Health-Related Quality of Life (HRQoL) for Surgically Treated Spinal Deformity in Young Adults: A Propensity Score-Matched Comparison Between African and US Populations.

STUDY DESIGN: Retrospective analysis of propensity score-matched (PSM) observational cohorts. OBJECTIVES: To evaluate and compare preoperative health-related quality of life (HRQoL) scores and radiographic measurements of young African and US adults with spinal deformity (ASD). SUMMARY OF BACKGROUND DATA: Young ASD patients in the United States are motivated more to correct coronal and sagittal plane deformities than to alleviate pain. Motivations for surgical correction in young ASD patients in Africa have not been previously investigated. METHODS: Retrospective review of two large databases of African and US patients with ASD. African patients who underwent ASD surgery were PSM by age, gender, and pelvic tilt with US patients. Preoperative radiographic parameters and HRQoL scores (ODI, SRS-22r, back/leg pain) were compared between cohorts. Pearson correlations used to evaluate relationships between radiographic parameters and HRQoL scores. RESULTS: Fifty-four US patients (average age 22.9 ± 4.9 years; 0% African American) and 54 African patients (24.6 ± 7.2 years) met inclusion criteria. Compared to the United States, African patients had significantly lower body mass index (21.1 ± 3.3 vs. 24.6 ± 7.2) and more severe scoliosis, coronal malalignment, and sagittal malalignment (p < .05). Africans also had significantly better Oswestry Disability Index (12.8 vs. 17.7), worse Scoliosis Research Society questionnaire (SRS-22r)-Appearance (2.5 vs. 3.2), SRS-Function (3.3 vs. 3.9), and SRS-Total (3.2 vs. 3.5) scores than US patients (p < .05). SRS-Appearance scores correlated with Cobb angles of the upper thoracic (r = -0.321), thoracic (r = -0.277), and thoracolumbar (r = -0.300) curves for US patients. For African patients, global sagittal alignment and C7 inclination correlated with SRS-Appearance (r = -0.347, -0.346, respectively). CONCLUSIONS: Young African ASD patients have significantly more severe deformity, less disability, and worse SRS-22r scores preoperatively than a matched cohort of US patients. Spinal deformity and associated poor self-image appear to be the major drivers of surgical intervention in this cohort. Global malalignment in African patients is most closely correlated with appearance scores and should be surgically addressed accordingly. LEVEL OF EVIDENCE: Level III.

Adult Native Septic Arthritis in an Inner City Hospital: Effects on Length of Stay.

The objective of this retrospective study was to assess what factors affected length of stay (LOS) in 183 adult patients with native septic arthritis. Diagnosis was based on a representative physical examination, fluid cell count/Gram stain, and organisms isolated from joint fluid culture. Data included demographics, comorbidities, laboratory results, treatment, and discharge times. Joint fluid cultures were positive in 55% (100 of 183) of the patients, and these patients were the subjects of this study. Blood cultures were taken for 65 patients and were positive in 54%; when positive, they were found to be the same as isolates from joint fluid analysis 91% of the time. Pathogens found in joint fluid analysis were as follows: methicillin-susceptible Staphylococcus aureus (MSSA), 44%; methicillin-resistant S aureus (MRSA), 21%; Streptococcus species, 14%; Pseudomonas, 10%; and other organisms, 11%. Surgical washout less than 24 hours from diagnosis affected LOS (12.25 vs 16.96 days for >24 hours; P<.05), but pathogen type and comorbid conditions did not. Average time for culture sensitivities was 4±1 days. Almost half of the patients had MSSA. Delays that could be controlled were getting an early diagnosis and expedient surgical washout of the joint. A lack of insurance and a requirement of intravenous antibiotics prolonged stay, whereas age, sex, and ethnicity did not. Waiting for bacterial sensitivities was a factor that could not be controlled. The authors believe that polymerase chain reaction or other technologies could lead to early diagnosis and expedient surgery. Effective oral antibiotics against resistant organisms would help the patients leave the hospital earlier. [Orthopedics. 2016; 39(4):e674-e679.].