Cell-Engineered Recombinant α-Synuclein: A Gage R&R Validated Protocol.

α-Synuclein (α-Syn) misfolding and its presence in Lewy bodies are observed in almost all Parkinson's disease (PD) patients. Basic biomedical research would benefit from a quick, low-cost approach to purifying α-Syn and developing in vitro and in vivo models for PD. Several research groups utilize PFF-based models, yet the production of α-Syn PFFs is inconsistent, resulting in nonconclusive findings. Some research laboratories prepare recombinant α-Syn (r α-Syn) by molecular cloning to overexpress α-Syn with various purifying techniques. Laboratory-to-laboratory protocols cause considerable variability and sometimes contradictory findings. PD researchers spend more on protein than solving α-Syn's riddles. This article uncovered a novel method for expressing and purifying r α-Syn validated through gage reproducibility and repeatability (Gage R&R). For the production of r α-Syn, we have employed the ability of a high-cell-density-based expression system to overexpress protein in BL21(DE3). A simple, high-throughput, nonchromatographical purification protocol has been devised to facilitate research with higher reproducibility, which was validated through Gage R&R. A crossover experimental design was utilized, and the purified protein was characterized using orthogonal high-end analytical methods, which displayed higher similarity between the isolated r α-Syn. Batch-to-batch variability was the least for produced protein and hence can be utilized for exploring the iceberg of PD.

Safety and Efficacy of 6.2 mm Patellar Button in Resurfacing Less than 20 mm Thin Patella: A Matched Pair Analysis.

PURPOSE: Restoring the native patellar thickness after patellar resurfacing provides optimal function of the knee after arthroplasty and minimises complications related to the patellofemoral articulation. The aim of this study was to assess the usefulness of a thin patellar button (6.2 mm) in patients with a patella thickness of less than 20 mm during total knee arthroplasty. MATERIALS AND METHODS: This is a retrospective case control study. A total of 54 female patients with an intraoperative patellar thickness of ＜20 mm, resurfaced with a patellar button of 6.2 mm in thickness were identified (group 1). They were matched with 54 patients with a patellar thickness of 20-23 mm, resurfaced with a patellar button of 8 mm (group 2), based on age, sex, body mass index, and deformity. A clinical and radiological evaluation was done at a minimum 2-year follow-up. RESULTS: The preoperative mean patellar thickness was 18.94±1.07 mm and was restored to 19.06±0.79 mm in group 1, as compared to 21.63±0.99 mm and 21.72±0.99 mm in group 2. The mean postoperative range of motion was 122.22°±9.25° in group 1 and 123.52°±8.72° in group 2 (p=0.13). No patellar bone or button related complications were observed in any patient in either group. CONCLUSIONS: The 6.2 mm thin patella is useful to restore the native thickness in patients with a patellar thickness of less than 20 mm without risk of button fracture, loosening or overstuffing.

Effect of Closed Suction Drain on Blood Loss and Transfusion Rates in Simultaneous Bilateral Total Knee Arthroplasty: A Prospective Randomized Study.

PURPOSE: Simultaneous bilateral total knee arthroplasty (TKA) is associated with excessive blood loss and morbidity arising from postoperative reduction in hemoglobin (Hb). The purpose of this prospective randomized study was to determine if drains have any effect on blood loss, postoperative reduction in Hb levels and transfusion rates compared to no drainage in simultaneous bilateral TKAs. MATERIALS AND METHODS: Two hundred and thirty patients who underwent simultaneous bilateral TKA by a single surgeon were randomly allotted to drain or no-drain group (n=115 in each group). Postoperative Hb level, blood loss volume and transfusion rate were compared between the two groups. RESULTS: The mean postoperative Hb level (p=0.38), blood loss volume (p=0.33) and transfusion rate (p=0.52) in the drain group were not significantly different compared to the no-drain group. No statistical difference was found in terms of complications, readmissions and mortality rates between the two groups. CONCLUSIONS: No significant difference was observed in the two groups with respect to blood loss and blood transfusion. Non-drainage does not offer an advantage over drainage with respect to conserving blood in simultaneous bilateral TKA.