The ability of a GroEL-based bio-layer interferometry (BLI) assay to detect structurally altered and/or aggregated species of pharmaceutically relevant proteins is demonstrated. Assay development included optimizing biotinylated-GroEL immobilization to streptavidin biosensors, combined with biophysical and activity measurements showing native and biotinylated GroEL are both stable and active. First, acidic fibroblast growth factor (FGF-1) was incubated under conditions known to promote (40 C) and inhibit (heparin addition) molten globule formation. Heat exposed (40 C) FGF-1 exhibited binding to GroEL-biosensors, which was significantly diminished in the presence of heparin. Second, a polyclonal human IgG solution containing 6-8% non-native dimer showed an increase in higher molecular weight aggregates upon heating by size exclusion chromatography (SEC). The poly IgG solution displayed binding to GroEL-biosensors initially with progressively increased binding upon heating. Enriched preparations of the IgG dimers or monomers showed significant binding to GroEL-biosensors. Finally, a thermally treated IgG1 monoclonal antibody (mAb) solution also demonstrated increased GroEL-biosensor binding, but with different Additional Supporting Information may be found in the online version of this article. Subhashchandra Naik and Ozan S. Kumru contributed equally to the work. kinetics. The bound complexes could be partially to fully dissociated after ATP addition (i.e., specific GroEL binding) depending on the protein, environmental stress, and the assay's experimental conditions. Transmission electron microscopy (TEM) images of GroEL-mAb complexes, released from the biosensor, also confirmed interaction of bound complexes at the GroEL binding site with heat-stressed mAb. Results indicate that the GroEL-biosensor-BLI method can detect conformationally altered and/or early aggregation states of proteins, and may potentially be useful as a rapid, stability-indicating biosensor assay for monitoring the structural integrity and physical stability of therapeutic protein candidates.

Background Plastic Surgeons and patients increasingly use social media. Despite evidence implicating its importance in Plastic Surgery, the large amount of data has made social media difficult to study. Objectives This study seeks to provide a comprehensive assessment of Plastic Surgery content throughout the world using techniques for analyzing large-scale data. Methods ‘#PlasticSurgery’ was used to search public Instagram posts. Metadata was collected from posts between December 2018 and August 2020. In addition to descriptive analysis, we created two instruments to characterize textual data: a multi-lingual dictionary of procedural hashtags and a rule-based text classification model to categorize the source of the post. Results Plastic Surgery content yielded more than 2 million posts, 369 million likes, and 6 billion views globally over the 21-month study. The United States had the most posts of 182 countries studied (26.8%, 566,206). Various other regions had substantial presence including Istanbul, Turkey, which led all cities (4.8%, 102,208). Our classification model achieved high accuracy (94.9%) and strong agreement with independent raters (κ= 0.88). Providers accounted for 40% of all posts (847,356) and included Physician (28%), Plastic Surgery (9%), Advanced-Practice-Practitioners and Nurses (1.6%), Facial Plastics (1.3%), and Oculoplastics (0.2%). Content between Plastics and non-Plastics groups demonstrated high textual similarity, and only 1.4% of posts had a verified source. Conclusions Plastic Surgery content has immense global reach in social media. Textual similarity between groups coupled with the lack of an effective verification mechanism presents challenges in discerning the source and veracity of information.

Background Retrospective studies evaluating tissue oximetry in a more recent cohort have shown superiority in flap outcomes. This study compares the use of tissue oximetry in a historical cohort to clinical observation and handheld doppler in a more recent cohort. We hypothesize that there is no benefit to using tissue oximetry. Methods A retrospective review was performed on patients who underwent abdominal‐based autologous breast reconstruction by five microsurgeons at an academic institution from 2009 to 2020. Method of postoperative flap monitoring was determined then operative details and complications were analyzed. Results 1367 flaps were reviewed; 740 flaps in 460 patients were monitored with clinical observation and tissue oximetry, and 627 flaps in 391 patients were monitored with clinical observation and handheld doppler. There were no statistical differences in ischemic (p = .59) or congestive complications (p = .41), flap salvage rates when exploring for venous or arterial compromise (p = .52), or early flap loss (p = .56). Although not significant, acute flap‐related return to the operating room was lower in the doppler group (4.6%) compared to the oximetry group (6.1%; p = .22). Flaps monitored with tissue oximetry had a statistical increase in length of stay (4.8 ± 1.4 days vs. 3.8 ± 1.6 days; p ≤ .001). The rates of late partial flap loss and fat necrosis were significantly higher in the oximetry group (2.6%, 19/740 vs. 0.3%, 2/740; p = .04) and (18.2%, 135/740 vs. 13.6%, 85/627; p = .02), respectively. Conclusions There is no statistical benefit to the use of tissue oximetry compared to handheld doppler in flap monitoring with regards to flap outcomes.

Background: Nipple reconstruction has been linked to patient satisfaction; however, there is debate about the validity of these findings in autologous breast reconstruction patients. This study hypothesized that satisfaction would increase with nipple reconstruction following autologous breast reconstruction. Methods: A comparison study was performed of autologous breast reconstruction patients. Patients completed a survey that included BREAST-Q and nipple satisfaction measures. A chart review identified reconstructive details. Results: A total of 191 patients completed the survey (48 percent response rate), with an average age of 53.7 ± 10.0 years and follow-up time of 2.8 ± 1.5 years. Nipple-areola complex reconstruction was completed in 33 percent of patients (63 of 191). Nipple-areola complex tattoos were used most frequently [n = 37 (58 percent)], followed by local flaps [n = 10 (16 percent)], free nipple-areola complex grafts [n = 9 (14 percent)], and a combination of local flaps and tattoos [n = 7 (11 percent)]. In comparison to women who did not undergo nipple-areola complex reconstruction, women who underwent any type of nipple reconstruction had a statistically higher BREAST-Q score for Sexual Well-Being (60 ± 24 versus 50 ± 22; p = 0.01), Postoperative Satisfaction with Breasts (65 ± 11 versus 61 ± 12; p = 0.01), and Satisfaction with Surgeon (97 ± 6 versus 93 ± 16; p = 0.009). The average nipple satisfaction score was 74 ± 19. There were correlations between the nipple satisfaction score and BREAST-Q scores for Sexual Well-Being (r = 0.50; p < 0.001), Psychosocial Well-Being (r = 0.43; p < 0.001), and Postoperative Satisfaction with Breasts (r = 0.43; p < 0.001). Conclusion: Reconstruction of the nipple-areola complex is an important part of autologous breast reconstruction, resulting in increased sexual well-being and satisfaction with reconstructed breasts.