Multiplexing the Quantitation of MAP Kinase Activities Using Differential Sensing.

Protein kinases are therapeutic targets for many human diseases, but the lack of user-friendly quantitative assays limits the ability to follow the activities of numerous kinases at once (multiplexing). To develop such an assay, we report an array of sulfonamido-oxine (SOX)-labeled peptides showing cross-reactivity to different mitogen-activated protein kinases (MAPKs) for use in a differential sensing scheme. We first verified using linear discriminant analysis that the array could differentiate MAPK isoforms. Then, using principal component analysis, the array was optimized based on the discrimination imparted by each SOX-peptide. Next, the activity of individual MAPK families in ternary mixtures was quantified by support vector machine regression. Finally, we multiplexed the quantification of three MAPK families using partial least squares regression in A549 cell lysates, which has possible interference from other kinase classes. Thus, our method simultaneously quantifies the activity of multiple kinases. The technique could be applied to other protein kinase families and the monitoring of diseases.

Postsurgical Pyoderma Gangrenosum After Penile Inversion Vaginoplasty: Case With Review of Diagnostic and Management Strategies.

Background: Postsurgical pyoderma gangrenosum (PSPG) is a highly uncommon and unpredictable wound healing complication. Rapid progression of ulcers at incisions can cause unfettered dehiscence. Most commonly, PSPG involves breast procedures; however, in this work, we detail a case of a patient who developed PSPG 10 days postoperatively after penile inversion vaginoplasty. Methods: The patient in this case underwent a penile inversion vaginoplasty with orchiectomy in the standard fashion. She had no risk factors for PSPG. Following an uncomplicated hospital stay, the patient developed difficulty with pain control and increasing serous drainage on the 10th postoperative day. On readmission, the patient was found to have developed large, mildly purulent ulcers throughout the perineal wound edges. On exam under anesthesia, the neovaginal canal was found to be patent and intact. The dehisced portions of the incisions were left open and redressed with occlusive bismuth-petrolatum dressing. Dermatology was promptly consulted with suspicion for PSPG. The patient was started on an 18-day prednisone taper with cyclosporine, along with doxycycline and ciprofloxacin. Results: After 5 days of immunosuppressive treatment, the ulcers visibly converted to healthy granulation tissue and were no longer actively purulent. Following another washout, the dehisced wound edges were reapproximated. At follow-up, the patient had no evidence of PSPG recurrence and continued dilating on schedule. Our patient recovered from PSPG without further complications and a satisfactory aesthetic result. Conclusions: This unique case highlights the importance of prompt dermatological consultation, immunosuppression, and avoidance of further pathergy in the setting of suspicion for PSPG.

Social Justice Education in CAATE-Accredited Post-Baccalaureate Athletic Training Programs.

Social justice education (SJE) prepares clinicians to provide patient-centered care. Our purpose was to assess SJE in CAATE-accredited, post-baccalaureate professional athletic training programs. We used a cross-sectional, online survey. Individuals (n=215) in their last year of post-baccalaureate professional education or those who were within 6 months post-graduation participated. The survey included closed and open-ended questions regarding types of education, topics of education, perceived confidence, and levels of agreement regarding identifying, addressing, and applying social justice concepts. Students from different cultural ethnicities reported having different educational experiences relative to their formal and informal instruction. Significant differences were identified between cultural ethnicity groups on perceptions of professors' knowledge (p<0.001), preceptors' knowledge (p<0.001), and agreement their program prepared them to address social justice issues (p<0.001). Two domains emerged from the open-ended responses: 1) social justice defined and 2) educational needs. Participants described social justice as equality, equity, and justice for minoritized people. Participants described SJE as not occurring or limited, and they expressed a desire for active practical experiences from heterogeneous and unbiased sources. As programs move to incorporate CAATE standards, more frequent and varied SJE is expected from minoritized students.

WORKPLACE CLIMATE FOR SEXUAL AND GENDER MINORITIES IN ATHLETIC TRAINING.

CONTEXT: Sexual and gender minorities (SGM) are individuals with sexual orientations, gender identities and/or expressions that differ from cultural norms. SGMs often face workplace discrimination and report decreased physical and emotional well-being from discrimination. OBJECTIVE: To explore the workplace climate of SGM athletic trainers (AT). DESIGN: Sequential mixed-methods study. SETTING: Web-based survey and interviews. PATIENTS OR OTHER PARTICIPANTS: Criterion sampling of SGM ATs (117 survey participants and 12 interview participants). DATA COLLECTION AND ANALYSIS: We modified the LGBTQ Inclusion Assessment and the Organizational Self-Assessment for the survey and developed a semi-structured interview script (scale-level content validity index=0.94). We used means, standard deviations, frequencies and the consensual qualitative research tradition to characterize participant responses. Trustworthiness was established through reflexivity (researchers checking bias throughout the research process), member-checking, multi-analyst triangulation, internal and external auditing. RESULTS: Participants indicated their workplace was inclusive (24, 20.5%), somewhat inclusive (29, 24.8%), not inclusive (14, 12.0%), or did not indicate at all (50, 42.7%). Respondents most often indicated they were unsure of the stage of change their organizations and organizational units were in addressing LGBTQPIA+ issues in the workplace as well as specific actions taken for inclusion. Two domains emerged from the interview data: safety and inclusion. The safety domain represents aspects of the workplace climate that make the participants feel safe including organizational initiatives (12/12), patient-centered policies (7/12), local and federal regulations (7/12), and signaling (12/12). The inclusion domain represents how the participants felt a sense of belonging to the organization through their own experience (12/12), through the experiences of their patients (9/12), and through an infrastructure designed for inclusion (12/12). Participants expressed both the affirmative and the negative feelings of safety and inclusion throughout their responses. CONCLUSIONS: Organizations must take both structural and cultural actions to address the issues of exclusion and lack of safety.

Quantification of ERK Kinase Activity in Biological Samples Using Differential Sensing.

The understanding of complex biological systems requires an ability to evaluate interacting networks of genes, proteins, and cellular reactions. Enabling technologies that support the rapid quantification of these networks will facilitate the development of biological models and help to identify treatment targets and to assess treatment plans. The biochemical process of protein phosphorylation, which underlies almost all aspects of cell signaling, is typically evaluated by immunoblotting procedures (Western blot) or more recently proteomics procedures, which provide qualitative estimates of the concentration of proteins and their modifications in cells. However, protein modifications are difficult to correlate with activity, and while immunoblotting and proteomics approaches have the potential to be quantitative, they require a complex series of steps that diminish reproducibility. Here, a complementary approach is presented that allows for the rapid quantification of a protein kinase activity in cell lysates and tumor samples. Using the activity of cellular ERK (extracellular signal-regulated kinase) as a test case, an array sensing approach that utilizes a library of differential peptide-based biosensors and chemometric tools was used to rapidly quantify nanograms of active ERK in micrograms of unfractionated cell lysates and tumor extracts. This approach has the potential both for high-throughput and for quantifying the activities of multiple protein kinases in a single biological sample. The critical advantages of this differential sensing approach over others are that it removes the need for the addition of exogenous inhibitors to suppress the activities of major off-target kinases and allows us to quantitate the amount of active kinase in tested samples rather than measuring the changes in its activity upon induction or inhibition.