Jamie E. Rayahin

Objective: Few strategies to improve pain outcome in knee osteoarthritis (OA) exist in part because how best to evaluate pain over the long term is unclear. Our objectives were to determine the frequency of a good pain experience outcome based on previously formulated OA pain stages and test the hypothesis that less depression and pain catastrophizing and greater self‐efficacy and social support are each associated with greater likelihood of a good outcome.Methods: Study participants, all with knee OA, reported pain stage at baseline and 2 years. Baseline assessments utilized the Geriatric Depression Scale, Pain Catastrophizing Scale, Arthritis Self‐Efficacy Scale, and Medical Outcomes Study social support survey. Using pain experience stages, good outcome was defined as persistence in or movement to no pain or stage 1 (predictable pain, known trigger) at 2 years. A multivariable logistic regression model was developed to identify independent predictors of a good outcome.Results: Of 212 participants, 136 (64%) had a good pain outcome and 76 (36%) a poor pain outcome. In multivariable analysis, higher self‐efficacy was associated with a significantly higher likelihood of good outcome (adjusted odds ratio [OR] 1.14 [95% confidence interval (95% CI) 1.04–1.24]); higher pain catastrophizing was associated with a significantly lower likelihood of good outcome (adjusted OR 0.88 [95% CI 0.83–0.94]).Conclusion: This stage‐based measure provides a meaningful and interpretable means to assess pain outcome in knee OA. The odds of a good 2‐year outcome in knee OA were lower in persons with greater pain catastrophizing and higher in persons with greater self‐efficacy. Targeting these factors may help to improve pain outcome in knee OA.

Cell lytic peptides are a class of drugs that can be used to selectively kill invading organisms or diseased cells. Several of these peptides have been identified as potential therapeutics. Herein, we report a novel process for purifying recombinant melittin, a cell lytic peptide that inserts into the membranes of cells causing cell lysis, from Escherichia coli. The process involves surfactant and low pH to solubilize melittin fusion proteins from the insoluble fraction of bacterial lysates. We are able to significantly improve purity of the final product and confirm the activity of the peptide. The process yields recombinant melittin that is effective when used to treat U‐87 MG glioma cells and inhibits growth of the gram‐positive pathogenic bacterium Streptococcus pyogenes. We demonstrate a method of repeated extraction of the insoluble protein fraction with mild detergent at a low pH that is able to generate a yield of pure, soluble melittin of ∼0.5–1 mg/L of E. coli culture. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1150–1157, 2013

Chemokine receptor-ligand interactions are important to support functioning of both normal and pathologic cells. The expression and function of chemokine receptors in epithelial ovarian carcinoma (EOC) is largely unknown. Here, we report that the lymphotactin receptor (XCR1) was expressed in primary and metastatic human epithelial ovarian carcinoma (EOC) specimens and cell lines. In contrast, expression of XCR1 was not detected in the normal ovary or in human normal ovarian surface epithelial cells. Our data indicate that XCL1 and XCL2 are either present in the malignant ascites or expressed by the ovarian carcinoma cells. The addition of lymphotactin (XCL1 and XCL2) stimulated migration and proliferation of XCR1-positive cells. Reduction of XCR1 expression in ovarian carcinoma cell line SKOV-3 resulted in abrogated diaphragm and peritoneal wall tumor formation and in reduced frequency of colonic, splenetic, and liver nodules in an in vivo xenograft mouse model. Taken together, our data suggest that XCR1 is expressed early during the course of tumorigenic transformation and contributes towards increased cell migration and proliferation, which can facilitate the prometastatic behavior of EOC cells. Mol Cancer Res; 10(11); 1419–29. ©2012 AACR.

Background: Many branches of biomedical research find use for pure recombinant proteins for direct application or to study other molecules and pathways. Glutathione affinity purification is commonly used to isolate and purify glutathione S-transferase (GST)-tagged fusion proteins from total cellular proteins in lysates. Although GST affinity materials are commercially available as glutathione immobilized on beaded agarose resins, few simple options for in-house production of those systems exist. Herein, we describe a novel method for the purification of GST-tagged recombinant proteins. Results: Glutathione was conjugated to low molecular weight poly(ethylene glycol) diacrylate (PEGDA) via thiol-ene “click” chemistry. With our in-house prepared PEGDA:glutathione (PEGDA:GSH) homogenates, we were able to purify a glutathione S-transferase (GST) green fluorescent protein (GFP) fusion protein (GST-GFP) from the soluble fraction of E. coli lysate. Further, microspheres were formed from the PEGDA:GSH hydrogels and improved protein binding to a level comparable to purchased GSH-agarose beads. Conclusions: GSH containing polymers might find use as in-house methods of protein purification. They exhibited similar ability to purify GST tagged proteins as purchased GSH agarose beads.

(2012). Hybrid nanocrystals: University of Kentucky US20060280680A1. Expert Opinion on Therapeutic Patents: Vol. 22, No. 3, pp. 341-348.