Comparative Analysis of Multiplex Platforms for Detecting Vitreous Biomarkers in Diabetic Retinopathy.

Purpose: To evaluate the feasibility of using the Proximity Extension Assay (PEA) platform to detect biomarkers in vitreous and to compare the findings with results obtained with an electrochemiluminescent (ECL) sandwich immunoassay. Methods: Vitreous samples from patients with proliferative diabetic retinopathy (PDR) and non-diabetic controls were tested using two different proteomics platforms. Forty-one assays were completed with the ECL platform and 459 with the PEA platform. Spearman's rank correlation coefficient (rs ) was used to determine the direction and strength of the relationship between protein levels detected by both platforms. Results: Three hundred sixty-six PEA assays detected the tested protein in at least 25% of samples, and the difference in protein abundance between PDR and controls was statistically significant for 262 assays. Seventeen ECL assays yielded a detection rate ≥ 25%, and the difference in protein concentration between PDR and controls was statistically significant for 13 proteins. There was a subset of proteins that were detected by both platforms, and for those the Spearman's correlation coefficient was higher than 0.8. Conclusions: PEA is suitable for the analysis of vitreous samples, showing a strong correlation with the ECL platform. The detection rate of PEA panels was higher than the panels tested with ECL. The levels of several proinflammatory and angiogenic cytokines were significantly higher in PDR vitreous compared to controls. Translational Relevance: This study provides new information on the yields of small-volume assays that can detect proteins of interest in ocular specimens, and it identifies patterns of cytokine dysregulation in PDR.

Giant lymphedema of the penis and scrotum: a case report.

Lymphedema of the penis and scrotum is a rare entity characterized by enlargement of the skin and subcutaneous tissue of the genital region due to lymphatic drainage impairment. This clinical condition is more frequent in tropical countries due to a higher incidence of filariasis, which, in turn, is the main etiology. We describe the case of a 33-year-old man with large lymphedema of the scrotum and penis due to an acute and chronic inflammatory process, foreign body granuloma, and marked hyalinization. Four consecutive surgical interventions were necessary to remove the great part of the affected tissue, which enabled satisfactory results and improved the patient's quality of life.

Use of a bioamplification assay to detect nonselective recombinants and assess the genetic stability of oncolytic adenoviruses.

Detection of nonselective adenoviruses in tissue- or tumor-selective oncolytic adenovirus preparations presents a technical challenge because of the conditionally replication-competent nature of oncolytic adenoviruses. Although quantitative PCR has been used extensively for detecting specific genes that are likely present in nonselective recombinants, the actual biological activity of nonselective genetic recombinants has not been demonstrated. Therefore, a bioassay that amplifies nonselective adenoviruses through multiple passages in nonpermissive cells was developed to detect biologically active nonselective recombinants using CG7870, a prostate-specific oncolytic adenovirus. The assay was sensitive, and its results were consistent with a quantitative PCR assay for four lots of CG7870. CG0070, a pan-tumor oncolytic adenovirus with no detectable wild-type-like recombinants by PCR, was subjected to a variation of this bioamplification assay using two different nonpermissive cell lines to both verify PCR results and assess its genetic stability under selection pressure. No evidence of the presence of biologically active nonselective recombinants was seen in the original material or after serial passaging in nonpermissive cells. Thus, this bioamplification assay is able to detect nonselective recombinants, and its results are consistent with quantitative PCR assays. A modified version of this assay is also useful for assessing the genetic stability of oncolytic adenoviruses that have no PCR-detectable recombinants.

The Src family kinase Fyn mediates signals induced by TCR antagonists.

FcR nonbinding anti-CD3 epsilon mAbs elicit partial TCR signaling that leads to T cell unresponsiveness and tolerance in vivo. In this study, the membrane-proximal events that promote T cell inactivation by FcR nonbinding anti-CD3 mAbs were examined. In the context of FcR nonbinding anti-CD3, TCR complexes did not aggregate and failed to translocate into glycolipid-enriched membrane microdomains. Furthermore, FcR nonbinding anti-CD3 mAbs induced tyrosine phosphorylation of the Fyn substrate Cbl, but not the ZAP-70 substrate linker for activation of T cells. Overexpression of Fyn, but not Lck, restored the mitogenicity of FcR nonbinding anti-CD3 in primary T cells. Taken together, these results suggest that Fyn mediates the partial signaling induced by TCR antagonists.