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      Sex-Based Analysis of Potential Inflammation-Related Protein Biomarkers in the Aqueous Humor of Patients With Diabetes Mellitus

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          Abstract

          Purpose

          To investigate whether men have higher inflammatory protein biomarker concentrations in their aqueous humor (AH) compared with women in groups of patients with varying levels of diabetic disease.

          Methods

          This cross-sectional study included AH specimens from 59 adult patients comprised of three groups: no diabetes mellitus (DM), DM without diabetic retinopathy (DR), and DM with proliferative diabetic retinopathy (PDR). Protein biomarker concentration values were quantified using a commercial proximity extension assay-based technique.

          Results

          Intersex comparisons of concentration values for each protein biomarker revealed no discoveries in patients with no DM or with PDR. In contrast, 24 discoveries were detected in patients with DM without DR. The mean concentration value for all 24 protein biomarkers was higher in men compared with women. Of these 24 proteins, 12 demonstrated a significant association with sex on multivariate linear regression analysis. The β coefficient results demonstrated a positive association between male sex and concentration value for all 12 of these proteins.

          Conclusions

          Higher AH concentration levels of several potential biomarkers, including chemokines, proteases, proteins involved in programmed cell death, and a T-cell surface protein, were detected in men with DM with no DR. These findings suggest that men may have a more inflammatory disease phenotype compared with women in this group of patients.

          Translational Relevance

          The findings of this study help explain differences in epidemiologic patterns of diabetic retinopathy development between men and women.

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          Most cited references29

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          Adaptive linear step-up procedures that control the false discovery rate

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            Homogenous 96-Plex PEA Immunoassay Exhibiting High Sensitivity, Specificity, and Excellent Scalability

            Medical research is developing an ever greater need for comprehensive high-quality data generation to realize the promises of personalized health care based on molecular biomarkers. The nucleic acid proximity-based methods proximity ligation and proximity extension assays have, with their dual reporters, shown potential to relieve the shortcomings of antibodies and their inherent cross-reactivity in multiplex protein quantification applications. The aim of the present study was to develop a robust 96-plex immunoassay based on the proximity extension assay (PEA) for improved high throughput detection of protein biomarkers. This was enabled by: (1) a modified design leading to a reduced number of pipetting steps compared to the existing PEA protocol, as well as improved intra-assay precision; (2) a new enzymatic system that uses a hyper-thermostabile enzyme, Pwo, for uniting the two probes allowing for room temperature addition of all reagents and improved the sensitivity; (3) introduction of an inter-plate control and a new normalization procedure leading to improved inter-assay precision (reproducibility). The multiplex proximity extension assay was found to perform well in complex samples, such as serum and plasma, and also in xenografted mice and resuspended dried blood spots, consuming only 1 µL sample per test. All-in-all, the development of the current multiplex technique is a step toward robust high throughput protein marker discovery and research.
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              Role of Inflammation in Diabetic Retinopathy

              Diabetic retinopathy is a common complication of diabetes and remains the leading cause of blindness among the working-age population. For decades, diabetic retinopathy was considered only a microvascular complication, but the retinal microvasculature is intimately associated with and governed by neurons and glia, which are affected even prior to clinically detectable vascular lesions. While progress has been made to improve the vascular alterations, there is still no treatment to counteract the early neuro-glial perturbations in diabetic retinopathy. Diabetes is a complex metabolic disorder, characterized by chronic hyperglycemia along with dyslipidemia, hypoinsulinemia and hypertension. Increasing evidence points to inflammation as one key player in diabetes-associated retinal perturbations, however, the exact underlying molecular mechanisms are not yet fully understood. Interlinked molecular pathways, such as oxidative stress, formation of advanced glycation end-products and increased expression of vascular endothelial growth factor have received a lot of attention as they all contribute to the inflammatory response. In the current review, we focus on the involvement of inflammation in the pathophysiology of diabetic retinopathy with special emphasis on the functional relationships between glial cells and neurons. Finally, we summarize recent advances using novel targets to inhibit inflammation in diabetic retinopathy.
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                Author and article information

                Journal
                Transl Vis Sci Technol
                Transl Vis Sci Technol
                tvst
                TVST
                Translational Vision Science & Technology
                The Association for Research in Vision and Ophthalmology
                2164-2591
                12 March 2021
                March 2021
                : 10
                : 3
                : 12
                Affiliations
                [1 ]University of California, San Francisco, Department of Ophthalmology, San Francisco, CA, USA
                [2 ]Zuckerberg San Francisco General Hospital and Trauma Center, Department of Ophthalmology, San Francisco, CA, USA
                Author notes
                Correspondence: Jay M. Stewart, University of California, San Francisco, Department of Ophthalmology, 490 Illinois Street, Floor 5, San Francisco, CA 94143-4081, USA. e-mail: jay.stewart@ 123456ucsf.edu
                Article
                TVST-20-3090
                10.1167/tvst.10.3.12
                7961117
                34003946
                0a4d1044-5d05-428c-a94f-ad08eb180f1c
                Copyright 2021 The Authors

                This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

                History
                : 26 January 2021
                : 13 October 2020
                Page count
                Pages: 8
                Categories
                Article
                Article

                sex,aqueous humor,protein biomarker,diabetes mellitus
                sex, aqueous humor, protein biomarker, diabetes mellitus

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