Towards hemerythrin-based blood substitutes: Comparative performance to hemoglobin on human leukocytes and umbilical vein endothelial cells

Abstract

Hemerythrin is a dioxygen-carrying protein whose oxidative/nitrosative stress-related reactivity is lower than that of hemoglobin, which may warrant investigation of hemerythrin as raw material for artificial oxygen carriers ('blood substitutes'). We report here the first biological tests for hemerythrin and its chemical derivatives, comparing their performance with that of a representative competitor, glutaraldehyde-polymerized bovine hemoglobin. Hemerythrin (native or derivatized) exhibits a proliferative effect on human umbilical vein endothelial cell (HUVEC) cultures, as opposed to a slight inhibitory effect of hemoglobin. A similar positive effect is displayed on human lymphocytes by glutaraldehyde-polymerized hemerythrin, but not by native or polyethylene glycol-derivatized hemerythrin.

Synthesis of structural analogues of hexadecylphosphocholine and their antineoplastic, antimicrobial and amoebicidal activity

“…Separation of lymphocytes from whole blood was made using a method improved by our laboratory [50], by density gradient centrifugation, using Histopaque solution. The lymphocyte ring separated between plasma and the separation solution was harvested using a pipette.…”

Protein-Based Blood Substitutes: Recent Attempts at Controlling Pro-Oxidant Reactivity with and Beyond Hemoglobin

“…It should be noted that addition of antioxidants like ascorbic acid, BSA, catalase, glucose, to poly-Hb does not appear to improve the performance to any significant extent. Tests in human lymphocytes (Figure 2b) show that after 24 h, Hb compounds did not inhibit significantly the lymphocyte viability [21]. Hemoglobin derivatives show a slightly inhibitory effect after 48 h, but not statistically significant.…”

“…Hemoglobin derivatives show a slightly inhibitory effect after 48 h, but not statistically significant. At 72 h a slight difference also begun to develop between native and polymerized Hb, in favor of the polymerized protein [21]. …”

Antitumoral and Immunomodulatory Effect of Mahonia aquifolium Extracts

“…The biologic system used for testing was the suspensions of human PBMC, obtained by venipuncture from a 25-year-old healthy male volunteer, who gave his informed written consent before the blood collection, according to the approvals from the Ethical Committee of the Institute of Oncology "Prof. Dr. Ion Chiricuta" from Cluj-Napoca (IOCN), Romania, member of OECI. CD4+ helper T cells, CD8+ effector T cells, CD19+ B cells, and CD14+ monocytes were separated as previously described [31] (see supplemental data (available here)).…”

“…To assess cytotoxicity serial dilutions from M. aquifolium extracts, stock solutions were prepared, in order to obtain 5 successive concentrations, from 20 μg/mL to 1 μg/mL. The cytotoxicity of the extracts was assessed using the MTS viability dye (CellTiter 96 Proliferation Assay, manufactured by Promega Corporation, Madison, WI, USA), as previously described [31] (see supplemental data).…”