Interactive Dynamics of Cell Volume and Cell Death in Human Erythrocytes Exposed to α-Hemolysin from Escherichia coli

α-hemolysin (HlyA) of E. coli binds irreversibly to human erythrocytes and induces cell swelling, ultimately leading to hemolysis. We characterized the mechanism involved in water transport induced by HlyA and analyzed how swelling and hemolysis might be coupled. Osmotic water permeability (Pf) was assessed by stopped-flow light scattering. Preincubation with HlyA strongly reduced Pf in control- and aquaporin 1-null red blood cells, although the relative Pf decrease was similar in both cell types. The dynamics of cell volume and hemolysis on RBCs was assessed by electrical impedance, light dispersion and hemoglobin release. Results show that HlyA induced erythrocyte swelling, which is enhanced by purinergic signaling, and is coupled to osmotic hemolysis. We propose a mathematical model of HlyA activity where the kinetics of cell volume and hemolysis in human erythrocytes depend on the flux of osmolytes across the membrane, and on the maximum volume that these cells can tolerate. Our results provide new insights for understanding signaling and cytotoxicity mediated by HlyA in erythrocytes.

Toxicological Activity of the Methanolic Leaf Extract of Some Medicinal Plants Used in Sokoto Township and Environs

Inspite of the availability of different antiseptic and antibiotics in most localities in some parts of the world, there is still a number of information on the usage of some local plants in various kinds of treatments of different ill-health conditions. Leaves of Ocimum basilicum, Leptadania hastata and Momordica balsamina are locally used by traditional birth attendants at pre and post-partum periods. The present study investigates the phytochemical compositions and toxicity of the leaf extracts of the plants against isolates of Listeria monocytogenes. Standard microbiological techniques and polymerase chain reaction was used to isolate and identify the bacteria. Phytochemical analysis was done and cytotoxicity of the extracts at different concentrations (MBC, OBC and LHC) were determined using human erythrocytes. Results of the phytochemical analysis revealed the presence of tannins, flavonoids, carbohydrates, alkaloids, terpinoids and glycosides in the studied extracts. Toxicity to erythrocytes, expressed as percentage hemolysis of only 17.27% (MBC1) was seen in one of the plants; M. balsamina. Similarly, the other extracts exhibited minimal toxicity to human erythrocytes (LHC1= 15.45%; OBC1= 7.6%). It was concluded that all the plant leaf extracts are safe for human consumption. Studies on the preparation, effective doses and side effects of these extracts in vivo are hereby recommended.

Redox Properties of Human Erythrocytes Are Adapted for Vitamin C Recycling

Ascorbic acid (AA; or vitamin C) is an important physiological antioxidant and radical scavenger. Some mammalian species, including homo sapiens, have lost the ability to synthetize AA and depend on its nutritional uptake. Erythrocytes from AA-auxotroph mammals express high amounts of the glucose transporter GLUT1. This isoform enables rapid uptake of glucose as well as dehydroascorbate (DHA), the fully oxidized form of AA. Here, we explored the effects of DHA uptake on the redox metabolism of human erythrocytes. DHA uptake enhanced plasma membrane electron transport (PMET) activity. This process is mediated by DCytb, a membrane bound cytochrome catalyzing extracellular reduction of Fe3+ and ascorbate free radical (AFR), the first oxidized form of AA. DHA uptake also decreased cellular radical oxygen species (ROS) levels. Both effects were massively enhanced in the presence of physiological glucose concentrations. Reduction of DHA to AA largely depleted intracellular glutathione (GSH) and induced the efflux of its oxidized form, GSSG. GSSG efflux could be inhibited by MK-571 (IC50 = 5 μM), indicating involvement of multidrug resistance associated protein (MRP1/4). DHA-dependent GSH depletion and GSSG efflux were completely rescued in the presence of 5 mM glucose and, partially, by 2-deoxy-glucose (2-DG), respectively. These findings indicate that human erythrocytes are physiologically adapted to recycle AA both intracellularly via GLUT1-mediated DHA uptake and reduction and extracellularly via DCytb-mediated AFR reduction. We discuss the possibility that this improved erythrocyte-mediated AA recycling was a prerequisite for the emergence of AA auxotrophy which independently occurred at least twice during mammalian evolution.

Potential Activity of Abrantes Pollen Extract: Biochemical and Cellular Model Studies

The aim of this study was to determine the grain composition and (poly)phenolic profile of pollen from Abrantes (Portugal), as well as its antioxidative and antidiabetic properties, and abilities to protect human erythrocytes against induced hemoglobin oxidation, lipid peroxidation, and hemolysis. The phytochemical profile of the Abrantes’ bee pollen revealed twenty phenolic compounds, identified by high-performance liquid chromatography with electrospray ionization mass spectrometry coupled with photodiode array detection. Among them, quercetin derivatives were the most abundant. Concerning the biological potential, the pollen extract showed notable capacity for 2,2-diphenyl-1-picrylhydrazyl, nitric oxide, and superoxide radicals, as well as for inhibition of α-glucosidase action, and protection of human erythrocytes against oxidative damage. Non-cytotoxic effects regarding the NHDF normal cell line, human adenocarcinoma Caco-2, and human liver HepG2 cells were observed. The results obtained contributed to further research on modes of action related to oxidative damage and metabolic health problems, to generate deeper knowledge of potential health-promoting effects to develop novel pharmaceutical drugs, nutraceuticals, and dietary supplements.

Evaluation of Leea rubra Leaf Extract for Oxidative Damage Protection and Antitumor and Antimicrobial Potential

Background. The leaves of Leea rubra contain an abundance of phenolic constituents and have medicinal uses as antipyretic and diaphoretic agents and are also used in the treatment of stomach ache, rheumatism, arthritis etc. In spite of the traditional uses, data on the scientific evaluation of the plant are not sufficient. So, the present study was designed to evaluate the protective role of the extract against oxidative damage to DNA and human erythrocytes as well as antitumor and antibacterial activities against some resistant bacteria. Methods. The protective activity of the ethyl acetate fraction (EAF) of the extract was investigated by evaluating the inhibition of oxidative damage of pUC19 plasmid DNA as well as hemolysis and lipid peroxidation damage to human erythrocytes induced by 2,2′-azobis-2-amidinopropane (AAPH). Antitumor activity was assessed by evaluating the percentage inhibition of cell growth, morphological changes of Ehrlich’s ascites carcinoma (EAC) cells, and hematological parameters. Antimicrobial activity was determined by the disc diffusion method against different resistant microorganisms. Results. EAF effectively inhibited AAPH-induced oxidative damage to DNA because it can inhibit the transformation of the supercoiled form of plasmid DNA to open circular and further linear form. The oxidative hemolysis caused by AAPH in human erythrocytes was inhibited by EAF extract in a time-dependent manner, and the production of malondialdehyde (MDA) was significantly reduced, which indicates the prevention of lipid peroxidation. In antitumor assay, 76% growth of inhibition of EAC was observed compared with the control mice ( p < 0.05 ) at a dose of 100 mg/kg body weight. Antimicrobial activity was evaluated against two pathogenic resistant microorganisms (Escherichia coli and Pseudomonas aeruginosa), and the highest antimicrobial activity was observed against Pseudomonas spp. Conclusion. EAF may have great importance in preventing oxidative damage to DNA, erythrocytes, and other cellular components as well as can be a good candidate in cancer chemotherapy and treating infectious diseases caused by antibiotic-resistant bacteria.