Erythrocytes lysis

Intracellular H2O2 is catalytically removed by catalase. The enzyme contains Fe(III) at its active site and is found in the cytosol of erythrocytes as well as the mitochondria and peroxisomes of most other cells. The concentration of catalase in rheumatoid synovial fluid is extremely low and may only be present as a result of erythrocyte lysis. 

In vivo heme is released into the plasma by erythrocyte lysis in the form of hemoglobin and by tissue trauma in the form of myoglobin, and both heme proteins are quickly oxidized to their ferric heme forms (methemoglobin and metmyoglobin) at the oxygen tension found in tissue capillary beds. 

Membrane permeabilization activity of peptides is currently measured by the use of artificial membrane bilayers, such as liposomes or erythrocytes. The hposome leakage assay can be performed by using spectrofluorimetry with a concentration-dependent quenching of a dye (calcein, carboxyfluorescein) encapsulated in liposomes. Disruption of hposomes in the presence of peptide-inducing leakage will lead to an increase in the fluorescence intensity of the liposome solution. Erythrocyte lysis assay is based on the absorption of hemoglobin, which can be measured once released into the extracellular medium upon erythrocyte lysis in the presence of peptide. 

Fig. 6.2. The FSC and SSC signals resulting from the cells in different blood preparations (whole peripheral blood whole peripheral blood after erythrocyte lysis and peripheral blood mononuclear cells [PBMCs] with granulocytes removed by density gradient centrifugation). The bottom panels indicate the five clusters into which the scatter signals fall.

The serum should be yellowish. A reddish color is indicative of erythrocytes lysis (hemolysis) which may interfere with clinical chemistry assays based on colorimetric values. If this happens, you may need to adjust the speed at which the blood is collected and processed or other steps that may cause sheer and red blood cell lysis. 

Eschbach E, Scharsack JP, John U, Medlin LK (2001) Improved erythrocyte lysis assay in microtitre plates for sensitive detection and efficient measurement of haemolytic compounds from ichtyotoxic algae. J Appl Toxicol 21 513-519... 

In one case, hemolysis started after the second dose of rifampicin, and the patient s blood contained rifampi-cin-dependent IgG and IgM antibodies, which caused erythrocyte lysis through an interaction with the I antigen on the erythrocyte surface (35). This antigen is also expressed on renal tubular epithelium and the hemolysis was accompanied by acute renal insufficiency. 

Brajtburg J, Elberg S, Schwartz DR, et al. Involvement of oxidative damage in erythrocyte lysis induced by amphotericin B. Antimicrob Agents Chemother 1985 27 172-6. 

Bierbaum, T J, Bouma, S R, and Fluestis, W-Fl, A mechanism of erythrocyte lysis by lysopho-sphatidylcholine, Biochim. Biophys. Acta 555 (1979) 102-110. 

Kellogg, E.W. and Fridovich, I. (1977). Liposome oxidation and erythrocyte lysis by enzymatically generated superoxide and hydrogen peroxide. ]. Biol Chem., 752, 6721-6728... 

Figure 9 Erythrocyte lysis caused by emulsion formulations. All data nornialized to 100% lysis. Key x control, amphotericin B emulsion/lecithin. Fugizone. amphotericin B emukion/Pluronic L92, amphotericin B emulsion/Pluronic FI27 (105).

Jimenez, L, Garrido, A., Bannach, R., Gotteland, M., Speisky, H. (2000). Protective effects of boldine against free radical-induced erythrocyte lysis. Phytother. Res., 14(5), 339-343. 

In the cell cultures neutrophils in RPMI-1640 medium were used. They were isolated from human blood by density gradient centrifugation and erythrocyte lysis. We used phor-bol 12-myristate 13-acetate to stimulate them by exchanging the medium in the wells. This chemical stimulant mimics the presence of a pathogen which leads to the chromatin release. The control culture did not receive the stimulating agent and was kept unstimulated. 

Sethu P, Anahtar M, Moldawer LL, Tompkins RG, Toner M (2004) Continuous flow microfluidic device for rapid erythrocyte lysis. Anal Chem 76 ... 

Erythrocyte hemolysis. The experiments were performed in PBS using a static method according to [12]. The degree of erythrocyte lysis due to sample activity was calculated as the hemolytic index %H = where Hb is the... 

Continuous flow microfluidic device for rapid erythrocyte lysis. Anal Chem 76 6247-6253... 

In a different line of research, we have proposed electrochemical manipulation of a single cell using either a bare or a conducting polymer film-coated microelectrode. In a previous report, we have shown with a three electrode system that erythrocytes burst on the surface of electrodes at an applied potential much lower than ca. 1.5 V vs. Ag/AgCl . The cause of erythrocyte breakdown remains unsolved. Electrical as well as physicochemical effects, caused by potential application, may induce erythrocyte lysis due to their susceptibility to breakdown by changes in pH, osmotic pressure, and so on. 

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