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Human Erythrocytes (human + erythrocyte)
Selected AbstractsNanoparticles: Investigations on the Structural Damage in Human Erythrocytes Exposed to Silver, Gold, and Platinum Nanoparticles (Adv. Funct.ADVANCED FUNCTIONAL MATERIALS, Issue 8 2010Mater. [source] Investigations on the Structural Damage in Human Erythrocytes Exposed to Silver, Gold, and Platinum NanoparticlesADVANCED FUNCTIONAL MATERIALS, Issue 8 2010P. V. Asharani Abstract Human erythrocytes or red blood cells (RBCs), which constitute 99% of blood cells, perform an important function of oxygen transport and can be exposed to nanoparticles (NPs) entering into the human body during therapeutical applications involving such NPs. Hence, the haemocompatibility of the Ag, Au, and Pt NPs on human RBCs is investigated. The parameters monitored include haemolysis, haemagglutination, erythrocyte sedimentation rate, membrane topography, and lipid peroxidation. The findings suggest that platinum and gold NPs are haemocompatible compared to Ag NPs. Erythrocytes exhibit significant lysis, haemagglutination, membrane damage, detrimental morphological variation, and cytoskeletal distortions following exposure to Ag NPs at a concentration of 100,µg,mL,1. Exposure of Ag+ to RBCs shows no lysis or deterioration, implying that the observed toxicity is solely due to NPs. The haemolyzed erythrocyte fraction has the ability to induce DNA damage in nucleated cells. Additionally, multiple pits and depressions are observed on RBC membrane following exposure to Ag NPs (50,µg,mL,1 onwards). Hence, it is apparent that Ag NPs exhibit toxicity on RBCs and on other cells that are exposed to NP-mediated haemolyzed fractions. [source] Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser-induced fluorescence detectionELECTROPHORESIS, Issue 3 2004Hua Zhang Abstract A method for monitoring amino acids in single erythrocytes is described. For intracellular derivatization, reagent fluorescein isothiocyanate (FITC) was introduced into living cells by electroporation. For an 8 ,m erythrocyte, the analytes were diluted by a factor of only 1.6. After completion of the derivatization reaction, a single cell was injected into the separation capillary tip and lysed there. The derivatized amino acids were separated by capillary electrophoresis, followed by laser-induced fluorescence detection. Nine amino acids were quantitatively determined, with amounts of amino acids ranging from 3.8,32 amol/single cell. [source] Malathion-induced oxidative stress in human erythrocytes and the protective effect of vitamins C and E in vitroENVIRONMENTAL TOXICOLOGY, Issue 3 2009Dilek Durak Abstract Malathion is an organophosphate (OP) pesticide that has been shown to induce oxidative stress in erythrocytes through the generation of free radicals and alteration of the cellular antioxidant defense system. We examined the effect of several different doses of malathion (25, 75, 200 ,M), or malathion in combination with vitamin C (VC; 10 ,M) or vitamin E (VE; 30 ,M), on the levels of malondialdehyde (MDA), and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities in human erythrocytes in vitro. Erythrocytes were incubated under various treatment conditions (malathion alone, vitamins alone, or malathion plus vitamin) at 37°C for 60 min, and the levels of MDA, and SOD, CAT and GPx activities, were determined. Treatment with malathion alone increased the levels of MDA and decreased SOD, CAT, and GPx activities in erythrocytes (P < 0.05). There were no statistical differences among VC-treated, VE-treated, or VC + VE-treated erythrocyes, as compared with nontreated control cells. Treatment of cells with malathion + VC, malathion + VE, or a combination of all three agents prevented malathion-induced changes in antioxidant enzyme activity and lipid peroxidation. However, this effect was seen only at low concentrations of malathion (25 and 75 ,M), and the combination of VC + VE had a more protective effect than VC or VE alone. These results indicated that the presence of vitamins at concentrations that are similar to the levels found in plasma have no effect on malathion-induced toxicity in erythrocytes at a concentration of malathion (200 ,M) that is typically used in pesticides. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2009. [source] Amiodarone Attenuates Fluoride-induced Hyperkalemia in VitroACADEMIC EMERGENCY MEDICINE, Issue 2 2003Mark Su MD Abstract Poisoning by hydrofluoric acid or fluoride salts results in hypocalcemia, hypomagnesemia, and hyperkalemia with subsequent cardiac dysrhythmias. In previous studies, quinidine attenuated fluoride-induced hyperkalemia in vitro, and enhanced survival in animals. Like quinidine, amiodarone is a potassium channel blocker, although amiodarone is more familiar to clinicians due to its recent inclusion in advanced cardiac life support (ACLS) protocols. Objectives: This in-vitro study of human erythrocytes was designed to determine whether amiodarone could attenuate fluoride-induced hyperkalemia. Methods: Six healthy volunteers each donated 60 mL of blood on three occasions. Each specimen was divided into 12 tubes, incubated at 37°C, and oxygenated with room air. An aqueous sodium fluoride (F,) solution was added to tubes 1,9. Incremental amounts of quinidine were added to tubes 1,4 (Q1,Q4) to attain calculated concentrations of 0.73 ,g/mL, 1.45 ,g/mL, 2.9 ,g/mL, and 5.8 ,g/mL, respectively. Incremental amounts of amiodarone were added to tubes 5,8 (A1,A4) to attain calculated concentrations of 0.38 ,g/mL, 0.75 ,g/mL, 1.5 ,g/mL, and 3.0 ,g/mL, respectively. Tubes 9,12 were controls for each of F,, amiodarone, quinidine alone, and no additive, respectively. Extracellular potassium concentration ([K+]) was followed, and an objective endpoint was defined as the rise in potassium concentration at 6 hours. Results: Fluoride produced a significant change in [K+] by 6 hours in all samples. Quinidine produced a J-shaped curve in its ability to attenuate the rise in [K+], with only one concentration, Q3, demonstrating significance versus tube 9 (control). Amiodarone also demonstrated a J-shaped dose,response effect, with statistical significance at A1, A2, and A3 versus tube 9 (control). There was no significant difference among the effective concentrations (Q3, A1, A2, and A3) of both drugs. Conclusions: In this in-vitro model using human blood, amiodarone and quinidine both attenuated F, -induced hyperkalemia. Further study is indicated to determine whether amiodarone enhances survival in F, -poisoned animals. [source] Changes in mu opioid receptors and rheological properties of erythrocytes among opioid abusersADDICTION BIOLOGY, Issue 2 2002ALLEN R. ZEIGER The high prevalence of anemia among chronic opioid users leads us to propose that chronic opiate use results in elevated mu opioid receptor levels on human erythrocytes and that these receptor changes may affect erythrocyte membrane properties. Blood samples from 17 opioid-dependent subjects (based on the Diagnostic and Statistical Manual of Mental Disorders, 4th edition or DSM-IV) and 15 drug-free controls were assayed for mu opioid receptors on erythrocytes using a flow cytometry immunoassay. Deformability and the hydration status of erythrocytes were studied by ektacytometry. Data were analyzed by independent t-tests, tests of correlation, chi square and cluster analyses. As expected, the percentage of erythrocytes from opioiddependent subjects with opioid receptors (opioid receptor levels) was significantly higher (47.4 ± 38.3%) than controls (22.8 ± 30.1%) (t = 2.01, df = 30, p < 0.05). Also, the opioid-dependent patients showed a wide variation in the percentage of erythrocytes bearing opioid receptors and data analyses of these patients showed two strongly defined clusters. One subgroup consisted of nine individuals with very high receptor levels (mean = 81.5%) while the other had eight patients with low receptor levels (mean = 9.1%) that were not significantly different than the receptor levels of controls. Ektacytometry of opioid dependent patients with high opioid receptor levels showed changes in rheological parameters of erythrocytes, such as deformability index and cellular hydration. For example, a positive correlation was observed between opioid receptor levels and deformability indices among opioid-dependent patients (r = 0.74, p < 0.005). Our findings indicate that the mu opioid receptor is present on human erythrocytes, although with considerable variation in receptor levels, and that the levels of this receptor are significantly elevated with chronic opioid exposure. Moreover, erythrocytes with high opioid receptor levels from chronic opiate users seem to have high deformability. This study may offer clues to the biological properties of peripheral blood cells that may be mediated by mu opioid receptors and lead to a better understanding of some of the clinical effects of opioid use. [source] Simulation study of methemoglobin reduction in erythrocytesFEBS JOURNAL, Issue 6 2007Differential contributions of two pathways to tolerance to oxidative stress Methemoglobin (metHb), an oxidized form of hemoglobin, is unable to bind and carry oxygen. Erythrocytes are continuously subjected to oxidative stress and nitrite exposure, which results in the spontaneous formation of metHb. To avoid the accumulation of metHb, reductive pathways mediated by cytochrome b5 or flavin, coupled with NADH-dependent or NADPH-dependent metHb reductases, respectively, keep the level of metHb in erythrocytes at less than 1% of the total hemoglobin under normal conditions. In this work, a mathematical model has been developed to quantitatively assess the relative contributions of the two major metHb-reducing pathways, taking into consideration the supply of NADH and NADPH from central energy metabolism. The results of the simulation experiments suggest that these pathways have different roles in the reduction of metHb; one has a high response rate to hemoglobin oxidation with a limited reducing flux, and the other has a low response rate with a high capacity flux. On the basis of the results of our model, under normal oxidative conditions, the NADPH-dependent system, the physiological role of which to date has been unclear, is predicted to be responsible for most of the reduction of metHb. In contrast, the cytochrome b5,NADH pathway becomes dominant under conditions of excess metHb accumulation, only after the capacity of the flavin,NADPH pathway has reached its limit. We discuss the potential implications of a system designed with two metHb-reducing pathways in human erythrocytes. [source] Effects of clotrimazole on transport mediated by multidrug resistance associated protein 1 (MRP1) in human erythrocytes and tumour cellsFEBS JOURNAL, Issue 24 2001Antonios Klokouzas Clotrimazole has been shown to have potent anti-malarial activity in vitro, one possible mechanism being inhibition of oxidized glutathione (GSSG) export from the infected human red blood cells or from the parasite itself. Efflux of GSSG from normal erythrocytes is mediated by a high affinity glutathione S-conjugate transporter. This paper shows that transport of the model substrate, 3 µm dinitrophenyl S -glutathione, across erythrocyte membranes is inhibited by multidrug resistance-associated protein 1 (MRP1)-specific antibody, QCRL-3, strongly suggesting that the high affinity transport is mediated by MRP1. The rates of transport observed with membrane vesicles prepared from erythrocytes or from multidrug resistant tumour cells show a similar pattern of responses to applied reduced glutathione, GSSG and MRP1 inhibitors (indomethacin, MK571) further supporting the conclusion that the high affinity transporter is MRP1. In both erythrocytes and MRP1-expressing tumour cells, MRP1-associated transport is inhibited by clotrimazole over the range 2,20 µm, and the inhibitory effect leads to increases in accumulation of MRP1 substrates, vincristine and calcein, and decreases in calcein efflux from intact MRP1-expressing human tumour cells. It also results in increased sensitivity to daunorubicin of the multidrug resistant cells, L23/R but not the sensitive parent L23/P cells. These results demonstrate that clotrimazole can inhibit the MRP1 which is present in human erythrocytes, an effect that may contribute to, though not fully account for, its anti-malarial action. [source] Energy metabolism and lipid peroxidation of human erythrocytes as a function of increased oxidative stressFEBS JOURNAL, Issue 3 2000Barbara Tavazzi To study the influence of oxidative stress on energy metabolism and lipid peroxidation in erythrocytes, cells were incubated with increasing concentrations (0.5,10 mm) of hydrogen peroxide for 1 h at 37 °C and the main substances of energy metabolism (ATP, AMP, GTP and IMP) and one index of lipid peroxidation (malondialdehyde) were determined by HPLC on cell extracts. Using the same incubation conditions, the activity of AMP-deaminase was also determined. Under nonhaemolysing conditions (at up to 4 mm H2O2), oxidative stress produced, starting from 1 mm H2O2, progressive ATP depletion and a net decrease in the intracellular sum of adenine nucleotides (ATP + ADP + AMP), which were not paralleled by AMP formation. Concomitantly, the IMP level increased by up to 20-fold with respect to the value determined in control erythrocytes, when cells were challenged with the highest nonhaemolysing H2O2 concentration (4 mm). Efflux of inosine, hypoxanthine, xanthine and uric acid towards the extracellular medium was observed. The metabolic imbalance of erythrocytes following oxidative stress was due to a dramatic and unexpected activation of AMP-deaminase (a twofold increase of activity with respect to controls) that was already evident at the lowest dose of H2O2 used; this enzymatic activity increased with increasing H2O2 in the medium, and reached its maximum at 4 mm H2O2 -treated erythrocytes (10-fold higher activity than controls). Generation of malondialdehyde was strictly related to the dose of H2O2, being detectable at the lowest H2O2 concentration and increasing without appreciable haemolysis up to 4 mm H2O2. Besides demonstrating a close relationship between lipid peroxidation and haemolysis, these data suggest that glycolytic enzymes are moderately affected by oxygen radical action and strongly indicate, in the change of AMP-deaminase activity, a highly sensitive enzymatic site responsible for a profound modification of erythrocyte energy metabolism during oxidative stress. [source] Sodium valproate inhibits glucose transport and exacerbates Glut1-deficiency in vitroJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 4 2005Hei Yi Wong Abstract Anticonvulsant sodium valproate interferes with brain glucose metabolism. The mechanism underlying such metabolic disturbance is unclear. We tested the hypothesis that sodium valproate interferes with cellular glucose transport with a focus on Glut1 since glucose transport across the blood-brain barrier relies on this transporter. Cell types enriched with Glut1 expression including human erythrocytes, human skin fibroblasts, and rat astrocytes were used to study the effects of sodium valproate on glucose transport. Sodium valproate significantly inhibited Glut1 activity in normal and Glut1-deficient erythrocytes by 20%,30%, causing a corresponding reduction of Vmax of glucose transport. Similarly, in primary astrocytes as well as in normal and Glut1-deficient fibroblasts, sodium valproate inhibited glucose transport by 20%,40% (P,<,0.05), accompanied by an up to 60% downregulation of GLUT1 mRNA expression (P,<,0.05). In conclusion, sodium valproate inhibits glucose transport and exacerbates Glut1 deficiency in vitro. Our findings imply the importance of prudent use of sodium valproate for patients with compromised Glut1 function. J. Cell. Biochem. © 2005 Wiley-Liss, Inc. [source] Imbalance of plasma membrane ion leak and pump relationship as a new aetiological basis of certain disease statesJOURNAL OF INTERNAL MEDICINE, Issue 6 2003G. Ronquist Abstract. The basis for life is the ability of the cell to maintain ion gradients across biological membranes. Such gradients are created by specific membrane-bound ion pumps [adenosine triphosphatases (ATPases)]. According to physicochemical rules passive forces equilibrate (dissipate) ion gradients. The cholesterol/phospholipid ratio of the membrane and the degree of saturation of phospholipid fatty acids are important factors for membrane molecular order and herewith a determinant of the degree of non-specific membrane leakiness. Other operative principles, i.e. specific ion channels can be opened and closed according to mechanisms that are specific to the cell. Certain compounds called ionophores can be integrated in the plasma membrane and permit specific inorganic ions to pass. Irrespective of which mechanism ions leak across the plasma membrane the homeostasis may be kept by increasing ion pumping (ATPase activity) in an attempt to restore the physiological ion gradient. The energy source for this work seems to be glycolytically derived ATP formation. Thus an increase in ion pumping is reflected by increased ATP hydrolysis and rate of glycolysis. This can be measured as an accumulation of breakdown products of ATP and end-products of anaerobic glycolysis (lactate). In certain disease entities, the balance between ATP formation and ion pumping may be disordered resulting in a decrease in inter alia (i.a.) cellular energy charge, and an increase in lactate formation and catabolites of adenylates. Cardiac syndrome X is proposed to be due to an excessive leakage of potassium ions, leading to electrocardiographic (ECG) changes, abnormal Tl-scintigraphy of the heart and anginal pain (induced by adenosine). Cocksackie B3 infections, a common agent in myocarditis might also induce an ionophore-like effect. Moreover, Alzheimer's disease is characterized by the formation of extracellular amyloid deposits in the brain of patients. Perturbation of cellular membranes by the amyloid peptide during the development of Alzheimer's disease is one of several mechanisms proposed to account for the toxicity of this peptide on neuronal membranes. We have studied the effects of the peptide and fragments thereof on 45Ca2+ -uptake in human erythrocytes and the energetic consequences. Treatment of erythrocytes with the ,1,40 peptide, results in qualitatively similar nucleotide pattern and decrease of energy charge as the treatment with Ca2+ -ionophore A23187. Finally, in recent studies we have revealed and published in this journal that a rare condition, Tarui's disease or glycogenosis type VII, primarily associated with a defect M-subunit of phosphofructokinase, demonstrates as a cophenomenon an increased leak of Ca2+ into erythrocytes. [source] Antitumour activity and specificity as a function of substitutions in the lipophilic sector of helical lactoferrin-derived peptideJOURNAL OF PEPTIDE SCIENCE, Issue 5 2003Nannan Yang Abstract A peptide L5 (PAWRKAFRWAWRMLKKAA), derived from the N -terminal ,-helical region of bovine lactoferrin (LFB 14,31), that is highly active against several tumour cell lines was reported earlier. In this study, a number of L5 analogues were designed in order to investigate how subsequent replacements of the aromatic amino acids in L5 with three amino acids representing different structural parameters influenced antitumour activity and tumour cell specificity relative to normal human cells. The Trp residues were substituted by Lys, Ile or Ala, while the Phe residue was substituted with Ala. The resulting peptides were investigated for their activity against prokaryotic cells, four tumour cell lines, human lung fibroblasts and human erythrocytes. Most of the peptides were highly active against both E. coli and S. aureus. The peptides were more active against the tumour cell lines than against normal eukaryotic cells but the activity against normal fibroblasts varied more among the peptides than did their antitumour activities. The results revealed that aromatic residues located opposite the cationic sector in L5 were more critical for antitumour activity than were aromatic residues located adjacent to the cationic sector. The biological responses for the peptides against tumour cell lines, fibroblasts, S. aureus (but not E. coli), were highly correlated with the amino acid descriptors used in our QSAR model. The result obtained from the QSAR study identified specific structural features that were important for lytic activity and membrane specificity. Certain structural properties in positions 3, 9 and 11 were shown to be important for antitumour activity, while additional structural properties in position 7 were found to be important with respect to tumour cell specificity. This information may offer a possibility for de novo design of an antitumour peptide with an improved therapeutic index. Copyright © 2003 European Peptide Society and John Wiley & Sons, Ltd. [source] PURIFICATION AND CHARACTERIZATION OF A LECTIN, BRYOHEALIN, INVOLVED IN THE PROTOPLAST FORMATION OF A MARINE GREEN ALGA BRYOPSIS PLUMOSA (CHLOROPHYTA) ,JOURNAL OF PHYCOLOGY, Issue 1 2006Gwang Hoon Kim When the coenocytic green alga Bryopsis plumosa (Huds.) Ag. was cut open and the cell contents were expelled, the cell organelles agglutinated rapidly in seawater to form protoplasts. Aggregation of cell organelles in seawater was mediated by a lectin,carbohydrate complementary system. Two sugars, N -acetyl- d -glucosamine and N -acetyl- d -galactosamine inhibited aggregation of cell organelles. The presence of these sugars on the surface of chloroplasts was verified with their complementary fluorescein isothiacyanate-labeled lectins. An agglutination assay using human erythrocytes showed the presence of lectins specific for N -acetyl- d -galactosamine and N -acetyl- d -glucosamine in the crude extract. One-step column purification using N -acetyl- d -glucosamine-agarose affinity chromatography yielded a homogeneous protein. The protein agglutinated the cell organelles of B. plumosa, and its agglutinating activity was inhibited by the above sugars. Sodium dodecyl sulfate polyacrylamide gel electrophoresis results showed that this protein might be composed of two identical subunits cross-linked by two disulfide bridges. Enzyme and chemical deglycosylation experiments showed that this protein is deficient in glycosylation. The molecular weight was determined as 53.8 kDa by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The N-terminal 15 amino acid sequence of the lectin was Ser,Asp,Leu,Pro,Thr,X,Asp,Phe,Phe,His,Ile,Pro,Glu,Arg,Tyr, and showed no sequence homology to those of other reported proteins. These results suggest that this lectin belongs to a new class of lectins. We named this novel lectin from B. plumosa"bryohealin." [source] Red Blood Cell Templated Polyelectrolyte Capsules: A Novel Vehicle for the Stable Encapsulation of DNA and ProteinsMACROMOLECULAR RAPID COMMUNICATIONS, Issue 6 2006Oliver Kreft Abstract Summary: A novel method for the encapsulation of biomacromolecules, such as nucleic acids and proteins, into polyelectrolyte microcapsules is described. Fluorescence-labelled double-stranded DNA and human serum albumin (HSA) are used as model substances for encapsulation in hollow microcapsules templated on human erythrocytes. The encapsulation procedure involves an intermediate drying step. The accumulation of DNA and HSA in the capsules is observed by confocal laser scanning microscopy, UV spectroscopy, and fluorimetry. The mechanism of encapsulation is discussed. Confocal fluorescence microscopy images of encapsulated TRITC-HSA (left) and dsDNA (right). Inserts demonstrate fluorescence profiles for both compounds. [source] Simultaneous separation of intracellular and extracellular lactate NMR signals of human erythrocytesMAGNETIC RESONANCE IN MEDICINE, Issue 2 2007Götz Kohler Abstract Intracellular/extracellular lactate (Lac) distribution has been determined before in human and animal erythrocytes (red blood cells [RBCs]) with various methods. However, all previous methods determine intra- and extracellular Lac separately or indirectly. Now, 13C-NMR spectroscopy has been used to monitor intra- and extracellular Lac simultaneously in intact RBCs. Isolated human RBCs were incubated with [3- 13C]-Lac, [3- 13C]-pyruvate (Pyr), and [1- 13C]-glucose (Gluc). A distortionless enhancement by polarization transfer (DEPT) sequence was used (TR = 3.3 s, N = 128) to monitor the 13C-NMR resonances in both compartments. The intra- and extracellular methyl group resonances of Lac and Pyr were clearly separated by 9.6 Hz and 7.0 Hz, respectively, under normoxic conditions due to the RBC chemical-shift effect. The results show that the chemical-shift effect of RBCs is convenient to monitor intra- and extracellular Lac simultaneously in intact RBCs under normoxic conditions. Magn Reson Med 58:213,217, 2007. © 2007 Wiley-Liss, Inc. [source] A flow cytometry-based assay for measuring invasion of red blood cells by Plasmodium falciparum,AMERICAN JOURNAL OF HEMATOLOGY, Issue 4 2010Amy K. Bei Variability in the ability of the malaria parasite Plasmodium falciparum to invade human erythrocytes is postulated to be an important determinant of disease severity. Both the parasite multiplication rate and erythrocyte selectivity are important parameters that underlie such variable invasion. We have established a flow cytometry-based method for simultaneously calculating both the parasitemia and the number of multiply-infected erythrocytes. Staining with the DNA-specific dye SYBR Green I allows quantitation of parasite invasion at the ring stage of parasite development. We discuss in vitro and in vivo applications and limitations of this method in relation to the study of parasite invasion. Am. J. Hematol., 2010. © 2010 Wiley-Liss, Inc. [source] Protective effect of resveratrol on markers of oxidative stress in human erythrocytes subjected to in vitro oxidative insultPHYTOTHERAPY RESEARCH, Issue S1 2010Kanti Bhooshan Pandey Abstract Resveratrol is a natural polyphenolic compound found largely in the skin of red grapes. Growing evidence suggests that resveratrol may play an important role in the prevention of many human diseases. Many of the biological actions of this polyphenol have been attributed to its antioxidant properties. The present study was undertaken to evaluate the effect of resveratrol on intracellular reduced glutathione (GSH) and membrane sulphydryl groups in erythrocytes subjected to oxidative stress in vitro by incubating with t-BHP (10 µm). The study was aimed to test the efficacy of the antioxidant effect of resveratrol on human erythrocytes. Subjecting erythrocytes to oxidative stress (in vitro) by incubating them with t-BHP (10 µm) caused a significant decrease in the intracellular GSH level and membrane ,SH content compared with basal values. Incubation of erythrocytes/membranes with resveratrol (1,100 µm final conc) resulted in significant protection against the t-BHP-induced oxidative stress as evidenced by the increase in GSH level and membrane ,SH content. It was observed that the effect of resveratrol is dose/concentration and time-dependent. Since resveratrol is naturally present in many fruits and vegetables, a diet rich in resveratrol may provide protection against degenerative diseases. Copyright © 2009 John Wiley & Sons, Ltd. [source] Protective effects of selected medicinal plants against protein degradation, lipid peroxidation and deformability loss of oxidatively stressed human erythrocytesPHYTOTHERAPY RESEARCH, Issue 4 2004S. M. Suboh Abstract The effects of seven medicinal plants including Artemisia herba-alba, Ferula hermonis, Hibiscus sabdariffa, Nigella sativa, Teucrium polium, Trigonella foenum-graecum, and Allium sativum on protein degradation, lipid peroxidation, erythrocyte deformability and osmotic fragility of erythrocytes exposed in vitro to 10 mM H2O2 for 60 min at 37 °C have been examined. Preincubation of erythrocytes with Nigella sativa and Allium sativum protected erythrocytes against protein degradation, loss of deformability and increased osmotic fragility caused by H2O2, while the other plants failed to protect erythrocytes against these damages. Artemisia herba-alba did not protect erythrocytes against lipid peroxidation, while Trigonella foenum-graecum unexpectedly increased lipid peroxidation of erythrocytes exposed to H2O2. Ferula hermonis, Hibiscus sabdariffa, Nigella sativa, Teucrium polium and Allium sativum protected erythrocytes against lipid peroxidation. The results indicate the importance of oxidatively damaged cellular proteins in compromising the rheologic behaviour of the erythrocytes, and that the medicinal plants which have anti-protein-oxidant activity (e.g. Nigella sativa and Allium sativum) could be rheologically useful, particularly in pathological conditions related to free radicals. Copyright © 2004 John Wiley & Sons, Ltd. [source] Histidine-stimulated divalent metal uptake in human erythrocytes and in the erythroleukaemic cell line HEL.92.1.7THE JOURNAL OF PHYSIOLOGY, Issue 2 2004F. Oakley The uptake of 65Zn by human erythrocytes was investigated in the presence of high (40 mm) and low (5 mm) concentrations of histidine and 0,500 ,m cobalt, nickel, manganese and zinc. Varying concentrations of metal mono- and bis-histidine complexes will be formed and the inhibition of 65Zn uptake could be correlated with the calculated complex concentrations to investigate competition between metals. For each metal, the calculated concentrations of bis-histidine complex giving 50% inhibition of 65Zn uptake were similar at both 5 mm and 40 mm histidine. Manganese,bis-histidine appeared to have a much higher affinity for the binding site than the other metal,bis-histidine complexes, which had similar affinities to each other. Studies of the inhibition of histidine-stimulated 54Mn uptake by the addition of manganese confirmed that manganese,bis-histidine does act as a substrate for the transporter in a similar fashion to the other metals studied. In addition, human erythroleukaemic cells (HEL cells) were used as a model for erythroid precursor cells. l -histidine, but not d -histidine, stimulated 65Zn uptake in a saturable fashion. The other metals competed with zinc in a similar manner to that seen in erythrocytes, and the affinity for manganese,bis-histidine was much greater than for the bis-histidine complexes of the other three metals. Both the capacity for metal transport per cell, and the affinity of the transporter for the metal,bis-histidine complexes, were much greater in the HEL cells than in the erythrocyte. It is suggested that histidine-stimulated metal transport may play a role in the supply of metals to maturing erythroid cells. [source] Structure of human erythrocyte NADH-cytochrome b5 reductaseACTA CRYSTALLOGRAPHICA SECTION D, Issue 11 2004Sachiko Bando Erythrocyte NADH-cytochrome b5 reductase reduces methaemoglobin to functional haemoglobin. In order to examine the function of the enzyme, the structure of NADH-cytochrome b5 reductase from human erythrocytes has been determined and refined by X-ray crystallography. At 1.75,Å resolution, the root-mean-square deviations (r.m.s.d.) from standard bond lengths and angles are 0.006,Å and 1.03°, respectively. The molecular structure was compared with those of rat NADH-cytochrome b5 reductase and corn nitrate reductase. The human reductase resembles the rat reductase in overall structure as well as in many side chains. Nevertheless, there is a large main-chain shift from the human reductase to the rat reductase or the corn reductase caused by a single-residue replacement from proline to threonine. A model of the complex between cytochrome b5 and the human reductase has been built and compared with that of the haem-containing domain of the nitrate reductase molecule. The interaction between cytochrome b5 and the human reductase differs from that of the nitrate reductase because of differences in the amino-acid sequences. The structures around 15 mutation sites of the human reductase have been examined for the influence of residue substitutions using the program ROTAMER. Five mutations in the FAD-binding domain seem to be related to cytochrome b5. [source] Structure of tetragonal crystals of human erythrocyte catalaseACTA CRYSTALLOGRAPHICA SECTION D, Issue 1 2001Martin K. Safo The structure of catalase from human erythrocytes (HEC) was determined in tetragonal crystals of space group I41 by molecular-replacement methods, using the orthorhombic crystal structure as a search model. It was then refined in a unit cell of dimensions a = b = 203.6 and c = 144.6,Å, yielding R and Rfree of 0.196 and 0.244, respectively, for all data at 2.4,Å resolution. A major difference of the HEC structure in the tetragonal crystal compared with the orthorhombic structure was the omission of a 20-residue N-terminal segment corresponding to the first exon of the human catalase gene. The overall structures were otherwise identical in both crystal forms. The NADPH-binding sites were empty in all four subunits and bound water molecules were observed at the active sites. The structure of the C-terminal segment, which corresponds to the last exon, remained undetermined. The tetragonal crystals showed a pseudo-4122 symmetry in molecular packing. Two similar types of lattice contact interfaces between the HEC tetramers were observed; they were related by the pseudo-dyad axes. [source] In vitro protective effect of Rhodiola rosea extract against hypochlorous acid-induced oxidative damage in human erythrocytesBIOFACTORS, Issue 3 2004Roberta De Sanctis Abstract Rhodiola rosea L. (Crassulaceae) is a plant living at high altitudes in Europe and Asia. Its roots have long been used in the traditional medical system of these geographical areas to increase the organism resistance to physical stress; today, it has become an important component of many dietary supplements. In this study we investigate the antioxidant capacity of the R. rosea aqueous extract evaluating its ability to counteract some of the main damages induced by hypochlorous acid (HOCl), a powerful oxidant generated by activated phagocytes, to human erythrocytes. Ascorbic acid was used as a reference substance because of its physiological HOCl-scavenging ability. Our study demonstrates that R. rosea is able to significantly protect, in a dose-dependent manner, human RBC from glutathione (GSH) depletion, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) inactivation and hemolysis induced by the oxidant. Furthermore, we demonstrate that R. rosea aqueous extract acts from the inside of the erythrocyte suggesting a probable involving of cell components. The protection on GSH afforded by the R. rosea extract with respect to ascorbic acid, occurred also if added 2 or 5 min. later than the oxidant, suggesting a more rapid or powerful effect. [source] Insight into the free-radical-scavenging mechanism of hydroxyl-substituent Schiff bases in the free-radical-induced hemolysis of erythrocytesCELL BIOCHEMISTRY AND FUNCTION, Issue 6 2007You-Zhi Tang Abstract This work aimed to explore the mechanism by which hydroxyl-substituent Schiff bases scavenge free-radicals. Thus, four Schiff bases, that is benzylidene aniline (BAN), 2-(phenyliminomethyl)phenol (BAH), 4-benzimidoylphenol (PBH) and 2-benzimidoylphenol (OBH), were applied to protect human erythrocytes against 2,2,-azobis(2-amidinopropane hydrochloride) (AAPH)-induced hemolysis. The results revealed that the OH attached to the ortho -position of methylene in Schiff base scavenges 1.46 radicals per molecule, the OH attached to the para -position of the N atom scavenges 2.94 radicals and the OH attached to the ortho -position of the N atom scavenges 3.63 radicals. In addition, four Schiff bases were used together with some familiar antioxidants, such as 6-hydroxyl-2,5,7,8-tetramethyl chroman-2-carboxylic acid (Trolox), L -ascorbic acid (VC), ,-tocopherol (TOH) and L -ascorbyl-6-laurate (VC-12) in AAPH-induced hemolysis of erythrocytes. It was found that, except for BAN+VC-12, BAH,+,VC-12, OBH,+,VC-12 and PBH+TOH, all the other combinations protected erythrocytes more perfectly than when used individually. This result demonstrated that a promotive protection existed between Schiff base and other antioxidants and this improved their ability to scavenge free-radicals. Finally, IC50 values of the aforementioned Schiff bases together with 2-((o -hydroxylphenylimino) methyl)phenol (OSAP) and 2-((p -hydroxylphenylimino)methyl)phenol (PSAP) were determined by reaction with two radical species, that is, 2,2,-azinobis(3-ethylbenzothiazoline-6-sulfonate) radical (ABTS+·) and 2,2,-diphenyl-1-picrylhydrazyl (DPPH). The results implied that the molecular framework of a Schiff base and an OH attached to the ortho -position of methylene were apt to reduce radicals, but the OH attached to the aniline ring in a Schiff base was prone to scavenge radicals directly. Copyright © 2006 John Wiley & Sons, Ltd. [source] The antioxidant effect of hydroxyl-substituent Schiff bases on the free-radical-induced hemolysis of human erythrocytesCELL BIOCHEMISTRY AND FUNCTION, Issue 2 2007You-Zhi Tang Abstract The major objectives of the present work were focused on assessing the antioxidant capacities of two hydroxyl-substituent Schiff bases, 2-((o -hydroxylphenylimino)methyl)phenol (OSAP) and 2-((p -hydroxylphenylimino)methyl)phenol (PSAP) either used alone or in combination with some familiar water-soluble antioxidants i.e. 6-hydroxyl-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) and L-ascorbic acid (VC), and lipophilic ones i.e. ,-tocopherol (TOH) and L -ascorbyl-6-laurate (VC-12). 2,2'-Azobis(2-amidinopropane hydrochloride) (AAPH). Induced hemolysis of human erythrocytes functioned as the evaluation experimental system in this research. The present findings showed that either OSAP or PSAP not only was an antioxidant with high activity in protecting erythrocytes against AAPH-induced hemolysis concentration-dependently, but can also protect erythrocytes by acting with Trolox, TOH, VC and VC-12 synergistically. Based on chemical kinetic deduction, the number of trapping peroxyl radicals, n, of the above-mentioned antioxidants can be calculated in relation to Trolox that traps two peroxyl radicals; thus, TOH can trap 3.83 peroxyl radicals, VC-12 traps 2.87 and VC can only trap 1.08. As for OSAP and PSAP, 8.71 and 13.7 peroxyl radicals can be trapped, respectively, indicating that they were the most efficient inhibitors against AAPH-induced hemolysis. Moreover, the total number of peroxyl radicals trapped by OSAP+Trolox, OSAP+TOH, OSAP+VC and PSAP+VC were higher than the sum of the above individual antioxidant used alone, demonstrating that a mutual promotive effect existed in the above mixed antioxidants. In contrast, owing to the fact that the total number of peroxyl radicals trapped by OSAP+VC-12, PSAP+Trolox, PSAP+TOH and PSAP+VC-12 were less than the sum of the above individual antioxidant used alone, a mutual antagonistic effect was suggested in these combinative usages. This information may be helpful in the pharmaceutical application of two Schiff bases. Copyright © 2005 John Wiley & Sons, Ltd. [source] In vitro effects of high glucose concentrations on membrane protein oxidation, G-actin and deformability of human erythrocytesCELL BIOCHEMISTRY AND FUNCTION, Issue 3 2005Halil Resmi Abstract The object of this study was to examine the effect of elevated in vitro glucose concentrations on protein modification and functional changes in human erythrocytes. Groups were exposed to 5,45,mM glucose concentrations. The time effect of any changes was also evaluated. In erythrocyte ghosts, protein glycation and oxidation were evaluated using spectrophotometric methods. G-actin was measured by a DNase I inhibition assay in cell lysates. Erythrocyte deformability was assessed using a cell transit analyser. At 24,h, a significant protein oxidation (at 25 and 45,mM glucose; p,<,0.05), and G-actin increase was observed for all concentrations (p,<,0.05). At 48,h, a significant increase in glycation (25 and 45,mM glucose; p,<,0.05), protein oxidation (p,<,0.05), and G-actin (p,<,0.05) was observed in all groups. A significant positive correlation was observed between glucose /protein oxidation, glucose/G-actin and protein oxidation/G-actin at 24 and 48,h. Our findings show that the oxidative effect of glucose on erythrocytes depends on concentration and incubation time. We also present the first evidence of increased G-actin in human erythrocytes exposed to high glucose concentrations as a diabetes model. Copyright © 2004 John Wiley & Sons, Ltd. [source] Reticulocyte binding protein homologues are key adhesins during erythrocyte invasion by Plasmodium falciparumCELLULAR MICROBIOLOGY, Issue 11 2009Tony Triglia Summary The Apicomplexan parasite responsible for the most virulent form of malaria, Plasmodium falciparum, invades human erythrocytes through multiple ligand,receptor interactions. The P. falciparum reticulocyte-binding protein homologue (PfRh or PfRBL) family have been implicated in the invasion process but their exact role is unknown. PfRh1 and PfRh4, members of this protein family, bind to red blood cells and function in merozoite invasion during which they undergo a series of proteolytic cleavage events before and during entry into the host cell. The ectodomain of PfRh1 and PfRh4 are processed to produce fragments consistent with cleavage in the transmembrane domain and released into the supernatant, at about the time of invasion, in a manner consistent with rhomboid protease cleavage. Processing of both PfRh1 and PfRh4, and by extrapolation all membrane-bound members of this protein family, is important for function and release of these proteins on the merozoite surface and they along with EBA-175 are important components of the tight junction, the transient structure that links the erythrocyte via receptor,ligand interactions to the actin,myosin motor in the invading merozoite. [source] Conformation and lytic activity of eumenine mastoparan: a new antimicrobial peptide from wasp venomCHEMICAL BIOLOGY & DRUG DESIGN, Issue 3 2004M.P. Dos Santos Cabrera Abstract:, Eumenine mastoparan-AF (EMP-AF) is a novel membrane active tetradecapeptide recently isolated from the venom of solitary wasp, Anterhynchium flavomarginatum micado. It was reported previously that EMP-AF peptide presented low cytolytic activities in human erythrocytes and in RBL-2H3 mast cells. In the present work, we observed that this peptide is able to permeate anionic liposomes, and in less extension also the neutral ones. We present evidences showing that the permeation ability is well correlated with the amount of helical conformation assumed by the peptides in these environments. This peptide also showed a broad-spectrum inhibitory activity against Gram-positive and Gram-negative bacteria. The permeability of liposomes and the antibiotic effect showed a significant reduction when C-terminus was deamidated (in acidic form). The removal of the three first amino acid residues from the N-terminus rendered the peptide inactive both in liposomes and in bacteria. The results suggest that the mechanism of action involves a threshold in the accumulation of the peptide at level of cell membrane. [source] PROTECTIVE ROLE OF A NOVEL ERYTHROCYTE-DERIVED DEPRESSING FACTOR ON BLOOD VESSELS OF RENOVASCULAR HYPERTENSIVE RATSCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 5-6 2007Huan Pang SUMMARY 1We have isolated a novel human erythrocyte-derived depressing factor (EDDF) that has a significant antihypertensive effect in various rat models of hypertension. The aim of the present study was to examine the mechanisms of action of EDDF on vascular function in two-kidney, one-clip (2K1C) renovascular hypertensive rats. 2The EDDF was prepared from human erythrocytes. Experiments were performed in 18 male Wistar rats. The vascular ring perfusion assay and a two-photon laser scanning fluorescence microscope (TMP) were used to evaluate the vascular contractile response. The effects of EDDF on phenylephrine (PE)- and noradrenaline (NA)-induced vascular contraction were evaluated in 2K1C hypertensive rats. The proliferation and DNA synthesis in vascular smooth muscle cells (VSMC) were determined using the [3H]-TdR (thymidine) incorporation and 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays. Flow cytometry, reverse transcription,polymerase chain reaction and western blots were used to measure cell cycle and apoptotic profiles, platelet-derived growth factor (PDGF)-A expression and the activity of extracelluar signal-regulated kinase (ERK)-1/2, as well as the expression of cyclin D1 and cyclin-dependent kinase (CDK) 4. 3At 10,5 g/mL, EDDF significantly decreased the PE- and NA-induced hypertensive vascular contraction. In addition, EDDF inhibited DNA synthesis in primary VSMC from 2K1C rats. The mRNA expression of PDGF-A in VSMC was twofold higher in 2K1C rats compared with control rats, whereas EDDF significantly inhibited the increment in PDGF-A mRNA expression. In addition, EDDF inhibited the phosphorylation of ERK1/2 and decreased the expression of cyclin D1 and CDK4; p21 (Cip1) levels were increased after treatment with EDDF. 4In conclusion, EDDF inhibits VSMC proliferation in 2K1C rats through G0/G1 cell cycle arrest. The effects may be mediated, in part, by enhanced expression of p21 (Cip1) and the inhibition of ERK1/2 phosphorylation and the expression of cyclin D1/CDK4 and PDGF-A. [source] | |||||||||||||||||||