We investigated the expression of aquaporin 1 (AQP1) in tissues from canines with an inherited anomaly that causes their erythrocytes to have high K+. Northern blot analysis revealed abundant AQP1 expression in lung and kidney, though little expression was found in spleen. Using anti-C-terminus for dog AQP1, abundant expression was shown in kidney, trachea, and eye, but little expression was shown in pancreas and cerebrum, indicating that AQP1 expression in canine tissues is similar to that noted in other mammals.
Animals ; Aquaporin 1/*metabolism ; Blotting, Northern ; Dogs ; Erythrocytes/*chemistry ; Immunoblotting ; Potassium/*analysis ; Viscera/metabolism

AIMTo study the relationship between erythrocyte filtration index (EFI) and cholesterol and Na+ -K+ -ATPase activity of erythrocyte membrane in early stage of severe burns.

METHODSThe EFI was measured by means of percolation, the erythrocyte membrane cholesterol level was measured by means of chemically modified electrode, and the membrane Na+ -K+ -ATPase activity by means of detection of Pi.

RESULTSIn early stage of severe burns, the cholesterol level and Na+ -K -ATPase activity of erythrocyte membrane were always higher than those of control group (P < 0.01), but the EFI was lower than that of control group (P < 0.01). The cholesterol level and Na+ -K+ -ATPase activity of the erythrocyte membrane were negative correlation with EFI (r cho= -0.871, r ATPase = -0.801, P < 0.01) nospectively.

CONCLUSIONThe cholesterol level and Na+ -K+ -ATPase activity of erythrocyte membrane play a key role in the change of EFI.

Adult ; Burns ; metabolism ; Case-Control Studies ; Cholesterol ; chemistry ; Erythrocyte Membrane ; chemistry ; Erythrocytes ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Sodium-Potassium-Exchanging ATPase ; metabolism

BACKGROUNDThe side effects of cyclosporine therapy include thromboembolic complications. However, the mechanisms underlying the hypercoagulable state induced by cyclosporine are not fully understood. Cyclosporine binds to red blood cells (RBCs) with a high affinity in circulation and alters the membranes of RBCs. Therefore, we propose that such alterations in RBCs membranes play a role in cyclosporine-induced coagulopathy and this disorder may be rectified by lactadherin, a phosphatidylserine binding protein.

METHODSRBCs from healthy adults were treated with various concentrations of cyclosporine. Procoagulant activity of the RBC membrane was measured by the single stage recalcification time and confirmed by detection of tenase and thrombin assembly through enzymatic assays. Inhibition assays of coagulation were carried out in the presence of lactadherin, annexin V or antitissue factor. Phosphatidylserine exposure was detected by flow cytometry and confocal microscopy through binding with fluorescein isothiocyanate (FITC)-labeled lactadherin as well as FITC annexin V.

RESULTSRBCs treated with cyclosporine demonstrated increased procoagulant activity. Cyclosporine treatment markedly shortened the clotting time of RBCs ((305 +/- 10) seconds vs (366 +/- 15) seconds) and increased the generation of intrinsic factor Xase ((7.68 +/- 0.99) nmol/L vs (2.86 +/- 0.11) nmol/L) and thrombin ((15.83 +/- 1.37) nmol/L vs (4.88 +/- 0.13) nmol/L). Flow cytometry and confocal microscopy indicated that cyclosporine treatment induced an increased expression of phosphatidylserine on the RBC membrane. Lactadherin was more sensitive in detecting phosphatidylserine exposure of the RBC membrane than annexin V. The modulating effect of procoagulant activity was concomitant with and dependent on phosphatidylserine exposure. Blocking of phosphatidylserine with lactadherin effectively inhibited over 90% of FXa generation and prothrombinase activity and prolonged coagulation time.

CONCLUSIONSProcoagulant properties of RBCs membranes resulting from phosphatidylserine exposure may play an important role in cyclosporine-induced thrombosis. Lactadherin can be used as a sensitive probe for phosphatidylserine detection. Its high affinity for phosphatidylserine may provide a new approach for the treatment of cyclosporine induced thrombogenic properties.

Adult ; Animals ; Annexin A5 ; chemistry ; Cattle ; Cell Membrane ; drug effects ; metabolism ; Cells, Cultured ; Cyclosporine ; pharmacology ; Erythrocytes ; drug effects ; metabolism ; Flow Cytometry ; Humans ; Membrane Glycoproteins ; chemistry ; Microscopy, Confocal ; Milk Proteins ; chemistry ; Phosphatidylserines ; chemistry ; metabolism ; Thrombosis ; chemically induced ; metabolism

Changes in microparticles (MP) from red blood cell (RBC) concentrates in the context of irradiation have not been investigated. The aim of this study was to evaluate how irradiation affects the number of MPs within transfusion components. Twenty RBC concentrates, within 14 days after donation, were exposed to gamma rays (dose rate: 25 cGy) from a cesium-137 irradiator. Flow cytometry was used to determine the numbers of MPs derived from RBC concentrates before and 24 hr after irradiation. The mean number of MPs (±standard deviation) in RBC concentrates was 21.9×10(9)/L (±22.7×10(9)/L), and the total number of MPs ranged from 2.6×10(9)/L to 96.9×10(9)/L. The mean number of MPs increased to 22.6×10(9)/L (±31.6×10(9)/L) after irradiation. Before irradiation, the CD41-positive and CD235a-positive MPs constituted 9.5% (1.0×10(9)/L) and 2.2% (263×10(6)/L) of total MPs, respectively. After irradiation, CD41-positive MPs increased to 12.1% (1.5×10(9)/L) (P=0.014), but the CD235a-positive MPs decreased to 2.0% (214×10(6)/L) of the total MPs (P=0.369). Irradiation increases the number of CD41-positive MPs within RBC concentrates, suggesting the irradiation of RBC concentrates could be associated with thrombotic risk of circulating blood through the numerical change.
Cell-Derived Microparticles/chemistry/*metabolism/radiation effects ; Erythrocytes/*cytology/radiation effects ; Flow Cytometry ; Gamma Rays ; Humans ; Membrane Glycoproteins/metabolism ; Metalloendopeptidases/metabolism ; Platelet Membrane Glycoprotein IIb/metabolism

Vibrio vulnificus cytolysin (VVC) has been implicated as one of the important virulence determinants of V. vulnificus that causes serious septicemia and wound infection. An attempt was made to investigate that VVC could act as a ligand which stimulates intracellular signaling systems. Cholesterol dose-dependently blocked VVC hemolytic activity through oli-gomerization of cytolysin. Among cholesterol derivatives including 7-dehydrocholesterol, cholesteryl esters, deoxycholate, and cholestane tested, only 7-dehydrocholesterol induced oligomerization as well as inactivation of VVC. These results show that oligomerization of VVC is completely dependent on three-dimensional structure of cholesterol where specific interaction of cholesterol at oligomerization sites of VVC is very selective. These findings support the idea that cholesterol which constitute many of cellular plasma membrane could be a receptor of VVC on plasma membrane of target cells.
Animals ; Bacterial Toxins/antagonists & inhibitors/chemistry/metabolism ; Cholesterol/chemistry/*pharmacology ; Cytotoxins/antagonists & inhibitors/*chemistry/*metabolism ; Dehydrocholesterols/chemistry/pharmacology ; Dose-Response Relationship, Drug ; Erythrocytes/drug effects ; Hemolysis/drug effects ; Mice ; Molecular Structure ; Signal Transduction ; Substrate Specificity ; Vibrio/*chemistry

After invasion of red blood cells, malaria matures within the cell by degrading hemoglobin avidly. For enormous protein breakdown in trophozoite stage, many efficient and ordered proteolysis networks have been postulated and exploited. In this study, a potential interaction of a 60-kDa Plasmodium falciparum (Pf)-heat shock protein (Hsp60) and Pf-calpain, a cysteine protease, was explored. Pf-infected RBC was isolated and the endogenous Pf-Hsp60 and Pf-calpain were determined by western blot analysis and similar antigenicity of GroEL and Pf-Hsp60 was determined with anti-Pf-Hsp60. Potential interaction of Pf-calpain and Pf-Hsp60 was determined by immunoprecipitation and immunofluorescence assay. Mizoribine, a well-known inhibitor of Hsp60, attenuated both Pf-calpain enzyme activity as well as P. falciparum growth. The presented data suggest that the Pf-Hsp60 may function on Pf-calpain in a part of networks during malaria growth.
Amino Acid Sequence ; Calpain/genetics/*metabolism ; Chaperonin 60/chemistry/genetics/*metabolism ; Erythrocytes/parasitology ; Humans ; Malaria, Falciparum/parasitology ; Molecular Sequence Data ; Plasmodium falciparum/chemistry/enzymology/genetics/*metabolism ; Protein Binding ; Protozoan Proteins/chemistry/genetics/*metabolism ; Sequence Alignment

OBJECTIVETo investigate the effect of Coptis chinensis on oxidative hemolysis of erythrocytes in mice.

METHODAcetylphenyhydrazine (APH)-induced oxidative hemolysis of erythrocytes in mice were used. The contents of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood, and malondialdehyde (MDA) of erythrocytes were measured. The activity of superoxide dismutase (SOD), reduced glutathione hormone (GSH), and glucose-6-phosp hate dehydrogenase (G-6-PD) of erythrocytes were also determined and the total-antioxygen capability (T-AOC) of blood was analyzed.

RESULTThe levels or amount of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood and MDA of erythrocytes were higher in APH (0.03 g x kg(-1))-induced mice than normal mice. The activity or content of SOD, GSH and G-6-PD was lower in APH-induced mice than in normal mice. Primaquine (0.058 g x kg(-1)) could aggravated the degree of elevated hemolysis of erythrocytes in APH-induced mice. C. chinensis (0.6 g x kg(-1) could deprssed significantly the elevated levels of indirect bilirubin in serum. The levels of free hemoglobin of blood plasma, indirect bilirubin of serum, reticulocytes of blood, the production of SOD and GSH and T-AOC were also decressed by C. chinensis (0.6 g x kg(-1)).

CONCLUSIONC. chinensis suppressed t he degree of hemolysis of erythrocytes in APH-induced mice due to the suppression of the production of lipid peroxidation and increasing of the activity of antioxidase of erythrocytes.

Animals ; Coptis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Erythrocytes ; drug effects ; metabolism ; Female ; Hemolysis ; drug effects ; Male ; Mice ; Oxidation-Reduction ; Phenylhydrazines ; adverse effects ; Plasma ; metabolism ; Random Allocation

OBJECTIVETo study the predominant calcium-antagonist components of Danshen injection.

METHODThe effects of danshensu, protocatechualdehyde and Danshen injection on calcium concentration in cytoplasm of erythrocytes were examined in vitro by the fluorescent Ca+ -chelator fura-2.

RESULTEither DS182 or PCAD can decrease in dose-dependent cytosolic free calcium concentration in human erythrocytes. They had additive effect when mixed, which was similar to Danshen injection.

CONCLUSIONDS182 and PCAD may be predominant calcium-antagonist components of Danshen injection.

Adult ; Benzaldehydes ; isolation & purification ; pharmacology ; Calcium ; metabolism ; Catechols ; isolation & purification ; pharmacology ; Cytoplasm ; metabolism ; Drug Synergism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocytes ; metabolism ; Female ; Humans ; Injections ; Lactates ; isolation & purification ; pharmacology ; Male ; Middle Aged ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo study the antioxidation activity and protective effect of ginger oil on DNA damage.

METHODChemical light assay was used to detect the oxygen radicals scavenging capacity of ginger oil. The erythrocyte oxidation damage was induced by H2O2. The effect of ginger oil on oxidative erythrocyte was observed by the colorimetric analysis assay, and the content of malondialdehyde (MDA) in rabit hepatocyte was measured. The anaylsis of DNA damage was made with single cell gel electrophoresis(SCGE) technique.

RESULTGinger oil might decrease light value compared with control group and inhibited erythrocyte oxidation damage. Compared with that in control group, the degress of DNA damage reduced significantly in the protected groups. Ginger oil might decrease the content of MDA remarkably and inhitibition rate was 48.16%.

CONCLUSIONGinger oil has dominantive protective effect on DNA damage induced by H2O2. Ginger oil might act as a scavenger of oxygen radical and might be used as an antioxidant.

Animals ; Antioxidants ; pharmacology ; DNA Damage ; drug effects ; Erythrocytes ; drug effects ; Ginger ; chemistry ; Hemolysis ; drug effects ; Humans ; Liver ; metabolism ; Lymphocytes ; drug effects ; Malondialdehyde ; metabolism ; Plant Oils ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Protective Agents ; pharmacology ; Rabbits ; Reactive Oxygen Species ; metabolism

BACKGROUND: Detection methods for ABO antibody (Ab) titers vary across laboratories, and the results are different depending on the method used. We aimed to compare titer values using different detection methods for the measurement of ABO Ab titers. METHODS: For ABO Ab detection, pooled group A or B red blood cells (RBCs) were reacted with each of 20 sera from blood groups A, B, or O without dithiothreitol treatment. The room-temperature (RT) incubation technique and the indirect antiglobulin test (IAT) were used in the tube test and gel card test. Flow cytometry (FCM) was performed by using anti-IgM and anti-IgG Abs. RESULTS: Regardless of the blood groups tested, the FCM assay with anti-IgM showed the highest titer compared to the tube test and gel card test with RT incubation in both. The tube test with IAT showed a higher titer than the gel card test with IAT (Gel-IAT) or FCM with anti-IgG in blood group A and B, while Gel-IAT showed the highest titer relative to the other tests, only for the anti-A Ab in blood group O. CONCLUSIONS: There were significant differences in the titers depending on the detection method used, and each method showed a different detection capacity for each ABO Ab depending on the ABO blood group tested. Therefore, caution should be exercised in interpreting ABO Ab titer results, taking into consideration the detection method used and the blood group.
ABO Blood-Group System/*immunology ; Adult ; Aged ; Agglutination Tests/instrumentation/*standards ; Antibodies/*analysis ; Antibodies, Anti-Idiotypic/analysis ; Erythrocytes/chemistry/metabolism ; Female ; *Flow Cytometry ; Humans ; Male ; Middle Aged ; Temperature