Leukotriene Release from Neutrophils of Patients on Hemodialysis with Cellulose Membranes

1992 ◽  
Vol 15 (2) ◽  
pp. 84-88 ◽  
Author(s):  
A. Jörres ◽  
D. Jörres ◽  
G.M. Gahl ◽  
E. Schulz ◽  
A. Mahiout
Keyword(s):  
Gradient Centrifugation ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Dialysis Patients ◽  
Healthy Donors ◽  
Significant Difference ◽  
Before And After ◽  
Ficoll Density Gradient Centrifugation ◽  
Cellulose Membranes

The role of cytokines in patients with chronic renal failure is currently under investigation. We therefore studied the release of leukotriene B4 (LTB4) from polymorphonuclear leukocytes (PMN) in stable dialysis patients treated with two different cellulose membranes, Cuprophan and Hemophan®, a modified cellulose with less complement activation. Six patients were treated for four weeks with Cuprophan then switched to Hemophan® for another four weeks. Before and after the last treatment of each period, PMN were separated from 20 ml heparinized blood by FICOLL density gradient centrifugation. Portions of 5x106 PMN were resuspended in Hanks' buffer and stimulated for 5 minutes with calcium ionophore A23187 (5 /μg/ml). LTB4 in cell supernatants was determined by specific radioimmunoassay. PMN from dialysis patients before HD released significantly (p<0.01) more LTB4 than healthy donors. No significant difference between pre- and post-dialysis values was observed with Cuprophan or Hemophan® dialyzers. Our data suggest that the acute effects of blood membrane interaction with either complement activating or non-activating dialyzers do not lead to changes in post-dialysis leukotriene metabolism, but leukotriene production is enhanced chronically in dialysis patients.

scholarly journals Human platelet fibrinogen: purification and hemostatic properties

Blood ◽  
1985 ◽  
Vol 66 (4) ◽  
pp. 808-815 ◽  
Author(s):  
TJ Kunicki ◽  
PJ Newman ◽  
DL Amrani ◽  
MW Mosesson
Keyword(s):  
Platelet Aggregation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Calcium Ionophore ◽  
Fluid Phase ◽  
Calcium Ionophore A23187 ◽  
Plasma Fibrinogen ◽  
Ionophore A23187 ◽  
Significant Difference

Abstract Conditions were developed in which 80% to 90% of platelet fibrinogen could be routinely purified in nondegraded form from the fluid phase of platelet suspensions stimulated with the calcium ionophore, A23187, in the presence of calcium, leupeptin, and prostaglandin E1. Fibrinogen was separated from other released proteins by chromatography on diethylaminoethanol (DEAE)-cellulose using a continuous pH and ionic strength gradient. Purified platelet fibrinogen, greater than 98% homogeneous by immunoelectrophoresis and sodium-dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE), consisted of intact A alpha, B beta and gamma A chains, but not gamma' chains, and was 95% to 96% clottable. Platelet fibrinogen was shown to compete for the binding of radiolabeled plasma fibrinogen to ADP-activated platelets in a manner identical to that of unlabeled plasma fibrinogen itself. Also, at equivalent protein concentrations, platelet and plasma fibrinogens supported platelet aggregation to an equivalent extent. Based upon these results, we conclude that there is no significant difference between platelet and plasma fibrinogen with respect to their size, their clottability, their affinity for the activated platelet fibrinogen receptor, or their capacity to support subsequent platelet aggregation.

scholarly journals Diverse stimuli engage different neutrophil extracellular trap pathways

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Elaine F Kenny ◽  
Alf Herzig ◽  
Renate Krüger ◽  
Aaron Muth ◽  
Santanu Mondal ◽  
...  
Keyword(s):  
Alternative Pathway ◽  
Group B Streptococcus ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Neutrophil Extracellular Trap ◽  
Ionophore A23187 ◽  
Chromosomal Dna ◽  
Healthy Donors ◽  
Nadph Oxidase Complex ◽  
Group B

Neutrophils release neutrophil extracellular traps (NETs) which ensnare pathogens and have pathogenic functions in diverse diseases. We examined the NETosis pathways induced by five stimuli; PMA, the calcium ionophore A23187, nigericin, Candida albicans and Group B Streptococcus. We studied NET production in neutrophils from healthy donors with inhibitors of molecules crucial to PMA-induced NETs including protein kinase C, calcium, reactive oxygen species, the enzymes myeloperoxidase (MPO) and neutrophil elastase. Additionally, neutrophils from chronic granulomatous disease patients, carrying mutations in the NADPH oxidase complex or a MPO-deficient patient were examined. We show that PMA, C. albicans and GBS use a related pathway for NET induction, whereas ionophores require an alternative pathway but that NETs produced by all stimuli are proteolytically active, kill bacteria and composed mainly of chromosomal DNA. Thus, we demonstrate that NETosis occurs through several signalling mechanisms, suggesting that extrusion of NETs is important in host defence.

scholarly journals Human platelet fibrinogen: purification and hemostatic properties

Blood ◽  
1985 ◽  
Vol 66 (4) ◽  
pp. 808-815
Author(s):  
TJ Kunicki ◽  
PJ Newman ◽  
DL Amrani ◽  
MW Mosesson
Keyword(s):  
Platelet Aggregation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Calcium Ionophore ◽  
Fluid Phase ◽  
Calcium Ionophore A23187 ◽  
Plasma Fibrinogen ◽  
Ionophore A23187 ◽  
Significant Difference

Conditions were developed in which 80% to 90% of platelet fibrinogen could be routinely purified in nondegraded form from the fluid phase of platelet suspensions stimulated with the calcium ionophore, A23187, in the presence of calcium, leupeptin, and prostaglandin E1. Fibrinogen was separated from other released proteins by chromatography on diethylaminoethanol (DEAE)-cellulose using a continuous pH and ionic strength gradient. Purified platelet fibrinogen, greater than 98% homogeneous by immunoelectrophoresis and sodium-dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE), consisted of intact A alpha, B beta and gamma A chains, but not gamma' chains, and was 95% to 96% clottable. Platelet fibrinogen was shown to compete for the binding of radiolabeled plasma fibrinogen to ADP-activated platelets in a manner identical to that of unlabeled plasma fibrinogen itself. Also, at equivalent protein concentrations, platelet and plasma fibrinogens supported platelet aggregation to an equivalent extent. Based upon these results, we conclude that there is no significant difference between platelet and plasma fibrinogen with respect to their size, their clottability, their affinity for the activated platelet fibrinogen receptor, or their capacity to support subsequent platelet aggregation.

Isolated canine mastocytoma cells: propagation and characterization of two cell lines

1986 ◽  
Vol 251 (6) ◽  
pp. C935-C944 ◽  
Author(s):  
S. C. Lazarus ◽  
R. DeVinney ◽  
L. J. McCabe ◽  
W. E. Finkbeiner ◽  
D. J. Elias ◽  
...  
Keyword(s):  
Mast Cells ◽  
Histamine Release ◽  
Cell Lines ◽  
Calcium Ionophore ◽  
Functional Change ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Significant Difference ◽  
Mastocytoma Cells ◽  
Dose Dependent

Five different dog mastocytoma tumors were successfully transplanted and propagated in BALB/c nude mice. Cells from two of these tumors were passaged serially through at least four generations of mice without morphological or functional change. The average yield from a 2-cm tumor harvested from a mouse was 1.2 +/- 2.8 X 10(9) mast cells with greater than 90% viability. Cells of one line were larger and more heavily granulated than the other, and contained 1.29 +/- 0.74 pg histamine/cell (mean +/- SD). Calcium ionophore A23187 and compound 48/80 caused dose dependent histamine release with no significant difference in release from generation to generation. The smaller cells contained 0.06 +/- 0.06 pg histamine/cell. Histamine release after calcium ionophore or compound 48/80 was dose dependent and unchanged through serial passages. Following passive sensitization antigen caused dose-dependent histamine release confirming the presence of IgE receptors on these cells. In both cell lines histamine release was inhibited by terbutaline, dibutyryl adenosine 3',5'-cyclic monophosphate, or isobutylmethylxanthine. These methods provide a morphologically and functionally stable population of nearly pure canine mast cells for biochemical and physiological studies.

Effects of bull, sperm type and sperm pretreatment on male pronuclear formation after intracytoplasmic sperm injection in cattle

1999 ◽  
Vol 11 (1) ◽  
pp. 59 ◽  
Author(s):  
Hong Wei ◽  
Yutaka Fukui
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Formation Rate ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Significant Difference ◽  
Bull Sperm ◽  
Chemical Pretreatments ◽  
Pronuclear Formation

This study investigated the effects of the bull, sperm type (dead, immotile or motile) and sperm pretreatment (i.e. mechanical (tail-cutting or tail-scoring) or chemical (heparin, heparin + caffeine, calcium ionophore A23187 or dithiothreitol)) on male pronuclear formation after intracytoplasmic sperm injection (ICSI) in cattle. Three experiments were conducted. In Experiment 1, spermatozoa from three bulls (A, B and C) were used for both ICSI and in vitro fertilization (IVF). The results were that sperm from bull B yielded a higher penetration/male pronuclear formation rate than that of bull C when used for IVF (89.6% v. 25.6%, P<0.01). However, when injected into oocytes by ICSI, sperm from bull C had a higher male pronuclear formation rate than that of bull B (34.6% v. 16.1%, P<0.05). The effects of sperm type and mechanical pretreatment were examined in Experiment 2. No significant difference was found in the male pronuclear formation rate when the three types of sperm were injected into oocytes. Tail-scored sperm achieved a higher male pronuclear rate than that of non-mechanically treated ones (38.2% v. 13.2%, P<0.005). In Experiment 3, chemical pretreatments were tested and compared. Higher male pronuclear rates, compared with the control, were obtained when sperm were pretreated with heparin + caffeine, calcium ionophore A23187 and dithiothreitol (48.2%, 62.5% and 64.5% v. 25.0%, P<0.05, 0.005 and 0.005, respectively). These results indicate that (1) there is a bull variation in male pronuclear formation with ICSI, and the male pronuclear rate by ICSI is not coincident with the results by IVF, (2) immobilization of a spermatozoon by tail-scoring before ICSI can improve the formation of the male pronucleus, and (3) an appropriate chemical pretreatment of spermatozoa is necessary to achieve a higher rate of male pronuclear formation.

scholarly journals Endothelial Dysfunction in Cerebral Vessels following Carotid Artery Infusion of Phorbol Ester in Rabbits: The Role of Polymorphonuclear Leukocytes

1994 ◽  
Vol 14 (6) ◽  
pp. 1078-1087 ◽  
Author(s):  
S. E. Akopov ◽  
R. Sercombe ◽  
J. Seylaz
Keyword(s):  
Endothelial Dysfunction ◽  
Carotid Artery ◽  
Sodium Nitroprusside ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Significant Difference ◽  
Left And Right ◽  
Endothelium Dependent Relaxation

The effect of 4β-phorbol-12β-myristate-13α-acetate (PMA) on endothelium-dependent and endothelium-independent vasoconstriction and vasodilation was studied in isolated segments of rabbit middle cerebral artery (MCA). Concentration-dependent responses of the left and right MCA to the constrictors KCl, noradrenaline, uridine 5′-triphosphate, serotonin, and histamine, as well as to the dilators acetylcholine, bradykinin, sodium nitroprusside, and calcium ionophore (A23187), were compared in control animals and after PMA injection into the left common carotid artery. In the control animals there was no significant difference in the responses of the left and right MCA to either the constrictors or the dilators studied. After PMA injection the endothelium-dependent relaxation in response to acetylcholine, bradykinin, and A23187 was reduced in the left MCA (PMA-injected side), whereas the effect of the endothelium-independent dilator sodium nitroprusside remained unchanged. Simultaneously greater contractile responses of the left MCA to serotonin and histamine were obtained. Neither infusion of l-arginine in vivo before the PMA injection nor incubation of the isolated MCA segments with l-arginine affected this difference in MCA reactivity. Platelet depletion did not change the PMA-induced reduction in the endothelium-dependent relaxation, whereas after leukocyte depletion this reduction practically disappeared. These results suggest that the PMA-induced brain microembolia causes acute endothelial dysfunction, which is possibly mediated by intravascular activation of leukocytes and is independent of nitric oxide synthesis from l-arginine. This phenomenon might play an important role in cerebral angiospastic disorders after intravascular activation of leukocytes in cerebral ischemia and reperfusion.

Protein changes in activated human platelets.

1984 ◽  
Vol 30 (12) ◽  
pp. 2078-2083 ◽  
Author(s):  
C S Giometti ◽  
N G Anderson
Keyword(s):  
Calcium Ionophore ◽  
Human Platelets ◽  
Calcium Ionophore A23187 ◽  
Cytoskeletal Proteins ◽  
Ionophore A23187 ◽  
Human Fibrinogen ◽  
Phosphorylated Form ◽  
Before And After ◽  
Molecular Masses ◽  
Dimensional Electrophoresis

Abstract Using two-dimensional electrophoresis, we mapped both the total and the cytoskeletal proteins of human platelets before and after activation with thrombin or the calcium ionophore A23187. Activation resulted in increased abundance of the phosphorylated form of myosin light chains with an approximate molecular mass of 20 kDa, decreased abundance of two proteins with molecular masses of approximately 18 and 25 kDa, and, in the case of activation with thrombin, the appearance of a new chain of protein spots (named "Thromb:1"). The latter, found associated with isolated detergent-insoluble cytoskeletons, reacted with antibody to human fibrinogen and thus were identified as gamma-gamma dimers of fibrin. The total number of proteins associated with the cytoskeleton increased after activation with either thrombin or A23187, but we observed some differences in which proteins were bound, and for how long.

Growth factors and the primary cilium in BALB/c 3T3 cells

1987 ◽  
Vol 45 ◽  
pp. 830-831
Author(s):  
R. W. Tucker ◽  
N. S. More ◽  
S. Jayaraman
Keyword(s):  
Growth Factors ◽  
Primary Cilium ◽  
Cultured Cells ◽  
Morphological Changes ◽  
Calcium Ionophore ◽  
Growth Stimulation ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
3T3 Cells ◽  
Microtubule Depolymerization

The mechanisms by which polypeptide growth factors Induce DNA synthesis in cultured cells is not understood, but morphological changes Induced by growth factors have been used as clues to Intracellular messengers responsible for growth stimulation. One such morphological change has been the transient disappearance of the primary cilium, a “9 + 0” cilium formed by the perinuclear centriole in interphase cells. Since calcium ionophore A23187 also produced both mitogenesis and ciliary changes, microtubule depolymerization might explain ciliary disappearance monitored by indirect immunofluorescence with anti-tubulin antibody. However, complete resorption and subsequent reformation of the primary cilium occurs at mitosis, and might also account for ciliary disappearance induced by growth factors. To settle this issue, we investigated the ultrastructure of the primary cilium using serial thin-section electron microscopy of quiescent BALB/c 3T3 cells before and after stimulation with serum.

Inhibition of Platelet Aggregation by Ethanol in Vitro Shows Specificity for Aggregating Agent Used and Is Influenced by Platelet Lipid Composition

1982 ◽  
Vol 48 (01) ◽  
pp. 049-053 ◽  
Author(s):  
C G Fenn ◽  
J M Littleton
Keyword(s):  
Platelet Aggregation ◽  
Lipid Composition ◽  
Saturated Fatty Acids ◽  
Calcium Ionophore ◽  
Cholesterol Content ◽  
Membrane Lipid ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Increased Sensitivity

SummaryEthanol at physiologically tolerable concentrations inhibited platelet aggregation in vitro in a relatively specific way, which may be influenced by platelet membrane lipid composition. Aggregation to collagen, calcium ionophore A23187 and thrombin (low doses) were often markedly inhibited by ethanol, adrenaline and ADP responses were little affected, and aggregation to exogenous arachidonic acid was actually potentiated by ethanol. Aggregation to collagen, thrombin and A23187 was inhibited more by ethanol in platelets enriched with saturated fatty acids than in those enriched with unsaturated fats. Platelets enriched with cholesterol showed increased sensitivity to ADP, arachidonate and adrenaline but this increase in cholesterol content did not appear to influence the inhibition by ethanol of platelet responses. The results suggest that ethanol may inhibit aggregation by an effect on membrane fluidity and/or calcium mobilization resulting in decreased activity of a membrane-bound phospholipase.

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