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Characterization of (Ca2+ + Mg2+) adenosine triphosphatase activity and calcium transport in boar sperm plasma membrane vesicles and their relation to phosphorylation of plasma membrane proteins.

Properties of (Ca2+ + Mg2+) adenosine triphosphatase (ATPase) in plasma membranes from boar epididymal spermatozoa are described. Enzyme activity is optimum at high pH and has a high affinity for Ca2+. It is not inhibited by the calmodulin antagonist trifluoperazine (TFP), but it is inhibited by low concentrations of Ca2+. Plasma membrane vesicles obtained by hypotonic lysis of intact sperm [mixed inside-out (IOV) and right side-out (ROV) vesicles] transport 45Ca2+ in the presence of oxalate. Similar to the Ca2+-stimulated Mg ATPase activity, transport is unaffected by TFP, but unlike the ATPase, transport is at an optimum rate near neutral pH and is completely inhibited by p-chloromercurphenylsulfonate (pCMS). When plasma membranes are labeled in the presence and absence of Ca2+ and Mg2+ with [gamma-32P]ATP, differences in the intensity of labeling and lability of bound 32P to alkali and hydroxylamine suggest that two polypeptides between 100-120K may be related to a transport ATPase. The addition of TFP at concentrations which stimulate net Ca2+ uptake in intact cells causes intense labeling of a single neutrally charged protein near 68K. These labeling patterns and the properties of (Ca2+ + Mg2+) ATPase identify particular plasma membrane proteins (PMPs) from the complex surface of these cells that may be involved in Ca2+-dependent functions and support the view that calmodulin is not directly involved in the regulation of ATP-driven Ca2+ efflux from boar spermatozoa.

Animals↗

Differential effects of staphylokinase, streptokinase and tissue-type plasminogen activator on the lysis of retracted human plasma clots and fibrinolytic plasma parameters in vitro.

A novel plasma clot lysis system was used to compare the fibrinolytic characteristics of staphylokinase, streptokinase and tissue-type plasminogen activator. 125I-fibrinogen-labelled human plasma clots were formed on needles and mechanically compressed after spontaneous retraction. This model is relatively resistant to lysis and differentiates between fibrin-specific and non-fibrin-specific plasminogen activators. The novel plasminogen activator, recombinant staphylokinase, produced high rates of clot lysis without markedly influencing fibrinogen, plasminogen and alpha 2-antiplasmin in the plasma containing the clots. At equimolar concentrations, streptokinase markedly depleted these parameters in plasma despite low clot lysis rates. Tissue-type plasminogen activator showed relatively high lysis rates at low concentrations, but at higher concentrations, plasminogen depletion caused a decrease in clot lysis. Staphylokinase can be characterised as a fibrin-specific and plasminogen-saving fibrinolytic agent with a high clot lysis potential.

Fibrinogen↗

Quantitation of adsorption and conjugation of plasma proteins by residual glutaraldehyde in fixed collagenous tissue with radioiodinated plasma proteins.

Residual glutaraldehyde (GA) in collagenous cardiovascular tissue prostheses after multiple saline rinses remains in the prostheses and accounts for adsorption and conjugation of a variety of plasma proteins. This may account for later beneficial or adverse effects. Human serum albumin (SA), gamma globulin (GG), and fibrinogen (FB) were iodinated with 125I using the iodogen-transfer technique. Bovine pericardium (PC) was fixed with 0.5% GA for 24 hr and rinsed to remove excess GA. Fresh and GA-fixed PC (FRPC, GAPC: 1 x 1 cm2), in triplicate, were incubated with 0.5-1.0 microCi of tracers in human, porcine, or bovine blood (2 ml) for a period of 0.5, 1, 2, and 3 hr and washed (5x) with saline. Maximum adsorbed proteins per unit weight of collagen (pmol/mg of PC, mean +/- SD) at 3 hr on FRPC and GAPC were quantified with a gamma counter. Fixed PC absorbed significantly more plasma proteins from blood than fresh PC. These conjugated plasma proteins are tightly bound to fixed PC. The adsorbed and conjugated plasma proteins for GAPC and FRPC have the same sequence: SA > GG > FB vs SA > GG > FB. Protein conjugation may affect the remodeling of collagenous cardiovascular tissue prostheses post implantation.

Adsorption↗

Reduced red cell membrane potential and acidification of the plasma in response to contrast materials. Time course of the alteration in plasma pH.

Radiographic contrast materials added to blood reduce the red cell membrane potential by balancing the internal impenetrable anions, hemoglobin and organic phosphates. In so doing, a redistribution of protons occurs such that plasma is acidified. The time course of acidification of plasma is measured in seconds, with a nadir of pH occurring 12 to 15 seconds after addition of Hypaque (1.5 to 3.0 ml/10 ml blood) and a half-time of acidification requiring about 6 seconds. The acidification process is slowed in part by an initial alkalosis due to Hypaque. The acidification of blood is more rapid after addition of Renografin (1.5 to 3.0 ml/10 ml blood) than after addition of Hypaque since the former solution is slightly acidic. The time course of plasma acidification indicates that a maximal reduction in blood pH may not occur in the capillaries of a regional circulation following injection of contrast materials into its afferent vessel, since the transit time of the contrast material may be less than the time required for maximal acidification of plasma.

Blood↗

A long-term study of plasma catecholamine levels and plasma renin activity in borderline hypertension.

Blood pressure and plasma catecholamine levels were followed up for a total of 7 years in 26 subjects with borderline hypertension and in 24 normotensive subjects. During the observation period, five subjects from the borderline hypertensive group became definitely hypertensive, 10 remained in the borderline hypertensive range and in 10, blood pressure fell below borderline hypertensive levels. Of the original 24 normotensive subjects, 15 remained in the normotensive range. In seven, blood pressure rose, but did not reach borderline hypertensive levels. Three subjects were lost to follow-up. In contrast with the subjects who remained normotensive and borderline hypertensive, those who became definitely hypertensive had consistently high plasma noradrenaline concentrations at rest. Plasma adrenaline levels and plasma noradrenaline concentrations during exercise did not differ, however. The study suggests that borderline hypertensive subjects who develop definite hypertension within a few years have consistently enhanced sympathetic activity.

Adult↗

Measurement of inactive renin in rat plasma: effect of nephrectomy and sialoadenectomy on the plasma concentration.

In this paper we describe a routine method of measuring inactive renin in rat plasma. The activation was performed by trypsin, at optimal concentration and incubation conditions. The trypsin treatment formed an interfering and high-performance liquid chromatography-verified tetradecapeptide-like material, which was removed before the assay by a simple batchwise use of a cation-exchange resin. The concentration of activated inactive renin was measured by an antibody-trapping method after the addition of exogenous angiotensinogen. Angiotensinogen was added in order to compensate for the trypsin destruction of angiotensinogen and in order to measure the parameter of renin concentration. The inactive renin concentration in plasma of conscious male rats was 0.48 +/- 0.13 Goldblatt units (GU) per litre (n = 38). This corresponds to 66% (range 42-92%) of the total renin concentration. Physiological experiments in conscious rats were initiated, demonstrating that nephrectomy decreased the inactive renin concentration from 0.45 +/- 0.14 to 0.27 +/- 0.05 GU/l after 24 h (n = 21; P less than 0.01). Submandibular sialoadenectomy decreased the plasma inactive renin concentration from 0.45 +/- 0.11 to 0.34 +/- 0.06 GU/l (n = 12; P less than 0.05) after 7 days. Combined sialoadenectomy and nephrectomy decreased the plasma inactive renin concentration from 0.45 +/- 0.11 to 0.24 +/- 0.06 (n = 12; P less than 0.01).

Angiotensinogen↗

Cumulative risks of early fresh frozen plasma, cryoprecipitate and platelet transfusion in Europe.

BACKGROUND: Injuries are a leading cause of death in the young. Of transfused patients in Sweden about 10% have injuries and 2% receive plasma and 0,2% platelets. Quality, safety, sufficiency and the optimal use of blood and blood components have been the focus for the Council of Europe since 1955 and Guidelines are published. In 2002 the European Community adopted the Directive 2002/98/EC with legally binding requirements for the quality and safety of blood and blood components, including hemovigilance systems. THE USE OF BLOOD COMPONENTS: Data on the use of blood components are regularly collected by the Council of Europe. The use per 1,000 inhabitants show wide variations. THE RISKS OF TRANSFUSION: The potential adverse effects of plasma and platelet components, listed in recently published monographs from the Council of Europe, have been compared with published data from hemovigilance systems. There are two principally different hemovigilance systems in Europe, and the reported overall rates show a fifty-fold difference. Severe transfusion reactions are rare or very rare. Transfusion related acute lung injury (TRALI) and sepsis due to inadvertent bacterial contamination are probably of particular inportance for trauma patients. These reactions are presumably insufficiently recognized and reported. CONCLUSION: Common definitions and systems for hemovigilance will be important for the evaluation of new safety measures. A commonly accepted definition of TRALI will aid in the differential diagnosis of transfusion reactions. Measures to further reduce the rate of transfusion reactions will render the risk-benefit ratio of appropiate transfusions more favorable.

Blood Component Transfusion↗

Beam-plasma coupling effects on the stopping power of dense plasmas.

The stopping power for ion beams in dense plasmas is investigated on the basis of quantum kinetic equations. Strong correlations between the beam ions and the plasma particles which occur for high ion charge numbers and strongly coupled plasmas are treated on the level of the statically screened T-matrix (binary collision) approximation. Dynamic screening effects are included using a combined scheme which considers both close collisions and collective effects. Applying this approach, the ion charge number dependence of the stopping power is determined. The result is a modification of the Z(2)(b) scaling law. In particular, the stopping power is reduced for strong beam-plasma coupling. Good agreement is found between T-matrix results and simulation data (particle-in-cell and molecular dynamics) for low beam velocities.

Journal Article↗

Plasma oscillations and expansion of an ultracold neutral plasma

We report the observation of plasma oscillations in an ultracold neutral plasma. With this collective mode we probe the electron density distribution and study the expansion of the plasma as a function of time. For classical plasma conditions, i.e., weak Coulomb coupling, the expansion is dominated by the pressure of the electron gas and is described by a hydrodynamic model. Discrepancies between the model and observations at low temperature and high density may be due to strong coupling of the electrons.

Journal Article↗

Complete spectrum of kinetic eigenmodes for plasma oscillations in a weakly collisional plasma.

Kinetic eigenmodes of plasma oscillations in a weakly collisional plasma, described by a collision operator of the Fokker-Planck type, are obtained in closed form for initial-value as well as for boundary-value problems. These eigenmodes, which are smooth and compose a complete discrete spectrum, play the same role for weakly collisional plasmas as the Case-Van Kampen modes do for collisionless plasmas.

Journal Article↗

Fusicoccin Binding to Its Plasma Membrane Receptor and the Activation of the Plasma Membrane H-ATPase: I. Characteristics and Intracellular Localization of the Fusicoccin Receptor in Microsomes from Radish Seedlings.

The characteristics of fusicoccin binding were investigated in microsomes from 24-h-old radish (Raphanus sativus L.) seedlings. The time course of fusicoccin binding depended on fusicoccin concentration: equilibrium was reached much faster at 10 nanomolar fusicoccin than at 0.3 nanomolar fusicoccin. Scatchard analysis of equilibrium binding as a function of fusicoccin concentration indicated a single class of receptor sites with a K(d) of 1.8 nanomolar and a site density of 6.3 picomoles per milligram protein. Similar values (K(d) 1.7 nanomolar and site density 7 picomoles per milligram protein) were obtained from the analysis of the dependence of equilibrium binding on membrane concentration at fixed fusicoccin concentrations. Fusicoccin binding comigrated with the plasma membrane H(+)-ATPase in an equilibrium sucrose density gradient: both activities formed a sharp peak (1.18 grams per milliliter) clearly distinct from that of markers of other membranes which all peaked at lower densities. The saturation profiles of fusicoccin binding and of fusicoccin-induced activation of the plasma membrane H(+)-ATPase, measured under identical conditions, were similar, supporting the view that fusicoccin-induced activation of the plasma membrane H(+)-ATPase is mediated by fusicoccin binding to its plasma membrane receptor.

Journal Article↗

Selective Inhibition of Active Uptake of Sucrose into Plasma Membrane Vesicles by Polyclonal Sera Directed against a 42 Kilodalton Plasma Membrane Polypeptide.

Several polyclonal sera were raised in rabbits and in mice against putative sucrose carrier proteins, i.e. a 42 kilodalton (O Gallet, R Lemoine, C Larsson, S Delrot [1989] Biochim Biophys Acta 978: 56-64) and a 62 kD (KG Ripp, PV Viitanen, WD Hitz, VR Fransceschi [1988] Plant Physiol 88: 1435-1445) polypeptide of the plasma membrane. The effects of these sera on the active uptake of sucrose and of valine into purified plasma membrane vesicles from sugar beet (Beta vulgaris L.) leaves and roots were studied. At a dilution of 1/50, the anti-42 kilodalton sera consistently inhibited sucrose uptake in plasma membranes from leaves or from roots. They had no effect on valine uptake. Under the same experimental conditions, the anti-62 kilodalton sera had no effect on active uptake of sucrose. The data further support the view that a 42 kilodalton polypeptide is a component of the transport system mediating sucrose uptake across the plasma membrane of plant cells.

Journal Article↗

Plasma lipids and physicochemical properties of the erythrocyte plasma membrane throughout pregnancy.

BACKGROUND: The physiopathological relevance of plasma lipid concentrations is supported by the observation that they might affect the physicochemical properties of the plasma membrane of circulating cells and might be crucial in the pathological conditions complicating pregnancy. METHODS: Plasma and erythrocyte membrane lipid composition, membrane fluidity and function [membrane-bound enzyme sodium/potassium adenosine triphosphatase (Na+/K+-ATPase) activity] were studied in 24 healthy women in the nonpregnant condition and at 12, 24 and 38 weeks' gestation. RESULTS: The plasma showed an increase in total and high density lipoprotein (HDL) cholesterol, triglyceride (TG) and phospholipid (PL) levels. In the erythrocyte membrane we found the cholesterol/phospholipid (C/PL) ratio increased and the saturated to unsaturated fatty acid (FA) ratio (Sat/Unsat) significantly reduced; fluorescence polarization showed an initial increase during pregnancy and a progressive decrease afterwards; Na+/K+-ATPase activity was progressively reduced. CONCLUSIONS: A generalized alteration in the composition and function of the maternal erythrocyte membrane is present during a physiological pregnancy, and these modifications may involve a more complex physiopathogenetic mechanism.

Case-Control Studies↗

Plasma cholinesterase and trophoblastic disease. Gestational trophoblastic disease and reduced activity of plasma cholinesterase.

A case of prolonged action of suxamethonium in a patient with gestational trophoblastic disease is reported. Postoperatively the patient was found to have markedly reduced plasma cholinesterase activity (363 IU/litre) with a normal cholinesterase phenotype. Consequently plasma cholinesterase activity and phenotype were measured in six other patients with the condition and these results compared with those of 22 patients with normal first trimester pregnancies undergoing therapeutic abortion. Plasma cholinesterase phenotype was normal in all patients studied. The activity was significantly decreased (p less than 0.05) from the normal range (620-1370 IU/litre) in all patients with trophoblastic disease. In the 22 patients with normal pregnancies, 14 had activity values in the abnormal range (less than 620 IU/litre) while the mean cholinesterase activity of the group as a whole was significantly decreased (561.8 IU/litre, p less than 0.05) below the normal range. These results confirm the presence of a decrease in plasma cholinesterase activity in early pregnancy and provide new evidence for a decrease in activity in a pseudopregnancy state.

Adult↗

Polymorphic plasma postalbumins of some domestic animals (pig PO2, horse Xk and dog Pa proteins) identified as homologous to human plasma alpha 1B-glycoprotein.

Pig, horse and dog plasma proteins, separated by horizontal polyacrylamide gel electrophoresis (pH 9.0) and electrophoretically transferred to nitrocellulose membranes, were tested for cross-reaction with antiserum to human plasma alpha 1B-glycoprotein (alpha 1B). The results showed that one previously reported polymorphic plasma postalbumin in each of these species (pig PO2, horse Xk and dog Pa protein) was homologous to human plasma alpha 1B. In the light of the previously known genetic linkages in these species, this implied: (1) alpha 1B gene is close linked to Phi, Pgd and Hal (halothane sensitivity locus) loci in pigs; and (2) alpha 1B gene is linked to ME1 and Phi loci in horses. This suggested that the alpha 1B gene may also be found to be closely linked to gene(s) controlling susceptibility to malignant hyperthermia in humans and other mammals.

Alpha-Globulins↗

The reliability of the mean normal prothrombin time of fresh plasmas and of the normal value from a lyophilized 'normal' plasma in prothrombin ratio determination.

The mean normal prothrombin time (MNPT) based on the fresh plasma of 20 healthy individuals is the recommended way to derive a normal value for the prothrombin ratio. This is generally believed to give a reasonable representation of the normal in the local population, but there may be difficulty in obtaining a good representative sample in some centres. The alternative of a uniform lyophilized 'normal' plasma has been suggested. The comparative value of a widely used commercial 'normal' plasma and the MNPT has been assessed in an international study at 37 centres. Two common thromboplastins were tested using the local coagulometer methods. The variability of the results and their relative performance in local thromboplastin calibration to derive the International Sensitivity Index (ISI) were assessed. The reliability of the two types of normal in local system ISI calibration with different lyophilized plasma calibrants has also been tested. When the lyophilized 'normal' was substituted for the MNPT in ISI determination no appreciable difference in the degree of correction for coagulometers effects on International Normalized Ratios was found.

Calibration↗

Quantification of leucocyte elastase and cathepsin G in plasma by a simple method: effect of elastase in plasma levels of D-dimer and thrombomodulin.

The purpose of this study was to determine levels of leucocyte elastase and cathepsin G in the plasma of patients in various pathological states, in which plasma increases or decreases in coagulation and fibrinolytic factors were seen. Simple methods were developed to measure the leucocyte proteinases and the results were correlated with conventional assays of coagulation and fibrinolytic factors. The total number of patients and total number of plasma samples examined were 340 and 1292, respectively. No correlation was observed between the plasma levels of elastase and cathepsin G, and plasminogen, fibrinogen and leucocyte counts. There was a weak overall correlation, however, between the leucocyte proteinases and each of the four parameters: D-dimer, thrombomodulin, antithrombin III and platelet count. There was a strong correlation between leucocyte proteinases and D-dimer and thrombomodulin in those patients with plasminogen levels within the normal range. Increased D-dimer levels, as well as plasmin, may suggest that elevated leucocyte proteinases contribute to elevated fibrinolytic mechanisms in these instances.

Biological Assay↗

Leucocyte-associated plasma proteins. I. Plasma proteins in supernatants and cell pellets during successive washings of human blood cells.

Sixteen plasma proteins were studied in the supernatants from 20 successive washings of human leucocytes by a new sensitive immunoelectrophoretic method. In the first few supernatants a fast and parallel decrease in concentration of all proteins took place, but for 11 proteins this was followed by a phase of slow decrease due to cellular release. Eight proteins could be quantitated throughout the 20 washings: prealbumin, albumin, orosomucoid, alpha 1-antitrypsin, transferrin, C3, haptoglobin, and IgG. These proteins and trace amounts of eight other plasma proteins were present in the cell lysate after the washings. In comparison with plasma, the leucocytes contained relatively higher concentrations of prealbumin, orosomucoid, alpha 1-antitrypsin, and haptoglobin than of albumin. In crossed immunoelectrophoresis these same four proteins were present as molecular variants differing from their plasma counterpart in electrophoretic mobility and morphology of the precipitate.

Animals↗