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Biomedical subjects

A Livne

Publications and source records attributed to A Livne.

At least 55 records · Page 3Linked to original sources
PubMed1987

Volume-regulating behavior of human platelets.

Human platelets exposed to hypotonic media undergo an initial swelling followed by shrinking (regulatory volume decrease [RVD]). If the RVD is blocked, the degree of swelling is in accord with osmotic behavior. The cells could swell at least threefold without significant lysis. Two methods were used to follow the volume changes, electronic sizing and turbidimetry. Changes in shape produced only limited contribution to the measurements. The RVD was very rapid, essentially complete in 2 to 8 minutes, with a rate proportional to the degree of initial cell swelling. RVD involved a loss of KCl via volume-activated conductive permeability pathways for K+ and anions, presumably Cl-. In media containing greater than 50 mM KCl, the shrinking was inhibited and with higher concentrations was reversed (secondary swelling), suggesting that it is driven by the net gradient of K+ plus Cl-. The K+ pathway was specific for Rb+ and K+ compared to Li+ and Na+. The Cl- pathway accepted NO-3 and SCN- but not citrate or SO4(2-). In isotonic medium, the permeability of platelets to Cl- appeared to be low compared to that of K+. After hypotonic swelling both permeabilities were increased, but the Cl- permeability exceeded that of K+. The Cl- conductive pathway remained open as long as the cells were swollen. RVD was incomplete unless amiloride, an inhibitor of Na+/H+ exchange, was present or unless Na+ was replaced by an impermeant cation. In addition, acidification of the cytoplasm occurred upon cell swelling. This reduction in pHi appeared to activate Na+/H+ exchange, with a resultant uptake of Na+ and reduction in the rate and amount of shrinking. Like other cells, platelets responded to hypertonic shrinking with activation of Na+/H+ exchange, but regulatory volume increase was not detectable.

Amiloride↗
PubMed1986

Lii-Nao countertransport and Li leak in erythrocytes are differentially affected by membrane enrichment with cholesteryl hemisuccinate.

Enrichment of erythrocytes with cholesteryl hemisuccinate caused a marked reduction in Li leak but did not change kinetic and thermodynamic properties of Lii-Nao countertransport of either normotensive persons or patients with essential hypertension. As cholesteryl hemisuccinate was shown to affect the membrane similarly to cholesterol, it is likely that the unique thermodynamic properties of erythrocyte Lii-Nao countertransport in essential hypertension are not caused by changes in cholesterol.

Cholesterol Esters↗
PubMed1986

Thermodynamic properties of erythrocyte Li efflux may facilitate detection of essential hypertension in pregnant women.

Thermodynamic properties of red cell lithium efflux were examined in pregnant women in relation to hypertension. Twenty-two normotensive women, 15 women with essential hypertension, and 27 with pregnancy-induced hypertension were studied. The rates of Li efflux at 37 degrees C in the three groups of pregnant women were similar and nondiscriminatory. The temperature dependence of the Li efflux, known to be uniquely modified in essential hypertension, allowed the differentiation of most (73%) of the pregnant women with essential hypertension as well. Among the women with pregnancy-induced hypertension, 63% showed a temperature-dependence pattern typical for normotensives, and they may be classified as patients with toxemia of pregnancy. The others (37%) showed a thermodynamic pattern of essential hypertension, but a follow-up study is required to ascertain whether they will indeed develop essential hypertension in the future.

Adult↗
PubMed1986

Specific impairment of ADP-induced platelet aggregation by cannabinoids.

The structure-activity relationship of the inhibition of ADP-induced platelet aggregation by cannabinoids was studied. A marked specificity was revealed: derivatives in which the two asymmetric centers have a configuration opposite to that of the natural cannabinoids and have a dimethylheptyl (DMH) side-chain were most inhibitory. It is concluded that the mode of inhibition includes a specific interaction of the cannabinoids with some membrane proteins, possibly including the receptor for ADP.

Adenosine Diphosphate↗
PubMed1985

Lysine binding to activated human platelets and its similarity to fibrinogen binding.

Platelet surface glycoproteins IIb-IIIa are considered to function as the binding site for fibrinogen. Fibrinogen binding is essential for platelet aggregation and several amines have been shown to inhibit this binding. The present study compares the binding properties of 125I-fibrinogen and [3H]lysine with platelets activated by the Ca2+ ionophore A23187. Many lines of similarities in the binding properties are apparent; however, several differences were also found. The similarities are listed below and the differences are pointed out in parentheses. Marked enhancement by platelet activation; deficiency of binding by thrombasthenic platelets lacking the glycoproteins IIb-IIIa; saturability (fibrinogen binding approaches saturation at more than 12 microM, within 10 min; lysine binding at more than 100 mM within 1 min); Ca2+-dependence (at 1 mM Ca2+ lysine binding is minute and fibrinogen binding is half-saturated); reversibility; the binding achieved within 10 min is exchangeable; dissociation depends upon time and external ligand concentration; inhibition by the oligoamines His-Lys and Lys4; inhibition by serum from a thrombasthenic patient who developed anti-glycoproteins IIb-IIIa antibodies; specificity; alanine neither binds to activated platelets nor inhibits fibrinogen binding; it thus appears that the lysine which associates with activated platelets is mostly bound onto the surface of the cells rather than being incorporated. Moreover, the major site of lysine binding seems to be the complexed glycoproteins IIb-IIIa.

Alanine↗
PubMed1985

Fibrinogen fragment D is a recognition site for release-related platelet aggregation.

The present study probes for the segment of the fibrinogen molecule which interacts with the platelet surface upon induction of release-related aggregation. The capability of platelets affixed with fibrinogen fragment D or fibrinogen fragment E to enhance aggregation of gel-filtered platelets (GFP) was compared with that of platelets affixed with fibrinogen. It is shown that release-related aggregation induced by either A23187, 10 microM ADP or thrombin is enhanced by the addition of fixed platelets bearing covalently bound fragment D by as much as 60-70% of the augmentation obtained by platelets bearing fibrinogen. On the other hand, fixed platelets bearing covalently-bound fragment E have no effect on the aggregation of GFP. It is concluded that fragment D bears the site for the interaction of fibrinogen with activated platelets, apparently, with surface-bound thrombospondin.

Adenosine Diphosphate↗
PubMed1984

Erythrocyte Lii-Nao countertransport system. Inhibition by N-ethylmaleimide probes for a conformational change of the transport system.

Human erythrocytes were treated by a series of SH-reagents, including maleimides, iodo compounds, mercurials and oxidizing agents. Rates of Li efflux into Na-rich medium, Li leak and Lii-Nao countertransport were then determined. Of the 13 different reagents studied, only N-ethylmaleimide, iodoacetamide and iodoacetate inhibited selectively the countertransport activity. The effect of the various reagents indicates that the sensitive SH-groups of the countertransport system are not externally exposed. N-Ethylmaleimide was used to probe for changes elicited by substrate cations in Lii-Nao countertransport. In Na- and Li-free medium, inhibition of Lii-Nao countertransport by N-ethylmaleimide of 35% was reached within 2 s. In Na or Li medium, maximal inhibition was twice as great, but was attained much more slowly, within 10 min. Kinetic data and Hill plot analysis indicate the involvement of two classes of SH-groups: one expressed in the various media with and without substrate cations, and an additional one, which becomes specifically available to N-ethylmaleimide in the presence of external Na or Li. The affinity of Na to the site promoting inhibition by N-ethylmaleimide (apparent Km = 12 mM) is higher than the affinity of Na to its external countertransport site (apparent Km = 25 mM, as reported by Sarakadi, B., Alifimoff, J.K., Gunn, R.B. and Tosteson, D.C. (1978) J. Gen. Physiol. 72, 249-265). Reactivity of N-ethyl[14C]maleimide was not modified by the media tested. It is concluded that external Na and Li cause a conformational change in the protein(s) of the countertransport system in human erythrocytes.

Biological Transport↗
PubMed1984

Platelet-platelet recognition during aggregation: distinct mechanisms determined by the release reaction.

Fixed platelets, bearing covalently bound fibrinogen, were previously shown to participate passively in aggregation induced by thrombin or A23187. The bound fibrinogen was specifically required for the interaction. The present study examines the role of the release reaction in controlling the passive participation of the fixed platelets in aggregation. Aggregation of fresh platelets induced by low ADP concentration (less than or equal to 2 microM) was not augmented by the fixed platelets, but was augmented when higher concentrations of ADP were employed. Fixed platelets failed to enhance aggregation of aspirin-treated fresh platelets induced by 10 microM ADP; the augmentation capability was reconstituted by a supernatant fraction from platelets activated by 10 microM ADP in the presence of EDTA. The binding of soluble fibrinogen to activated fresh platelets occurred regardless of the release reaction, but the interaction of fragmented fixed platelets bearing bound fibrinogen with aggregating fresh platelets was apparent only when release took place. It is concluded that fibrinogen covalently bound to fixed platelets is selectively recognized by released compound(s).

Adenosine Diphosphate↗
PubMed1984

Inhibition of platelet aggregation and endogenous lectin activity by oligoamines.

Amino sugars and basic amino acids inhibit platelet aggregation and the activity of the endogenous platelet lectin, yet, relatively high concentrations (approximately 30 mM) are required for the inhibition. If cooperative interactions are involved in these platelet surface activities, oligomers of primary amines should be more potent inhibitors than their individual component amines. Accordingly, series of oligomers of basic amino acids, of the polyamines (putrescine, spermidine and spermine) and of aliphatic diamines differing in chain length were tested for potency of inhibition of platelet aggregation and endogenous platelet lectin activity. Indeed, oligoamines were much more potent inhibitors of platelet aggregation than their corresponding monomers or shorter oligomers, more than accountable by an additive effect. For example, 60, 3 and 0.3 mM were needed for 50% inhibition of platelet aggregation by lysine, (lys)3 and (lys)5, respectively. A similar pattern was observed for the effect of the oligoamines on the activity of the endogenous platelet lectin. The inhibition of platelet aggregation by spermine is competitive, since the effect of a given dose of spermine decreased with increasing platelet concentration. Neither inhibition of platelet-inducer interaction nor Ca2+ insufficiency explain the inhibitory effects of the oligoamines. The results are consistent with the hypothesis that cooperative surface interactions underlie platelet aggregation and platelet lectin activity. The cooperative effects may reflect the formation of patches or clusters of positively charged groups on the surface of activated platelets.

Amino Acids↗
PubMed1984

The erythrocyte membrane in essential hypertension. Characterization of the temperature dependence of lithium efflux.

Erythrocytes of most patients with essential hypertension are distinguished by a typical pattern of temperature-dependence of Li efflux. In the present study we have attempted to characterize this unique temperature response. Measurements of Li efflux into Na medium and Lii-Nao countertransport were conducted simultaneously at finely spaced temperature intervals with increments of 1 to 2 degrees C in the range of 10-40 degrees C. The Arrhenius plots for the efflux in Na medium and for Lii-Nao countertransport in erythrocytes of both normotensives and hypertensives were biphasic with slopes representing apparent energies of activation of about 28 and 8 kcal/mol below and above the 'break', respectively. However, the 'break' in the Arrhenius plot appeared at distinctly different temperatures: 30 degrees C for normotensives and 20 degrees C for hypertensives. The Li efflux was resolved into N-ethylmaleimide-sensitive and -insensitive components. The sensitive component exhibited a typical biphasic temperature response, with the characteristic 'break': at 30 degrees C for normotensives and at 20 degrees C for hypertensives. In contrast, the N-ethylmaleimide-insensitive component was alike in normotensives and hypertensives. It is concluded that: (a) the unique temperature dependence of Li efflux in erythrocytes of hypertensives results from a localized modification in the membrane; (b) the N-ethylmaleimide-sensitive component represents a protein moiety which distinguishes between the erythrocyte membrane of normotensives and hypertensives; (c) the expression of the temperature dependence as judged by the sharp transition in slope (within 1 to 2 degrees C), apparently reflects the cooperative involvement of membrane lipids, associated with the Li efflux system.

Biological Transport↗
PubMed1984

Thrombospondin plays a role in platelet-platelet recognition during release-related aggregation.

Fixed platelets, bearing covalently bound fibrinogen, participate passively in aggregation of fresh platelets when the aggregation process is release related (G. Agam and A. Livne, Thromb Haemostasis 51:145-149, 1984). Inhibition of the release by aspirin abolishes the capability of the fresh platelets activated by 10 microM ADP to interact with the fixed platelets. A supernatant fraction from fresh platelets activated by 10 microM ADP (releasate) reconstitutes the interaction. Purified thrombospondin (TSP) replaces the releasate. Moreover, anti-TSP antibodies abolish the reconstituting effect of the releasate. It is concluded that TSP plays a role in the molecular mechanism of platelet-platelet recognition during release-related aggregation.

Adenosine Diphosphate↗
PubMed1984

Li efflux in erythrocytes of pregnant women: comparison of rates and temperature dependence for detection of hypertension.

Two determinants of lithium efflux in erythrocytes were compared, in relation to pregnancy: (a) efflux rates at 37 degrees C; (b) efflux temperature dependence, expressed by the 'break' of Arrhenius plots. Eighteen women were studied both at term and after delivery. While efflux rates were changed markedly, from 0.87 +/- 0.07 to 0.56 +/- 0.05 mmol/ (IRBC h) at term and post-partum, respectively, the characteristic break temperature of each woman remained essentially constant during and after pregnancy. The property of temperature dependence is more suitable than efflux rates for differentiation of hypertension during pregnancy.

Adult↗
PubMed1983

Unique profile for erythrocyte membrane acetylcholinesterase in hereditary spherocytosis.

Acetylcholinesterase of human erythrocytes from healthy donors and from patients with hematological disorders was analysed in a search for differential membrane parameters. Two substrates were used to estimate the exposure of acetylcholinesterase active site in the membrane: phenylacetate, a hydrophobic substrate, to determine total enzyme activity, and acetylcholine, an ionic substrate, to measure the externally reactive enzyme. The sensitivity of acetylcholinesterase to added stearic acid was also analysed. Three categories of the disorders studied were discerned: (a) The erythrocyte acetylcholinesterase profile was indistinguishable from normal control in beta-thalassemia minor and groups of patients with autoimmune hemolytic anemia or congenital dyserythropoietic anemia type II. (b) A marked decline in acetylcholinesterase with both substrates and reduced sensitivity to stearic acid were exhibited by the erythrocytes of paroxysmal nocturnal hemoglobinuria, beta-thalassemia major and other autoimmune hemolytic anemia and congenital dyserythropoietic anemia type II patients. Normal erythrocytes, either aged or pretreated to 50 degrees C, also showed similar characteristics. (c) Hereditary spherocytosis was singly differentiated by an elevated acetylcholinesterase activity with acetylthiocholine and by a vastly diminished sensitivity to stearic acid, while activity with phenylacetate was equal to control. This distinct profile may reflect the unique organization of the erythrocyte membrane in hereditary spherocytosis.

Acetylcholinesterase↗
PubMed1983

Passive participation of fixed platelets in aggregation facilitated by covalently bound fibrinogen.

The role of fibrinogen in interplatelet recognition during aggregation was examined by combining two cell types: fresh platelets (in limiting density) activated by thrombin or A23187, and formaldehyde-fixed platelets, bearing cross-linked fibrinogen. The fixed platelets did not aggregate by themselves, nor with resting platelets, but were capable of interacting with activated platelets and of participating passively in aggregation. The participation, expressed by enhanced aggregation, was assayed by the conventional turbidometric traces and by cosedimentation of fixed 3H-platelets with aggregates of fresh platelets. Platelet suspensions, prepared without special means to avert spontaneous activation, retained plasma fibrinogen to the extent of 50 micrograms/ml of a suspension containing 10(8) platelets, and the derived fixed platelets participated in aggregation, independently of added fibrinogen. The capability of such fixed platelets to participate in aggregation was sensitive to proteolytic digestion and to massive acetylation. When platelet separation was aided by apyrase or aspirin, PGE1 and gel filtration, the residual plasma fibrinogen was limited to 0.4 micrograms/ml of 10(8) platelet suspension. The derived fixed platelets were incapable of participating in aggregation unless fibrinogen was added prior to fixation. The affixed fibrinogen could not be replaced by soluble fibrinogen or affixed albumin. It is concluded that fibrinogen, which binds to platelets upon activation or is linked to them covalently, is a recognition site for platelet-platelet interaction during aggregation.

Apyrase↗
PubMed1982

Impairment of platelet aggregation by Echis colorata venom mediated by L-amino acid oxidase or H2O2.

Echis colorata bites cause impairment of platelet aggregation and hemostatic disorders. The mechanism by which the snake venom inhibits platelet aggregation was studied. Upon fractionation, aggregation impairment activity and L-amino acid oxidase activity were similarly separated from the crude venom, unlike other venom enzymes. Preparations of L-amino acid oxidase from E. colorata and from Crotalus adamanteus replaced effectively the crude E. colorata venom in impairment of platelet aggregation. Furthermore, different treatments known to inhibit L-amino acid oxidase reduced in parallel the oxidase activity and the impairment potency of both the venom and the enzyme preparation. H2O2 mimicked characteristically the impairment effects of L-amino acid oxidase and the venom. Catalase completely abolished the impairment effects of the enzyme and the venom. It is concluded that hydrogen peroxide formed by the venom L-amino acid oxidase plays a role in affecting platelet aggregation and thus could contribute to the extended bleeding typical to persons bitten by E. colorata.

Amino Acid Oxidoreductases↗
PubMed1982

The erythrocyte membrane in essential hypertension. Modified temperature-dependence of Li+ efflux.

The rate of ouabain-resistant Li+-efflux was studied in erythrocytes of normal controls and of patients with essential hypertension. Despite variability in rate, erythrocytes from normotensive persons revealed a uniform pattern of temperature dependence of the efflux, with two slopes (K = 9.4 and 19.1 kcal/mol, respectively) and a transition at about 25 degrees C. Erythrocytes from the patients showed both a higher rate of Li+ efflux and significant changes in the temperature response, with essentially a single slope (Ka = 14 kcal/mol). The data indicate localized changes in the membrane organization of hypertensive erythrocytes, involving lipid-protein interaction.

Acetylcholinesterase↗