Search PubMed⌕ Search

Biomedical subjects

P Ganguly

Publications and source records attributed to P Ganguly.

At least 55 records · Page 3Linked to original sources
PubMed1980

Evidence for multiple mechanisms of interaction between wheat germ agglutinin and human platelets.

Wheat germ agglutinin induced aggregation and secretion of serotonin from human platelets in plasma. This aggregation of platelets was blocked by ethylenediaminetetraacetate, azide or prostaglandin E1. The secretion of serotonin was not affected by ethylenediaminetetraacetate but was inhibited by prostaglandin E1. Thus, wheat germ agglutinin acts on platelets in plasma as a true aggregating agent. Washed platelets showed increased light transmission when treated with the lectin which was not blocked by ethylenediaminetetraacetate or prostaglandin E1. The capacity to inhibit platelet clumping by the above agents was restored if plasma was added back to the cell suspension. Washed platelets did not release serotonin under the conditions of cell clumping. Thus, in contrast to platelets in plasma, washed platelets are agglutinated by the lectin. Platelets fixed in formaldehyde were not agglutinated by the lectin in the aggregometer but agglutination was observed in the microtiter plate. This agglutination may be mediated by interplatelet bridging. The results show that the same agent may act on platelets by different mechanisms depending on the state of the cell and its environment.

Azides↗
PubMed1980

Differential effect of thrombin on the growth of human fibroblasts.

The ability of thrombin to alter the growth of human skin fibroblasts was studied under a variety of experimental conditions. In agreement with previous reports, we obtained a moderate level of cell growth in confluent cultures using 0.5-8.0 U/ml of thrombin. In subconfluent cultures, the effect was strikingly different and was found to be dependent upon the time in culture when the enzyme was added. Cultures exposed to thrombin 24 h after subculturing showed growth stimulation several days later. In contrast, thrombin added at the time of cell plating produced a complete block of DNA synthesis and cell growth that lasted for at least 3 d. Cells exposed to thrombin under these conditions were morphologically altered and smaller. These thrombin-induced effects were reversible and could be completely prevented by pretreatment of the enzyme with hirudin before it was added to the culture medium. Growth inhibition and altered morphology were found to be the result of changes generated in the growth medium by thrombin and could be blocked by higher serum concentrations. The results of this study indicate that thrombin's influence on cell growth can be stimulatory or inhibitory and suggest that the state of the cell surface determines the response.

Blood↗
PubMed1980

Binding of thrombin to cultured human fibroblasts: evidence for receptor modulation.

Previous work from this laboratory has indicated that thrombin's influence on cell growth can be negative as well as positive. Addition of enzyme to actively growing or confluent cultures of human skin fibroblasts produced growth stimulation, whereas cultures receiving thrombin at the time of subculture displayed inhibited DNA synthesis and mitosis. The specific binding of [125I]thrombin to cells under stimulatory and inhibitory conditions has been studied. Fibroblasts receiving enzyme at subculture bound about two times more [125I]thrombin than those processed in the same way several hours later. The apparent dissociation constant for both groups was approximately 1.5 x 10(-8) M. In each case binding was saturable, although cells receiving enzyme at subculture showed a much higher rate of binding. Experiments were conducted in which enzyme was added to cells at various times after subculture. It was found that the ability of these fibroblasts to specifically bind [125I]thrombin decreased progressively over a 2-h period after subculture and then remained constant for at least 24 h. Evidence is also presented indicating that the binding of [125I]thrombin in both experimental groups was inversely dependent upon the culture density. The biological effects of elevated thrombin binding in cells receiving enzyme at subculture were examined. It was found that inhibited DNA synthesis and altered cellular morphology were directly to this parameter. This study suggests that fibroblasts may possess cryptic thrombin receptors that become exposed during subculture or after injury in vivo. These possibilities and the relationship of cell shape to the availability of thrombin receptors are discussed.

Cell Count↗
PubMed1979

Interaction of lectins with human platelets. Effects on platelet stimulation by thrombin and ristocetin.

Wheat germ agglutinin induced aggregation and secretion of fresh platelets. Aggregation, but not secretion of serotonin by platelets in plasma, by the lectin was inhibited by 5 mM EDTA. Further, the lectin-induced stimulation of fresh platelets was blocked by prostaglandin E1. Thus, this lectin stimulates platelets by a mechanism which closely mimics thrombin activation and is independent of intercellular crosslinking. Lentil lectin did not stimulate platelets. Each platelet contained about 6 . 10(-5) binding sites for the lectins with an apparent dissociation constant of 3.0 . 10(-7) M. Wheat germ agglutinin, which binds mainly to glycoprotein I (Mr 150 000), increased the subsequent binding of thrombin to fixed platelets while lentil lectin was without effect. It appears that thrombin and wheat germ agglutinin bind to independent but interacting sites. Wheat germ agglutinin, but neither thrombin nor lentil lectin, inhibited the agglutination of platelets by ristocetin. Further, rat platelets were not aggregated by either ristocetin or wheat germ agglutinin. It appears that the interaction sites of ristocetin and wheat germ agglutinin on platelets are overlapping.

Animals↗
PubMed1979

Thrombin receptors of human platelets: thrombin binding and antithrombin properties of glycoprotein I.

Washed human platelets were solubilized and the proteins were separated by preparative gel electrophoresis in the presence of sodium dodecyl sulphate. The gel was cut into slices and the effect of the eluted proteins on the clotting of fibrinogen by thrombin was evaluated. The isolate from only one gel slice strongly inhibited the clotting of fibrinogen. The prolongation of the clotting time was dependent on the concentration of the protein and reached a plateau around 5 microgram. Gel electrophoresis of this isolate showed a prominent glycoprotein with an apparent Mr=150 000. Gel filtration studies with [125I]thrombin showed that the protein isolate bound a significant amount of thrombin which could be displaced with unlabelled thrombin. Another preparation from the same gel or purified gamma-globulin did not bind thrombin or prolong the clotting time of fibrinogen. Glycoprotein I was isolated from human platelets by affinity chromatography on lectin-Sepharose columns. The isolated glycoprotein prolonged the clotting of fibrinogen and bound [125I]thrombin which could be displaced by unlabelled thrombin. It is proposed that the high affinity receptor of thrombin on human platelets is glycoprotein I. In addition, the antithrombin activity of intact platelets is due to binding of thrombin to this glycoprotein.

Antithrombins↗
PubMed1979

Interaction of thrombin with mammalian platelets.

In this study we have explored the interaction of thrombin with platelets from human and rat. Compared to human platelets, rat platelets suspended in plasma required a higher concentration of thrombin for aggregation. This difference in sensitivity to thrombin was maintained when platelets were washed and suspended in buffer. Platelets from both mammals bound thrombin and showed a similar number of binding sites. However, the apparent dissociation constant of thrombin binding for rat platelets was approximately 15-fold higher than that for human. Thus, the decreased aggregation response of rat platelets may be due to a reduced binding of thrombin to its receptor. It is known that the thrombin receptor is located on the platelet surface. Gel electrophoresis of platelets followed by specific staining as well as fluorography showed significant differences in the surface glycoproteins of human and rat platelets. Human platelets showed labeled components corresponding to 210,000 and 160,000 daltons, whereas rat platelets showed glycoproteins with molecular weights of 240,000 and 190,000. A 135,000-dalton component was present in platelets from both sources. These results suggest that either or both glycoproteins of 210,000 and 160,000 daltons may be involved in the interaction of thrombin with human platelets.

Animals↗
PubMed1978

Defective binding of thrombin to platelets in myeloid leukaemia.

Platelets from patients with myeloid leukaemia showed reduced aggregation with collagen or thrombin. These platelets also had a lower capacity to bind thrombin. This lower thrombin binding is due to a decrease in the total quantity of receptors available and not because of a change in the affinity. In the presence of the patients' plasma, the aggregation behaviour of normal platelets induced by thrombin as well as the clotting time of fibrinogen remained unchanged. The results suggest that the platelet dysfunction in myeloid leukaemia is partially due to a membrane defect involving the thrombin receptors which leads to an impaired induction of the initial stimulation.

Adolescent↗
PubMed1977

Cross-linking of actin and fibrin by fibrin-stabilizing factor.

Fibrin-stabilizing factor is known to cross-link fibrin monomers. This study was undertaken to explore whether a similar interaction might occur between actin and fibrin. Purified actin and fibrinogen were labeled with 131I and 125I, respectively. The interaction of actin and fibrin, under different conditions, was evaluated by gel electrophoresis. The data show that actin and fibrin are cross-linked in the presence of fibrin-stabilizing factor.

Actins↗
PubMed1977

Binding of thrombin to functionally defective platelets: a hypothesis on the nature of the thrombin receptor.

The initial step in the interaction of thrombin with human platelets is binding of thrombin to specific structures on the platelet surface. Data are presented which show that platelets from thrombasthenic patients bind thrombin similar to controls. In contrast, platelets from a patient with Bernard-Soulier syndrome had lower thrombin binding capacity. It is suggested that glycoprotein one (GP-I) on the platelet surface might be the receptor for thrombin.

Blood Platelet Disorders↗