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Biomedical subjects

S V Pizzo

Publications and source records attributed to S V Pizzo.

At least 271 records · Page 15Linked to original sources
PubMed1981

Clearance and binding of two electrophoretic "fast" forms of human alpha 2-macroglobulin.

These studies explore the role of conformational change and exposed carbohydrate residues in the clearance of alpha 2-macroglobulin-trypsin (alpha 2M-T) complexes in the mouse. Human alpha 2-macroglobulin (alpha 2M) was purified and demonstrated to be homogeneous in the electrophoretic "slow" form. Two conformationally altered derivatives, alpha 2M-T and alpha 2-macroglobulin-methylamine (alpha 2M-MeNH2), were prepared and demonstrated to exist in the electrophoretic "fast" form. Radiolabeled alpha 2M-T and alpha 2M-MeNH2 were cleared rapidly with a half-life of 2-4 min following injection into mice. Radiolabeled native alpha 2M, however, remained in the circulation with a half-life of several hours. Both alpha 2M-T and alpha 2M-MeNH2 bound specifically to mouse peritoneal macrophages at 4 degrees C and occupancy of receptor sites increased with increasing time and radioligand concentration. Excess amounts of unlabeled alpha 2M-T or alpha 2M-MeNH2 cross-completed with trace amounts of the other in both clearance studies and binding assays, indicating that both derivatives were removed by the same receptor pathway. The clearance and binding of alpha 2M-T and alpha 2M-MeNH2 were not inhibited by excess amounts of unlabeled asialoorosomucoid, fucosyl-bovine serum albumin, mannosyl-BSA, or N-acetylglucosaminyl-BSA. Our results indicate that the clearance pathway removing alpha 2M-T complexes from the circulation recognizes a fundamental conformational change in alpha 2M secondary to protease binding, which can also be induced by exposure to methylamine. Therefore, other chemical or physical alterations that occur in alpha 2M upon binding trypsin, apart from the conformational change also present in alpha 2M-MeNH2, do not seem necessary for the recognition of alpha 2M-T by cells in the clearance pathway. In addition, this pathway appears distinct from several systems already described mediating clearance of glycoproteins through recognition of terminal galactose, fucose, N-acetylglucosamine, or mannose on oligosaccharide side chains.

Animals↗
PubMed1981

Fibrinolytic response and oral contraceptive associated thromboembolism.

A case-control study of fibrinolytic activity was conducted comparing 12 women with a recent history of thromboembolism while taking oral contraceptives and 28 matched female controls without a history of thromboembolism. All subjects had stopped using oral contraceptives at least 12 months prior to study. A new assay, recently developed in this laboratory, was used to evaluate the mean fibrinolytic response to venous occlusion in both cases and controls. The fibrinolytic response of all subjects was stratified into quartiles as previously described. While the controls segregated as expected, all 12 cases occurred in subjects whose fibrinolytic response fell in the first two quartiles with 8 of the 12 subjects having first quartile responses. The mean fibrinolytic response for all controls was 12.3 units while cases showed a mean response of only 3.9 units. The above data supports and extends our recent suggestion that low fibrinolytic response may signal a natural predisposition to venous thromboembolism which could be triggered by use of synthetic estrogens. However, these data do not support the use of this assay as a screening test for oral contraceptive related risk to thromboembolism since at least 50% of the control population also segregate in the first two quarters.

Caseins↗
PubMed1981

Simple and sensitive assay employing stable reagents for quantification of plasminogen activator.

A simple and sensitive indirect assay for quantifying plasminogen activator using [3H]-casein as substrate is described. The assay has been used to measure plasminogen activators from various sources including bacteria, cultured cells, and human plasma. The assay is linear with respect to concentration of plasminogen activator and time of incubation and is independent of concentration of plasminogen. The assay can routinely be used to quantify as few as 8 X 10(-3) international units ml-1 of streptokinase. Results obtained with the [3H]-casein assay correlate well with those obtained by the fibrin plate method (R = 0.83 by Spearman's ranked co-efficient of correlation). The reagents are extremely stable and easily stored. The ease with which the assay can be performed gives the clinical laboratory the potential to test large numbers of patients upon short notice and within a short period of time.

Animals↗
PubMed1981

Endothelial cell surface expression and binding of factor VIII/von Willebrand factor.

Factor VIII/von Willebrand factor (FVIII/vWF), a glycoprotein molecular complex found in human plasma, has been demonstrated by cell membrane fluorescence to be present on the surface of cultured human umbilical cord vein endothelial cells. The endothelial nature of these cells was established by electron-microscopic studies that revealed the presence of Weibel-Palade bodies in virtually all cells cultured. A newly developed radioreceptor assay was used to detect FVIII/vWF in the medium taken from these endothelial cell cultures; FVIII/vWF concentration in the medium samples increased with time in culture. FVIII/vWF binding studies showed no significant FVIII/vWF-specific binding to endothelial cell surfaces and did not corroborate a previous report suggesting a FVIII/vWF-specific receptor on human umbilical cord vein endothelium. The presence of FVIII/vWF on endothelial cell membranes and the lack of receptor-mediated binding suggests that the FVIII/vWF either has been absorbed non-specifically to the cell surface or is an integral part of the endothelial cell membrane.

Blood Coagulation Factors↗
PubMed1981

A radioreceptor assay for quantitating plasma factor VIII/von Willebrand's protein.

A sensitive and precise radioreceptor assay for determining plasma levels of human factor VIII/von Willebrand's factor (FVIII/vWF) has been developed by taking advantage of the FVIII/vWF receptor sites on human platelets. Paraformaldehyde-fixed platelets, which were processed and then stored, retained FVIII/vWF binding activity and therefore could be used as a convenient source of receptors. The human plasma samples to be tested were initially filtered on 4% agarose columns to concentrate the FVIII/vWF protein in the void volume and to remove the factor(s) that interferes with the assay. The percent recovery of FVIII/vWF in the pooled eluent was measured by the recovery of added trace 125I-FVIII/vWF. The coefficients of intra- and interassay variation were 6% and 10%, respectively. The plasma FVIII/vWF concentrations determined by the assay for pooled normal plasma, hemophilia A plasma, and plasmas from two patients with von Willebrand's disease were 16.3 +/- 0.5, 52.6 +/- 1.5, 6.8 +/- 0.8, and 3.2 +/- 0.2 microgram/ml, respectively. The range of plasma FVIII/vWF concentrations varied between 8.3 microgram/ml and 24.9 microgram/ml for 10 normal adults. The plasma FVIII/vWF concentrations determined by the radioreceptor assay correlated well with levels measured by the ristocetin-induced platelet aggregation method, thus demonstrating the functional relevancy of the radioreceptor assay for plasma FVIII/vWF.

Blood Coagulation Factors↗
PubMed1980

Physical conditioning augments the fibrinolytic response to venous occlusion in healthy adults.

The effects of a 10-week physical-conditioning program on fibrinolytic activity at rest and after stimulation by venous occlusion were studied in 69 healthy adults 25 to 69 years old. Physical conditioning was documented by treadmill performance, and fibrinolysis was measured with a newly developed radioenzymatic assay. Whereas fibrinolysis declined at rest from 16.2 +/- 1.3 to 11.4 +/- 0.8 units (mean +/- S.E.M.) (P = 0.0017), the increment in fibrinolysis produced by venous occlusion was increased from 21.7 +/- 2.9 to 33.8 +/- 4.7 units (P = 0.0037). This augmentation was most marked in women, persons with low initial levels of stimulated fibrinolysis, and persons with low initial physical fitness. We conclude that physical conditioning can enhance the augmentation of fibrinolytic activity that occurs in response to venous occlusion. Enhanced fibrinolysis in response to thrombotic stimuli could be an important mechanism in the beneficial effect of habitual physical activity on the risk of cardiovascular disease.

Adult↗
PubMed1980

Blood coagulation and idiopathic thromboembolism among fertile women.

A case-control study of fibrinolytic activity and antithrombin-III (AT-III) was conducted comparing 16 women within the childbearing ages who had a recent history of idiopathic thromboembolism to 29 age- and race-matched female controls without a history of thromboembolism. All subjects had stopped using oral contraceptives at least 12 months prior to study. No difference in AT-III was evident between cases and controls. The mean fibrinolytic response to venous occlusion in cases was significantly lower; however, baseline fibrinolysis was essentially identical. These data demonstrated a bimodal distribution of fibrinolytic response among controls. By dichotomizing fibrinolytic response between the two modes in the controls, we estimated that the rate of low fibrinolytic response was 6 times as frequent in cases as controls. Thus, low fibrinolytic response may signal a natural predisposition to thromboembolism which could be triggered by use of synthetic estrogens.

Adult↗
PubMed1979

Platelet receptors for human Factor VIII/von Willebrand protein: functional correlation of receptor occupancy and ristocetin-induced platelet aggregation.

Previous studies of von Willebrand disease indicate that a deficiency of blood clotting Factor VIII/von Willebrand factor (FVIII/vWF) activity is responsible for the failure of platelets to participate fully in the initial stages of hemostasis. We have recently identified specific FVIII/vWF binding sites on platelets, suggesting that the interaction of these sites with FVIII/vWF may be functionally important in the development of platelet clumps. We have now studied how different ristocetin concentrations, various known platelet aggregation inhibitors, and the exposure of platelets to proteases affect the ability of platelets to bind FVIII/vWF and to form aggregates. Our results demonstrate a highly significant linear correlation between the degree of FVIII/vWF receptor binding and the extent of ristocetin-induced platelet aggregation. Because neither FVIII/vWF binding nor platelet aggregation occurs after platelets are exposed to low concentrations of proteases, the FVIII/vWF receptors must be in the platelet membrane. We conclude that the interaction between FVIII/vWF protein and its receptors on the platelet membrane is an important mechanism by which platelet aggregation occurs during primary phase hemostasis.

Binding, Competitive↗