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Biomedical subjects

A R Giles

Publications and source records attributed to A R Giles.

71 records · Page 4Linked to original sources
PubMed1984

Factor V (Quebec): a bleeding diathesis associated with a qualitative platelet Factor V deficiency.

Studies were performed on a French-Canadian family afflicted with a bleeding disorder exhibiting an autosomal dominant inheritance pattern and a severe bleeding diathesis after trauma. Clinical laboratory coagulation tests were unimpressive; the only persistent abnormalities include mild thrombocytopenia and moderately reduced Factor V clotting activities. Some individuals had prolonged Stypven times when platelet-rich plasma was used, suggesting that their platelets could not support functional prothrombinase complex assembly. Detailed studies were performed by use of plasma and isolated, washed platelets from a sister and brother. Bioassay data indicate that both individuals had Factor V activities of approximately 40 and 36% of normal, respectively. A comparison of the Factor V radioimmunoassay and bioassay data on the brother's plasma indicated that the circulating amount of Factor V functional activity was low relative to Factor V antigen concentration (approximately 65-75%). In both individuals, the platelet Factor V functional activities were extremely low (2-4%) relative to antigen levels present as determined by radioimmunoassay. These discrepancies between Factor V activities and antigen concentration do not appear to be due to an unstable Factor V molecule or to the presence of a Factor V or Factor Va inhibitor or inactivator. Kinetics of prothrombin activation by use of purified clotting factors indicated that thrombin-activated platelets from both individuals supported prothrombinase complex assembly identical to controls in the presence of added purified Factor Va. Consequently, their bleeding diathesis appears to reflect their platelet, rather than their plasma, Factor V activity. These results suggest that platelet Factor V is an essential component in maintaining stable and prolonged hemostasis after trauma.

Adult↗
PubMed1984

Studies of Factors V and VIII:C in an animal model of disseminated intravascular coagulation.

An experimental animal model of disseminated intravascular coagulation (DIC) induced by the co-infusion of coagulant-active phospholipid and activated Factor X (Factor Xa) is described. The infusion of Factor Xa at a dose of 6.6 X 10(-12) mol/kg with phosphatidylcholine/phosphatidylserine (PCPS) lipid vesicles at a dose of 4.0 X 10(-8) mol/kg was associated with significant falls in the levels of fibrinogen and Factors V and VIII, and a bleeding diathesis developed. Assays of Factors V and VIII were performed by a one-stage prothrombin time and activated partial thrombin time system, respectively. In additional experiments, the effect of the same dose combination of Factor Xa/PCPS on Factor V kinetics was studied by preinfusing 125I-labeled Factor V. After Factor Xa/PCPS infusion, Factors VIII and V were reduced at 2 min by 90 and 50% of the preinfusion levels, respectively, and at 1 h by 80 and 75%, respectively. During the same period, there was little change in the total circulating radioactivity. Autoradiography indicated small but detectable levels of circulating proteolytic products of Factor V that comigrated with peptides obtained by the incubation of Factor V with Factor Xa and activated protein C. The majority of radioactivity remained associated with the intact single-chain precursor Factor V. These observations suggested maintenance of the precursor pool after the onset of DIC. This was confirmed by performing two-stage assays of Factors V and VIII, whereby each was completely converted to the active cofactor, i.e., Va and VIII:Ca, by preincubation of the test sample with thrombin before assaying in a one-stage system as before. The Factor V levels assayed by the two-stage procedure did not change appreciably over 1 h. The Factor VIII levels fell but corrected within 1 h at a time when the level measured by a one-stage assay remained depressed. These results indicate that in the dog, infusion of Factor Xa/PCPS induces changes characteristic of DIC, and this is associated with the appearance of Factor V peptides characteristic of the expression of Factor Xa and activated protein C-like activities. The differences noted between the one-stage and two-stage assays suggest that the one-stage assay is measuring the activated fraction of each cofactor and not the total level of the available precursor for each activated species. The results suggest a close correlation between the activated fraction of both cofactors and the hemostatic abnormality that occurs in DIC.

Animals↗
PubMed1984

Development of factor VIII:C antibodies in dogs with hemophilia A (factor VIII:C deficiency).

Classic hemophilia A (factor VIII:C deficiency) was diagnosed in a miniature Schnauzer dog and a breeding program established. Inbreeding and crossbreeding produced 16 hemophilic animals. All were initially treated with canine cryoprecipitate, as required, for sporadic hemorrhagic events. Five animals developed potent antibodies to canine factor VIII:C. All were the offspring of obligate carriers, resulting from the mating of a hemophilic purebred miniature Schnauzer male to a normal female Brittany spaniel. The mean age at first treatment and factor VIII exposure at the time of inhibitor development was 10.3 wk and 286.3 U, respectively. The remaining hemophilic animals have not developed antibodies, despite receiving a mean factor VIII dosage of 1.5 X 10(3) U. This group includes animals derived from a mating between the same purebred miniature Schnauzer hemophilic male and a purebred miniature Schnauzer carrier female. In each case, the antibodies recognize both canine and human but not porcine VIII:C. They are non-precipitating IgG immunoglobulins. Following inhibitor development, infusion of canine cryoprecipitate was hemostatically ineffective and factor VIII:C recovery at 30 min was negligible. Infusion of a concentrate of porcine factor VIII resulted in a correction of the hemostatic defect and optimal factor VIII:C recovery. All animals receiving porcine factor VIII:C subsequently developed antibodies to this protein. The chance occurrence of this complication should facilitate further studies directed at elucidating the pathogenesis and management of hemophilia complicated by the development of antibodies to factor VIII:C.

Animals↗
PubMed1983

Plasmapheresis therapy in rheumatoid arthritis. A controlled, double-blind, crossover trial.

Twenty-six patients with rheumatoid arthritis (average age, 57 years; average duration of disease, 11 years) who were unresponsive to antiinflammatory and slow-acting antirheumatic drug therapy were entered into a controlled, double-blind, crossover study to assess the efficacy of plasmapheresis therapy. All patients received 10 true and 10 sham aphereses as outpatients and continued to take their usual drugs. Twenty patients completed the study, and six were withdrawn--three because of poor venous access. Standard clinical and laboratory measures were assessed by personnel blinded to the therapy administered. Paired t-test analysis of seven clinical measures failed to show significant differences between the true and sham procedures (P = 0.36 to 0.96), although transient, mild improvement did occur during both cycles of apheresis, probably because of a placebo response. Significant reductions in the erythrocyte-sedimentation rate, rheumatoid factor titer, and levels of hemoglobin, IgM, and C3 occurred only with true therapy (P = 0.001, 0.01, 0.03, 0.045, and 0.005, respectively). We conclude that plasmapheresis does not have clinical benefit in chronic rheumatoid arthritis, in spite of impressive laboratory changes.

Adult↗
PubMed1982

Stroma free human platelet lysates potentiate the in vivo thrombogenicity of factor Xa by the provision of coagulant-active phospholipid.

A stroma free platelet lysate supernatant (PLS) was prepared by repeatedly freezing and thawing a human platelet suspension separated from platelet rich plasma by gel filtration. The material was potently thrombogenic in a stasis model in rabbits, but only when combined with a purified preparation of factor Xa. The dose of factor Xa used was not thrombogenic when given alone. Initial evaluation suggested the presence of a factor V activator when PLS shortened the clotting time of normal but not a V deficient plasma. Subsequent evaluation, in a system employing purified components of the prothrombinase complex and a marker of thrombin generation, dansyl-arginine-N-(3-ethyl-1,5-pentanediyl)amide (DAPA), failed to confirm the presence of a factor V activator. Further evaluation in this system demonstrated that the procoagulant activity related to the content of coagulant-active phospholipid. Thin layer chromatography confirmed the presence of phospholipids known to have coagulant activity. These components were isolated by a lipid extraction procedure and the phospholipid replacing activity in the in vitro assay with DAPA was quantitatively retained. The extracted material was non-thrombogenic in vivo but augmented the thrombogenicity of a purified preparation of Xa.

Animals↗
PubMed1982

A canine model of hemophilic (factor VIII:C deficiency) bleeding.

A model of bleeding due to clotting factor deficiency has been developed in dogs. Normal and hemophilic (factor VIII:C deficient) animals were used. Bleeding was induced in lightly anesthetized animals by severing the apex of the nail cuticle using a guillotine device. In normal animals, bleeding usually ceased spontaneously after 2-8 min. In contrast, in hemophilic animals, bleeding continued for up to 20 min and necessitated either cauterization or the application of topical thrombin to achieve hemostasis. Pretreatment of the hemophilic animals with canine cryoprecipitate corrected the cuticle bleeding time to within the range noted for normal animals. The method is simple and reproducible and has the advantage that a number of observations can be made sequentially on the same animal. Rebleeding of the cauterized cuticle of the hemophilic animals did not usually occur. This model has considerable potential for the preclinical testing of products considered to bypass or replace factor VIII:C in patients with acquired inhibitors of factor VIII:C and may be adapted to the study of other mechanisms involved in normal and abnormal hemostasis.

Animals↗
PubMed1982

The coagulant-active phospholipid content is a major determinant of in vivo thrombogenicity of prothrombin complex (Factor IX) concentrates in rabbits.

In vitro evaluation of prothrombin complex concentrates in a thrombin generation assay, using DAPA and purified components of the prothrombinase complex, demonstrated significant levels of coagulant-active "phospholipid replacing" activity. Quantification of this activity showed a significant correlation (r = 0.8747, p less than 0.01) with thrombogenicity measured in vivo in a stasis model in rabbits. Extracted lipid material retained full phospholipid replacing activity in the vitro assay. Thin-layer chromatographic characterization confirmed the presence of phospholipids with known coagulant activity in vitro. In vivo, the extracted material was nonthrombogenic but augmented the thrombogenicity of purified factor Xa. Substitution of a synthetic coagulant-active phospholipid (phosphatidylcholine-phosphatidylserine lipid vesicles) for the extracted phospholipid produced a similar augmentation of a factor-Xa-induced thrombogenicity in vivo. It is concluded that the coagulant-active phospholipid content of prothrombin complex concentrates is a major determinant of thrombogenicity but requires the presence of activated clotting factors for its expression in vivo.

Animals↗
PubMed1981

Drug-induced thrombocytopenia is associated with increased binding of IgG to platelets both in vivo and in vitro.

Thrombocytopenia is a common serious adverse effect of drug treatment. A variety of in vitro diagnostic techniques to confirm the diagnosis are available, but the majority lack sufficient sensitivity to detect all cases of drug-induced thrombocytopenia. We studied 19 patients with suspected drug-induced thrombocytopenia and demonstrated that platelet-associated IgG (PAIgG) was elevated in all at the time of thrombocytopenia, and PAIgG returned to normal levels as the thrombocytopenia resolved. In the majority of patients, the platelet count rapidly returned to normal after the drug was discontinued; however, in six patients, the thrombocytopenia persisted well beyond the period of time that the offending drug would be expected to be cleared from the blood. In 13 patients, serum obtained after recovery was used to identify the drug responsible for the thrombocytopenia in an in vitro assay. In all cases, the addition of the drug historically associated with the thrombocytopenic episode was associated with an increased binding of IgG to control platelets. For uncertain reasons, the concentration of drug required to increase the in vitro binding of IgG to test platelets was often more than the concentration usually achieved in vivo. Wider application of these techniques may provide better understanding of the clinical characteristics and mechanisms responsible for drug-induce thrombocytopenia.

Adult↗
PubMed1980

Comparison of two direct assays for platelet-associated IgG (PAIgG) in assessement of immune and nonimmune thrombocytopenia.

A number of methods have been developed to measure platelet-associated IgG (PAIgG). The antiglobulin consumption assay directly quantitates IgG on the platelet and is sensitive and specific. A fluorescent anti-IgG assay has recently been described and has the advantage of simplicity. We compared the results of these two PAIgG assays in immune and nonimmune thrombocytopenia and nonthrombocytopenic controls. The antiglobulin consumption assay was negative in 61 of 62 and the fluorescent negative in 54 of 62 assays in nonthrombocytopenic controls, and they were negative in 11 of 13 and 8 of 13 assays, respectively, in nonimmune thrombocytopenic patients. The antiglobulin consumption assay was positive in 54 of 58 and the fluorescent positive in 24 of 58 assays of patients with immune thrombocytopenia (ITP and SLE). The overall sensitivity and specificity of the antiglobulin consumption assay was 94% and 95% and of the fluorescent assay was 44% and 82%.

Blood Platelets↗