alpha-chymotrypsin and Blood-Coagulation-Disorders

Topics: Amyloidosis; Animals; Arteriosclerosis; Arthritis, Rheumatoid; Blood Coagulation Disorders; Blood Coagulation Factors; Chymotrypsin; Chymotrypsinogen; Diabetes Mellitus; Emphysema; Fibrinolysis; Gastrointestinal Diseases; Humans; Kallikreins; Kidney Diseases; Muscular Dystrophies; Oligopeptides; Peptide Hydrolases; Substrate Specificity; Swine

Fibrinogen Milano XII was detected in an asymptomatic Italian woman, whose routine coagulation test results revealed a prolonged thrombin time. Fibrinogen levels in functional assays were considerably lower than levels in immunologic assays. Polymerization of purified fibrinogen was strongly impaired in the presence of calcium or ethylenediaminetetraacetic acid (EDTA). Two heterozygous structural defects were detected by DNA analysis: Aalpha R16C and gamma G165R. As seen previously with other heterozygous Aalpha R16C variants, thrombin-catalyzed release of fibrinopeptide A was 50% of normal. Additionally, the release of fibrinopeptide B was delayed. Immunoblotting analysis with antibodies to human serum albumin indicated that albumin is bound to Aalpha 16 C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of plasmin digests of fibrinogen Milano XII in the presence of calcium or EDTA showed both normal and novel D1 and D3 fragments. Further digestion of abnormal D3 fragments by chymotrypsin resulted in degradation products of the same size as the fragments derived from normal fibrinogen. SDS-PAGE analysis under reducing conditions showed no difference between normal fibrinogen and fibrinogen Milano XII or between their plasmic fragments. Circular dichroism analysis revealed a shift in the mean residual ellipticity and a significant reduction of the alpha-helix content in the variant D3 fragment. It is concluded that the Aalpha-chain substitution is mainly responsible for the coagulation abnormalities, whereas the substitution in the gamma-chain induced a conformational change in the D3 fragment.

Topics: Aged; Blood Coagulation Disorders; Calcium; Chymotrypsin; Circular Dichroism; DNA; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Female; Fibrinogen; Fibrinogens, Abnormal; Fibrinolysin; Fibrinopeptide A; Fibrinopeptide B; Heterozygote; Humans; Immunoblotting; Kinetics; Mutation; Peptide Fragments; Protein Conformation; Sequence Analysis, DNA; Serum Albumin; Thrombin; Thrombin Time

To show whether direct proteolysis of coagulation factors may play a role in patients with so-called consumption coagulopathy, granulocytic neutral proteases in the plasma of patients with acute myelocytic leukemia and septicemia were assayed by one- and two-dimensional Laurell electrophoresis. Complexes between serum alpha1-antitrypsin and elastase-like granulocytic protease could be demonstrated in those patients with acute myelocytic leukemia and septicemia who also had moderate or severe coagulation defects. Despite the presence of a high antiprotease potential, addition of the elastase-like enzyme to normal plasma resulted in coagulation defects in vitro comparable to those seen in the patients. These results and the ability of the elastase-like protease to destroy isolated clotting factors suggested that in certain types of coagulation factor deficiencies direct proteolysis rather than consumption of clotting factors due to disseminated intravascular coagulation may be operational.

Topics: alpha 1-Antitrypsin; Blood Coagulation Disorders; Chymotrypsin; Electrophoresis, Agar Gel; Factor XIII; Fibrin Fibrinogen Degradation Products; Granulocytes; Humans; Immunoelectrophoresis, Two-Dimensional; Leukemia, Myeloid; Leukocytes; Pancreatic Elastase; Peptide Hydrolases; Sepsis

Anticoagulated whole blood from patients and control subjects was circulated through an annular perfusion chamber in which the fibrillar collagen of alpha chymotrypsin-digested subendothelium and intact subendothelium were exposed. The blood flow conditions corresponded to those in arteries (830 sec-1 wall shear rate). Platelet surface interaction was measured morphometrically. Decreased adhesion to fibrillar collagen associated with normal spreading and normal adhesion-induced formation of platelet thrombi was found with blood of patients with von Willebrand's disease and the Bernard Soulier Syndrome, indicating a defect in the initial attachment reaction of platelets with collagen. Platelets of patients with thrombasthenia did normally adhere to the collagen fibrils and also lost their subcellular organelles during this reaction, but they totally failed to adhere to each other. In storage pool disease platelet thrombus formation was consistently inhibited whereas adhesion and spreading was inhibited in some patients and normal in others. In contrast adhesion was always normal after ingestion of aspirin which consistently caused a marked inhibition of platelet thrombi. These findings correspond -- in essence -- to those previously described on intact subendothelium. However, the observed defects are more pronounced on the fibrillar collagen than on intact subendothelium.

Topics: Animals; Aspirin; Blood Coagulation Disorders; Chymotrypsin; Collagen; Humans; Platelet Adhesiveness; Platelet Aggregation; Rabbits; Rheology; von Willebrand Diseases

Topics: Acute Disease; Blood Coagulation Disorders; Chymotrypsin; Granulocytes; Humans; Leukemia; Leukocytes; Pancreatic Elastase; Peptide Hydrolases; Sepsis

Topics: Birth Weight; Blood Coagulation Disorders; Chymotrypsin; Female; Gestational Age; Humans; Infant, Newborn; Infant, Newborn, Diseases; Obstetric Labor Complications; Pregnancy; Trypsin Inhibitors; Umbilical Veins

Topics: Adolescent; Adult; Blood Coagulation Disorders; Chymotrypsin; Diagnosis, Differential; Female; Humans; Male; Methods; Peptide Hydrolases; Shock, Hemorrhagic; Trypsin Inhibitors

Topics: Adolescent; Adult; Blood Coagulation Disorders; Chymotrypsin; Clinical Enzyme Tests; Diagnosis, Differential; Female; Fibrinolysis; Humans; Male; Pregnancy; Shock, Hemorrhagic; Trypsin

Topics: Blood Coagulation Disorders; Chymotrypsin; Electrophoresis; Fibrin; Fibrinogen; Humans; Immunodiffusion; Immunoelectrophoresis; In Vitro Techniques; Papain; Peptide Hydrolases; Peptides; Plasminogen; Streptokinase; Trypsin

Topics: Aminocaproates; Aminocaproic Acid; Blood Coagulation Disorders; Chymotrypsin; Enzyme Inhibitors; Fibrinolysin; Fibrinolysis; Hemorrhage; Kallikreins; Trypsin