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PubMed1993

The antiphospholipid-thrombosis syndromes. Fact, fiction, confusion, and controversy.

Anticardiolipin antibodies and the lupus anticoagulant are strongly associated with thrombosis and appear to be the most common of the acquired blood protein defects causing thrombosis. Although the precise mechanism(s) whereby antiphospholipid antibodies alter hemostasis to induce a hypercoagulable state remain unclear, several theories have been advanced. Because the aPTT is unreliable in patients with lupus anticoagulant and is not usually prolonged in patients with anticardiolipin antibodies, definitive tests, such as ELISA for IgG, IgA, and IgM anticardiolipin antibodies and the dRVVT (followed by cephalin correction for confirmation) for lupus anticoagulant, should be immediately ordered when suspecting antiphospholipid syndrome in persons with otherwise unexplained thrombotic or thromboembolic events or fetal wastage syndrome. The laboratory diagnosis of APL-T syndrome is summarized in Figure 1.

Antibodies, Anticardiolipin↗
PubMed1976

The effect of increased contact activation time on the activated partial thromboplastin time.

With the kaolin-cephalin activated partial thromboplastin time technic, the plasmas of persons who have Fletcher factor deficiency have shown considerable shortening of clotting times when contact activation has been lengthened from 3 (PTT-3) to 10 minutes (PTT-10). The authors demonstrate that in plasma of most normal individuals, and in coagulopathies of other sorts, only slight shortening usually occurs. Abnormal shortening occurs in plasmas of a few otherwise normal people, the "slow activators," and patients receiving coumarin drugs, who have greatly prolonged prothrombin times. Longer activation may produce greatly prolonged PTT's in plasmas containing heparin in relatively high concentrations. The authors discuss the significance of these findings.

Blood Coagulation Disorders↗
PubMed1980

Factor X assays using chromogenic substrate S-2222.

Factor X was assayed using chromogenic substrate S-2222 for four patients with severe factor X deficiency and for nine patients with homozygous or heterozygous factor X Friuli disorder. Factor X Friuli disorder is characterized by the presence of an abnormal factor X that is normally activated by Russell's viper venom, but is not activated by tissue thromboplastins. The levels of factor X found in factor X deficiency varied between 2 and 10% of normal and therefore were higher than those found in the same plasmas using "clotting" methods (1% or less than 1% of normal). The levels of factor X found in homozygous factor X Friuli patients varied between 4 and 11% of normal, and therefore were practically identical to those found by means of clotting methods that employed tissue thromboplastins (7-9% of normal). These values were definitely lower than those obtained using a Russell's viper venom and cephalin mixture as thromboplastin (82-92%). A similar pattern was observed for patients heterozygous for the abnormality. These findings indicate that "amidolytic" methods are not necessarily identical to clotting methods. Furthermore, they indicate that substrate S-2222 is not specific for factor X.

Blood Coagulation Tests↗
PubMed1994

Reversal of post-reperfusion coagulopathy by protamine sulphate in orthotopic liver transplantation.

We report details of two liver transplant procedures in which post-reperfusion coagulopathy was reversed by administration of protamine sulphate. Both procedures were uncomplicated until about 30 min after reperfusion of the graft, when the cut surfaces began to ooze blood. Failure of coagulation was confirmed by thrombelastography and in both cases routine coagulation tests revealed a prolonged kaolin cephalin clotting time (KCT). A heparin-like effect was suspected. Protamine sulphate 50 mg was given i.v., resulting in cessation of bleeding and normalization of the thrombelastogram and KCT.

Adult↗
PubMed1990

Prevalence and biology of endosymbionts of fleas (Siphonaptera: Pulicidae) from dogs and cats in Alachua County, Florida.

A study was conducted to determine the prevalence and biology of endosymbionts in local populations of fleas collected from dogs and cats in Alachua Co., Florida. Four hundred three Ctenocephalides felis (Bouché), 194 Pulex simulans Baker, and 44 Echidnophaga gallinacea (Westwood) were examined. Fleas were collected from 52 dogs and 51 cats. From 1 to 20 fleas were dissected from each host. A variety of microorganisms and metazoa was observed, including a baculovirus, gram-negative bacteria, rickettsia-like organisms, amoebae, trypanosomatid flagellates, cephaline gregarines, and microsporidia. Microfilariae of the dog heartworm, Dirofilaria immitis, entomophilic nematodes, and metacestodes of the tapeworm Dipylidium caninum were also observed.

Animals↗
PubMed1981

Essentiality of dietary phosphatidylcholine for the survival of juvenile lobsters.

The inclusion of phosphatidylcholine (PC) in a purified diet is essential for the survival of juvenile lobsters. Attempts to substitute this substance with its alkaline hydrolysis products, fatty acids, an emulsifier or other phospholipids (cephalin, phosphatidylinositol) were not successful. The effectiveness of the PC ingredient in reducing mortality related to a molt death syndrome is dependent upon the source of the compound. Those PC molecules comprised of polyunsaturated fatty acids, were most effective. The lobster's nutritional requirement for PC is probably the result of a low rate of biosynthesis and the physiological role of PC is more probably associated with hemolymph lipoproteins and the transport of lipids, possible cholesterol.

Animal Nutritional Physiological Phenomena↗
PubMed1975

Hepatocarcinogenesis by diethylnitrosamine in rats fed high dietary levels of lipotropes.

The effects of high dietary levels of lipotropes on the carcinogenic activity of diethylnitrosamine (DENA) in rats were studied. All animals given DENA, with or without a dietary supplement, developed hepatocellular carcinomas. The mean survival times of all groups of rats dying with hepatocellular carcinomas after DENA treatment were determined. Choline, betaine, and folic acid consistently exerted no significant effect on those mean survival times. In rats receiving approximately equal to 2 mg DENA/day, methionine administration led to a slight but significant increase in the mean survival time of the carcinogen-treated rats, whereas vitamin B12 significantly lowered the survival time; neither substance altered the mean survival times of those treated with only 1.0 mg DENA/day. On the other hand, ethanolamine decreased the mean survival times of rats given 1.0 mg DENA daily, but had no effect on animals receiving 2 ml/day of carcinogen. Dimethylthetin (sulfur analogue of betaine), methotrexate, lecithin, and cephalin exerted no effect on the carcinogenic activity of DENA. The administration of dimethylthetin and betaine along with DENA led to markedly increased liver weights in animals dying of hepatocellular carcinomas, when compared to liver weights of animals treated with DENA alone.

Animals↗
PubMed1991

Coagulation studies and fistula blood flow during erythropoietin therapy in haemodialysis patients.

An increased incidence of fistula thrombosis has been reported in haemodialysis patients treated with recombinant human erythropoietin (rHuEpo). The present study sought to investigate this problem by measuring fistula blood flow, blood viscosity, and a variety of tests of coagulation and haemostasis in a group of ten haemodialysis patients treated with rHuEpo. Fistula blood flow did not alter during the first 12 months of rHuEpo despite a significant increase in whole-blood viscosity. Bleeding time improved in all ten patients after 4 months of therapy, and this improvement was maintained at 12 months. There were no significant changes in one-stage prothrombin time, kaolin cephalin clotting time, whole-blood clotting time, prothrombin consumption index, plasma fibrinogen factor VII, factor VIII, antithrombin III, or platelet aggregability to ADP over the first 4 months of rHuEpo. In contrast, protein C decreased from 84.3 to 66.4% (P less than 0.01) and protein S from 124.1 to 68.3% (P less than 0.001) over the first 4 months. By 8 and 12 months, the concentrations of these substances had returned to pretreatment values. The levels of protein C and S attained at 4 months are known to predispose to thrombosis, and it is possible that this effect may contribute to the increased incidence of fistula thrombosis observed in haemodialysis patients treated with rHuEpo.

Adult↗
PubMed1989

Isolation and characterization of an anticoagulant protein from human placenta.

An anticoagulant protein was purified from the EDTA extract of human placental tissue. The purified protein had a molecular weight of 73,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis under both reducing and non-reducing conditions. Because this protein had the ability to bind phospholipids such as phosphatidylserine, phosphatidylinositol, and cardiolipin in the presence of Ca2+, this protein was designated as calphobindin II (CPB-II). CPB-II prolonged the clotting time of normal plasma when coagulation was induced by tissue factor, cephalin and ellagic acid or recalcification, but did not affect thrombin-initiated fibrin formation. CPB-II also inhibited the activation of prothrombin by the complete prothrombinase complex or factor Xa-phospholipid-Ca2+ but not that by phospholipid-free factor Xa. In addition, CPB-II had an inhibitory activity against phospholipase A2.

Amino Acids↗
PubMed1979

Purification of Streptomyces chromofuscus phospholipase D by hydrophobic affinity chromatography on palmitoyl cellulose.

Phospholipase D [phosphatidylcholine cholinehydrolase, EC 3.1.4.4] excreted from Streptomyces chromofuscus was purified from the culture supernatant by precipitation with acetone and column chromatographies on palmitoylated gauze (Pal-G), DEAE-cellulose, and Sephadex G-150 with an overall recovery of 46% and 1000-fold increase in specific activity. The purified enzyme preparation showed a single band on sodium dodecyl sulfate (SDS) polyacrylamide disc gel electrophoresis. The enzyme had a molecular weight of about 50,000 by gel filtration on Sephadex G-150 or about 57,000 by SDS-polyacrylamide disc gel electrophoresis and an isoelectric point (pI) of pH 5.1 on isoelectric focusing. The enzyme hydrolyses lecithin, lysolecithin, sphingomyelin, and cephalin; the relative reaction velocities and Km's for choline-phospholipids were 87% and 1.43 mM for lecithin, 100% and 1.67 mM for lysolecithin, and 22% and 0.56 mM for sphingomyelin. The enzymatic reaction was optimal at pH 8, and its velocity was appreciably increased by either detergent (Triton X-100, deoxycholate), Ca2+ or both detergent and Ca2+. Diethyl ether stimulated the enzymatic activity by 30%; SDS and EDTA inhibited the activity. Bovine serum albumin, Triton X-100, and lipids (lecithin, lysolecithin, phosphatidic acid, lysophosphatidic acid, palmitic acid, and oleic acid) inhibited adsorption of the purified enzyme onto palmitoyl cellulose (Pal-C) and affected both the enzyme activity and stability: albumin and Triton X-100 increased the activity and enhanced the heat-stability; lysophospholipids decreased the activity but other lipids increased the activity; all the lipids lowered the heat-stability. The enzyme adsorbed on Pal-C was active, although its activity was about one-ninth of that of free enzyme, and was protected from heat-inactivation. Thus this enzyme appears to possess a hydrophobic site distinct from its catalytic site and to be adsorbed onto Pal-C through the hydrophobic site. Albumin, Triton X-100, and lipids seem to bind to the hydrophobic site and to have an appreciable effect on the enzyme activity and stability.

Cations, Divalent↗
PubMed1991

Haemostatic changes in long-distance runners and their relevance to the prevention of ischaemic heart disease.

Haemostatic changes may explain the paradoxical observations that regular exercise helps to prevent ischaemic heart disease but the risk of myocardial infarction and sudden death is actually increased during exercise. This study measured relevant haemostatic variables in 100 athletes before and after races of 10-26.2 miles duration and compared resting levels in athletes with 25 non-exercising controls. Prothrombin time, kaolin cephalin clotting time, fibrinogen, factor VII, factor VIII clotting (one and two stage), von Willebrand factor antigen, euglobulin clot lysis time, fibrin degradation products, full blood count, mean platelet volume, and platelet aggregation to collagen, adrenalin and adenosine diphosphate were measured. The immediate post-race results showed the familiar rise in platelet count and factor VIII clotting but there was no evidence of consumption or thrombin modification of factor VIII clotting. Platelet aggregation to adrenalin was reduced after the race and fibrinolysis was increased (P less than 0.05). The athletes at rest showed no significant differences from controls in their coagulation factor levels but showed increased fibrinolytic activity and reduced platelet aggregation to adrenalin (P less than 0.05). These results suggest a hypocoagulable rather than a hypercoagulable state during running and are consistent with the epidemiological evidence that such exercise is beneficial in the prevention of ischaemic heart disease.

Adult↗
PubMed1991

Potential thrombogenicity of heat-treated prothrombin complex concentrates in Haemophilia B.

We have studied the potential thrombogenicity of two different heat-treated prothrombin complex concentrates (PCC) in patients with Haemophilia B. Seven patients were studied on nine separate occasions. Four of the patients had chronic hepatitis C (HCV) associated liver disease and three were HIV-antibody positive. The PCCs were Profilnine (Alpha Therapeutics, Thetford, UK) and 9A (Bio-Products Laboratory, Elstree, UK) and the dose administered ranged from 35 to 60 U/kg. Blood samples were taken on ten separate occasions; twice before the infusion and at 15, 40, 60, 75 and 120 min and 4, 8 and 24 h after the infusion of PCC. Investigations included prothrombin time, kaolin cephalin clotting time, thrombin time, fibrin(ogen) degradation products, factor VIII, factor IX, antithrombin III and fibrinopeptide A (FPA). Fibrinopeptide A rises were seen following two of six infusions of 9A and one of three infusions of Profilnine. On all three occasions the rise in FPA was transient, returning to baseline levels within 120 min. Plasma beta-thromboglobulin (BTG) was assayed in three patients and in one patient, the rise in FPA was followed by an increase in BTG. No other changes were observed and there were no clinical features of disseminated intravascular coagulation. Our results indicate that even with normal clinical doses of PCC, intravascular thrombin generation can occur in patients with Haemophilia B. However, this effect is inconsistent both with respect to PCC batch and patient, but may occur in the absence of HIV infection and HCV liver disease.

Blood Coagulation↗
PubMed1996

Experimental cerebral venous thrombosis: evaluation using magnetic resonance imaging.

Diffusion-weighted (DWI), dynamic contrast-enhanced (perfusion imaging), and conventional spin-echo magnetic resonance imaging (MRI) were applied to characterize the pathophysiology of cerebral venous thrombosis (CVT) in the rat. We induced CVT by rostral and caudal ligation of the superior sagittal sinus (SSS) and injection of a thrombogenic cephalin suspension. The resulting pathology was monitored in an acute and long-term study group. Evans blue and hematoxylin-eosin staining was performed for comparison with MRI data. A subgroup of animals was treated with i.v. tissue plasminogen activator (t-PA). Successful thrombosis of the SSS was confirmed by macropathology or histopathology in all rats. Parenchymal lesions as shown by MRI, however, were present only in animals with additional involvement of cortical cerebral veins (11 of 18 rats). The early pathology was clearly detected with the DWI. The apparent diffusion coefficient declined to 56 +/- 7% of control value at 0.5 h and slowly increased to 84 +/- 8% by 48 h. Perfusion imaging showed parasagittal perfusion deficits. Treatment with t-PA partially resolved the hyperintensity on DWI. Evidence of blood-brain-barrier disruption was observed 2 to 3 h after induction of CVT. In conclusion, experimental CVT is characterized by early cytotoxic edema closely followed by vasogenic edema. The t-PA treatment partially reversed the DWI signal changes consistent with regional tissue recovery, as shown by histopathology. These results encourage the use of cytoprotective drugs in addition to anticoagulant or thrombolytic therapy.

Animals↗
PubMedDate not supplied

Prevention by ticlopidine of platelet consumption during extracorporeal circulation for heart surgery and lack of effect on operative and postoperative bleeding.

We studied the effects of ticlopidine, a platelet antiaggregant drug, on platelet consumption during and after extracorporeal circulation (ECC) and on the operative and postoperative blood loss in a double-blind, placebo-controlled trial on 20 patients who underwent open-heart surgery for implantation of a valvular prosthesis. We monitored the changes in platelet count during and after ECC, the need for platelet transfusions to compensate for excessive consumption, and the operative and postoperative blood loss. We also followed the cephalin-kaolin time, the prothrombin time, the fibrinogen level, the fibrin/fibrinogen degradation products, and the euglobulin lysis time. Ticlopidine effectively reduced operative and post-ECC thrombopenia and prolonged the bleeding time. Apart from these changes, no differences in coagulation tests were observed between the ticlopidine-treated group and the placebo group. No side effects were observed. There was no significant difference between the groups concerning operative and postoperative blood loss, indicating that ticlopidine does not induce a hemorrhagic diathesis. Ticlopidine therapy may be of value during the first postoperative days because it prevents clotting on valvular prosthesis and complications such as systemic embolization. However, more trials and controlled studies are needed before such therapy can be recommended for routine use.

Adult↗
PubMed2001

Hepatic dysfunction after cardiac surgery in children.

OBJECTIVE: The objective of this study was to analyze the incidence and significance of hepatic dysfunction after cardiac surgery in children. DESIGN: Prospective, observational study. SETTING: Pediatric intensive care unit of a university hospital. PATIENTS: The study consisted of 232 children ranging in age from newborn to 17 years with no history of liver disease. MEASUREMENTS AND MAIN RESULTS: Aspartate aminotransferase (AST), alanine aminotransferase (ALT), gammaglutamyltranspeptidase (GGT), alkaline phosphatase, total and conjugated bilirubin, blood glucose, urea, creatinine, and coagulation studies were determined at admission, at 24 and 48 hrs, and at 7 days. Hepatic dysfunction was taken as an ALT of > 100 IU/L or a moderate or high hepatic score. The statistical study included bivariate analysis and multivariate logistic regression to study the risk factors for hepatic dysfunction. Twenty-one patients (9%) showed an ALT > 100 IU/L, and 29.3% had a moderate or high hepatic score. A relationship was found between hepatic dysfunction and the type of cardiopathy (D-transposition of the great arteries and coarctation of the aorta), shock, the administration of dopamine or epinephrine, renal insufficiency, the presence of pulmonary changes (pulmonary edema, atelectasis, pulmonary hypertension, hypoxemia), hematologic disturbances (prothrombin time, kaolin-cephalin time, fibrinogen, and platelets), and the need for a greater number of transfusions of packed cells, plasma, and platelets. Compared with 7.6% of the rest of the patients (p <.001), 38% of patients with an ALT > 100 IU/L died. The hepatic score of those patients who died was 4.2 (2.3)-higher than that of the survivors at 1.5 (1.8), (p <.001). Shock and renal insufficiency were the factors most significantly related to the development of hepatic dysfunction. CONCLUSIONS: Hepatic dysfunction is an uncommon complication in children after cardiac surgery. This complication is related mainly to hemodynamic disturbances and renal insufficiency and is an indicator of poor prognosis.

Journal Article↗
PubMed1983

Epidural analgesia and anticoagulant therapy. Experience with one thousand cases of continuous epidurals.

One thousand lumbar epidural blocks in 950 patients undergoing vascular surgery are reported. All patients were receiving oral anticoagulants pre-operatively. Mean thrombotest (TT) was 19.3% (normal range 70-130%). During surgery intravascular heparin was administered. At the end of surgery, the kaolin cephalin clotting time (KCCT) was 68 (+/- 0.8) seconds (normal range 35-60 seconds), and partial thromboplastin time (PTT) was 536 (77.9%, normal control of 100%). Despite the anticoagulant therapy, no side effects were observed in any patient which could be related to haemorrhage or haematoma formation in the epidural space. It is concluded that, provided adequate precautions are taken, epidural analgesia can be safely used in patients receiving anticoagulant therapy.

Aged↗
PubMed1987

ORG 10172: a low molecular weight heparinoid anticoagulant with a long half-life in man.

ORG 10172 is a heparinoid with mean molecular weight 6500 daltons. Intravenous bolus injections of ORG 10172 were compared with placebo and heparin injections in 91 separate studies in 83 healthy male subjects. 6400 units ORG 10172 produced a mean maximum change of 14.7 s in kaolin cephalin time (c.f. greater than 120 s for 5000 units heparin). Changes in prothrombin time were minimal (1.6 s for 6400 units ORG 10172 and 4.5 s after 5000 units heparin). A dose-related increase in bleeding time occurred after ORG 10172 and at high doses (greater than 3200 units) some secondary bleeding, which was never serious, occurred at between 1 and 4 h after incision. A dose-dependent reduction in ex vivo platelet adhesiveness was found at 10 min after ORG 10172 injection. ORG 10172 promoted a much smaller release of lipoprotein lipase as compared with heparin. The effect of ORG 10172 on plasma factor Xa activity (one measure of its action) was described by a biexponential decay with a mean distribution half-life of 2.34 (s.e. mean 0.16) h and mean disposition half-life of 17.6 (s.e. mean 1.1) h. It thus has a much longer duration of effect than heparin. There was a linear relationship of plasma anti-Xa response to increasing dose although there was some variability only partly explained by differences in body weight or surface area. ORG 10172 administration by bolus intravenous injection was well tolerated and there was no evidence of adverse effects on clinical chemistry or haematology tests.

Adolescent↗
PubMed1985

The importance of thrombin inhibition for the expression of the anticoagulant activities of heparin, dermatan sulphate, low molecular weight heparin and pentosan polysulphate.

The effects of standard heparin, three low molecular weight derivatives of heparin, dermatan sulphate and pentosan polysulphate on the intrinsic coagulation pathway were compared in order to evaluate the contributions of the anti-factor Xa and anti-thrombin activities to their anticoagulant activities. The anticoagulant potency was measured by the ability of each sulphated polysaccharide to inhibit the generation of thrombin activity in plasma. Similarly, the ability of the six sulphated polysaccharides to enhance the rates of inactivation either factor Xa or thrombin in defibrinated plasma containing calcium chloride and cephalin were also determined. Standard heparin was the only sulphated polysaccharide that could equally inhibit thrombin generation and enhance the inactivation of factor Xa and thrombin by plasma. Dermatan sulphate and pentosan polysulphate were more effective as inhibitors of thrombin generation than potentiators of factor Xa inactivation. The two smallest derivatives of heparin, which had high anti-factor Xa (but low antithrombin) activity, were the poorest inhibitors of thrombin generation. Our results therefore suggest that only sulphated polysaccharides that enhance the inactivation of thrombin by plasma and/or inhibit the generation of thrombin activity in plasma are good anticoagulants. These two activities of sulphated polysaccharides appear to be good predictors of the relative antithrombotic potency in vivo.

Anticoagulants↗