bluuebunny/crossref_metadata_2025 huggingface dataset:
CANCER CHEMOTHERAPY AND THROMBOSIS
Authors: Levine M , Arnold A , Kelleher L , Lord S , Hryniuk W , Hrish J , Gent M
Published: August 2018

Malignant disease is recognized as a risk factor for venous thromboembolism. A number of recent reports have suggested that cancer chemotherapy may contribute to this risk, but it was not possible to separate the role of chemotherapy from the effects of the malignant disease. We are conducting a randomized trial to determine the optimal duration of adjuvant chemotherapy in women with Stage II breast carcinoma. These ambulatory patients, with negligible tumour burden, receive either 12 weeks of chemo-hormonal therapy (cyclophosphamide, methotrexate, 5 fluorouracil, vincristine, prednisone, adriamycin and tamoxifen) or 36 weeks of chemotherapy (cyclophosphamide, methotrexate, 5 fluorouracil, vincristine and prednisone). This study has provided us with an opportunity to evaluate the thrombogenic effects of chemotherapy since patients in the 12 week group, while off chemotherapy, can be compared directly to the patients in the other group who are still on chemotherapy. This allows the confounding influence of the malignant process to be circumvented. All patients undergo screening tests for thrombosis (impedance plethysmography and Doppler ultrasound) and routine clinical assessments. Suspected venous thrombosis is confirmed by venography and suspected pulmonary embolism by either pulmonary angiography or high probability ventilation perfusion scanning. There have been 11 episodes of venous thromboembolism to date among 191 patients of whom 164 have completed the first 36 weeks of study. There were 3 episodes in each group during the first 12 weeks. During the subsequent 24 weeks there have been no events in the group whose treatment was stopped and 5 events in the group still on treatment (p 0.03). These findings demonstrate that chemotherapy per se is an important risk factor for venous thromboembolism in patients with malignant disease.
SUBCUTANEOUS ENOXAPARINE (LOVENOXR) VERSUS PLACEBO FOR PREVENTING DEEP VEIN THROMBOSIS (DVT) AFTER TRANSURETHRAL PROSTATECTOMY (TUP)
Authors: LE GAGNEUX F , STEG A , LE GUILLOU M
Published: August 2018

The aim of this study was mainly to evaluate the risk of bleeding, and the efficiency of Enoxaparine, a low-molecular-weight-heparin, in preventing DVT in patients undergoing TUP.

89 patients (mean age : 67.5 years + 1.3), undergoing TUP, were included in a randomized, double blind study. Patients with a major risk of thromboembolism were excluded. 44 patients received one daily subcutaneous (SC) injection of 60 mg of Enoxaparine ; 45 patients received placebo. All the patients received the first injection 12 hours before operation.

Red cell transfusions requirements were not significantly different between the two groups : 18 % of patients in the Enoxaparine group and 13 % of patients in the placebo group received red cell transfusions (p = 0.57). The amount of red cell units required was 3.3 units ± 0.9 in the Enoxaparine group and 2.5 U ± 0.8 in the placebo group (p = 0.51). The urethral catheters were removed on the 4th post operative day in each group.

There was no significant difference in daily hemoglobin levels between the two groups.

No DVT occurred : 125I fibrinogen scanning was negative in all patients but two : in these two patients (one in each group), DVT was not confirmed by a radiographic phlebography. No pulmonary embolism occurred.

Enoxaparine, begun 12 hours before operation, however injected at high dose (60 mg/24 hrs), is safe in patients undergoing TUP. No significant bleeding complication occurred in the Enoxaparine group comparing with the placebo group. Red cell transfusions requirements were the same in both groups. There was no DVT in our patients.

Enoxaparine (LOVENOXR) - PHARMUKA S.F.
PREVENTION OF DEEP VEIN THROMBOSIS (DVT) AFTER TOTAL HIP REPLACEMENT (THR) by ENOXAPARINE (LOVENOXR) : ONE DAILY INJECTION OF 40 MG VERSUS TWO DAILY INJECTIONS OF 20 MG
Authors: PLANES A , VOCHELLZ N , MANSAT C
Published: August 2018

Venous thromboembolism is a common complication in patients undergoing THR. In a previous open study, Enoxaparine, a low-molecular-weight-heparin, in a dose of 40 mg/24 hrs by SC injection, had been shown to be efficient in preventing DVT in these patients. This treatment was not associated with an increased risk of bleeding. The present trial compares the efficiency and the risks of bleeding of two regimens : treatment A (2 daily subcutaneous (SC) injections of 20 mg of Enoxaparine) and treatment B (1 daily SC injection of 40 mg of Enoxaparine).

118 patients, over 40 years, with a non traumatic hip disease, requiring THR, were included in a randomized, double blind trial. 59 patients received the treatment A. 59 patients received the treatment B. In both groups administration of 40 mg of Enoxaparine was begun 12 hours before operation. Patients were treated for 12-15 days, until bilateral ascending phlebography (BAP) had been completed.

Lower limbs BAP were performed in 114 patients. The frequency of DVT is low and is not significantly different between the two regimens : a DVT was detected in 1 of 57 patients who received the treatment A and in 6 of 57 patients who received the treatment B (p = 0.11) . No pumonary embolism occurred in the 114 patients.

There was no serious bleeding complication, and the two groups are not significantly different on this point. 3 patients in each group had an important hematoma of the thigh. None required a surgical treatment. Red cell transfusion requirements were 3.88 U ± 1.71 in the group A and 3.5 3 U ± 1.06 in the group B (p = 0.20). There was no significant difference in daily hemoglobin levels between the two groups.

One daily injection of 40 mg of Enoxaparine was as effective as two daily SC injections of 20 mg of Enoxaparine in preventing DVT, in patients undergoins THR. The frequencies of bleeding complications were the same in each group.

Enoxaparine (LOVENOXR) - PHARMUKA S.F.
HAEMOSTASIS CHANGES FOLLOWING TOTAL HIP REPLACEMENT IN A RANDOMIZED TRIAL WITH LMW HEPARIN (KABI 2165) AND ADJUSTED DOSE STANDARD HEPARIN
Authors: Berruyer M , Dechavanne M
Published: August 2018

70 patients were randomized to 3 groups. Group I : patients received 5,000 IU calciparineR subcutaneously given 2h before operation, twice a day for 3 days and then, doses were adjusted ty heparin levels. Groups II and III : 2,500 IU Kabi 2165 were given instead of calciparineR, but in group III, a single daily dose of 5,000 IU Kabi 2165 was administered from the third day. Fibrino-peptide A (FPA), FPA generation, D-dimers were measured in plasma by ELISA methods. Anti Xa activity, Xa generation, heparin cofactor II (HC II), antithrombin III (AT III) were assayed in plasma using amidolytic methods. Serial measurements were done : pre-operatively, 2 and 8h after the first heparin injection and on the 7th post-operative day, before heparin administration. Pre-operatively, the mean level of D-dimers was higher (p<0.05) in 9 patients with bleeding complications after surgery (0.83 ± 0.57 pg/ml) compared to other patients (0.42 ± 0.33 pg/ml) 2h after heparin injection : 1) Xa activity was lower (p<0.001) in group I (<0.05 IU/ml) than in groups II and III (0.137 ± 0.1 IU/ml) ; 2) inhibition of Xa generation decreased (p<0.001) in group I (11.7 ± 8%) compared to that in groups II and III (25.8 ± 13%) ; 3) inhibition of FPA generation was lower (p<0.05) in group I (27.1 ± 32.2%) than in groups II and III (45.7 ± 29.2%) ; 4) diminution of D-dimers under heparin (0.37 ± 0.28 pg/ml) was only significant (p<0.05) in groups II and III. 8h after the first heparin injection, in all groups : 1) D-dimers increased (p<0.001) compared to pre-operative values ; 2) AT III and HC II diminished (p<0.001) ; 3) no anti Xa activity and no inhibition of Xa and FPA generation were found. On the 7th day : 1) 4 patiertts with deep vein thrombosis had higher FPA than other patients ; 2) AT III but not HC II fell in group I (p<0.05) ; 3) no anti Xa activity and no inhibition of Xa and FPA generation were observed in all groups. In conclusion : 1) although clinical results are similar in the 3 groups, Xa activity, Xa generation, FPA generation change differently according to the heparin used ; 2) the occurence of deep vein thrombosis is followed by an FPA increase and bleeding complications are correlated with a high level of pre-operative D-dimers.
A DOSE RANGING STUDY DESIGNED TO ASSESS THE SAFETY AND EFFICACY OF A LOW MOLECULAR WEIGHT HEPARIN FRAGMENT (FRAGMIN) AS PROPHYLAXIS AGAINST DEEP VEIN THROMBOSIS
Authors: Kakkar V V
Published: August 2018

A preliminary study has reported that a single daily injection of 2500 and 5000 U of Fragmin was equally effective in preventing deep vein thrombosis1 (DVT). All the patients who developed DVT (4.9%) had undergone surgery for malignant disease. However, small numbers of patients were investigated and therefore detailed subject analysis was not possible. In the present study, 1000 patients scheduled to undergo elective major abdominal, urological or gynaecological surgery were investigated. They received one of three regimens; 5000 (Group I), 2500 (Group II) or 1500 (Group III) U of Fragmin daily as a single injection. The 125I-fibrinogen uptake test was used to detect DVT. Operative and postoperative blood loss, transfusion requirements, fall in postoperative haemoglobin and frequency of haematoma formation were recorded to assess the safety of three regimens. An analysis of variance was carried out using a computerized statistical package. Isotopic DVT was detected in 60 patients (6.7%). 11.1% of patients with malignant disease and 5.8% with benign disease developed DVT (p 0.01). Data were also analyzed in respect of age and there was a highly significant increase in the frequency of thromboembolic disease in patients over sixty years (10.1%) compared to 2.6% in the under 60 group (p 0.001). Significant differences were observed in the frequency of bleeding complications in the three groups and these were related not only to underlying disease process but also the type of surgery being performed. The detailed results will be presented and a safe and effective regimen for different groups of patients will be defined.

1. Efficacy and Safety of Two Regimens of Low Molecular Weight Heparin Fragment (Fragmin) in Preventing Postoperative Venous Thrombolism. Kakkar W, Kakkar S, Sanderson RM, Peers CE. Haemostasis 16(Suppl.2) 1986, pp 19-24.
THE SAFETY AND EFFICACY OF A LOW MOLECULAR WEIGHT HEPARIN (FRAGMIN) IN THE PREVENTION OF DEEP VEIN THROMBOSIS IN MEDICAL PATIENTS: A RANDOMIZED DOUBLE-BLIND TRIAL
Authors: Poniewierski M , Barthels M , Poliwoda H
Published: August 2018

The safety and efficacy of 2500 anti-Factor Xa U of a low molecular weight heparin (Kabi 2165, Fragmin) subcutaneously once a day, and 5000 IU of standard unfractionated Heparin (KabiVitrum, Stockholm) subcutaneously twice daily as thromboprophylaxis was compared in 200 medical patients in a randomized double blind trial. According to the risk of DVT the patients were stratified before randomization in a high and low risk group. The high risk group consisted of 100 patients mainly with malignant diseases and/or previous history of thromboembolism, the low risk group of 100 patients with mainly myocardial infarction and/or coronary heart disease. The prophylaxis was given for seven to ten days. In 192 consecutive patients the clinical status and thermographic screening for DVT (leg temperature profiles, DeVeTherm) were daily evaluated. In two cases of suspected DVT and one case of suspected PE, the following phlebography or pulmonary scintigraphy were found to be negative. In the high risk group, one patient treated with Fragmin having a central venous catheter developed on day 10 symptoms of an arm vein thrombosis. There were no bleeding complications observed in either of the two treatment groups. Two patients with trombocytopenia (25.000 and 22.000/pl) due to chemotherapy and underlying malignant disease were successfully treated with Fragmin without developing any bleeding complications. In eight patients during Fragmin prophylaxis invasive diagnostic methods as heart catheterization, gastroscopy, bronchoscopy or spinal puncture were performed without noticing any bleeding events. 2500 anti-Factor Xa U of Fragmin gave plasma levels by anti-Factor Xa assay (S-2222, Kabi) of mean 0,1 U/ml when blood was sampled three to four hours after the subcutaneus application. There was no accumulation during the treatment periode observed.

This study suggests that 2500 anti-Factor Xa U of Fragmin once daily is as safe and effective as 5000 IU of standard heparin twice daily in these medical patients. Especially in patients who need prophylaxis for a long time eg. with malignant disease, the once daily injection is welcomed.
INTER-INDIVIDUAL PHARMACOKINETIC VARIATIONS AFTER INTRAVENOUS (IV) AND SUBCUTANEOUS (SC) INJECTION OF CY 216 IN HEALTHY SUBJECTS
Authors: Boneu B , Houin G , Rostin M , Montastructure J L , d’Azemar P , Bayrou B
Published: August 2018

We investigated the pharmacokinetic parameters and their inter individual variations of a low molecular weight heparin (LMWH) derivative (CY 216, Fraxiparine R, Choay). In a cross-over study, 100 anti Xa IC u/kg were injected in 12 healthy volunteers, either by IV or SC route, at one week interval. The pharmacological effects were followed on 12 serial citrated samples for 24h: - anti factor Xa (AXa) activity (chromogenic assay calibrated against CY 216); - APTT and thrombin clotting time prolongation. The main pharmacokinetic parameters (elimination half-life (T|); clearance (cl); distribution volume (Vd); bioavailability F (SC/ IV)) were calculated from the anti Xa activity curves using con-, ventional methods. The results (mean, range) indicated below confirm some classical properties of CY 216: poor anticoagulant effect (APTT-TT), even after IV injection, longer half-life than . standard Heparin (SH), distribution volume similar to plasma volume, excellent bioavailability of the drug.

We also emphasize important inter-individual variations between volunteers, as known for the pharmacological effects of SH in vitro and ex-vivo. From those results it could be assumed that, as for SH, close monitoring of treatment with LMWH would be suitable with higher dosages, after validation of the correlations between those biological tests and clinical results.

* anti Xa IC u : anti factor X activated Institut Choay unit.
ORG 10172 CAUSES INCREASED URINARY BLEEDING AFTER TRANSURETHRAL PROSTATECTOMY (TURP)
Authors: Gallus A , Murphy W , Nacey J , Magnani H
Published: August 2018

Standard anticoagulant prophylaxis is rarely used after TURP because it is believed to increase urinary bleeding. Org 10172 prevents experimental thrombosis and causes less bleeding than heparin in animals. We therefore evaluated its influence on urinary blood loss after TURP in a double blind, placebo controlled, study, which examined the effects of three Org 10172 regimens (750 antiXa u/day, 500 u bid, 750 u bid), given subcutaneously. Pilot studies have shown that 750 u bid prevents venous thrombosis (VT) after thoracotomy. At each dose level, 10 patients received Org 10172 and 6 placebo. Because patients had spinal anesthesia, they received their first injection of trial drug after anesthesia was established and immediately before surgery. Bleeding was estimated by measuring urine hemoglobin (Hb) and volume. The three dosage groups all showed trends towards increased postoperative urine bleeding in Org 10172 treated patients, but no apparent increase in blood loss with increasing dose, so they were pooled for analysis:

Hence, Org 10172 in doses which are likely to be needed for VT prevention causes a moderate increase in urine bleeding after TURP.
EFFECT OF PENTOSAN POLYSULPHATE IN MINIPIGS WITH EXPERIMENTAL PULMONARY EMBOLI
Authors: Klöoking H-P , Sedlarik K
Published: August 2018

In anaesthetized minipigs emboli were induced by reinjection of electrically activated autologous blood. The animals were then infused with either saline or pentosan polysulphate (SP 54) for three hours; 2 mg SP 54/kg body weight injected as bolus followed by intravenous infusion of 1 mg SP 54/kg body weight per hour. Before and during infusion of SP 54 or saline, we determined haemodynamics, degree of pulmonary arterial occlusion, blood gases and plasminogen activator levels. As a result of treatment with SP 54 a decrease in mean pulmonary arterial pressure, an increase in blood flow of pulmonary artery, and an increase in plasminogen activator activity was seen, compared with control animals who received saline infusion only. The efficacy of SP 54 treatment was demonstrated by a mean haemodynamlc and blood flow improvement of about 50 %, three hours after starting the experiment.
THE THROMBOTIC COMPLICATIONS OF TWO GROUPS OF PATIENTS WITH DIFFERENT INR THERAPEUTIC RANGES. THE NECESSITY OF INTENSE ORAL ANTICOAGULANT TREATMENT
Authors: Kontopoulou-Griva Ir , Spiliotopoulou J , Digenopoulou L , Georgopoulos J
Published: August 2018

One of the reasons why oral anticoagulants fell into disrepute is the absence of internationally acceptable standarised procedures for controlling the level of anticoagulation. This deplorable situation resulted in over and under coagulation and uncertainty in the therapeutic range. The International Normalised Ratio (INR) can safely be applied in patients on oral anticoagulants.

We present two Groups of patients under long term anti coagulation, mainly because of prosthetic heart valves that have recently been added to our outpatients clinic. These patients were till then attended by two cardiologists with different attitudes on the intensity of the anticoagulant treatment. The thromboplastin reagent used is that of ox origin and the results are expressed on INR.

The Group A with 32 patients had at the time that we started attending them an INR x = 1,80 ± 0,48 and a daily dose of acenocoumarol x = 1,65 ± 0,51.

The Group B with 49 patients had an INR x = 2,75 ± 0,51 and a daily dose of acenocoumarol x = 2,52 ± 1,53.

Seven patients of the Group A referred thrombotic complications, while three patients of the Group B referred transiant thrombotic complications.

The statistical analysis with the t-test of the INR between the two Groups is p<0,001 while that of the thrombotic complication with the x2 is p<0,05.

The introduction of the INR and the acceptance by the medical people of the necessity of the intense oral anticoagulant treatment especially on high risk patients with mechanical heart valves as is the majority of the presented patients, will minimize the thromboembolic complications without high risk of bleeding.
PROTEIN C RESPONSE TO INDUCTION AND WITHDRAWAL OF ORAL ANTICOAGULANT TREATMENT
Authors: Schofield K P , Thomson J M , Poller L
Published: August 2018

Protein C (PC) activity and antigen levels have been related to clotting activities of factors VII and X during the induction and withdrawal periods of oral anticoagulant treatment. Both factor VII and PC activities fell rapidly during a gradual induction regime of nicoumalone in six consecutive patients but factor VII showed a more rapid and much more marked depression than PC. In contrast reductions in factor X were much slower. PC antigen although depressed rapidly at the initiation of treatment did not subsequently fall to the same degree as PC activity, The ratio of activity to antigen became progressively smaller.

In six further serial patients discontinued from long-term treatment with nicoumalone (mean duration 12-6 months) there was a reversal of the pattern, but with two important differences. Firstly, there was evidence of an excessive rise (“rebound”) of factor VII compared with the steady state levels in these patients; and secondly there was an unexpectedly slow return of PC activity and antigen to normal levels after the oral anticoagulant was withdrawn (levels were still below normal on day 4). Factor X also showed a slow rate of increase, similar to PC activity recovery. These observations lend support to gradual withdrawal of oral anticoagulants after a period of long-term administration. The results suggest that after discontinuation of long-term oral anticoagulants patients may have increased coagulability up to four days.
NORMOTEST. A SIMPLE METHOD FOR DETERMINATION OF FACTOR VII-PHOSPHOLIPID COMPLEX.
Authors: Dalaker K , Janson T L , Prydz H
Published: August 2018

We have previously described a factor Vll-phospholipid complex (the phospholipase C(PLC)-sensitive component of factor VII) in plasma from pregnant women (1,2) as well as in some men at high risk for cardiovascular disease (3), and in male survivors of acute myocardial infarction (4).

Factor VII present in such complexes is markedly activated, and this activation is reversible upon treatment with PLC, which hydrolyses phospholipids to diglycerides.

We report here that a highly significant positive correlation was found when the PLC-sensitive component of factor VII was tested in a spesific factor VII test system and in Normotest (Nycomed A/S, Oslo, Norway) (p<0,0001). The Normotest system, which is sensitive for coagulation activities within the normal range, was not influenced by PLC when the enzyme was added immediately before the coagulation reaction was started.

We conclude therefor that Normotest is well suited for determination of the factor Vll-phospholipid complex in plasma.

References.

K. & Prydz, H. Br J Haematol 56, 233-241, 1984

Dalaker, 1984. Dalaker, Dalaker, Dalaker,

K. Br J Obstet Gynecol 93, 17-21, 1986.

K. et al. Br J Haematol 61, 315-322, 1985. K. et al. Submitted.
MONOCYTE STIMULATION IN BLOOD EXPRESSED BY INDUCED THROMBOPLASTIN SYNTHESIS IS CONTROLLED BY THE RELEASE OF ARACHIDONIC ACID AND THE FUNCTION OF PLATELETS.
Authors: Osteurd B , Olsen J O , Wilsgard L
Published: August 2018

Inhibitors of phoapholipaae A2, block the release of arachidonic acid (20:4) in the cell membrane. Adding such an inhibitor, dibromoacetophenone (20µM) to hepar-inized blood incubated with LPS for 2 hours, blocked totally the induction of thromboplastin synthesis. Liposomes prepared from soyalecithin, containing 60* linoleic acid (18:2) had no stimulatory effect by themselves, but enhanced the stimulating effect of LPS up to 10 fold. When the liposomes were added to the blood samples 0,15,30,60 and 90 min after the LPS had been added, a time dependent response of the liposomes was seen. Blood samples incubated with LPS for 2 hours but only exposed to liposomes for 30 min had monocytes with a thromboplastin activity of 30x10/10-3cells as compared to an activity of 152x10 /10-3 cells in the monocytes of blood incubated with LPS and liposomes for 2 hours. Although the linoleic acid (18:2) is metabolized to arachidonic acid (20:4), it may be more likely that the effect of liposomes is exerted by a mechanism whereby the fatty acid 18:2 is preventing arachidonic acid from being reacylated. This will cause more 20:4 to be free and metabolized to give products required for monocyte activation.

A tremendous difference in response to monocyte stimulation between different individuals has been observed. Recently we found that this phenomenon could partially be explained by very active platelets in those with high cell stimuli response. Thus, when platelet rich plasma (PRP) from a high responder was incubated with white cells from a low responder followed by incubation with LPS, there was a drastic increase in monocyte response as compared to the samples where PRP from the low responder was incubated with white cells of its own plus LPS. PRP from a low responder combined with white cells of a high responder resulted in a low response of the monocytes to LPS stimuli.

It is concluded that fatty acids and the activity function of platelets may play a central role in monocyte function.
ACQUIRED FACTOR X DEFICIENCY IN MULTIPLE MYELOMA:A COMPLETE RESPONSE TO CHEMOTHERAPY.
Authors: Kos M , Geibler K , Ratheiser K , Pabinger I , Korninger Ch , Lechner K
Published: August 2018

A 64 year old women without any previous history of bleeding diathesis presented with bone pain and gastrointestinal bleeding. An isolated severe factor X deficiency (factor X activity 0.5%, factor X antigen less than 12.5%) was found. No inhibitor that inactivated factor X in vitro or interfered with factor X assay could be demonstrated. Substitution therapy with a prothrombin complex preparation containing factor X (PPSB Biotest) was given. Factor X recovery in the first 2 days was lower than expected (below 20%) and half life of factor X was shortened (150 minutes). Subsequently, a diagnosis of multiple myeloma (light chain myeloma, type kappa) was made. Amyloidosis was excluded by electronmicroscopic examination of rectum biopsies. Chemotherapy according to the M2 protocol (Case et al) was initiated. Factor X recovery improved dramatically within 2 weeks and there was a continuous increase of factor X activity and antigen during chemotherapy. After 6 courses a complete haematological remission (less than 5% plasma cells in the bone marrow, disappearance of light chains) was obtained and factor X activity and antigen returned to normal.

Isolated factor X deficiency is a wellknown complication of amyloidosis. To our knowledge, this is the first case of factor X deficiency in multiple myeloma without amyloidosis. The complete normalization of factor X after successful chemotherapy indicates that plasma cell proliferation may have been the cause of the factor X deficiency. Binding of factor X to plasma cells containing light chains could be a possible explanation, and we are currently examining this hypothesis.
HOMOZYGOUS FACTOR XII CONGENITAL DEFICIENCY: STUDY OF 10 NEW FAMILIES.
Authors: Castaman G , Rodeghiero F , Ruggeri M
Published: August 2018

Sporadic cases of thromboembolic events have been reported in patients with congenital factor XII deficiency and a relationship with a reduced intrinsic fibrinolysis has been suggested.

We report here the results of clinical and laboratory investigations in 10 new families comprising 15 homozygotes (age 16-72) and 14 heterozygotes (age 18-65).

In homozygotes, kaolin-activated-PTT was indefinitely prolonged and F XII activity and antigen were undetectable, whereas functional assays . of high molecular weight kininogen ahd kallikrein yielded normal values. Intrinsic fibrinolytic activity - assayed on fibrin plate by measuring lysis zones determined i. by euglobulin fraction, obtained in presence of dextran sulphate and flufenamate (Blood activator inventory test, Kluft 1979) - was reduced in all homozygous pts. to about 50% of normal (range 15-70%; normal range 80-120%); normal values were observed in all heterozygotes. Basal extrinsic fibrinolytic activity (measured after addition of Cl-inhibitor) was absent or minimal as in normal controls. None of our patients showed evidence of thrombotic diathesis.

In conclusion, our study demonstrates that a reduced intrinsic fibrinolysis, as assayed by blood activator inventory test, is a common finding in F XII deficiency. The absence of thrombotic diathesis in our cases suggests that, this defect is probably devoid of any clinical significance.
INTERACTIONS OF DYSFUNCTIONAL Cl-INHIBITORS FROM PATIENTS WITH TYPE II HEREDITARY ANGIONEUROTIC EDEMA (HANE) WITH ACTIVATED HAGEMAN FACTOR (FACTOR XIIa).
Authors: Donaldson V H , Mitchell M D B H
Published: August 2018

Type II HANE is characterized by a deficiency of Cl-inhibitor (Cl-INH) activity in serum which is associated with a dysfunctional inhibitor protein having a normal or increased quantity o|_the antigenic properties of normal serum Cl-inhibitor. Dysfunctional Cl-INH proteins were purified from members_of eight different kindred with Type II HANE and compared to normal Cl-inhibitor with respect to their inhibitory activity directed against the amidolytic and clot-promoting properties of purified activated Hageman factor. All but one dysfunctional Cl-inhibitor blocked the amidolytic activity of ellagic acid-activated Hageman factor; all eight blocked the clot-promoting activity of Hageman factor activated in solutions of sulfatides and BSA. The inhibition _of amidolytic activity was equal to or greater than that of normal Cl-INH (Donaldson, et al., 3. Clin. Invest. 75:124,1985). The impairment of the specific Hageman factor coagulant activity of activated Hageman factor by six^f the eight dysfunctional inhibitors was less than that of the normal Cl-inhibitor, although readily measured. Dysfunctional Cl-inhibitor proteins were also heterogeneous with respect to their formation of stable complexes and their susceptibility to cleavage by Hageman factor activated with BSA-sulfatides when analyzed in SDS-gel electrophoresis. Although these observatons cannot be directly applied to in vivo pathophysiologic changes in plasma, dysfunctional Cl-inhibitors do have the potential of regulating activated Hageman factor.
ANTITHROMBOTIC PROPHYLAXIS THROUGH MODIFICATION OF FIBRIN NETWORK STRUCTURE:A NEW APPROACH.
Authors: Dhall D P , Shah A G , Nair C H
Published: August 2018

Morphometric analysis of electron micrographs have demonstrated that diameters of fibrin strands follow a bimodal distribution comprising a major network of thicker fibres with a minor network interspersed and made up of very fine fibres. The relationship between the two networks is not fixed and invariant. It will be shown that the two networks are highly responsive to changes in the physiological conditions of clotting. Such a system has biological adaptability and tends to result in fibrin which is more suited to serve its varying roles in haemostasis, in limiting sepsis, in promoting neovascularization and in acting as a scaffolding for wound repair.

The response of the two network system to dextran, an agent widely used and well known for its antithrombotic properties, has been examined. It will be shown that in vitro dextran increases fibrin fibre thickness, reduces permeability and tensile strength of networks developed in plasma. Similar changes were found to follow when dextran is infused in clinical dosage in man. It was found that the increase in the thickness of major network fibre is at the expense of protein in the minor network. Such means of modulating distribution of fibrin fibre diameter within fibrin networks provide a new approach to antithrombotic prophylaxis.
SPECIFICITY OF MONOCLONAL ANTIBODIES (MABS) TO FIBRINOGEN/FIBRIN MOLECULE USING THE AVIDIN-BIOTIN IMMUNOPEROXIDASE STAINING OF FROZEN SECTIONS OF HUMAN FIBRIN
Authors: Houranieh A , Kudryk B , Davis G L , Ito R K , Statland B E , Yunis E J
Published: August 2018

To study the effect of Factor XHIa mediated cross-linking of fibrin on the specificity of MAb/TD-1 (specific for the D-dimer of the DD/E region of human cross-linked [XL] fibrin, MAb/T2Gls (specific for the amino terminal region of β chain, β 15-42 and Fibrin II [ des AABB fibrinogen ]) and MAb/l-8C6 (specific for fibrinogen, Fibrin I [ des AA fibrinogen ] and B8 1-42) an Avidin Biotin Immunoperoxidase staining of frozen sections of human fibrin was used. The three monoclonals were applied as first antibodies in the Vectastain ABC Kit ( Vector Laboratories, Burlingame, CA ). Clots were prepared from Imco fibrinogen using 0.015 M Na2HPO4, 0.075 M NaCL, pH 6.35. Where applicable, EDTA 0.01 M was used in the buffer. Bovine thrombin 1 u/ml was added and the clots were incubated overnight at 20°C. Frozen sections were prepared and stained. Kabi grade (L) fibrinogen was also used, but failed to clot in the presence of EDTA. The results of the comperative study were as follows:

The positive reaction with T2Gls in both clots preparations is due to the presence of the. neoepitope in cross-linked and noncross linked Fibrin II. The specificity of TD-1 for cross-linked fibrin is supported by a positive reaction in the normal clot, while the reaction was very weak in the presence of EDTA. Due to the cleavage by thrombin of Bβ (Argininel4-Glycinel5) MAb/l-8C6 was negative in both clots. These results indicate, that the Avidin-Biotin Immunoperoxidase staining of frozen sections of human fibrin, is useful for the evaluation of monoclonal antibodies specificity to fibrinogen/fibrin molecule.
THE SIGNIFICANCE OF VON WILLEBRAND FACTOR ANTIGEN (vWFAg) AS AN ENDOTHELIAL MARKER DURING HAEMODIALYSIS
Authors: Rylance P B , Gordge M P , Dood N J , Weston M J
Published: August 2018

It has been suggested that platelet activation during haemodialysis (HD) is associated with damage to the endothelium which might contribute to increased atheroma seen in HD patients. We have therefore measured vWFAg (by radial immunodiffusion) as an endothelial marker during HD and B-thromboglobulin (BTG) (by RIA) as a marker of platelet activation with various anticoagulant regimes: unfractionated heparin (UFH) (Leo), low molecular weight heparin (LMWH) (Choay CY 216), regional citrate anticoagulation, prostacyclin (PGI2) (5ng/kg/min) and the combination of PGI2 + UFH.

Platelet activation occurred (as shown by a progressive rise of BTG) during HD with UFH, LMWH and citrate, but not with PGI2 or PGI2 + UFH. Pre-dialysis vWFAg levels were elevated in HD patients (2.37 ± 1.16 (SD) U/ml) compared with healthy controls (1.15 ±0.34 U/ml, p<0.001). Plasma concentrations of vWFAg increased during HD with UFH (26 ± 13* rise after 120 mins, p<0.001), but not during HD with LMWH, citrate, PGI2 or PGI2 + UFH. UFH infusion without HD did not result in vWFAg rise nor did UFH or IMWH interfere in vitro with the vWFAg assay. Thus PGI2 prevented both platelet activation and release of vWFAg. With IMWH and citrate there was no increase of vWFAg despite evidence of platelet activation.

Elevated pre - dialysis vWFAg concentrations in HD patients may reflect chronic endothelial injury or alternatively may be due to impaired clearance of vWFAg in uraemia. The results obtained during HD suggest that vWFAg release occurred only in the presence of both platelet activation and high molecular weight heparin chains. Whether the release of vWFAg reflects disruption and damage to the endothelium or stimulation and displacement of vWFAg remains uncertain.
PROSTACYCLIN AND THROMBOXANE A2 GENERATION IN VIVO IN MAN -EFFECT OF LOW DOSE ASPIRIN
Authors: Kyrle P A , Eichler H G , Lechner K
Published: August 2018

The effect of a low-dose aspirin regimen on platelet and vascular prostaglandin metabolism was studied in vivo in man. In a double-blind placebo-controlled cross-over study, 7 healthy male volunteers were treated with aspirin (35 mg.day−1 or placebo for 7 days. After a washout period of 2 weeks, the subject were crossed to the alternate treatment. 12 hours after the last dose of aspirin or placebo formation of thromboxane A2 (TxA2) and prostacyclin (FGI2) was measured in blood emerging from a standardized injury of the microvasculature made to determine bleeding time. TxA2 and PGI2 were measured as their stable degradation products, thromboxane B2 (TxB2) and 6-keto-prostaglandin F1α (6-keto PGF1α), using radioimmunoassay procedures. When subjects were treated with placebo, there was a rapid and substantial generation of both TxA2 and PGI2 at the site of plate-let-vessel wall interaction. This was reflected by an increase of TxB2 from 2.8±1 ng/ml and of 6-keto-PGF1α from 38.6±14.6pg/ml in the first minute to 4.5±0.6 ng/ml TxB2 and 154±50 pg/ml 6-keto-PGF1α after 4 minutes. Low-dose aspirin caused a significant inhibition of both TxA2 and PGI2 generation in bleeding time blood as represented by 65-92% and 81-84% inhibition of TxB2 and 6-keto-PGF1α Respectively throughout the 4 minute study period. We conclude that (a) rapid activation of both platelet prostaglandin metabolism and vascular PGI2 biosynthesis occurs at the site of platelet-vessel wall interaction and (b) low-dose aspirin results in a significant inhibition of platelet and vascular cyclo-oxygenase activity. Thus, our data fail to confirm the concept of a differential effect of low-dose aspirin on platelet and vascular prostaglandin synthesis in vivo in man.
STUDIES ON THE ROLE OF THROMBOXANE IN THROMBIN-INDUCED PULMONARY INSUFFICIENCY IN THE RAT
Authors: Sandler H , Gerdin B , Saldeen T
Published: August 2018

The pathophysiology of the acute pulmonary damage that may occur in association with sepsis, hemorrhagic shock and microembolism seems to involve the activity of cyclooxygenase derived arachidonate metabolites. In the rat, pulmonary microembolism due to infusion of thrombin together with inhibition of fibrinolysis has been found to induce pulmonary insufficiency with similarities to the clinical adult respiratory distress syndrome. The infusion of thrombin results in systemic hypotension, pulmonary hypertension and platelet aggregation, and subsequently, with a certain dealy in time, to increased pulmonary vascular permeability.

The main purposes of this investigation were to study the release of TxA2 and PGI2 following thrcmbin-induced pulmonary microembolizaticn in the rat and to examine the effects of a thromboxane synthetase inhibitor (DazoxibenR) on pulmonary arterial and systemic mean arterial pressure and vascular permeability.

During infusion of thrcmbin in rats pulmonary arterial pressure rose from 15 ± 2 to 35 ± 3 rmHg and mean arterial pressure fell from 120 ± 6 to 49 ± 27 mmHg. Plasma thromboxane B2 (TxB2) increased from 0.3 ± 0.004 to 3.6 ± 0.5 ng/ml. Ninety minutes later the lung weight and albunin concentration in the lung were increased (2.21 ± 0.13 g and 22.7 ± 4.7 mg/g) compared with controls (1.12 ± 0.14 g and 8.5 ± 0.9 mg/g). Dazoxiben reduced the elevated pulmonary arterial pressure and the elevated plasma TxB2 concentration following infusion of thrombin. Ninety minutes after infusion of thrcmbin, the in vitro synthesis of TxB2 in lung tissue was increased. Dazoxiben and a specific rabbit anti-rat neutrophil serun reduced this synthesis of TxB in vitro. The lung weight (2.18 ± 0.20 g) and lung albumin concentration (21.4 ± 3.4 mg/g) was not affected by Dazoxiben. The results indicate that TxA2 in an important mediator of the pressure changes in the early phase after infusion of thrombin and that neutrophils are associated with thromboxane formation in the lung tissue. Rats pretreated with the antineutrophil serum had less pulmonary weight (1.69 ± 0.28 g) and the albumin concentration in the edema fluid was decreased (17.3 ± 3.6 mg/g), suggesting that polymorphonuclear leukocytes contributed to the pulmonary dysfunction. Dazoxiben when incubated with thrcmbin in a chromogenic substrate system had a linear, dose-related anti-thrombin effect. At a Dazoxiben concentration of 2.4 mM only a few percent of the thrombin activity remained.
THROMBOXANE PRODUCTION BY PATHOLOGICAL VESSELS AND ITS POSSIBLE CONTRIBUTIION TO THE DEVELOPMENT OF THROMBOSIS
Authors: Saldeen P , Saldeen T
Published: August 2018

It has been proposed that a decreased synthesis of PGI2 may be a critical factor in the development of thrombosis on the basis that generation of PGI2 may under ly the ability of normal vessels to resist platelet adhesion. Vascular synthesis of thromboxane A2 (Tx) has been believed to be nonexistent or of very slight degree and of no physiological or pathological importance. Recent studies, however, have shown that other soirees of Tx than platelets may exist. Veins from patients with deep venous thrombosis, atheromatous arteries from patients, autopsy cases and rabbits with atheromatosis, and umbilical vein grafts inserted in the carotid arteries of sheep, all was shown to produce increased amounts of thromboxane, compared to normal vessels. Platelets was probably not the source of Tx since these cells were not found in many of the vascular walls microscopically and sheep platelets are known to produce insignificant amounts of Tx. Leukocytes were often found in the walls of these vessels, and activated leukocytes may e.g. by release of active oxidant species, stimulate endothelial cells to produce an increased amount of thromboxane. This increased production of thromboxane may be of importance for induction of arterial thrombosis, which was prevented experimentally by inhibitors of thromboxane synthetase but may be of less importance for the development of venous thrombosis. The findings suggest that in prevention of arterial thrombosis, thromboxane synthesis should be blocked not only in platelets but also in the vascular wall,which requires higjner doses of inhibitor.
EFFECTS OF THROMBOXANE PATHWAY ANTAGONISTS ON HEPARIN-INDUCED PLATELET AGGREGATION (H-IPA) AND TESTING FOR HEPARIN-INDUCED THROMBOCYTOPENIA (HIT)
Authors: Brace L D , Fareed J , Hoppensteadt D
Published: August 2018

We have reported that heparin and heparin fractions can induce platelet aggregation (PA) in a substantial number of normal healthy drug-free donors. H-IPA was shown to depend upon the molecular weight and concentration of the heparin preparation used, but its mechanism remains unknown. Therefore, we performed experiments with antagonists of the thromboxane pathway to determine whether arachidonic acid metabolites contribute to H-IPA. When indomethacin or 13-azaprostanoic (a thromboxane receptor anatagonist) was added to the PRP of donors whose platelets had been shown to aggregate in response to heparin, H-IPA was completely inhibited: heparin (bovine or porcine) caused 75% PA, while pretreatment with indomethacin or 13-APA reduced the response to 6%. Similarly, if the same donors ingested 650 mg aspirin 3 hours prior to phlebotomy, the PA response to heparin was reduced to approx. 10%. These results demonstrate that at least part of the mechanism of H-IPA is mediated through thromboxane generation. However, the mechanism by which'heparin stimulates thromboxane production in platelets remains unknown.

In some patients, heparin is known to induce an immune response that causes severe thrombocytopenia (HIT) and is associated with arterial and venous thrombosis. Fratantoni, et al. (Blood 45:395-401, 1975) have introduced a PA method for the diagnosis of HIT. We have used a modification of this method to show that the PA observed when heparin is added to a mixture of normal donor PRP and HIT patient’s serum or plasma can be inhibited by antagonists of the thromboxane pathway. When normal donor PRP was pretreated with indomethacin or 13-APA and then mixed with serum from a HIT patient (290 uL PRP:160 uL serum), the PA response to heparin was reduced from 75% to 10% or less. Similarly, if the PRP donors ingested 650 mg aspirin prior to phlebotomy, PA in the HIT test was reduced from 75% to 10% or less. Thus, the interaction of heparin with the antibody and platelets causes thromboxane generation and leads to PA. Cyclo-oxygenase specific antiplatelet drugs and inhibitors of thromboxane generation may be useful in the clinical management of HIT and H-IPA.
RAPID ONSET INHIBITION OF AFRICAN GREEN MONKEY PLATELETS AFTER INTRAVENOUS ADMINISTRATION OF A MARINE OIL LIPID EMULSION
Authors: Ward M , Pavlina T , Butchin R , Johnson R , Cotter R
Published: August 2018

To evaluate how rapidly and to what extent a lipid emulsion rich in n-3 fatty acids could alter platelet function, six male juvenile African Green Monkeys (4-6 kg) were given a 6-hour IV continuous infusion of a 10% marine oil (MO) lipid emulsion (5 ml /kg/hr). Following a 21-day washout period, the same monkeys were given a similar infusion of a 10% soybean oil (SO) lipid emulsion (TRAVAMULSION®, Travenol Labs). Blood samples were collected pre-infusion, and at 6, 12, and 24 hours following initiation of infusion, upon which the following were measured: whole blood platelet aggregation and thromboxane B2 release following collagen activation, platelet count, and platelet total fatty acid composition (pre-infusion and 24 hrs only). Lipid Emulsion Fatty Acid Composition: mg/ml(% total F.A.) Both emulsions elicited comparable reductions in both platelet aggregation and thromboxane B2 release immediately following infusion (6 hr). Platelet aggregation response after MO was significantly less than that after SO at both 12 (pc.001) and 24 hrs (p<.001), and thromboxane B2 release was significantly less after MO vs SO at 24 hrs (p<.03). Platelet counts remained unchanged after both treatments. Platelet total fatty acid analyses revealed significant increases in % total F.A. for C20:5 [1.87(pre) vs 4.79(24hr); p<.005] and for C22:6 [1.09(pre) vs 3.15(24hr); p<.001] and significant decreases in % total F.A. for C18:2 [8.94(pre) vs 7.77(24hr); p<.05] and C20:4 [22.6(pre) vs 19.6(24hr) p<.05], following infusion of M0. Following infusion of SO, the % total F.A. change in C22:6 was the only one of significance [0.85(pre) vs 1.25(24hr); p<.05]. This was attributed to the C18:3 in the SO lipid emulsion. Whereas the IV infusion of an n-6 rich lipid emulsion has little effect upon platelet fatty acid composition and function, similar administration of an n-3 rich lipid emulsion markedly reduces platelet function and effects a significant increase in the n-3/n-6 fatty acid ratio of the platelets.
VITAMIN K1 IN HUMAN MILK
Authors: Kries R V , Shearer M J , McCarthy P T , Haug M , Harzer C
Published: August 2018

Fatal vitamin K deficiency haemorrhage has been observed in breast fed babies. Though the incidence of vitamin K deficiency haemorrhage seems to be low in exclusively breastfed babies in Germany, subclinical vitamin K deficiency is by far more common as demonstrated in recent studies. Vitamin K concentrations in human milk are lower than in cow's milk and infant formula, however, nothing is known about the factors determinating the vitamin K1 concentrations in human milk. Vitamin K1 concentrations in human milk were studied during the first five weeks of lactation with respect to a) stage of lactation, b) interindividual differences, c) relationship of vitamin K1 to other lipids, and d) influence of oral supplements of vitamin K1 given to the mother. Milk samples from 9 mothers were collected on day 1,3,5,22,29 and 36 of lactation using standarized techniques.

a) Vitamin K1 concentrations in colostral milk, day 1-5 (median 1,8 ng/ml) were significanctly higher than in mature milk, day 22-36 (median 1,1 ng/ml) (Wilcoxon U-Test p< 0,01). These changes during the course of lactation must be considered for estimation of the vitamin K supply in breastfed babies.

b) Vitamin K concentrations both for colostral and mature milk were found to vary widely: colostral milk 0,6-4,4 ng/ml, mature milk 0,4 - 2,8 ng/ml.

c) For colostral milk regression analyses revealed good correlations of vitamin K1 to cholesterol but none to total lipid and phospholipids, whereas no correlation to either lipid was observed for mature milk. Cholesterol appears to have a role in vitamin K1 secretion into colostral milk.

d) Vitamin K1 concentrations of maternal milk were influenced by oral supplements given to the mother. Even with a dose of 100 μ vitamin K1 (similar to the dose which may be ingested with a meal) a twofold increase of the vitamin K1. content of breast milk was observed. These data suggest that mutritional factors may influence the vitamin concentration in human milk. Vitamin K supplements for breastfeeding mothers on vitamin K1 poor diets could improve the vitamin K supply of these babies.