bluuebunny/crossref_metadata_2025 huggingface dataset:
ABNORMAL STRUCTURE OF VON WILLEBRAND FACTOR IN MYELOPROLIFERATIVE SYNDROME IS ASSOCIATED WITH EITHER THROMBOTIC OR BLEEDING DIATHESIS
Authors: López-Fernández M F , López-Berges C , Martín R , Pardo A , Romos F J , Batlle J
Published: June 2019

The multimeric and subunit patterns of plasma von Willebrand factor (vWF) were analyzed in eight patients with myeloproliferative syndrome (MS) in order to investigate the possible existence of heterogeneity in the "in vivo" proteolytic cleavage of the protein, previously observed in this entity. Six patients lacked large vWF multimers, five of them having normal bleeding times (BT) and clinically documented episodes of thrombotic origin, whereas one patient had long BT and bleeding symptoms. Seven patients showed an increase 176 kDa subunit fragment while the 189 kDa polypeptide was increased in only one. In addition, another patient (and prior to any therapy) showed the presence of a new fragment of aproximately 95 kDa which disappeared after Busulfan therapy. The collection of blood from these patients with proteinase inhibitors did not correct the abnormalities.

The infusion of DDAVP to two patients with abnormal vWF was accompanied by: the appearence of larger vWF multimers which disappeared rapidly from plasma; an increase in the relative proportion of the satellite bands of each multimer and a further increase in the 176 kDa fragment. These data show some heterogeneity in the vWF abnormality present in MS which may be related in part to a variable degree of proteolysis of vWF ocurring "in vivo" rather than "in vitro", and which may be associated to either a bleeding or (even) a thrombotic diathesis. They also suggest that despite the presence of abnormal, already proteolyzed vWF, DDAVP-enhanced proteolysis occurs in MS to a similar extent as described in normal individuals.
STUDIES ON THE MULTIMERIC COMPOSITION OF VON WILLEBRAND FACTOR IN CHRONIC MYELOCYTIC LEUKEMIA
Authors: Tatewaki W , Takahashi H , Hanano M , Takizawa S , Hattori A , Shibata A
Published: June 2019

In chronic myeloproliferative disorders, thrombohemorrhagic complications occur occasionally in association with high platelet count. These patients may have acquired platelet dysfunction, hut no apparent relationship with clinical symptoms has been found. Recently, the abnormalities of the multimeric composition of von Willebrand factor (vWf) have been shown in some patients with essential thrombocythemia and polycythemia vera. We studied the multimeric composition of vWf in 31 patients with chronic myelocytic leukemia (CML); 20 in chronic phase (CP) and 11 in blast crisis (BC). vWf multimer was analyzed by SDS-1.2% HGT agarose gel electrophoresis, followed by visualization with an immunoperoxidase staining. Small (multimer band 1-5), medium (band 6-10) and large (band ≥ll) multimers were calculated by densitometer scan. Relative amount of large multimers was 15.5±1.6% (meantSD) in normal controls, 10.1±5.3% in CP and 15.0±5.5% in BC. CML patients in CP (but not in BC) had smaller amount of large multimers than normal controls (p<0.00l). Marked decrease in large multimers (<5$ of total vWf) was observed in 5 patients (25%) in CP and one patient (9%) in BC. The amount of large multimers was positively correlated with ristocetin cofactor/vWf:Ag ratio (p<0.0l) and granulocyte counts (p<0.05), and was negatively correlated with platelet counts (p<0.00l). We conclude that some patients with CML, especially in CP, lack large multimers. The negative correlation between the amount of large multimers and platelet counts suggests that large multimers are consumed in high platelet count states.
LARGE SCALE PREPARATION OF VON WILLEBRAND FACTOR BY AFFINITY CHROMATOGRAPHY
Authors: Newman J , Farb D
Published: June 2019

Treatment of bleeding in von Willebrand's disease usually consists of the infusion of cryoprecipitate or plasma. DDAVP is effective in some patients. Commercial concentrates for the treatment of Hemophilia A are ineffective as a source of von Willebrand Factor (vWF) replacement in von Willibrand's disease presumedly because of the absence of higher molecular weight forms of vWF protein. A vWF concentrate obtained during the course of preparation of an affinity purified Factor VIII may provide an alternative therapeutic agent without impacting the available Factor VIII supplies. The process used for the preparation of a highly purified Factor VIII concentrate (MonoclateTM, Armour Pharmaceutical Co.) from cryoprecipitate includes an affinity chromatography step which separates vWF/Factor VIII complex from other proteins in cryoprecipitate using an anti-vWF monoclonal antibody (C. A. Fulcher & T. S. Zimmerman, 1982). Factor VIII is then dissociated from the vWF remaining on the column and is eluted immediately by 3M sodium thiocyanante (NaSCN) as a step in the regeneration of the column. Unless the NaSCN is rapidly removed from the elutriant, the vWF activity as measured by platelet agglutination is destroyed. We have taken the NaSCN eluate and processed it immediately over an in-line G-10 or G-25 Sephadex column which removes the NaSCN while the vWF is eluted with a buffered isotonic solution. Alternatively, the vWF has been precipitated from the NaSCN by ammonium sulfate or polyethylene glycol. VWF prepared by any of these methods retains platelet agglutinating activity and has a distribution of vWF multimers similar to those of vWF in normal plasma.

The potency of vWF prepared from cryoprecipitate by this process is 20 units/ml and the specific activity is 20 units/mg. In several fractionations of kilogram amounts of cryoprecipitate, vWF was isolated and subjected to lyophilization and heating at 68° for 30 hours without loss of bioactivity. Drug development in progress suggest that the combined effect of affinity chromatography and exposure to NaSCN may negate the need for a heating procedure to reduce the risk of viral transmittance.
CALPAIN AND ELASTASE ARE NOT RESPONSIBLE FOR THE VON WILLEBRAND FACTOR FRAGMENTS IN NORMAL PLASMA AND IIA VON WILLEBRAND DISEASE
Authors: Berkowitz S D , Nozaki H , Titani K , Murachi T , Zimmerman T S
Published: June 2019

Recent evidence suggests that proteolysis plays an important role in some forms of inherited and acquired von Willebrand disease (vWD). Using monoclonal epitope mapping, we have examined the proteolysis of the von Willebrand factor (vWF) subunit with platelet calcium activated neutral protease (CANP) and human leukocyte elastase and found that they are not responsible for the proteolytic cleavage sejen in normal individuals and IIA vWD. Previously we have shown that in vivo proteolysis of vWF is a normal event with a small but consistent proportion of plasma vWF being composed of 189, 176, and 140 kD fragments cleaved from the 225 kD subunit. In IIA vWD the proportion of cleaved vWF is increased. Because calcium activated neutral protease (CANP, calpain) and one or more enzymes released from polymorphonuclear leukocytes are known to proteolyze vWF in vitro with resultant loss of large multimers similar to that seen in IIA vWD, they have been suggested as being responsible for the proteolysis in vivo. We have now digested highly purified vWF with porcine CANP I and II and performed monoclonal epitope mapping on the resulting fragments. We found no difference in the size, location, and quantity of the fragments produced by calpain I versus calpain II. New fragments were detected of approximately 200, 170, 150, and 125 kD. There was no evidence for generation of the native fragments. Mapping of the 170 and 150 kD calpain-cleaved fragments revealed them to be from different parts of the molecule than the native 176 and 140 kD fragments. Digestion of vWF with human leukocyte elastase produced new fragments at 210/205, 190, 165, 145/140, and 130/125 kD. No generation of native fragments was detected. Monoclonal epitope mapping of the 190 and 145/140 kD elastase-cleaved bands proved that they come from opposite ends of the vWF molecule than the native 189 and 140 kD fragments, respectively. Therefore, CANP and human leukocyte elastase do not produce the proteolyzed fragments present in normal and IIA vWD and probably do not cause the loss of large multimers seen in that disorder.
ASYMPTOMATIC vWD VARIANT WITH ABSENT RISTOCETIN ACTIVITY BUT PRESERVED BOTROCETIN ACTIVITY AND A DAUGHTER WITH TYPE III (HOMOZYGOUS) vWD
Authors: Hill F G H , Williams C W , Enayat S M , Darbyshire P J
Published: June 2019

The propositus, female aged 1 year, was investigated because of severe bruising. Bleeding time (BT) was in excess of 20 mins, VIIIC <0.01 u/ml and vWF Ag and ristocetin cofactor absent. Family studies showed:-

Multimeric analysis was not possible on the propositus, but was normal in the plasma of all other family members. The father of the propositus, however, had an abnormal multimer pattern on lysed platelets, in that the faint low molecular weight doublet is absent and a dense band in the position of the lower band of the doublet with anodal and cathodal trailing.

The maternal grandmother and mother appear to have asynpto-matic type I vWD and the father possibly has asymptomatic type I together with an alteration in the biologically vWF site with loss of the ristocetin site. This abnormality is not seen in his parents and his daughter possibly has type III vWD. The propositus' father although having some similarities to the patient of Howard et al., is different in other aspects.

1. Howard MA, Salem HH. et al. (1982) Variant von Willebrand's disease Type B - Revisited. Blood, 60: 1420-1428.
SEVERE VON WILLEBRAND'S DISEASE WITH ABNORMAL PLATELET AGGREGATION
Authors: Ihara A , Kobayashi Y , Aramitsu Y , Hara Y , Fujimura K , Kuramoto A
Published: June 2019

A 17-year-old boy with a life long history of easy bruising, epistaxis, subcutaneous hematoma and prolonged bleeding time from minor injuries, was initially diagnosed as having von Willebrand's disese, when he presented epistaxis at 2 years of age. Since his initial diagnosis, he has been treated with cryoprecipitate on many occasions to correct his bleeding tendency. The laboratory findings in the patient and his family are summarized in the table. The patient's mother, father and one brother have not complained any bleeding tendency.

Multimeric analysis of vWF using SDS agarose gels showed absence of large multimer in the patient's plasma, decreased large multimer in mother's and brother's plasma. When the patient's PRP was tested for aggregation and release of ATP by ADP, epinephrine and collagen, using lumi aggregometer, platelet aggregation was abnormal as shown by disaggregation and almost no ATP activity was detectable in the supernatant. ATP contents of patient's and father's platelet was about 50% of normal platelets. Washed platelets of the patient in normal plasma did not aggregate normally, but washed normal platelets in the patient's plasma aggregate normally. No inhibitor of vWF could be demonstrated in the patient's plasma. Clot retraction was normal. These findings suggest that our patient has inherited vWD from materal side and the platelet aggregation defect, probably a kind of storage pool disease, from the paternal side of the family.
PROTEOLYTIC DEGRADATION OF VON WILLEBRAND FACTOR AFTER DDAVP ADMINISTRATION IN NORMAL INDIVIDUALS
Authors: Batlle J , López-Fernández M F , López-Berges C , Berkowitz S D , Ruggeri Z M , Zimmerman T S
Published: June 2019

The infusion of l-Deamino-8-D-Arginine Vasopressin (DDAVP) in normal individuals is followed by an increase in factor VIII/von Willebrand factor in plasma and by the appearance of larger multimers of von Willebrand factor (vWF) than those seen in the resting state. Since the larger multimers are rapidly cleared and proteolysis is known to cause disaggregation of large multimers, we evaluated the degree of vWF proteolysis after DDAVP. DDAVP was infused into eight normal adult volunteers and the relative proportions of the intact 225 kDa subunit and the 189, 176 and 140 kDa fragments were compared before and at different times after DDAVP infusion. The relative proportion of the 176 kDa fragment was increased while that of the other species was decreased, indicating that proteolytic fragmentation had occurred. However, plasmin did not appear to be responsible because the vWF fragments characteristically produced by this enzyme could not be detected. Concomitant analysis of vWF multimeric structure showed that these changes were accompanied by an increase in the relative proportion of the satellite bands suggesting that they were ptoteolytically generated. Proteolysis may explain, at least in part, rapid clearance of larger vWF multimers released by DDAVP.
EFFECTIVENESS OF DDAVP IN PATIENTS WITH VON WILLEBRAND'S DISEASE WITH SEVERELY REDUCED BASAL LEVEL OF F VIII/VWF AND NORMAL PLATELET CONTENT(TYPE I, PLATELET NORMAL) UNDERGOING TOOTH EXTRACTION
Authors: Rodeghiero F , Castman G , Di Bona E
Published: June 2019

DDAVP has been used for hemorrhage prevention during dental extraction in patient (pts.) with von Willebrand's disease (vWd), usually of type I. However, only about 20 cases have been reported. Moreover, themajority of pts. had basal level ofVIII:C and vWf equal to or higher than 20% so that dental procedures could probably have been performed safely also without any specific treatment.

We report the experience with DDAVP for hemorrhage prevention during dental extraction in 14 pts. withtype I vWd (confirmed byplasma multimeric analysis, Prof. Mannucci, Milan) and severely reduced basal level of VIII:C (mean 12.9 ± 3 UI%; range 8-18 UI%) and vWf (< 3 UI%), measured as ristocetin cofactor. VWf:Ag and vWf platelet content (UI%/109 platelets; platelets separated by Ficoll-Hypaque and lysed with TritonX) was in normal range (17-35 UI%and 22-68% in pts.; 17-37 UI% and 24-62 UI% in 18 controls). After DDAVPinfusion (0.4 μg/ml), VIII:Cand vWf activities increasedrespectively 8.5 folds (range 6.4-11.8) and 28.2 folds (range 16-40). Return to basal levels occured with an half-life of60 -80 minutes for all the activities. Bleeding time, prolongedin 8 pts., was ormalizedin 7 cases and shortened in 1. All the pts.referred of previous hemorrhage after tooth extractionperformedin the past. A total of 50 extractions were performed (3.67 teeth/pts). Mild bleeding occured in only 2pts.3-5 days after extraction and waspromptly stopped by repeating DDAVPinfusion.

In conclusion, DDAVP has proved effective for dental extraction in type I vWd pts. with severely reduced VIII:C and vWf activities in plasma but normal platelet content.
Immunoglobulins of HIV positive and negative haemophiliac children treated with different concentrates
Authors: Köhler-Vajita K , Grütler L , Bidlingmaier F
Published: June 2019

Aim of this investigation is the quantitative comparison of theimmunoglobulins G, A, M in haemophiliac children treated with different types of factor VIII concentrates.

Patients and methodsSince May 1984 22 patients with haemophiliaA were investigated. Instead of the more often analysed parameterof the cellular immunity now we want to draw the attention to the immunoglobuline values, especiallyof the HIV negative group of patients treated exclusively with Haemate HS Behringwerke in comparison with other differently treated HIV positive and negative patients.

Results The group of patients treated exclusively with Haemate HS Behringwerke (in solution heatsterilized) shows completely normal immunolobuline levels and a safe HIV negativity.

Patients treated with conventional, later with heat treated (inlyophilized form) concentrates can be divided into HIV positive and negative groups. HIV positive heamophiliacs show pathological IgG and border line IgA and IgM values. HIV negative patients have IgG, A levels within the normal limits -but clearly higher than the Haemate group. A correlation withliver enzyme status (hepatitis B, NANB) could not be found.
ANTIBODIES TO HUMAN IMMUNODEFICIENCY VIRUS (HIV) IN HAEMOPHILIA AND VON WILLEBRAND'S DISEASE AND IN THEIR HETEROSEXUAL PARTNERS
Authors: Nosari A M , Barbarnano L , Caimi T M , Strinchini A , Muti G , Branbo B , decataldo F , Baudo F
Published: June 2019

The prevalence of antibodies to HIV in Haemophiliacs varies in different series: the latest figures (19636) are 12-94%, the lower values referring to patients with the mild form or treated by blood bank products. Serum conversion dates back to 1978 in USA and to 1980 in Europe. Antibodies to HIV in heterosexual partners are reported in percentage from 0-24%. Our population of patients:

ALL VON WILLEBRANND'disease-patients were treated by crycoprecipitates and are all antibodies to HIV negative lymphocyte subsets: CD4 × 109/1 (x) 863; CD4/CD8(x)1.148.

10 hetrosexual partners of anti HIV positive heamophiliacs were tested:

Stored samples of prozen plasam of 19 haemophiliacs were also tested:p patients were found anti HIV positive ;1 in 1980;2 in 1982;2 in 1994.
AIDS IN A COHORT OF HEMOPHILIACS
Authors: Kamradt T , Niese D , Brackmann H J , Musch E , Steinbeck A
Published: June 2019

780 hemophiliacs are treated at the Institut fiir experimentelle Hamatologie, University of Bonn. Of these, 61% (475 patients) are infected with HIV.

From 1982 through January 1987, 15 of the HIV-infected hemophiliacs developed AIDS. It seems alarming, that 9 patients had theirprimary manifestations of AIDS in 1986 and one in January 1987. The primary manifestations of AIDS were:

We describe here the clinical features of the hemophilic AIDS patients and discuss the differences in clinical manifestation and prognosis compared to AIDS-patients belonging to other risk groups.

These data are discussed on the background of other clinical, immunological and epidemiological findings in our large cohort of HIV-infected hemophiliacs.
THROMBOCYTOPENIA IN ACQUIRED IMMUNE DEFICIENCY SYNDRGME(AIDS)-RELATED COMPLEXES:RESOLUTION DURING HERPES VIRAL INFECTION
Authors: WAJIMA T
Published: June 2019

Immune thrombocytopenia has been recognized as a major hematologic manifestation associated with the acquired immune deficiency syndrome(AIDS) and AIDS-related complexes. The mechanism of thrombocytopenia in human immune deficiency virus infection is probably multifactorial.. The role of platelet-associated immuno-globulin(PAIgG)and circulating immune complexes(CIC) in mediating thrombocytopenia is controversial.. We experienced a case in which immune thrombocytopenia in AIDS-related complexes resolved during herpes zoster infection. A 37 y.o. white homosexual male with AIDS-related complexes and thrombocytopenia presented a 4-day history of painful, violaceous, non-pruritic vesicles which started on the right arm and hand, which then progressed to the chest wall, abdomen, back, and left arm. Peripheral lymphadenop-athy and splenomegaly was not noted. On admission WBG 13.5 Hct 40.8, platelet 46,000, Tzanck smear of vesicles revealed herpetic type giant cells. HTLV-III pos., Helper/suppressor T-cell ratio 0.3, Total protein 7.9 gm%, . Alb 3.95gm% IgG 1740 mg%, IgA 157 mg IgM 91.2 mg, Monospot neg., HBsAg Neg., HBsAb pos., HBcAb pos., VDRL neg. Before this admission, immune thrombocytopenia was documented by increased levels of PAIgG, CIC, bone marrow magakaryocytic hyperplasia, peripheral thrombocytopenia with giant platelets, absence of splenomegaly, and response to prednisone. He was treated with Acyclovir 250 mg/m2 1-hr infusion Q8h for 9 days ;to control the spread of his herpes infection. Recovery of thrombocytopenia wasobserved during herpes zo.ster infection. The platelet count rose to 158,000 and sustained over 4- weeks. Duringnormalization of platelet count thelevel of GIC (assayed by Raji cells, reference ranges, less than 12) droppedfrom 300 to less than 12 and PAIgG(fluorescence-activated flow cytometric assay, reference ranges, less than 1.5 RF) was markedly decreased from 90 to 2.9 When herpes infection had subsided the platelet count again decreased. These findings suggestthat PAIgG and GIG were contributing factors to immune thrombocytopenia and that herpes viral infection and Acyclovir altered this immunologically mediated thrombocytopenia in AIDS-related complexes.
EXPERIMENTAL STUDIES CONCERNING THE VIRUS SAFETY OF PASTEURIZED FIBRINOGEN
Authors: Heimburger N , Fuhge P , Hilfenhaus J , Kumpe G , Karges H E
Published: June 2019

Fibrinogen is available for substitution in afibrinogenaemic patientssince about 4 decades. However, it soon turned out that those concentrates bear a high risk of transmitting serum hepatitis. Over many years it was not possible to produce safe concentrates of fibrinogen. Hence, the therapy with this protein was limited to vital indications. We have now succeeded to stabilize fibrinogen inaqueous solution for pasteurization over 10 to 20 hours at 60°C.

The efficacy of the virus inactivation was tested using various animal viruses. Following results were obtained.

Tests in chimpanzees for hepatitis B safety revealed that this procedure inactivates and eliminates 105.2 CID50 of hepatitis B virus; HIV experiments are going on.

By immunizing rabbits with the pasteurized fibrinogen and absorption of the antiserum obtained with the unpasteurized product, an exposition of neoantigens during heating in aqueous solution could be excluded. This result could be further confirmed using passive cutaneous anaphylaxis.

The coagulability of the pasteurized fibrinogen is unchanged, compared to not pasteurized material. It iseasily soluble and can be used for both, i. v. infusion and as a tissue adhesive. The clinical tolerability is very good.
PROPERTIES AND VIRUS SAFETY OF A PASTEURIZED ANTITHROMBINIII-CONCENTRATE
Authors: Karges H E , Fuhge P , Heimburger N
Published: June 2019

The possible occurance of up to now unknown human pathogenic viruses makes it necessary to reconsider thestrategies for the preparation of each plasma protein concentrate used for substitution therapy. Even up to now safe products may be contaminated by infectious particles in the near future. Hence, each procedure for the preparation of such proteins should critically be checked for the elimination of contaminating proteins and should contain an inactivation step for pathogenic agents.

Since about 4 decades the pasteurization of albumin in presence of stabilizers has been found to be a safe and mild method to inactivate infectious agents. Using the stabilizersglycine and sucrose, we have now been able to pasteurize antithrombin III without the usual changes in molecular properties.

The product is more than 98 % pure and contains only traces of contaminating proteins. The pasteurization does not alter the electrophoretic behavior of the molecule in membrane electrophoresis (ME) and polyacrylamidgel electrophoresis (PAGE). The reactivity with heparin is nearly unchanged as tested by heparin cofactor activity and twodimensional immunoelectrophoresis. No neoantigen formation due to the pasteurization could be detected.

Using model viruses the efficacy of the pasteurization step has been tested. The following results have been obtained: HIV ≥106.7 (1), CMV ≥ 104.5 (2), HSV ≥106.8 (4), Poliomyelitis virus ≥106.9 (4); the number in brackets represent the time in hours necessary to totally inactivate the given virus titer.
FIBRONECTIN CONTENTS AND LEVELS IN BLOOD COMPONENTS DURING STORAGE
Authors: Müller N , Velten U
Published: June 2019

Fibronectin has been proposed to have an antithrombotic effect, protecting against platelet and fibrinogen consumption after injury. For the supply of platelets the possibility of extending platelet storage is important forthe management of platelet logistics. This study was designed to determine the effect of storage on the contents and levels of fibro-nectin (FN) in whole blood and components suchas packed RBCs, PRPs and platelet concentrates (PC) in two different plastics. For care of critically ill patients the FN present in components often used in large amounts could supplement the use of purified FN as a source of this opsonic protein. FN protein was assayed using an electroimmunoassay as well as a turbidimetric assay for quantitative determination at 2 day intervals during storage of CPDA-1 stabilized red cells at 4° C for 35 days and daily during end-over-end rotational storage of platelets at 22° C in conventional plastic containers (I) and trimellitate plasticised polyvinylchlo-ride bags (II) (F-763 Biotest). Moreover platelet functional tests, fibrinogen, F XIII and F VIII-complex were tested. FN levels in red cell componentsgradually decreased during storage until to 40% of the initial levels. Platelets maintained a concentrationof 404 ±70 ug/dl (I) and 378±66 ug/dl(II). There were no significant differences between the values determined in the two differentbags over the 8-days storage period. This study demonstrate the stabilityof FN protein during storage and formore effective use of limited donor resources the FN content of each of these products should be considered when determing the dose of FN for replacement therapy in critically ill patients with FN depletion following trauma and surgery.
VARIABLE EXPRESSION OF ALLOANTIGENS IN PLATELET COHORTS OF DIFFERENT MEAN DENSITY:AN EFFECT OF AGING IN VIVO
Authors: Pereira J , Cretney C , Aster R H
Published: June 2019

Platelets differ widely in size and density, but the relationship of this heterogeneity to platelet age and function is not established. Published evidence suggests that platelet alloantigens of the HLA and PlA systems may be acquired by or releasedfrom platelets in the circulation. We therefore studied expression of HLA, PlAl, and other markers in platelet cohorts of high density (HD) and low density (LD) separated on a linear, isoosmotic arabinogalactan gradient. HD and LD cohorts contained 11-14% of total platelets and did not differ significantly in mean cell volume. Alloantibodies reactive with antigens P1A1, Baka, and HLA-A2 were used to saturate alloantigen sites. Surface markers were quantified (Human Immunol. 15:251, 1986) with radiolabeled monoclonalprobes specific for HLA A, B, C antigens (W6/32), the Fc fragment of IgG (Hb-43) and the glycoprotein IIb/IIIa complex (AP-2).

As shown in the Table, HD platelets carry significantly more PlAl (located on GPIIIa) and significantly less HLA than LD platelets. However, HD and LD cohorts express the same number of GPIIb/IIIa and Baka (located on GPIIb) molecules. These findings are consistent with preferential loss of HLA molecules from HD- platelets in the circulation or acquisition by LD platelets. The variable expression of P1A1 in HD and LD cohorts is apparently due to a conformational change in GPIIIa, rather than acquisition or loss of the GPIIIa molecule, because total GPIIb/IIIa was thesame in the two platelet populations. Whether antigen differences in HD and LD platelets are determined at the time of platelet production or result from aging of platelets in the circulation is under investigation.
STUDIES ON THE ANTITHROMBOTIC ACTION OF A LOW MOLECULARWEIGHT HEPARIN OBTAINED FROM BEEF MUCOSAL ORIGIN AND PEROXIDATIVE CLEAVAGE
Authors: Bianchini P , Nonn R , Fareed J , Walenga J M , Kumar A
Published: June 2019

We have studied a low molecular weight heparin (LMWH) obtained by acontrolled peroxidative depolymerization of beef mucosal heparin (OP 2123, Opocrin, Corlo, Italy). This product was found to be significantly different from other LMWHs in that it exhibits the same COO-/SO2- ratios as unfractionated heparin, contains reducing end groups composed of 2-sulfated iduronic acid or 6-disulfated glucosamine and retains an identical structural integrity as that of native heparin. As opposed to most other LMWHs the oligosaccharide components of OP 2123 consist of homogeneous progressive units. In addition, the relative amount of AT-IIIaffinity components in OP 2123 were 20-30% less than other LMWHs. OP 2123 has a mean molecular weight of 6200 daltons with a potency of 90 anti-factor Xa U/mg and 68 USP U/mg. This agent produced strong antithrombotic actions in a rabbit stasis model against both an activated prothrombin complex and a prothrombin complex concentrate/Russell's viper venomcombination (ED50:(IV) 30-70 ug/kg;(SC) 0.6-1.5 mg/kg). The antithrombotic effects were comparable to other LMWHs in normal rabbits: however, in AT III depleted rabbits (immunodepleted and y thrombin depleted), OP 2123 produced stronger antithrombotic effects than most other LMWHs. The in vitro systems in contrast to other LMWHs, CP 2123 produced stronger inhibitory effects in AT III depleted plasma as measured by fibrinopeptide A generation and amidolytic anti-factor Xa and anti-factor Ila methods. The relative heparin cofactor II activity as measured by amidolytic method was also found to be higher than with most LMWHs. These results suggest that OP 2123, unlike most LMWHs, non AT III mediated actions play a major role in themendiation of the antithrombotic actions.
DO APTT AND ANTI Ila ACTIVITY PERMIT THE PREDICTION OF THE EFFICIENCY OF HEPARIN THERAPY IN PATIENTS WITH PROXIMAL DEEP VEIN THROMBOSIS?
Authors: Villians Ph , Bouvier J L , Le Corff G , Elias A , Juhan-vague I , Serradimigni G
Published: June 2019

One hundred and forty two patients admitted for proximal deep venous thrombosis (PDVT)from 01.01.85 to 12.20.86 were treated with Na-heparin. After an initial bolus (100 units/kg), a continuous intravenous infusion was started (500 units/kg/24h). Heparin activity in daily drawn blood samples was determined by two different assays = APTT (general diagnostic) and anti Ila activity (clotting method). Doses of heparin were adjusted to maintain APTT ratio between 2 and 3 .

A phlebography was performed for each patient at DO and D10. Two groups of patients with PDVT were identified : -Group I:n=14 increased thrombosis -Group II :n=26 partly or fully cleared thrombosis

No difference in localization or etiology of PDVT was found between the two groups. Thrombocytopenia was excluded in the two groups by platelet counts (D0, D5, D10).

Daily mean values and mean of mean values were analyzed in each group for APTT ratio andanti Ila activity :

Statistical analysis (student's T test) shows no difference between the two groups. It can be concluded that mean values of APTT and anti IIa activity in therapeutic range are not predictive of heparin efficiency for the treatment of PDVT
THE MOLECULAR WEIGHT DEPENDENT EFFECTS OF HEPARIN ON THROMBOPLASTIN ACTIVATED PROTEASES
Authors: Emanuele R M , Lormeau J C , Choay J , Fareed J
Published: June 2019

Five different molecular weight (M. W.) fractions of heparin (M. W.'s 23,000 ; 17,450 ; 13,300 ; 9,000 and 5,100) were prepared by gel-filtration. Screening of these fractions in global and amidolytic assaysat a concentration of 2.5 ug/ml revealed a dependence on M. W. for the potency response. In the APTT, TT, Heptest, amidolytic anti Xa and Ila assays, an increase in potency was observed with increasing M. W.. This relationship remained consistent to 13,300 M. W. after which no further increases in potency were observed. However, when the same fractions were screenedfor their effect on prothrombin time (PT), a different M. W. dependent pattern was observed. In this assay, a continued increase in potency was observed for the 17,450 and 23,000 M. W. fractions. At a concentration of 20 ug/ml, the 23,000 M. M. fraction produced the largest prolongation of the PT while the 5,100 M. W. fraction showed the least effect. The correlation between M. W. and potency in the PT assay was 0.98. No M. W. dependent potency response was observed whenidentical PT.'s were performed in plasma deficient in antithrombin-III. Further testing of the M. W. fractions using a Ca++/thromboplastin activated fibrinopeptide-A generation test resulted in a potency response similar to that observed in the PT. To more precisely investigate this M. W. dependent effect upon thromboplastin activation an amidolytic assay using factor VII-thromboplastin to convert factor X to Xa wasdeveloped. The inhibition of Xa in the presence of the M. W. fractions and antithrombin III was used as the assay endpoint. As with previous assays involvingthromboplastin activation, potency increased with increasing M. W. from 5,100 through 23,000. These observations coupled with the failure of amidolytic anti Xa assays to show increased potency beyond 13,000 M. W., suggest that heparin of high M. W. may beacting at the level of thromboplastin or factor VII / VIIa.
LONG TERM PROPHYLAXIS OF THROMBOEMBOLISM IN OUTPATIENTS WITH LOW MOLECULAR WEIGHT HEPARIN
Authors: Harenberg J , Leber P , Raedsch R , Zimmermann R
Published: June 2019

The prophylaxis of thromboembolism with oral anticoagulants and low dose heparin is established. However, bleeding episodes and other side effects may occur. Alternative compunds for further anticoagulation do not exist so far. We report, therefore, of the experiences of 66 outpatients who were treated with low molecular weight heparin up to nineteen months.

All patients had major bleeding on conventional anticoagulants. Further anticoagulation was strongly indicated because of recurrent thromboembolism, artificial heart valves, dilative cardiomyopathy or heart valve diseases with artrial fibrillation. The dose ranged from 2.500 to 15.000 aXa units once daily subcutaneously. The LMW heparin Tedelparin (Kabi 2165) was used. The dose was chosen according to the risk ofbleeding or thromboembolism, and body weight of eachpatient. In patients with high bleeding risks (esophagiai varices) the average dose was 100 units/kg bodyweight and in patients with high risk of thromboembolism (artificial heart valves) the average dose was140 units/kg bodyweight. The dose was adjusted according to the aXa activity and aPTT inhibition: 2-4 hours after the s.c. administration 0,3 - 0,6 heparin units/ml or 0,6 - 1,0 units/ml in the above mentioned groups should be reached respectively, and the aPTT should be not more than 10 sec above the upper normal limit.

Four of 66 patients experienced rethrombosis, 2 of them were non- compliant. Six of 66 patients had minor haemorrhages, 0 major or fatal bleeding occured. Bleeding was more frequent when the aXa activity increased to more than 1 unit/ml. With a dose reduction of 20 % no recurrence of bleeding was seen and antithrombotic effect was still maintained.

The data demonstrate that LMW heparin can be safely and effectively used for long term anticoagulation in patients with major haemorrhage due to cönventional anticoagulants.
RANDOMIZED DOUBLE BLINDED STUDY OF NORMAL AND A LOW MOLECULAR WEIGHT HEPARIN IN GENERAL MEDICAL PATIENTS
Authors: Harenberg I , Kallenbach B , Martin U , Zimmermann R
Published: June 2019

Low molecular weight (LMW) heparin has been proven effectively and safely at low doses with one injection per day in the postoperative medicine. This hasnot been confirmed so far in general medical patients requiring prophylaxis of thromboembolism. We report here of the results of a double blinded prospective randomized trial with normal and LMW heparin.

Two hundred patients were included into the study after having given informed consent. They received 3 × 5.000 units sodium pig intestinal mucosa heparin (MW12.000) or 1 × 1.500 aPTT units (MW 3.800, Sandoz AG, Niimberg, FRG) for 10 days. The development of thromboses was screened by clinical investigation, impedance plethysmography and Doppler sonography of the femoral vein at days 1,3,5,7 and 10. The incidence of haemorrhage was assessed by clinical examination, hemoglobin and hematuria at the same time intervals and factor Xa inhibition, aPTT, thrombin inhibition, antithrombin III and thrombocyte count were measured.

The following incidences occuredin the heparin / LMW heparin treated groups: clinically evident thrombosis 4 / 3 (phlebography being indicated only in 1 patient of the LMW heparin group and documenting freshthrombosis), pulmonary embolism 0 / 0, local allergy1 / 0, fatal bleeding complication 2 / 0, mortality 2 / 3 (mortality was not due to thromboembolism or treatment related complications in the LMW heparin group, one patient being antopsied). Local hematomas were 3 times more frequent in the heparin group (p <0,05).

It is concluded that one daily injection of low doses of LMW heparin is as effective as normal heparin in preventing thromboembolism in hospitalized patients in general medicine.
SAFE DOSING OF LMW HEPARIN IN HIP SURGERY
Authors: Hedner U , Mätzsch T , Berggvist D , Fredin H , Østergaard P
Published: June 2019

An enzymatically degraded low molecular weight heparin (mean mol wt 4900 dalton, LOGIPARIN, NOVO In-dustri, Denmark) was given s.c. in a dose of 35 Xal u/kg b.w. once daily to 10 patients undergoing elective hip surgery in order to study the plasma levels of Xal and Hal activity and with special regard to safety. The Xal activity in plasma was ≤ 0.24 Xal u/ml and the Hal activity ≤ 0.043 Hal u/ml. No bleeding complications were observed, and no increase in per- and postoperative blood loss was seen in the present study. It is therefore postulated that plasma levels of the Xal and Hal activities within the range obtained are safe with regard to bleeding complications for the LMW heparin used and in the dose administered. No accumulation was seen, which may contribute to the safety. No phlebographically verified thrombi were registered but the antithrombotic efficacy of the dose regimen used remains to be demonstrated in a larger number of patients under controlled conditions, a study that is well under way.
ASSESSMENT OF DIAGNOSIS OF DEEP VENOUS THROMBOSIS OF THE LOWER LIMBS USING ESTIMATION OF CLINICAL PROBABILITY
Authors: Krahenbuhl B , Sheybani E , Bounameaux H
Published: June 2019

The value of a diagnostic test (or test combination) depends not only on its sensitivity and specificity but also on the prevalence of the disease. In the present study, we have re-assessed the value of the clinical diagnosis in a group of 45 consecutive patients suspected of deep venous thrombosis (DVT) of the lower limbs. The clinical probability (CP) of the diagnosis was estimated on a clinical basis alone (history and clinical examination) by trained physicians (2-month training in an Angiology Unit). Afterwards, the diagnosis of DVT was established using the combination of Doppler ultrasounds and venous occlusive plethysmography. Venography was performed when the non invasive techniques were inconclusive.

DVT was found in 14 patients (31 %). In the 21 patients in whom CP was ⩽ 0.20, there was no DVT. In the 10 patients in whom CP was ⩾ 0.80, DVT was confirmed in 9 cases. When CP was between 0.21 and 0.79, a DVT was found in 5 out of 14 patients.

Thus, when CP is ⩾ 0.80 or ⩽ 0.20, further investigation does not seem to provide additional diagnostic information. The estimation of CP by a trained physician allows to define a subgroup of patients in whom the clinical diagnosis is valuable.

Two third of the patients in our series belong to this subgroup. In one third of the patients, CP was intermediate and non invasive tests or venography are necessary.

Since the range of clinical probability in which clinical diagnosis is reliable will depend on both the physician and the recruitment of the patients (medical or surgical, ambulatory or hospitalized), it must be determined in each center.
SERIC ANGIOTENSIN CONVERTING ENZYME : AN ENDOTHELIAL CELL MARKER (application to thrombo-embolic pathology)
Authors: Drouet L , Baudin B , Baumann F Ch , Caen J P
Published: June 2019

As a blood marker of endothelium, we investigated the seric activity of Angiotensin Converting Enzyme (ACE) at rest and after stimulation either by local venostasis or dDAVP infusion. dDAVP did not induce any significant change in ACE contrarily to venostasis. Searching for an endothelial abnormality implicated in the genesis of Deep Vein Thrombosis (DVT) we applied the local venostasis test to patients affected by recurrent DVT. Patients, divided in 3 groups (group I : documented history of recurrent DVT, group II : only one DVT or recurrent superficial venous thrombosis, group III : history of arterial thrombo embolism), and controls were screened for ACE as well as for plasmatic fibrinolytic activity and von Willebrand factor (vWF) level. Two types of abnormalities of seric ACE activity were found : low basal level in group I and low response to venostasis in groups I and III : group II did not differ from controls. This suggests an endothelial lesion participating to the etiology of some recurrent DVT and supports the measurement of seric ACE to discriminate some patients at high risk of DVT. Measures of fibrinolytic and ACE activities are not redundant since the two types of ACE abnormalities were not individually encountered in the same patients and were independent from abnormalities of the fibrinolytic system
COMPARISON BETWEEN INITIAL AND ACHIEVED ANTICOAGULATION LEVEL WITH PHLEBOGRAPHIC EVOLUTION IN PATIENTS WITH DEEP VEIN THROMBOSIS TREATED BY DIFFERENT HEPARINS
Authors: FAIVRE R , KIEFFER K , DUCELLIER D , BILSTEIN F , BASSAND J P , MAURAT J P
Published: June 2019

Effectiveness of standard heparin (SH) in patients (pts) with deep vein thrombosis (DVT) remains contreversial in regard with the subcutaneous (sc) or intravenous administration route.

To determine the relation between initial (day 1 after the beginning of treatment) and achieved anticoagulation level with base-line to control (at day 10) phlebo-graphic score variations, we carried out a randomized study including 68 pts with acute (less than 2 weeks) DVT treated either by SH, 500 ui/kg/day or low molecular weight heparin (LMWH, CY 222, institut Choay, 750 u anti-Xa IC/kg/day), both given by 2 daily sc injections for 10 days. Plasma was collected at the middle (at 8 a.m) of 2 sc injections to evaluate aPTT (CK prest, STAGO) and anti-Xa activity (Stachrom, Stago). The results show that : 1/ Thrombus reduction was similar (thrombolysis more than 30%, 65% in SH pts and 64% in LMWH pts, p NS) ; 2/ in SH pts, phlebographic score variations were correlated neither with achieved anticoagulation level (mean aPTT of day 3, 5, 10) nor initial day 1 aPTT (R=-.33 and R = .03 respectively) ; 3/ in CY 222 pts, no modification of aPTT was observed during the 10-day treatment, and phlebographic score variations were correlated neither with initial nor achieved anti-Xa activity (R=.06 and R=.24 respectively) ; 4/ only 2 SH pts extended DVT and 1 patient in each group developed a recurrent pulmonary embolism in spite of a well initial or achieved anticoagulation level (aPTT >1.5 the control value in SH pts).

In conclusion, SH- or LMWH-related anti coagulation (or antithrombotic) effect is certainly not the sole prognosis parameter for well achieving venous thrombolysis in patients with DVT.
ANABOLIC STEROID AND INTRAPULMONARY HEPARIN IN THE PREVENTION OF POSTOPERATIVE DEEP VEIN THROMBOSIS
Authors: Zawilska K , Tokarz A , Psuja P , Szymczak P , Kawczyński S , Zozulińska M , Turowiecka Z , Sowier J
Published: June 2019

150 patients over 40 years old undergoing major abdominal surgery were divided into 3 groups:

1/ group I - receiving a single injection of long acting anabolic steroid /nandrolone phenylpropio-nate, 50 mg intramusculary/ a day prior to surgery 2/ gropup II - receiving the same dose of anabolic steroid plus a single dose of heparin /800 U/kg of body weight/ intrapulmonary a day prior to surgery 3/ group III - receiving only a single dose of heparin /800 U/kg of body weight/ intrapulmonary a day prior to surgery.

The deep vein thrombosis /DVT/ was detected using the 125 I-fibrinogen test. The occurence of DVT was:

in group I - 14%

in group II - 4%

in group III - 8%

There were no detectable haemorrhagic complications in patients of group I and III, in 6% of patients of group II a sgliht increase of intraoperative bleeding and/or wound hematoma appeared.

We conclude that prophylaxis of DVT in the postoperative period with the single dose of anabolic steroid and intrapulmonary heparin is an effective, safe and easy to handle procedure.
HEMOSTASIS AND HEMORHEOLOGY AFTER ISCHEMIC STROKE IN PATIENTS WITHOUT CARDIOPATHY AND OTHER LOCALISATION OF ATHEROSCLEROSIS
Authors: Boisseau M R , Bonnet J , Freyburger G , Desbordes P , Brottier L , Orgogozo J M , Bricaud H
Published: June 2019

Hemorheological parameters (whole blood filtration, whole blood viscosity, plasma viscosity and erythrocyte deformability by ektacytometry) and fibrinolysis parameters (Euglobulin Lysis Time ELT, t-PA activity, and Plasminogen ActivatorInhibitor -PAI- before and after Venous Occlusion Test -VOT-) were measured in 20 stroke patients. All these patients had a crerebrovascular accident (CVA) localised to the carotid arterial tree. They has no signs of heart disease and were without risk factors for atherosclerosis (high blood pressure, diabetes) ; they were investigated in the week following the CVA. They were divided into 4 groups : 1/ transcient ischemic accident, 2,3,4/ size of infarct classified from results of C. T. Scan carried out 2 or 3 days after the stroke (2/small +, 3/moderate ++, 4/large +++).

For the hemorheological parameters we noted only a slight increase in whole blood filtration (27±8“ vs 21±2”, p<0,05).

The ELT was however significantly increased in these patients (258 ±57 min. vs 133 ±33,p<0,001)suggesting impairement of fibrinolysis. The following points were noteworthy : the ELT return to normal after to VOT,there was a relationship between stroke severity and the PAI levels and the viscosity at low shear stress. These two parameters were dramatically impaired in the most severely affected patients. Transcient ischemic accidents had comparable features to the CVA of moderate size.

The extend of the disorders of the hemorheological disorders and the abnormalities in fibrinolysis seemed to be related to the severity of the involvement in patients with ischemic stroke with no evidence of heart disease or atherosclerosis in other systems.
HEM0RRHE0L0GY AND KETANSERIN
Authors: De Crée J , Geukens H , Demoen H , Verhaegen H
Published: June 2019

Red blood cell (RBC) filtration in platelet rich plasma (PRP) and platelet poor plasma (PPP) was equally decreased (p < 0.0001) in 120 patients with acute myocardial infarction (AMI) as compared to a control group. In a double-blind experiment 2 groups of 30 patients with AMI received an acute oral dose of 60 mg of ketanserin, a serotonin (5-HT) antagonist at 5-HT2-receptors, or placebo. Ketanserin treatment improved RBC filtration in PRP with an average increase of 30%. A similar experiment using PPP showed a significant increase of 10%. Filtration of plasma improved after ketanserin treatment in PRP, but not in PPP. Cross-exchange experiments showed the ketanserin-induced improvement of RBC filtration in PRP and PPP to be also plasmadependent. 5-HT in vitro at 10−9M deteriorated RBC filtration in PPP (p < 0.05), and ketanserin in vitro at 10−7M counteracted this phenomenon (p < 0.001). Finally we found that the effect of a subacute treatment with ketanserin on the filtration of RBC Suspensions, enriched with a constant amount of white blood cells (WBC), was more pronounced than on control RBC suspensions of patients with AMI.

These results indicate that the impaired RBC filtration, reported in vascular diseases may be dependent on a subtle interaction between platelets, WBC, RBC and plasma. Treatment with ketanserin is capable to interrupt this vicious circle of rheological disturbances at different levels, first of all, by improving RBC deformability, but also by counteracting the platelet mediated effects on RBC and by favourably influencing the physical properties of WBC and so preventing clogging phenomena. Serotonin probably plays a pivotal role in these cascade of events and therapy with ketanserin might be of clinical value in diseases where microcirculatory flow is compromised.