bluuebunny/crossref_metadata_2025_split huggingface dataset:
IMAGING OF THROMBI USING ANTI-PADGEM ANTIBODIES SPECIFIC FOR ACTIVATED PLATELETS
Authors: Palabrica Theresa M , Furie Barbara C , Konatam Marvin A , Brockway Barbara , Aronovitz Mart , Furie Bruce 
Published: August 2018

PADGEM protein is a platelet-specific alpha grannie membrane protein that is translocated to the plasma membrane during platelet activation and secretion. Monoclonal EC4 and polyclonal anti-PADGEM antibodies are specific for PADGEM on activated platelets. Because PADGEM is internal in unstimulated platelets* these antibodies do not bind to resting platelets. Since activated platelets are concentrated in thrombi, we used radiolabeled anti-PADGEM antibodies to image thrombi in baboons.In vitro: Dacron graft material incubated with thrombin-activated platelet-rich plasma and 131I-KC4, 131I-anti-PADGEM, 131I-nonimmune IgG or 131-BSA bound 95%, 70%, 9% and 1% of the radiolabel, respectively, after exhaustive washing.In vivo: Imaging experiments were carried out in baboons with an external Dacron shunt between the femoral artery and vein. Gamma camera images following 123I-anti-PADGEM infusion (1.0 mCi; 300 μg) demonstrated intense uptake in the thrombus induced by the Dacron vascular graft within 10 minutes. The graft:blood pool activity ratio was 33:1. Control experiments with 123I-nonimmune IgG showed minimal uptake.
123I-anti-PADGEM cleared the circulating blood pool with an initial half-disappearance time of 2-6 minutes. The primary routes of metabolism were hepatic uptake, dehalogenation and urinary excretion of free iodine. There was no evidence that the anti-PADGEM antibodies had any adverse effects on the normal processes of hemostasis in any of these experiments.Anti-PADGEM antibodies, both monoclonal and polyclonal, are directed against activated platelets in thrombi or in areas of tissue injury; they do not bind to resting platelets in the circulation. Radioimmunoscintigraphy with these agents provides a novel approach to the noninvasive detection and localization of thrombi in vivo, with potential application to human disease. Furthermore, linkage of these antibodies to fibrinolytic enzymes offers a strategy for the specific lysis of thrombi in vivo.
IMMUNOCHEMICAL ANALYSIS OF FACTOR VIII (FVIII) IN PLASMA AND HEAT-TREATED CONCENTRATES
Authors: Edwards J S , Kembal-Cook G , Barrowcliffe W T 
Published: August 2018

Using the method of Weinstein et al. (Proc.Natl.Acad.Sci. USA# 78, 5137-41, 1981) the FVIII polypeptide distribution in wet and dry heated concentrates, a monoclonal-purified concentrate and fresh plasma was examined. Samples were incubated for 2 hours at 37°C (in the presence of polyethyleneglycol 4000 to aid complex formation) with 125I-Fab' fragments prepared from a polyclonal human anti-FVIII :C antibody. The complexes were elec-trophoresed in a 3-9% polyacrylamide gradient gel, in the presence of SDS, under non-reducing conditions and visualised by autoradiography.Fresh plasma slowed a range of peptide bands of apparent M.Wt. 80-280 kD, wicn a major band at 280 kD. FVIII concentrates showed a similar range of bands and, for one manufacturer's product (product E), an additional strong band of 40-50 kD. The proportion of total EVIII antigen in the 280 kD band was estimated by densitometry to be 20-40% in concentrates, can-pared with 65% in fresh plasma. Severe haemophilic plasma had no bands, confirming the specificity of the technique. FVIII antigen in 'wet' heated concentrates was shown to be more degraded (increase in low molecular weight forms) than in dry heated concentrates.Fresh plasma incubated at 37°C for 24 hours shewed increased amounts of FVIII antigen in a low molecular weight form (90 kD).Treatment of concentrates and plasma with thrombin resulted in a change of the peptide band pattern, which was dependent upon thrombin concentration and incubation time. Loss of the 280 kD band and intensification of a 90 kD band was observed, which correlated with an increase in FVIII :C by one-stage assay. Further proteolysis resulted in a band of inactive material of 40-50 kD, with identical mobility to the band seen in product E. FVIII :C activity in product E was higher by one-stage than by two-stage assay, and these results suggest more extensive thrombin degradation in this product.The results show that the molecular form of FVIII in concentrates is dependent upon storage of plasma, the methodof concentrate preparation and the type of heat treatment.
BLOOD BANK PRODUCTION OF HIGH YIELD, HIGH PURITY, HEAT TREATED F VIII CONCENTRATE FROM HEPARINIZED BLOOD
Authors: Wallevik K , Inger-slev J , Stenbjerg Bernvil S 
Published: August 2018

Applying trivial blood bank equipment we have developed a method to produce a heat-treated factor VIII concentrate from heparinized blood with an over-all yield between 40 and 50%, a specific activity around 2 i.u./mg and a low fibrinogen content (3-4 g/1). Electrophoretic multimer analysis of the von Wille-brand factor reveals a band pattern close to that of native plasma. The in vivo recovery is 99±25% and t/2 11.4±2.4 h (n=5). The method includes 3 precipitations in commercial "blood-bag" systems: (1) The classical cryoprecipitation, (2) Precipitation at 0°C of "cold insoluble globulins" including F VUI/vWf, (3) Precipitation of non F VIII/vWf related proteins at 10°C, pH 7.0 and specified ionic conditions.The F VIII rich supernatant is stabilized by amino acids and citrate, freeze dried, and heated to 68°C for 24 h.In our blood bank with ∼40,000 donations per year we have in a year produced ∼1,4 mill i.u. of the High Purity F VIII concentrate. This means that around 16,000 donations per year are collected in heparin, and that F VIII depleted plasma which we deliver for clinical use, either as liquid plasma or as a freeze-dridd product, contains solely heparin (∼4 i.u./ml) as anticoagulant.The method is technically simple which together with the high yield and high purity of the F VIII concentrate makes the production economically profitable. The method is applicable in areas with limited technical resources.We find it also important that F VIII concentrate, manufactured locally from healthy voluntary donors with personal attachment to their blood bank reveals the lowest risk for transmission of diseases to the patients.
RHEOLOGY AND ISCHAEMIA
Authors: Dormandy J 
Published: August 2018

While the previous presentations have dealt with the experimental evidence linking flow patterns and shear stress to thrombosis and atherogenesis, this presentation will concentrate on the clinical evidence linking Theological abnormalities to macro and micro-circulatory ischaemia. Whole blood viscosity undoubtedly influences blood flow along larger vessels as suggested by Poiseuille. The two important determinants of whole blood viscosity are the red cell concentration and plasma fibrinogen.There is overwhelming epidemiological evidence that the red cell concentration is a primary risk factor for the development of essential hypertension, myocardial, cerebral and leg ischaemia. It is also a secondary risk factor in patients who already had a clinical episode of ischaemia in any of these territories. There is similar evidence in relation to a high plasma fibrinogen.Furthermore therapeutic haemodilution or defibrinogenation are increasingly used in the prevention and treatment of ischaemia.Haemorheological factors in the microcirculation are probably equally important but more difficult to assess clinically. The concept of a 'vicious viscous spiral' in ischaemic tissue is gaining increasing support. Whatever the initial cause of the ischaemia, important secondary haemorheological changes occur which can perpetuate or aggravate the ischaemia. Most of the changes known to occur locally in ischaemic tissues such as hypoxia, acidosis, release of serotonin and platelet activation have been shown to impair the deformability of blood cells. This will be particularly important if the perfusion pressure is also decreased and may result in capillary plugging and uneven distribution of flow in the microcirculation. Abnormal blood cell filtrability, just as whole blood viscosity, has been shown to be associated with acute as well as chronic ischaemia in most territories. Furthermore there is a correlation between the magnitude of the haemorheological changes measured and the subsequent clinical course of the patient following an ischaemic injury.The newest aspect of haemorheology to attract clinical attention is the role of the white cell in ischaemia. Epidemiological as well as recent experimental and clinical studies suggest that the Theologically activated white cell may be the most dangerous component of blood in terms of perpetuating and extending tissue ischaemia.The assessment of the Theological properties of blood should form an integral part of studies looking at the causes and possible therapy of all forms of acute and chronic ischaemia.
CARRIER DETECTION IN JAPANESE FAMILIES WITH HAEMOPHILIA A USING FACTOR VIII GENE PROBE(F8A) AND ST 14-1 PROBE
Authors: Nishino M , Nishimura T , Naka H , Mikami S , Yoshioka A , Fukui H 
Published: August 2018

Recently, the gene structure for human F.VIII protein was clarified, and F.VIII DNA probes have been used for carrier detection and prenatal diagnosis ofhaemophilia A. In order to make sure that the phenomena are universal, we have analysed the RFLPs of F.VIII gene in 16 Japanese families with haemophilia A, including a female	haemophiliac case, using an intragenic F.VIII DNA probe(F8A) and an extragenic(linked) DNA probe(Stl4-1).The probe F8A revealed two variant bands after digestion by Bel I. Of normal 60 X chromosomes (females) examined, about 85% bore the 879-bp fragment and 15%the 1165-bp fragment. Five of sixteen mothers of hemophiliacs, definite carriers, were found to be heterozygous for Bel I polymorphism. Since the relationship between Bel I alleles and hemophilia gene has been identified in the 5 families in which the mothers were heterozygous, we could diagnose the carrier status of two women whose brothers are hemophiliacs. Onthe other hand, we could identify that one "haemophilic woman" with less than 10% of F.VIII:C was a carrier status when we analysed the Bel I alleles in theother members of the family.The probe DNA(ST 14-1) revealed seven variant bands ranging from 5.5 kb to 3.4 kb after digestion by Taq I. In 6 out	of 16 families, the RFLPs of ST 14 locus were informative for carrier detection.From these data, it was concluded that the Bel I polymorphism of	F.VIII	gene and the Taq I polymorphism of ST 14 locus were informative for carrier detection in 8 out of 16 families with haemophilia A
THE RELATIVE EFFICACY OF GENETIC ANALYSIS AND COAGULATION TESTING IN THE DIAGNOSIS OF CARRIERS OF HEMOPHILIA A
Authors: Lillicrap D , Giles A R , Holden J J A , White B N 
Published: August 2018

This study has assessed the relative benefits of restriction fragment length polymorphism (RFLP) linkage and coagulation testing in the diagnosis of carriers of hemophilia A. 221 samples from 55 families have been studied for intragenic and flanking RFLPs. All samples were tested for the Factor VIII intragenic Bell RFLP and for the flanking marker St 14.	83% of obligate carrier females were heterozygous at oneor both of these two polymorphicsites. However, only38% of these women were heterozygous at the intragenic site and might safely be offered prenatal diagnosis using this marker for the hemophilia mutation. Carrier diagnosis was obtained in 52% of 81 potential carriers tested. Diagnosis wasbased on intragenic RFLP information in only 48% of these cases. Genetic diagnosis was possible in 27 atrisk women from families with no prior history of hemophilia. Four of these women were diagnosed as carriers on the basis of a gross Factor VIII gene deletion and the remaining 23 women were identified as non-carriers by the Bell (11) and Stl4 (12) RFLP data. 39 women remained undiagnosed after gene analysis studies. 23 of these women were female relatives of sporadic hemophiliacs and thus RFLP	segregation analysis was inappropriate. A further 9 potential carriers were undiagnosed because of homozygosity in key individuals in their families. In 31 potential carriers we have quantitated Factor VIII:C (one stage assay) and vWf:Ag (Laurell and ELISA) and derived probabilities for carrier status. In 3 women there was conflicting genetic and coagulation data. Meanwhile, in 12 undiagnosed women from sporadic families, carrier diagnostic probabilities	of > 0.9 were obtained. These studies indicate that optimal carrier detection for hemophilia A requires more intragenic and closely linked RFLPs and the continuance of coagulation testing to assist women from sporadic families.
DETECTION OF MUTATIONS IN HEMOPHILIA A
Authors: Higuchi M , Kochhan L , Schwaab R , Brackmann H H , Egli H , Olek K 
Published: August 2018

Hemophilia A (HA) is a x-linked bleeding disordercaused by lack or abnormality of factor VIII:C. Because of the heterogeneity of the clinical picture thedisease might result from many different molecular lesions.We examined 202 patients of HA from 160 families by restriction analysis with Taq I, Msp I and EcoR I using three subcloned cDNA fragments of factor VIII:C.We detected 13 mutations within the factor VI11:C gene. All of these patients suffer from the severe form of HA.Table I shows six deletions which we characterized. We also identified seven point mutations on four different exons using restriction enzyme Taq I. The results are shown in Table II.That means in approximately 3.8% of the patients we found deletions and in 4.4% we found point mutations.This confirms the results of Youssofian et al (1986):These authors consider the CpG-dimer as a mutation hotspot in the factor VI11:C gene.
THE ORIGIN OF THE MUTATION IN FAMILIES WITH AN ISOLATED CASE OF HAEMOPHILIA A
Authors: Bröcker-Vriends A H J T , Briët E , Dreesen J C F M , Bakker E , van de Kamp J J P , Pearson P L 
Published: August 2018

Approximately one third of the patients with haemophilia appears to have no affected relatives. The proportion of cases due to a new mutational event as well as the gamete origin of the mutation has been much debated. The objective of this study was to define the origin of the mutation in families with an isolated case by DNA analysis. We investigated 22 families with an isolated case of haemophilia A. Intragenic (Bell, Xbal) and extragenic (BglII/DX13, Taql/Stl4) RFLPs were investigated for. If necessary, paternity was tested by DNA fingerprint patterns obtained with the 33.15 mini satellite probe.In seven of the 22 families it could be demonstratedthat the abnormal X-chromosome of the haemophiliac was derived from the normal maternal grandfather. In six of these 7 families the mother of the patient had a high probability of carriershio on the basis of clotting factor VIII assays, so that, probably, the mutation had occurred in the paternal gamete. As a consequence, carriership could be excluded for aunts, nieces and more distant female relatives of the patient.In three families the abnormal X-chromosome was derived from the maternal grandmother, while, sofar, in the remaining 12 families no conclusions as to the origin of the mutation could be drawn.In contrast with earlier findings, these results illustrate that at least thirty percent of the isolatedcases of haemophilia A are due to a recent de novo mutation.
EXTRINSIC PATHWAY INHIBITOR (EPI) DURING ELECTUVE SUGERY : A COMPARISON WITH OTHER COAGULATION INHIBITORS
Authors: Sandset P M , Anderson T R , Ødegaard O R , Larsen M L 
Published: August 2018

Hypercoagulation after surgical trauma is probably induced by tissue thromboplastin (TP) released into the circulation. EPI, in conjuction with activated factor x (FXa), is a potent inhibitor of the IP-factor VII complex. EPI levels (chromogenic substance assay) were compared to other inhibitors; antithrombin (AT), heparin cofactor II (HCII), andprotein C (PC) in patients undergoing cholecystectomy(n=4), hip prosthesis operation (n=5),and aortic grafts operationa(n=5) Mean AT and PC levels parallelled the decrease in albumin. HCII decreased more suggesting a real consumption ofsurgery. The changes in EPI levels depended on the type ofsurgery. In hip surgery, the decline in EI levels was marked and parallelled HCII. In constrast to the other inhibitors, EPI levels stayed low over the first week after surgery:In cholecystectomy, changes were less marked and all inhibitors behaved similar. During aortic operations, EPI increased from mean 110% preoperatively to 252% peroperatively. It decreased to 86% the first day after operation. It then increased similar to the other inhibitors, but the level stayed higher than expected from the preoperative value. The patients received 3000 IU heparin peroperatively.In conclusion, hip operations produce a sustained drop in EPI activity.In aortic operations injection of heparin induced a dramatic, shortlived increase peroperatively followed by a restoration to high normal levels.
LCNG-TEFM SUBSTITUTION WITH Cl-INACTIVATOR IN PATIENTS WITH HEREDITARY AND ACQUIRED Cl-INH DEFICIENCY AND LIFE-THREATENING ANGIOEDEMA
Authors: Bork K , Witzke G 
Published: August 2018

Patients with hereditary car acquired cl-inactivator (Cl-INH) deficiency suffer frcm repeated episodes of oedema of the skin and internal organs, which often constitute a threat to life (laryngeal, pulmonary and brain oedama). The treatment which is rapidly effective in the other, much more common, forms of angio-oedema (idiopathic, allergic or anaphylactoid, often associated with urticaria) has little or no effect in these cases. This applies particularly to treatment with antihistamin agents and corticosteroids. However, in patients with hereditary angiooedana (HAE) Danazol is effective for long-term prophylaxis and the substitution of Cl-INH for acute treatment. In isolated cases long-term prophylaxis, which is generally reliable, cannot be used because of intolerability reactions. Moreover, it is ineffective in the oedema of acquired Cl-INH deficiency.As other forms of therapy did not achieve the desired response, long-term substitution with a concentrate of Cl-inactivator (Behringwerke AG, Marburg) was carried out in 2 patients (H.W., 47 yrs, male, hereditary angiooedana, 9 siblings died from angiooedana; W.K., 55 yrs, male, professional truipeter, suffering from angiooedana of unknown origin for 5 years, without other underlying disorders). The initial values wereSubstitution was carried out according to the clinical symptoms. Patient H.W. required 500 U Cl-inactivator every 4th day and patient W.K. 1000-1500 U every 5th day until. During substitution therapy (which in pat. H.W. has so far been carried out for 8 months and in pat. W.K. for 6 months) there was a rise in Cl-INH and C4 with an almost total absence of clinical syirptans. No undesirable effects were observed.
KALLIKREIN-KININ SYSTEM IN PATIENTS WITH SHOCK. COMPARISON BETWEEN SEPTIC AND CARDIOGENIC SHOCK
Authors: Martínez-Brotóns F , Oncins J R , Mestres J , Amargós V , Reynaldo C 
Published: August 2018

Alterations of the kallikrein-kinin system (KKS) consistent with activation and increased consumption have been reported in septic patients and it has been suggested that this activation could contribute to the development of septic shock. As similar alterations have been found in other critically ill patients, many of them prone to shock, we wonder if activation of the KKS could be a consequence rather than a cause of the hemodynamic instability. To answer this question we compared 12 patients with septic shock (8 fatal) with 10 cases of cardiogenic shock secondary to myocardial infarction (8 fatal) as a model of non septic shock. In adition 25 episodes of uncomplicated sepsis and 10 intra-intensive care unit controls were studied. A functional measure of factor XII, high molecular weight kininogen (HMWK) (coagulative methods), prekallikrein (PK), Cl-inhibitor (Cl-INH), α2-macroglobulin (α2-M)- Antithrombin III (AT-III) and kallikrein inhibitor activity (KIA) (chromogenic methods) was performedRESULTS: Patients with septic shock, specially in fatal cases, showed a highly significant decrease in activities of factor XII (P<0.001), PK (P<0.0001), HMWK (P<0.005), α2-M (P<0.001), AT-III (P<0.0001) and KIA (P<0.005). Cl-INH activity was increased in uncomplicated sepsis (P<0.001) but came back to normal or was slightly decreased in septic shock. Components and inhibitors of the KKS were within normal limits in all patients with cardiogenic shock.Our findings support the idea of a contribution of the KKS to the development of septic shock but this system neither seems to play a role in the pathogenesis of cardiogenic shock nor to be altered as a consequence of it.
MODERATE DECREASE OF FACTOR VII AND PROTEIN C WITHOUT OCCURRENCE OF HEPATIC VENO-OCCLUSIVE DISEASE AFTER ALLOGENEIC BONE MARROW TRANSPLANTATION IN 18 PATIENTS TREATED WITH LOW DOSE HEPARIN
Authors: CARON C , JOUET J P , HIMPENS J , HIVES P , GRUSON H , GOUDEMAND J 
Published: August 2018

Decrease of factor VII (F VII) and protein C (PC) has been said to allow an early detection of hepatic veno-occlusive disease (VOD) (VILMER, Path. Biol. 1986, 34 : 79), that represents a serious complication of bone marrow transplantation (BMT). In this purpose, F VII (activity) and PC (antigen) have been measured in 18 patients (aged 9 to 45 yr- m : 26 yr) who underwent allogeneic bone marrow graft for chronic myelogenous leukemia (9 cases), acute lymphocytic leukemia (7 cases) acute myelogenous leukemia (2 cases). All patients received as preparation for BMT total body irradiation (mean dose = 10 Gy) along with cyclophosphamide (120 mg/Kg). All were given low dose heparin (100 UI/Kg/24 hr) from days -7 to +30. None of the patients developed VOD but graft-versus-host disease occurred in 13 out of them between days 18 and 52. Moreover, coagulation studies performed from days 1 to 28 detected a moderate decrease of F VII and PC (maximum on day 11). These parameters were normalized on day 28. The level of the other vitamin K-dependent factors was not significantly changed.So the moderate decrease of F VII and PC found in the post-graft period was not associated with hepatic VOD. However, as none of the patients developed this complication, these results do not exclude that a major decrease of these parameters could serve as an early diagnosis of VOD. On the other hand, a prophylactic effect of low dose heparin cannot be ruled out.
A STUDY OF THE EARLY HAEMOSTATIC CHANGES FOLLOWING RUSSELL'S VIPER BITE IN HUMANS
Authors: Than Than , Ei Han Khin , RA Hutton , Lwin Mvint , Nu Swe Tin , DA Warrell 
Published: August 2018

Amongst its many actions, Russell's viper (RV) venom activates factors X and V and enhances fibrinolysis, leading to defibrination which contributes to the clinical sequelae of RV bite. Early administration of antivenom may be life-saving, but not all of those bitten become sufficiently envenomed to require treatment. In an attempt to predict at an early stage those subjects who will progress to defibrination, we have serially monitored the haemostatic changes in 20 bite victims using the PT, APTT, thrombin time, platelet count, assays for factors X and V and fibrinogen and fibrin(ogen) degradation products (FDP).In five patients, no evidence of defibrination was seen at any time and none of these developed obvious clinical symptoms. In a further six subjects, slight prolongation of the PT (16-21/14s), APTT (39-51/38s) and thrombin time (16-25/14s) occurred concomitantly with a moderate fall in factor X (20-80%), factor V (30-66%) and fibrinogen (0.6-2.Og/1), but FDP never exceeded 40ug/ml. In the remaining nine subjects who all eventually defibrinated completely, moderate coagulation factor deficiency and thrombocytopenia developed as early as 1-2h after bite. The most pronounced and consistent changes were a rise in FDP to above 80ug/ml (80-640ug/ml) and a fall in factor V (2-50%), these results being obtained on admission, 1-12h after bite. We conclude that an FDP level of 80ug/ml or more is highly suggestive of impending defibrination and could be regarded as a criterion for commencing antivenom therapy.