bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Results of Synovectomies and Other Orthopedic Corrective Surgery in Patients with Hemophilia A and B
Authors: Scharrer I. , Willert H.G.
Published: May 2019

Since 1970 twenty orthopedic corrective operations were performed in patients with hemophilia A and B without complications. The bleeding frequency was markedly reduced after the operations. The mobility of the joints improved distinctly after surgery. Based on our results we shall present the special indications and techniques for the operations, the substitution plans during and after surgery and a physiotherapy program after the operations.
Low Dose Factor Replacement in the Hemophiliac
Authors: Kelly P. , Boutaugh M. , Penner J.
Published: May 2019

The standard dose of factor administered for treatment of acute bleeding in hemophiliacs is usually 20 units per Kg. To assess the effectiveness of lower doses, 103 hemophilic campers were randomly assigned and treated with 3 different dosages, i.e. 7–9 u/kg, 11–15 u/kg, and 15–17 u/kg, for treatment of soft tissue and joint bleeds. Patients with inhibitors, and those with bleeding into cricical areas were excluded from the study. It was found that, when administered early, the 7–9 u/kg dose was as effective as doses in the 15–17 u/kg range for treatment of spontaneous bleeding in the extremities.

In summary, our experience with a large number of hemophilic youngsters attending a summer camp would indicate that a low dose therapeutic approach (7–9 u/kg) is effective, and permits most of the patients to participate fully in camp activities.
A Demonstration of Physical Therapy
Authors: Weissman Jerry , Zottoli Carolyn
Published: May 2019

It has been generally accepted that physical therapy has a role to play in the total habilitation and rehabilitation of the hemophillic patient. These types of services are readily provided within the framework of a comprehensive hemophilia clinic.

All patients are advised as to physical fitness and recreational type activities. In addition, most patients are on a home program of specific exercises directed at specific orthopedic problems. Physical therapy is also available through our out-patient clinic when it is felt that the assistance and supervision of a physical therapist is required. An evening clinic is held to accommodate those working or attending school. In-patient physical therapy is most commonly reserved for those patients who have undergone an orthopedic procedure though, on occasion, patients have been admitted solely for an intensive physical therapy program.

The concepts underlying the exercise programs are similar to the ones used in managing arthropathies of other etiologies. Special exercise techniques will be demonstrated.
Long Term Results of Physical Therapy
Authors: Dietrich Shelby L.
Published: May 2019

Physical therapy has been one of the three key elements in treatment of hemophilic musculoskeletal problems at Orthopaedic Hospital for the past 15 years. Prompt treatment of bleeding episodes with plasma concentrates and conservative orthopaedic treatment are the other basic elements of care. Active exercise programs, (including appropriate sports), are prescribed to maintain limbs in their optimum physical condition to withstand stresses and strains of normal activities. These programs are similar in concept to “conditioning” programs for athletes. When musculoskeletal weakness or deformity results from bleeding problems, prescriptive physical therapy is an essential part of rehabilitation. Review in 1976 of mobility status of 227 hemophilic patients ages 20 through 73 under active care at Orthopaedic Hospital reveals 16 patients (7%) had signigicantly impaired mobility, using crutches, canes, or wheelchairs for assistive devices and 93% are both mobile and appliance free. Although it is impossible to isolate physical therapy from the other parts of the treatment regimen as the most significant cause of the finding it is our clinical impression that physical therapy, applied both prophylactically and therapeutically in active exercise programs, contributes significantly to the continued mobility of the hemophilic patient. Long-term prospective studies are urgently needed to document the effect of physical therapy and muscular “fitness” on frequency and severity of hemarthroses and on hemophilic degenerative joint disease.
Total Joint Replacement in the Hemophilias
Authors: Duthie Robert B.
Published: May 2019

Haemoarthrosis making up 85% of all bleeds in the haemophilic, most commonly affects the knee, the elbow and the ankle joint. However, it is the arthropathy in the hip joint which has produced the most serious disability requiring total joint replacement. Major reconstructive surgery in the haemophiliac is based upon the following principles: (i) healing of soft tissues must occur per primum. (ii) the surgical technique must be standard without any need for revision or secondary procedures. (iii) the factor replacement therapy programme should be of sufficient amount and duration to cover both healing and mobi1ization. (iv) the presence of antibodies to either Factor VIII or IX is an absolute contradiction. Sixteen haemophilic patients have received a bicomponent total hip replacement.
Effects of Self Hypnosis in the Management of Hemophilia in Children
Authors: Olness K.N. , Singher L.J.
Published: May 2019

A group of families with hemophiliac children were given the opportunity to learn techniques of self hypnosis to alleviate anxiety, discomfort, pain, and hopefully to reduce their requirement for replacement therapy. These families have met on a monthly basis for twenty-four months. Initially, children and later their parents, were taught standard techniques for inducing self hypnosis. Children were specifically taught to control perceptions of pain and discomfort and were taught to give themselves suggestions to reduce bleeding. Parents were given suggestions for generalized relaxation.

Children, as young as three years, clearly demonstrated their ability to selectively turn off or reduce their perception of pain in designated extremities. Their unsolicited statements indicated increased comfort during procedures and their belief that they were able to reduce the number and/or duration of bleeding episodes.

We have concluded that this beneficial adjunct therapy in hemophilia is readily learned and followed through by hemophiliac children and their families.
Clinical Severity, Personality Characteristics and Maternal Attitudes in Hemophiliac Children
Authors: Ackerman L.L. , Verrecchio J. , Charney D.S. , Strickler E.M.
Published: May 2019

The interrelationships of severity of hemophilia, personality characteristics, and maternal attitudes were evaluated in 21 hemophiliac boys and 17 of their mothers. The 8 to 11 year old boys were given the Children’s Personality Questionnaire and those between 12 and 18, the Junior-Senior High School Personality Questionnaire. The mothers were administered the Mother-Child Relationship Evaluation which measured maternal attitudes of four scales; Acceptance (A), Overprotection (OP), Overindulgence (OI), and Rejection (R).

Results were analyzed by comparing children with Factor VIII levels >1% (N=10) to those whose levels were <1% (N=ll) as determined on more than one occasion. There were no significant differences between the two groups in terms of personality characteristics or maternal attitudes.

Results were also analyzed by comparing those children (N=6) who had accepting mothers (A> 60, OI < 30, OP < 30, R < 30) with those of non-accepting mothers (N=ll). The children of the non-accepting mothers (mean Factor VIII levels 3.0%) were significantly more sober (P<.02), and internally restrained (P <.01) and tended to be less intelligent and less emotionally stable than the children of accepting mothers (mean Factor VIII level 1.5%).

These findings suggest that personality characteristics may be more closely related to maternal attitudes than severity of disease. Future studies should identify and focus on determinants of maternal attitudes.
The Successful Treatment of Aquired Inhibitors Against Factor VIII.
Authors: Brackmann H.-H. , Etzel F. , Hofmann P. , Fgli H.
Published: May 2019

From the 582 inhibitor tested hemophilia A patients 19 were found to have an inhibitor against factor VIII. In 2 years of study the observation has been made, that a specific therapy combination of an activated prothrombin complex (Fraction Feiba) with high dosages of factor VIII concentrate can reduce the inhibitor titer to sero. A modified method of Bethesda was used for the detection of inhibitors. An inhibitor concentration of 1 U/ml was determined when the patients plasma inactivated 75% of that activity of factor VIII in the incubation mixture after the incubation time at 37° C. The combination therapy dosages for Fraction Feiba range from 30 to 120 U/kg body-weight and for factor VIII concentrate from 40 to 150 U/Kg bodyweight. The dosages are given 2 to 3 times a day in cases of acute bleeding episodes, and 3-7 times a week durincr regular prophylactic treatment to remove the inhibitor. In 11 of our 19 inhibitor patients, the inhibitor titer was reduced to sero on the basis of this combination therapy. High and low responsers were recorded, the highes response being 147 U/ml. The remaining (8) patients are still undergoing treatment with this procedure.
Sgot Elevations and Hepatitis B Infections in Hemophiliacs
Authors: Eyster E. , Haverstick J. , Plotkin S.
Published: May 2019

SGOT, Hepatitis B surface antigen (Ag) and antibody (Ab) were performed at yearly or twice yearly intervals for 1-2 years on 75 hemophiliacs receiving an average of 40,000 Factor VIII units per patient per year. Of 42 receiving >10,000 VIII units on 5 or more days per year, 38 (90%) had SGOT elevations. In 30 (71%), abnormalities were recurrent. Of 33 receiving less Factor VIII, only 10 (30%) had recurrent SGOT elevations; all 10 had either Ab or persistent Ag.

Fifteen patients received only cryoprecipitate. In 3/5 receiving >50 bags on 5 or more days per year, SGOT elevations persisted; one had persistent Ag and one had Ab. None of the 10 receiving less cryo had persistent SGOT elevations; 5 had Ab.

Sixty-three (84%) were Ab pos. Recurrent SGOT elevations were seen in 35/63 (55%), including 7 with Ab titers >1000. Seven patients were Ab neg; none had an ↑ SGOT. All but one with agammaglobulinemia had received <50 transfusions per lifetime and <5000 VIII units on <3 days that year. Five of the 7 had received only cryo. Five patients (6%) were persistently Ag pos; all had persistent SGOT elevations.

It was concluded that SGOT elevations in hemophiliacs were related to the frequency and intensity of Factor VIII infusions, and were regularly seen in patients receiving >10,000 VIII units or >50 bags of cryo on 5 or more days per year. Patients with Ab frequently had SGOT elevations while those with no Ab usually did not. Ab (or Ag) was found in 58/60 patients who received concentrate compared to 10/15 who received cryo only (p <.005).
Collected Experience of the Pennsylvania Hemophilia Program
Authors: Shapiro S.S. , Eyster M.E. , Lewis J.
Published: May 2019

The Pennsylvania Hemophilia Program was initiated in March 1973, with the establishment of 9 Hemophilia Centers throughout the state. From an initial enrollment of 150, the number of patients has grown to 669 as of October 1976. Of these, 491 have Hemophilia A and 91 have Hemophilia B, a prevalence rate of 4.2 and 0.76 per 100,000, respectively in the total state population of some 11,800,000. A total of 210 patients (36%) with Hemophilia A or B are on home therapy programs. Two hundred fifty-five patients with Hemophilia A (52%) have severe disease, of whom 160 (63%) are on home therapy. Thirty-six patients with Hemophilia B (40%) have severe disease, of whom 22 (61%) are on home therapy. The remaining patients are treated in-center as necessary. Thirty-seven patients (7.5%) with Hemophilia A have inhibitors to Factor VIII, while only 1 of 91 patients with Hemophilia B has an inhibitor to Factor IX. Total Factor VIII and Factor IX usage for hemophiliacs in the past year was 15,040,000 and 1,282,000 biological units, respectively. At current prices, this represents $1.5 million for Factor VIII and approximately $150,000 for Factor IX. The average annual use of Factor VIII in severe Hemophilia A, excluding surgery, was 44,300 units/patient for patients on home therapy and 32,000 units/patient for patients on Center therapy. These figures are roughly comparable when corrected for patient age (14% of home therapy patients but 28% of Center therapy patients under the age of 10). These observations suggest that the actual prevalence rates of Hemophilia A and B are lower than previously quoted, that more patients with milder disease exist than expected and that home and Center therapy require equal product usage.
Elective Synovectomy in Hemophilia
Authors: Bennetts Geni A. , Myers Marilyn G. , Ewald Brian A.
Published: May 2019

A total of twenty-one elective synovectomies and three repeat elective synovectomies, as stimulated by the work of Storti, et al., were performed on elbows, knees, and ankles of six patients with Classical Hemophilia (circulating antihemophiliac globulin factor VIII < 1%) from 1970-1976. Decision to perform surgery was based on recurrent bleeding into a joint, decrease in joint function, and radiographic evidence of joint destruction. The surgical protocol established included preoperative therapy with commercial Antihemophiliac Globulin Concentrate (AHG) and epsilon aminocaproic acid. Surgical synovectomy was followed by postoperative management which included parenteral therapy with AHG and epsilon aminocaproic acid, joint drainage and, in addition, interarticular epsilon aminocaproic acid and antibiotics with mobilization. When indicated, manipulation under anesthesia combined with a regimen of physical therapy was carried out. Initial results were encouraging, but enthusiasm after up to six years follow-up has been tempered by recurrence of hemarthroses and further radiographic deterioration in several of the synovectomized joints. This has necessitated the three aforementioned repeat synovectomies.
The Use of Fibroseal in Hemophilic Cysts - A Case Report
Authors: Anderle K. , Böhler Wien N. , Baudo F. , de Cataldo F. , Silvello L.
Published: May 2019

Based on the good results achieved with a fibrin-sealing-system consisting of a high concentrated fibrinogen-containing plasma fraction, thrombin, and calcium chloride in animal experiments and in patients suffering from bone tumors, this sealing method was used for the first time in a severe hemophiliac. Due to a massive, fractured hemophilic cyst in the proximal part of the right femur this patient had been unable to walk for months.

Combining homologous and autologous bone transplantation, fibroseal, and osteo synthesis, it was possible to avoid amputation. At present, the patient is following a rehabilitation program.

The result in this case and experience gained from earlier studies indicate that the use of fibroseal for bone reconstruction offers a new approach to orthopaedic problems in hemophilia patients.
Hepatitis B Surface Antigen Testing in Plasmapheresis Donors
Authors: McAuley Clyde B.
Published: May 2019

It is well recognized that the safety of biological products prepared from blood, such as ProfilateR, a freeze-dried factor VIII concentrate, may be increased by testing for hepatitis B surface antigen.

Abbott Laboratories has been routinely screening all plasma donations for hepatitis B surface antigen since January, 1971. Originally, the screening was performed using the second generation counter-immunoelectrophoresis technique, AUS-tect. In April, 1973, the counter-immunoelectrophoresis testing was replaced with the more sensitive third generation radioimmunoassay technique, Aus-RIA, for routine hepatitis B surface antigen testing. The most sensitive current radioimmunoassay technique, Aus-RIA II, has been used since February, 1975. This presentation will review the results of six years of testing experience. The data indicate that the incidence of positive donors in the commercial plasmapheresis system is 1.4 per 1,000, similar to that reported for a volunteer blood program.
Observations on a Change from “Altruistic” to” “Assurance” Blood Donors
Authors: Morrison F.S. , Lotz L.
Published: May 2019

Differences in positive Hepatitis testing results have been noted in different blood donor populations. A recent change in our policy for blood donors provided an opportunity to relate hepatitis antigen results to motivation. A blood center previously used donors whose only motivation was the opportunity to give blood for the community where it was needed (“altruistic”). The center then changed to an approach which provides an insurance policy to each volunteer covering all his blood costs in the ensuing year (“assurance”). Certain changes in results of blood testing have become apparent. In the four month period preceding the policy change, 3,364 usable units were drawn at the center. Of these units, six were rejected in the laboratory because of a positive STS (0.178%) and seven were positive by RIA for HBAg (0.208%). After the publically announced change in policy, 3,084 usable units were drawn in the ensuing four months. From this donor pool, 18 units were rejected for positive STS (0.58%) and 16 for a positive test for. HBAg (0.518%). These changes represent a statistically significant difference in the incidence of positive serologies for syphilis and for hepatitis, in the donor populations. We suggest that the motivation of blood donors may be influenced by the type of compensation offered and that an all-volunteer blood procurement program appears the safest course.
Four Years’ Follow-Up Study of Liver Function of Haemophiliacs
Authors: Panicucci F. , Baicchi U. , Sagripanti A.
Published: May 2019

By physical and biochemical examinations 112 patients with severe haemophilia A or B and 24 with mild to moderate forms were studied to determine the incidence of liver damage, which may have resulted from commercial concentrate therapy.

Data of the follow-up study during a period of 4 years showed transiently elevated values of SGOT and SGPT in 100 patients, persistently elevated values in 14 and normal values in 22 patients. Test for hepatitis B antigen was positive in 9 patients. Hepatomegaly, splenomegaly or both were found in 15 patients. Acute hepatitis with jaundice was reported by 23 patients, 17 of whom still showed abnormal enzyme tests. Chronic active hepatitis was very strongly suggested in 5 patients, who on repeated occasions showed, in addition to elevated SGOT and SGPT, abnormal values of serum albumins, immunoglobulins, alkaline phosphatase and Normotest. The majority of patients were free from clinical liver disease.
High Molecular Weight Fibrinogen Complexes in Patients with History of Myocardial Infarction and Cerebrovascular Disorders
Authors: Neri Serneri G.G. , Gensini G.F. , Abbate R. , Prisco D. , Mugnaini C. , Favilla S.
Published: May 2019

The increased turnover of fibrinogen and decreased platelet survival observed in many patients with history of myocardial infarction (MIP) and in patients with chronic cerebrovascular disorders (CVP) (Neri Serneri et al 1970, Harker and Slichter 1972) could suggest a hypercoagulable state. We investigated 28 MIP, 23 CVP and 31 controls for circulating high molecular weight fibrinogen complexes (HMWFC) by gel-filtration (agarose 4%, 100–200 m, column 1.5 × 90 cm, buffer Tris-Cl-citrate pH 7.6, flow 13 ml/hour, recording of OD at 280 um) of plasma beta-alanine precipitate. HMWFC are eluted in a peak at an alution volume corresponding to the void volume of the column, at which volume globular proteins of MW over 1 million are eluted. HMWFC concentration was in the controls 2.98±1.52% of the fibrinogen eluted, in MIP 8.27±2.9 % (P<0.0l) and in CVP 7.48±1.9 % (P<0.01). When HMWFC concentration was higher than 6-7%, PAA electrophoresis of the eluted complexes (after mercaptoethanol reduction) allowed to detect gamma-gamma dimers, so indicating the cross-linkage of HMWFC. Heparin treatment (12,500 U × 2) markedly lowered the concentration of HMWFC and made gamma-gamma dimers undetectable. These results indicate that in MIP and in CVP a hypercoagulability frequently exists.
Soluble Fibrinogen-Fibrin Complexes in Obstetrical Conditions
Authors: McKillop C.A. , Howie P.W. , Forbes C.D. , Prentice C.R.M.
Published: May 2019

Soluble fibrinogen-fibrin complexes isolated by 6% agarose gel filtration (Bio-Gel A5m), were identified by the staphylococcal clumping test for the void volume polymers and radial immunodiffusion for the lower molecular weight oligomers. Women taking the oral contraceptive pill had significantly increased oligomer levels compared to non-pill controls;whilst in normal pregnancy there were small increases in both polymer and oligomer concentrations. In pre-eclampsia a marked increase in both types of soluble complex was found. This did not simply reflect the combination of hypertension and pregnancy, as soluble complex levels in pregnant women with essential hypertension did not differ from those in normal pregnancy. In pregnancies with intrauterine growth retardation there was also a small but significant increase in oligomer concentration compared with normal pregnancy.

While these results may simply reflect differing degrees of hypercoagulability, they could suggest increased local intravascular coagulation within the placenta in intrauterine growth retardation and disseminated intravascular coagulation in pre-eclampsia.
Thrombin Time Dilution Test: A Simple Method to Control Heparin Therapy
Authors: Pizzuto J. , Reyna M.P. , García-Méndez S. , Morales M.R.
Published: May 2019

A modification of thrombin time (TT) is described, allowing quantitative measurement of plasma heparin activity during therapy. When heparin-containing plasma shows prolonged TT, TT is corrected by diluting with platelets rich plasma (PRP) thus finding the adequate heparin activity; the latter is estimated when prolonged TT of test plasma is corrected by PRP in the proportion between 1:4 and 1:8 (±20%). Results of 30 PRP samples studied in vitro by adding different amounts of heparin showed that samples diluted 1:4 and 1:8 did not correct prolonged TT (P<0.05) when they contained respectively, more than 0.2 or 0.3 U of heparin/ml. Therefore within these levels of heparin one should place the adequate dosage of heparin when using this method. Patients treated with 100 U of heparin/Kg every 6 h were thus studied: 52 without previous coagulation defects and 22 with disseminated intravascular coagulation (DIC). The results in the first group showed good heparin effect in 29 cases, overdosage in 12 and un derdosage in 11. Hemorrhage ocurred in 5 of the overdosage group. In the cases of DIC, 4 with underdosage of heparin did not improve; in 11 out of 18 with adequate levels of heparin, both hemorrhage and coagulopathy dissapeared and the other 7 who had more severe complications did not improve. To sum up, the diluted TT is a very useful test to assess heparin therapy even in patients with previous coagulation abnormalities.
Heparin in Acute Myocardial Ischemia: Dose and Source Dependent Effects
Authors: Saliba M.J.
Published: May 2019

Because conventional anticoagulant doses of nonspecified source heparin used in myocardial ischemia had produced controversy, studies with larger doses of heparin from intestine and lung source were initiated. In dogs, control occlusion for 20 minutes of the left anterior coronary artery with epicardial electrocardiographic recordings, was followed after one hour of reperfusion with a second occlusion plus a 60,000 unit heparin infusion. The ischemic S-T segment elevations on control occlusions (64.5 ± 8.5 mV)were lowered 84% in intestinal heparin added occlusions (10.4 ± 3.0, p < .00005). Lung heparin lowered S-T segments a lesser 36% (p < .008). The difference between intestinal and lung heparin was significant (p < .0007). Myocardial adenosine triphosphate levels at 90 minutes of occlusion were 31% higher in intestinal heparin dogs than in lung (p < .01). At 24 hours, myocardial necrosis was 32% less, and myocardial creatine Phosphokinase levels were 15% higher, in dogs with 110,000 total units of intestinal heparin added, than in control dogs. Lung heparin has not been tested. Thus, large dose of both heparins significantly modified myocardial ischemia, and intestinal was significantly better than lung. Lung and intestinal heparin are probably functionally and structurally different. Then, in 18 acute myocardial infarction patients, anticoagulant doses of intestinal heparin significantly lowered S-T segments, but did not clearly improve cardiac enzyme levels. Optimal human heparin dose is probably larger than conventional anticoagulant dose. These studies may explain partially the controversy regarding heparin use in myocardial ischemia, and through increased dose of specific source heparin produce new therapy. They may eplain other paradoxical findings with heparin in other studies. More animal studies seem indicated prior to large dose heparin use in humans.
Membrane Glycoprotein Loss from Circulating Platelets: Inhibition by Dipyridamole and Aspirin
Authors: George J.N. , Lewis P.C.
Published: May 2019

Platelet senescence during circulation has been suggested by evidence for the decreased size and hemostatic effectiveness of older platelets. We previously reported that when platelets are labeled simultaneously with [125I]-diazodiiodosulfanilic acid (I), which reacts with surface exposed plasma membrane glycoproteins, and 51cr, which reacts with soluble cytoplasmic material, the disappearance of I is more rapid than Cr (J Lab Clin Med, 88:247, 1976). We postulated that platelet surface material may be lost during reversible contact interactions in the process of hemostasis. To test this hypothesis we studied the survival of doubly labeled platelets in 5 rabbits treated with dipyridamole (D) (5 mg/kg IV at 0.5, 3 and 23 hours after infusion of labeled platelets) and aspirin (ASA) (25 mg/kg IV at 0.5 hours after infusion) in comparison with 13 control rabbits (untreated or given the drug diluent fluids in the same regimen). The diluent fluids had no effect. D and ASA had no effect on Cr disappearance but prolonged the disappearance rate of I so that it was equal to Cr. In control rabbits, the T½ of Cr (33.3 + 7.9 (SD) hours) was greater than I (19.5±3.6, ± < 0.001). In treated rabbits, the T½ of Cr (36.0 ± 5.3) was not different from that of I (30.8 ± 8.1, p > 0.2). The T½ of both Cr and I in the treated rabbits were greater than that of I in the control rabbits (p < 0.001) but not different from the T½ of Cr in the control rabbits (p > 0.5). We conclude that platelet surface glycoprotein loss is a continuous process in the normal circulation, possibly related to the changes of platelet senescence. The effect of D and ASA suggests that the membrane glycoprotein loss may occur during hemostatic encounters.
Movements of Calcium Ions and Their Role in the Activation of Platelets
Authors: Massini P. , Käser-Glanzmann R. , Lüscher E.F.
Published: May 2019

The increase of the cytoplasmic Ca-concentration plays a central role in the initiation of platelet activation. Four kinds of movements of Ca-ions are presumed to occur during this process: (a) Ca-ions liberated from membranes induce the rapid shape change. (b) Vesicular organelles release Ca-ions into the cytoplasm which initiate the release reaction, (c) The storage organelles, called dense bodies, secrete their contents including Ca-ions to the outside during the release reaction, (d) At the same time a rearrangement of the plasma membrane occurs, resulting in an increase in its permeability for Ca-ions as well as in an increase in the number of Ca-binding sites.

Since most processes occurring during platelet activation are reversible the platelet must be equipped with a mechanism which removes Ca-ions from the cytoplasm. A vesicular fraction of platelet homogenate indeed accumulates Ca actively. This Ca-pump is stimulated by cyclic AMP and protein kinase; it might be involved in the recovery of platelets after activation.
Genetic Variants of Hemophilia B.
Authors: Roberts H.R. , Chung K.S. , Goldsmith J.C.
Published: May 2019

The purpose of this report is to describe genetic variants of hemophilia B. Most variants have been distinguished on the basis of clinical severity of their disease, as well as immunological, functional, and biochemical characterization of the Factor IX molecule. They have been classified according to the degree of cross-reactivity of the Factor IX molecule with specific homologous and heterologous anti-Factor IX antibodies using both inhibitor neutralization and radioimmunoassay techniques. Some hemophilia B variants have a Factor IX molecule that cross-reacts completely with anti-Factor IX antibodies. Other hemophilia B patients have a Factor IX molecule that has no detectable cross-reaction with anti-Factor IX antibodies. Still other hemophilia B patients show reduced levels of cross-reacting material. We have extensively studied one hemophilia B variant with a Factor IX molecule that shows complete cross-reaction with anti-Factor IX antibody. The Factor IX from this patient has been isolated and has been shown to have 5% clotting activity and 100% antigenic activity. This variant Factor IX molecule shows delayed activation in the presence of partially purified Factor XIa and Ca2+ although it is otherwise similar to normal human Factor IX in terms of molecular weight and the number of α-carboxyglutamic acid residues per molecule. Other Factor IX variants have also been characterized and will be discussed. For example, hemophilia BM variants, unlike the usual type of hemophilia B, show prolonged ox-brain prothrombin times. These patients have an abnormal Factor IX molecule that is recognized antigenically, but not functionally.
Symposium on Antithrombin III: Introductory Remarks.
Authors: Shapiro S.S.
Published: May 2019

Interest in Antithrombin III has increased greatly since Abildgaard’s isolation of this protein and his demonstration that it is identical with the heparin cofactor. It is now clear that Antithrombin III plays a central role as an inhibitor of many serine proteases, including activated coagulation factors, kallikrein and plasmin. Heparin enhances the neutralizing activity of Antithrombin III towards each of these enzymes, from several-fold to more than 1,000-fold. Although Antithrombin III represents only a small fraction of the total protease neutralizing capacity of human blood, variation in the level of this protein seems to correlate with some clinical thromboembolic or “hypercoagulable” states. Variations in Antithrombin III levels occur regularly in several disease states, including disseminated intravascular coagulation. Heterozygosity for Antithrombin III is associated, in adult life, with a distinct tendency to thrombosis.

This symposium will be concerned with methods of measuring Antithrombin III and the correlation of its levels with disease states, the mechanism of action of this inhibitor and the role of heparin in its enhancement, and the thromboembolic tendency seen in hereditary Antithrombin III deficiency.
The Involvement of Factors XII & XI in the Platelet Coagulant Activity
Authors: Connellan J.M. , Bowden S. , Castaldi P.A.
Published: May 2019

Platelets provide coagulant activities, other than the Phospholipid Platelet factor 3, which can be released from the platelet. In Platelet Poor Plasma, the initiation of the intrinsic pathway is proposed to proceed via factors Xll & XI as well as 2 other factors, Prekallekrein and Kininogen. The role of these factors in the initiation of coagulation in Platelet Rich Plasma is under investigation. Walsh, (Br.J. Haemat. 22. 393, 1972) and Muntz (Haemostasis in press 1977), have shown that platelets devoid of factor Xll will still produce a coagulant activity when Platelet Rich Plasma is incubated with collagen. However, we have recently shown that this reaction requires the presence of factor XI. Factor XII has been purified by the method of Saito (Circ.Res.34,641,1974) , and factor XI has been purified by the method of Connellan, Castaldi and Muntz. (Haemostasis, in press 1977).

Specific antibodies were developed in rabbits and the IgG from the antiserum was coupled to Sepharose. These factors could be extracted from plasma by the immobilized antibodies and could then be eluted by the use of alkali or guanidine HCL, thus resulting in a rapid purification of these factors.

Platelets were washed free of loosely bound clotting factors and provided significant amounts of coagulant activity when ruptured by either freezing and thawing or by glycerol treatment. Platelets from a factor XI deficient patient also provided this activity. The platelet activity was extracted by passage through both Sepharose-heparin column and Sepharose-anti-factor XI column;the activity appeared to be identical to plasma factor XI.
Identification of a Prothrombin Derivative in Human Urine
Authors: Bezeaud A. , Aronson D. , Ménaché D. , Guillin M.C.
Published: May 2019

Urine samples of 13 normal subjects and 18 patients with constitutional coagulation defects were screened for the presence of prothrombin derivatives using specific rabbit antisera to human prothrombin, prothrombin Fragment 1 and Prethrombin 2. Patients comprised 15 severe and one moderate factor VIII deficiency, one factor VII deficiency and one mild factor XI deficiency. Immunological techniques included immunodiffusion, counter electrophoresis, Immunoelectrophoresis, crossed Immunoelectrophoresis and electroimmunoassay.

Antiprothrombin antiserum reveals in normal urine a prothrombin derivative which is in complete identity with Fragment 1, in partial identity with prothrombin and in non identity with Prethrombin 1. Other characteristics such as adsorption onto tricalcium phosphate and electrophoretic mobility are also those of Fragment 1. Similar results are obtained in the urine of patients with contitutional coagulation defects. Trace amounts of another prothrombin related antigen are also shown following concentration of normal urine.

These results indicate that prothrombin Fragment 1 is a constituent of human urine, whether this constituent appears following glomerular filtration of a plasmatic Fragment 1 or following tubular degradation of prothrombin is not known.
The Induction of Platelet Aggregates in Healthy Subjects by Venousocclusion
Authors: Chamone D.A. F. , Vermylen J.
Published: May 2019

Circulating platelet aggregates have been observed in various clinical conditions (Wu and Hoak, Lancet, 1974, ii, 924). Using a slightly modified method, we have found that platelet aggregates can be induced in vivo in healthy subjects.

Nine volunteers (7 males, 2 females, age 23-38 years) were studied. Blood was drawn from an antecubital vein of one arm immediately before and of the other arm after twenty minutes of occlusion midway between systolic and diastolic pressure. The ratio of the platelet count in platelet-rich plasma (PRP) obtained from blood collected on forma lin-EDTA to that from blood collected on EDTA only was 0.934 + 0.028 (mean ± S.E .) before and 0.768 ± 0.033 after occlusion (p < 0.001 ). Spontaneous aggregation in PRP, measured as percent increase in light transmission during 10 minutes of stirring in the a gg re gome ter, was 4 .20 ± 1.17 before and 3 .80 + I .69 after occlusion (p > 0 .1).

This system may help elucidate some of the mechanisms involved in the generation of circulating platelet aggregates. It may also constitute a simple set-up for the in vivo evaluation of drugs affecting platelet function.
Collagen-Induced Reduction in Rat Circulating Platelet Count: Influence of Platelet Function Inhibitors
Authors: Holmes I.B.
Published: May 2019

The effect on circulating platelet count of repeated intravenous infusions of collagen fibrils was measured in male OFA Sprague-Dawley rats (400-550 g). Citrated blood was pumped from the left carotid artery of anaesthetized animals, via a siliconized double-lumen cannula, into the manifold of a Technicon Autocounter, for continuous registration of platelet count. Native collagen fibrils (Collagenreagent ‘Horm’) were infused intravenously for 1 min at 15 min intervals. Successive increasing collagen doses (20-320 pg/kg) induced dose-dependent reduction in platelet count, measured as absolute platelet number disappearing from the circulation. Repeated infusion of collagen 160 pg/kg produced constant, partially reversible, reduction in platelet count. Several known inhibitors of platelet aggregation were investigated in the described test system. Collagen effects were inhibited in a dose-dependent manner to a maximum of 50-60 %, and drug activity was thus quantitated on the basis of dose producing 30 % inhibition (ID30): prostaglandin E1 (1.6 pg/kg/min i.v. infusion), SH-869 (1.1 mg/kg i.v.), aspirin (33.1 mg/kg p.o.), proquazone, a new non-steroidal antiinflammatory compound (5.0 mg/kg p.o.). That part of the collagen response not inhibited might be attributed to the initial phase of platelet adhesion to collagen, known to be relatively refractive to platelet function inhibitors.
Platelet and Fibrin Deposition on Subendothelium: Opposite Dependence on Blood Shear Rate
Authors: Baumgartner H.R.
Published: May 2019

The rate of platelet deposition on subendothelium (adhesion + adhesion-induced aggregation) from anticoagulated blood is shear rate (~ v/d) dependent and thus increases with increasing blood flow velocity (v) and decreasing vessel diameter (d). To investigate concomitant fibrin deposition, subendothelium of rabbit aorta was exposed to native blood under controlled flow conditions (Schweiz. Med. Wschr. 106, 1367 (1976)). Native blood was circulated by a roller pump for 3 min from a carotid artery through an annular perfusion chamber (maintained at 37°c) into a jugular vein of a rabbit at flow rates of 5, 20 and 40 ml/min, producing wall shear rates at the exposed subendothelial surface of 500, 2000 and 4000 s-1, respectively. Subendothelium and adhering blood elements were immediately fixed by perfusion of glutaraldehyde avoiding any blood-air interphase. Platelet interaction with subendothelium and fibrin deposition were determined morphometrically. At a shear rate of 500, 2000 and 4000 s-1, 23±3, 43±4 and 66±3% (mean ± SE) of the surface were covered with platelets and 67±12, 26±12 and 10±4% with fibrin, respectively. The corresponding values for adhesion-induced aggregation were 12±3, 38±6 and 56±5% in native and about 4, 17±5 and 29±4% in citrated (15 mM) blood indicating that even a low citrate concentration inhibits adhesion-induced aggregation. Similar results were obtained with human blood drawn from a cubital vein.

This is direct evidence that fibrin deposition on subendothelium predominates at low (veins), platelet adhesion and aggregation at high (arteries + small vessel) shear rates.
Properties of Plasminogen Activators in Circulating Human Blood and Human Cadaveric Veins
Authors: Mackie Michael , Bennett Bruce , Ogston Derek
Published: May 2019

The properties of 3 partially purified plasminogen activators prepared from human blood have been compared with activator prepared from cadaveric veins and urokinase.

Circulating plasminogen activator present in blood after venous occlusion of the arm has been partially purified by gel filtration on Sephadex G-200 in phosphate buffer containing 0.15 M NaCl followed by gel filtration of active fractions on Sephadex G-200 in buffer containing 1 M NaCl.

Plasminogen activator generated from lysine-sepharose treated plasma after activation of Hageman factor by kaolin has been partially purified by elution of the activator from kaolin by 0.5 M NaCl and gel filtration on Sephadex G-200; the activator precipitated by dextran sulphate from the euglobulin supernatant prepared from lysine-sepharose treated plasma has been partially purified by similar gel filtration techniques.

Cadaveric plasminogen activator has been prepared by ammonium sulphate precipitation of the washout from cadaveric veins followed by sequentail filtration on Sephadex G-200 in (a) low and (b) high concentrations of NaCl. The properties of these 4 plasminogen activators are compared with those of urokinase in terms of their stability, susceptibility to protease inhibitors and molecular size.
The Effect of Prostaglandins G2, D2, E2, E1 and Arachidonic acid on Thrombus Formation in Vivo
Authors: Westwick J. , Lewis G.P.
Published: May 2019

Arachidonic acid (AA) and prostaglandin (PG) G2 have been shown to he precursors of both pro-aggregatory and anti-aggregatory agents in vitro. If PGG2 is produced in thrombotic and inflammatory situations, it is important to know its effects on thrombus formation in vivo. Mural thrombus formation was induced in the arterioles (40-70 μm) of the hamster cheek pouch by combining micro-electrical damage with perivascular application of ADP (10-6M).

PG or vehicle was applied perivascularly, followed 30 sec and 1 min later by electrical micro-damage and application of ADP (lOM). The vessel was observed and thrombus formation was quantitated by timing the adherence of thrombi for the following 10 nriruEach animal served as its own control and results were expressed as % difference (mean - s.e.) from control. PGGs, AA and PGE]_ produced a dose-related (12.5 - 1250 ng) inhibition (lO ± 8% - 90 ± 15%) of thrombus formation.

Both PGG2 (Lewis, Vestwick & Williams, Br.J.Pharmac., 1977, in press) and AA induce a short-lasting vasoconstriction followed by vasodilatation. However, another potent vasodilator, PGE1, in a low Jose (125 ng) potentiated (49 - 20%) while high doses (1250 ng) produced a weak inhibition (15 ± 10%) of thrombus formation. PGD2 had little activity up to a concentration of I25O ng.

These results demonstrate that AA and PGG2 can be converted to anti-thrombotic agents in vivo when applied perivascularly. Since PGD5 and PGE2 were not anti-thrombotic, it is possible that the observed effect was due to generation of prostacyclin.
Degradation of Sulphated Glycosaminoglycans at the Surface of Cultured Human Endothelial Cells by a Platelet Heparitinase
Authors: Busch C. , Glimelius B. , Wastesson Å , Westermark B.
Published: May 2019

The non-thrombogenic property of the endothelial cell surface is a prerequisite for maintainance of blood circulation. The nature of this property is poorly understood. Recent advances in culturing techniques of endothelial cells in vitro may facilitate studies of the surface biochemistry. Human endothelial cells (EC) isolated from umbilical veins were shown to synthesize and secrete sulphated glycosaminoglycans (GAG). The recent finding of a platelet enzyme capable of degrading heparin sulphate (HS) raised the question:Can platelet lysate or a purified heparitinase detach and degrade endothelial HS? EC cultured in the presence of 35S-sulphate, produce 35S-labelled GAG which was isolated from the incubation medium from a cell associated trypsin labile pool and from a cellular pool not liberated by trypsin. After 48 hours of incorporation about 95% of 35S-GAG was found in the medium fraction, 5% in the trypsin fraction and negligible amounts in the cell fraction. In the trypsin pool (“surface fraction”) heparin sulphate comprised about 85%, while the remaining 15% consisted of chondroitin sulphate and/or dermatan sulphate. Incubation of 35S-labelled EC with platelet lysate or a partially purified preparation of the enzyme from the same source caused a marked release of cell-surface associated HS to the incubation medium as oligosaccharides. These effects could be ascribed to heparitinase activity and may alter the properties of the EC-surface and influence the interaction between these cells on one hand and blood cells or plasma components, e.g., coagulation factors on the other.
Hemostatic Factor Consumption in Tumor Dogs and Response of Platelet Function Inhibitors
Authors: Slichter S.J. , Weiden P. , Storb R.
Published: May 2019

Twenty dogs with a variety of untreated naturally occurring tumors [lymphoma (6), soft tissue sarcomas (6), adenocarcinoma of the prostate (2), mast cell tumor (2), malignant melanoma (2), osteogenic sarcoma (1) and intraductal breast CA (1)] hadCr labeled platelet and 125I fibrinogen survivals measured to determine the effects of malignancy on the hemostatic mechanism. Seven dogs with limited tumors had platelet survivals of 5·5 days ±0.6 (normal 5.6 days ± 0.6) and fibrinogen survivals of 2.9 days ± 0.5 (normal 2.7 days ± 0.4). In three dogs who died of extensive disease, platelet survivals averaged 1.8 days ±0.6 and fibrinogen survivals 1.2 days ± 0.8. Platelet and fibrinogen turnovers were both 2.5 x normal. In 10 dogs with intermediate disease who lived long enough to be studied serially, baseline platelet survivals averaged 2.8 days ± 1.4 and fibrinogen survivals 2.5 days ± 0.6. Since platelet survivals were disproportionately reduced in this group, antiplatelet drugs were given (dipyridamole, SH 1117 or RA 233). Repeat studies showed a prolongation of platelet survival in 5 of 10 animals studied (4.0 days ± 1.0). Responders were 3/5 sarcomas, 1/4 lymphomas and 1/1 adenocarcinoma of the prostate. These data suggest that consumption of clotting factors is related to the extent of the underlying malignancy and response to platelet function inhibitors is governed by the type of malignancy. Since platelet and fibrinogen consumption have similarly been found in metastatic human tumors (Ann. NY Acad. Sei. 230:252, 1974), this animal model may represent an opportunity to explore the value of antithrombotic therapy in maintaining hemostatic factors and preventing thromboembolic complications.
Plasma and Platelet Antiplasmins: Radial Diffusion in Fibrin(OGEN)
Authors: Andes W.A.
Published: May 2019

An antiplasmin assay has been developed utilizing radial diffusion of plasma from wells cut in plasmin-enriched, fibrinogen-agarose plates. After diffusion the fibrinogen is clotted. Zones of fibrin protected from background fibrinolysis develop as the result of plasma antiplasmin activity. The sizes of the fibrin antiplasmin zones are proportional to the concentration of plasma placed in the wells. A pooled plasma standard was taken to contain 100% antiplasmin activity. Antiplasmin activity of 52 normal subjects varied from 64 to 132%. Washed platelets (bovine albumin gradient) contained 1-5% antiplasmin activity. Using antisera to precipitate individual inhibitors, physical methods of separation, and electrophoresis of plasma in agarose, several different proteins were found to have antiplasmin activity. Thus α1-macro-globulin contributed 56%, α1-antitrypsin 20%, antithrombin III 2%, and other proteins 22% of the total antiplasmin activity.1 One millilitre of whole plasma neutralized 7.0 CTA units of plasmin. Using this assay, patients with haemophilia treated with fibrinolytic inhibitors had significantly increased antiplasmin levels.
Determination of a Fast Acting Plasmin Inhibitor in Plasma from Patients with Tendency of Thrombosis and Increased Fibrinolysis
Authors: Teger-Nilsson A-C. , Gyzander H. , Myrwold H. , Noppa H. , OLsson R. , Wallmo L.
Published: May 2019

Antiplasmin activity has been determined by means of the plasmin specific trlpeptlde substrate H-D-Vai-L-Leu-L-Lys-p-nltroanlllde (S-2251)It Is concluded that the method mainly measures the antiplasmin described by Collen and coworkers ( Thromb. Res. 7, 245, 1975), although α2-macroglobulln may interfere to a small extent. Different categories of patients with tendency to thrombosis and Increased fibrinolysis have been studied During normal mlddlesized operations antiplasmin decreased slightly, but increased postoperatively. Antiplasmin likewise decreased during deliveries, and Increased In th PuerperiumPatients with acute deep vein thrombosis had normal antiplasmin activity at the time for diagnosis, but antiplasmin Increased later during the course of eventsStreptokinase treated patients showed a pronounced and rapid drop In antiplasmin activity, which was normalized as soon as the streptokinase administration was InterruptedPatients with liver cirrhosis had lower antiplasmin activity than normals , and so had patients with disseminated Intravascular coagulation.