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Biomedical subjects

H R Gralnick

Publications and source records attributed to H R Gralnick.

At least 91 records · Page 5Linked to original sources
PubMed1985

Benefits of danazol treatment in patients with hemophilia A (classic hemophilia).

Danazol, an attenuated androgen, was given intermittently in three patients with hemophilia A. The rise in factor VIII activity with danazol treatment was associated with a fivefold decrease in hemorrhagic episodes and plasma concentrate product utilization when compared with similar periods without the drug. Four other patients with moderate hemophilia A who received danazol for 14 days had an increase of 400% to 850% in their factor VIII levels. Of four patients without detectable levels of factor VIII who were treated with danazol, two had elevations of the factor VIII level, to 3% and 4%, respectively. Danazol appears to be an effective treatment for patients with hemophilia A.

Adult↗
PubMed1985

Normal human blood density gradient lymphocyte subset analysis: I. An interlaboratory flow cytometric comparison of 85 normal adults.

An interlaboratory flow cytometric comparison of several commercially available human lymphocyte subset reagents was undertaken in three different laboratories. Fresh Hypaque-Ficoll purified blood mononuclear cells were stained at 4 degrees C or 22 degrees C. Direct or indirect surface immunofluorescence was carried out at all sites using an EPICS V flow cytometer. Fullbright, 10-micron fluorescent polystyrene microspheres were used for optical alignment and standardization. A log integral fluorescent histogram gated on forward and right angle scatter was collected on 1-2 X 10(4) cells for each reagent and the proportion, of positive cell determined for each reagent. With the exception of one reagent, anti-B1, which showed an approximately twofold variation, all three laboratories showed remarkable agreement. Thus there was no significant difference noted for the following reagents: OKT4, CCT4, Leu 3a, Leu 2a, OKT8, or CCT8. We attribute these findings to the availability of quality reagents, precision instrumentation, and a standard lymphocyte preparation.

Adult↗
PubMed1985

Abnormal coagulation results in patients with Hodgkin's disease.

Case records of 177 patients admitted with Hodgkin's disease were reviewed to assess the frequency and significance of coagulation abnormalities. Prolongation of the prothrombin time, activated partial thromboplastin time, or thrombin time occurred in 56 patients, 32 percent of all evaluable cases. The most frequent clotting abnormalities involved the prothrombin time, which was increased in 43 patients (24 percent). Prothrombin time prolongation correlated with bulky or advanced disease as defined by stage (p = 0.001), constitutional symptoms (p less than 0.0001), massive mediastinal involvement (p = 0.02), and elevated alkaline phosphatase levels (p less than 0.0001). Abnormal coagulation test results followed the course of disease, normalizing with tumor regression and reappearing during relapse. Despite the surprising incidence of abnormal coagulation results, bleeding complications were reported in only two cases. Patients undergoing invasive procedures in the presence of clotting abnormalities fared no worse than those in whom procedures were cancelled. There is no evidence that complete staging evaluation should be comprised because of these abnormal test values. Extensive hematologic testing revealed no single mechanism to explain the coagulation factor disorders found in Hodgkin's disease.

Adult↗
PubMed1985

In vitro correction of the abnormal multimeric structure of von Willebrand factor in type IIa von Willebrand's disease.

Type IIa von Willebrand's disease (vWd) has been characterized by the absence of the largest and a reduction in the intermediate-sized multimers of the plasma and platelet von Willebrand factor (vWf) and by the diminished response of the platelet-rich plasma of these patients to ristocetin. Other recently demonstrated abnormalities include the presence of an abnormal triplet structure of vWf. We have studied the plasma and platelets from three patients with this form of vWd and have found that both their plasma and platelets manifest the previously described abnormalities. Because of the heterogeneity of the multimeric structure of the vWf in these patients, we considered the possibility that postsynthetic events may have modified the vWf. When blood was collected in 5 mM EDTA or 5 mM EDTA/leupeptin/N-ethylmaleimide, the abnormal multimeric structure of the plasma and platelet vWf was partially normalized in that the intermediate and the largest vWf multimers were increased, the abnormal multimer structure was no longer as apparent, and the fastest migrating band (an abnormality seen only in the type IIa vWd plasma and platelets) disappeared. The enzymatic activity responsible for this degradation can be classified as a calcium-dependent protease. Studies of normal radiolabeled vWf incubated with platelet lysates from normal subjects and these patients revealed that the patients' platelets did not contain increased amounts of calcium-dependent protease activity as assessed by degradation of normal vWf. These data suggest that patients with type IIa vWd synthesize an abnormal vWf protein that is susceptible to in vitro proteolytic degradation and that proteolytic degradation can play a significant role in the phenotypic expression of vWd by modifying the plasma and platelet vWf multimeric structure.

Blood Coagulation Factors↗
PubMed1985

Inhibition of cold-promoted activation of the prothrombin time studies of new siliconized borosilicate collection tubes in normals and patients receiving warfarin.

Studies of the prothrombin time in normals and patients receiving oral anticoagulant therapy revealed a partial inhibition of the in vitro cold-promoted activation of Factor VII and shortening of the prothrombin time when blood was collected in a new commercial siliconized borosilicate tube (Becton-Dickinson, Rutherford, NJ, 6418 and 6419). The data collected with normal blood indicated that the cold-promoted activation was almost totally inhibited, while the patients had only a mean reduction of 50% of the cold-promoted activation. In pregnant patients there was no inhibition of the cold-promoted activation of the prothrombin time or Factor VII using the new siliconized tubes.

Blood Specimen Collection↗
PubMed1985

IgA inhibitor to factor VIII/von Willebrand factor.

A 60-year-old Black female presented with a haemorrhagic diathesis and an acquired factor VIII/von Willebrand factor (VIII/vWf) inhibitor. This inhibitor was classified as an IgA immunoglobulin and was active not only against factor VIII coagulant (VIII:C) activity but also against plasma von Willebrand factor (vWf). The purified IgA also interacted with normal platelets to inhibit ristocetin-induced platelet aggregation (RIPA). In contrast, studies with haemophilia A plasma and platelets revealed that the inhibitor did not react significantly with these plasmas or platelets. The significant differences in the inhibition of vWf assay both of the plasma and the platelets of the haemophilia A patients suggests that part of the haemorrhagic diathesis may be related not only to the inhibition of VIII:C but also to interference with platelet function. In addition, these studies suggest that there may be significant differences in the factor VIII-related antigen (VIII R:Ag) on platelets in haemophilia A patients compared to normal.

Blood Coagulation Factors↗
PubMed1985

Criteria for the diagnosis of acute leukemia of megakaryocyte lineage (M7). A report of the French-American-British Cooperative Group.

For the diagnosis of M7, the bone marrow aspirate shows a leukemic cell infiltrate that comprises 30% or more of all cells. These cells are identified as being of megakaryocyte lineage by the platelet peroxidase reaction on electron microscopy or by tests with monoclonal or polyclonal platelet-specific antibodies. Myelofibrosis or increased bone marrow reticulin are a prominent aspect in most patients with M7. In patients with increased reticulin, the bone marrow sample may be difficult to obtain and the counts done on the marrow films may be misleading. In these patients, the diagnosis of M7 should be based on excellent bone marrow biopsy sections that show an excess of blasts and, at times, increased numbers of maturing megakaryocytes; and on the presence of unequivocal megakaryoblasts in the peripheral blood or bone marrow (or both) as shown by immunologic techniques.

Acute Disease↗
PubMed1985

Effect of calcium on the availability of platelet von Willebrand factor.

The binding of a polyclonal immune-purified rabbit anti-human von Willebrand factor (VWF) antibody to activated and nonactivated normal human platelets was studied in the presence and absence of extracellular calcium. Extracellular Ca++ in the incubation buffer results in no increase in the amount of antibody bound to the surface of unstimulated platelets when compared with that bound in the absence of Ca++. Alpha thrombin stimulation of the platelets increases antibody binding to the platelets in both the presence and absence of extracellular Ca++; however, the increase is approximately four times as great in the presence of extracellular Ca++. The amount of VWF detected in the incubation supernatants of unstimulated platelets is similar in either the presence or absence of Ca++, and this amount is increased without demonstrating a Ca++ dependency after thrombin stimulation. Measurement of thrombin-induced beta thromboglobulin (beta TG) release reveals some parallel between anti-VWF antibody binding and beta TG release. However, the amount of beta TG release after thrombin stimulation is independent of extracellular Ca++, whereas the amount of bound anti-VWF is markedly increased in the presence of Ca++. These studies demonstrate that thrombin induces an increase in the surface exposure of platelet VWF even in the absence of extracellular Ca++. The data also suggest that there may be a quantity of platelet VWF that can become exposed on the platelet surface by an action of thrombin that is independent of the Ca++-dependent granule release reaction induced by thrombin.

Animals↗
PubMed1984

Acute megakaryoblastic leukemia in childhood.

Routine morphologic and cytochemical study of the leukemic cells of a 13-month-old child did not permit definitive determination of either a lymphoid or a myeloid cell origin. However, ultrastructural cytochemical analysis revealed platelet peroxidase (PPO) reactivity. The malignant cells expressed PPO in both unfixed and fixed preparations. PPO was observed in the perinuclear space and reticulum but not in the Golgi saccules. In a 1% glutaraldehyde, 1% formaldehyde solution known to inhibit PPO but not myeloperoxidase, no reaction product was observed. The demonstration of PPO activity in these undifferentiated cells established their megakaryoblastic lineage. Further studies including DNA flow cytometry confirmed that these cells were megakaryoblasts. This study demonstrates that megakaryoblastic malignancy may occur as an acute leukemia of childhood and emphasizes the value of ultrastructural cytochemistry in cases of acute leukemia which defy routine classification. Abnormalities of chromosome 21 were present in our patient as in four previous cases of megakaryoblastic leukemia in childhood. The nonrandomness of chromosome 21 involvement in this disease should therefore be considered.

Acute Disease↗
PubMed1984

Circulating heparan sulfate anticoagulant in a patient with a fatal bleeding disorder.

We have identified a circulating, heparin-like anticoagulant in a patient with multiple myeloma (IgG4 lambda) who had serious clinically evident bleeding that contributed to his death. Purification of the patient's circulating coagulation inhibitor was accomplished by ammonium sulfate concentration, anion exchange chromatography, and affinity chromatography on protamine sulfate. Analysis of the purified inhibitor showed that it was a proteoglycan that comigrated with heparan sulfate on lithium acetate-agarose-gel electrophoresis and that it contained 39 per cent L-iduronic acid. Control samples of heparan sulfate and heparin contained 29 and 68 per cent L-iduronic acid, respectively. Functional coagulation studies revealed that the purified inhibitor had cofactor activity with antithrombin III that could be abolished by prior incubation with protamine sulfate or platelet factor 4. Recognition of the existence of this or of other similar inhibitors in bleeding patients is important because of the potential for treatment with agents such as protamine sulfate and platelet factor 4, which neutralize the anticoagulant effects of proteoglycans.

Blood Coagulation↗
PubMed1984

Inhibition of the cold activation of Factor VII and the prothrombin time.

Normal whole blood, collected and stored in borosilicate or commercial siliconized borosilicate tubes at 4 degrees C, undergoes a time-dependent activation of Factor VII and a shortening of the prothrombin time (PT). Addition of inhibitors to Factor XII fragments (corn Hageman factor inhibitor, CHFI) or to activated Factor XII (Cytochrome-C) inhibited in a concentration-dependent way the cold-promoted activation of Factor VII and the shortening of the PT. When celite was added to whole blood in polypropylene tubes at 4 degrees C, Factor VII was activated and the PT shortened. Preincubation of celite with CHFI or Cytochrome-C prevented the activation of Factor XII and the subsequent activation of Factor VII and the PT. These studies demonstrate the importance of Factor XII in the cold activation of Factor VII and shortening of the PT and indicate that inhibitors to activated Factor XII or XII fragments are useful in inhibiting in vitro shortening of the PT. These findings suggest that the development of a collection tube that prevents contact activation also would inhibit adequately the cold-promoted activation of the PT and Factor VII.

Aprotinin↗
PubMed1984

Fibrinogen competes with von Willebrand factor for binding to the glycoprotein IIb/IIIa complex when platelets are stimulated with thrombin.

Two monoclonal antibodies--one that blocks ristocetin-induced platelet binding of von Willebrand factor to glycoprotein Ib and one that blocks adenosine diphosphate-induced binding of fibrinogen to the glycoprotein IIb/IIIa complex--were used to assess the binding site(s) for von Willebrand factor when platelets are stimulated with thrombin or adenosine diphosphate (ADP). Neither agonist induced binding of von Willebrand factor to glycoprotein Ib. ADP and thrombin induced von Willebrand factor binding exclusively to the glycoprotein IIb/IIIa complex. The results of the site of binding of von Willebrand factor with thrombasthenic platelets were consistent with the data obtained with the monoclonal antibodies and normal platelets. Human fibrinogen caused complete inhibition of thrombin-induced von Willebrand factor binding to normal platelets at concentrations considerably below that found in normal plasma. We conclude that thrombin induces very little binding of exogenous von Willebrand factor to platelets at normal plasma fibrinogen levels.

Adenosine Diphosphate↗
PubMed1984

The oligosaccharide of human thrombin: investigations of functional significance.

Functional properties of the carbohydrate chain of human thrombin were examined by quantitating the activity of the enzyme before and after partial removal of its oligosaccharide by exoglycosidases. The following activities were studied: fibrinogen clotting, factor VIII coagulant (VIII:C) activation, inhibition by defibrinated plasma (anti-thrombin-III and alpha 2-macroglobulin), binding to polymerized fibrin, and stimulation of platelet release and aggregation. In general, the published information about the activity of native thrombin in these interactions was confirmed, though differences were observed in the association constants describing thrombin binding to fibrin. Partial deglycosylation had no apparent effect on any of these activities. It is concluded, therefore, that the oligosaccharide of human thrombin is located outside the major protein and platelet-binding regions of the molecule.

Antigens↗
PubMed1984

Collagen-factor VIII/von Willebrand factor protein interaction.

Factor VIII/von Willebrand factor (FVIII/vWF) protein interaction with collagen was studied by incubating plasma or purified FVIII/vWF with purified type I fibrillar collagen. Collagen adsorbed FVIII/vWF activities in a similar time and concentration-dependent manner from normal plasma, plasmas from classical and variant type von Willebrand's disease (vWD), and from purified FVIII/vWF. Incubation with denatured collagen or fibrin, produced in the presence or absence of fibronectin, showed no adsorption of FVIII/vWF. Examination of the multimeric structure of the remaining unadsorbed FVIII/vWF protein by agarose gel electrophoresis and autoradiography showed that the largest multimers had been adsorbed to the collagen. Studies of the adsorbed FVIII/vWF protein when eluted from collagen showed that it complemented the alterations in multimeric structure observed in the supernatants following collagen exposure. The multimeric structure of normal plasma following collagen adsorption resembled that of unadsorbed type IIb plasma; however, the collagen-adsorbed normal plasma did not produce enhanced ristocetin-induced platelet aggregation ( RIPA ). This phenomenon, therefore, must not be due solely to absence of large multimers from type IIb FVIII/vWF protein. The adsorbed multimers of FVIII/vWF protein may act as a subendothelial collagen-platelet bridge to promote primary hemostasis.

Adsorption↗
PubMed1983

The carbohydrate of human thrombin: structural analysis of glycoprotein oligosaccharides by mass spectrometry.

The carbohydrate structure of human thrombin has been determined by direct probe mass spectrometry of the oligosaccharides released by trifluoroacetolysis from the asialo glycoprotein. The free oligosaccharides were studied as permethylated and N-trifluoroacetylated oligosaccharide alditols. The structure was confirmed by sequential exoglycosidase digestion of intact thrombin and sugar and methylation analysis of the oligosaccharides by gas-liquid chromatography-mass spectrometry. The results indicate the following structure: (formula; see text) with Fuc present on only about 50% of the oligosaccharides.

Carbohydrate Sequence↗