bluuebunny/crossref_metadata_2025 huggingface dataset:
Impaired factor X and prothrombin activation associated with decreased phospholipid exposure in platelets from a patient with a bleeding disorder
Authors: Rosing J , Bevers EM , Comfurius P , Hemker HC , van Dieijen G , Weiss HJ , Zwaal RF
Published: October 2019

Platelets from a platelet factor 3-deficient patient, which was first described by Weiss et al (Am J Med 67:206, 1979), were found to be equally impaired in their ability to promote factor X and prothrombin activation. Compared to normal platelets, the patient's platelets showed upon stimulation with thrombin plus collagen a much slower generation and a considerably lower level of platelet prothrombin- and factor X-converting activities. Treatment of stimulated platelets with phospholipases revealed a decreased exposure of negatively charged phospholipid at the outer surface of the patient's platelets, relative to control's. We suggest that the combined impairment of prothrombin- and factor X-converting activities in this patient is due to a defect in the mechanism by which phosphatidylserine becomes exposed at the outer surface of stimulated platelets.
Studies on Platelets
Authors: STEFANINI MARIO , CHATTERJEA JYOTI B. , DAMESHEK WILLIAM , ZANNOS LEDA , SANTIAGO ENRIQUE PEREZ , Salomon Lucy
Published: October 2019

1. Direct transfusions of platelet-rich polycythemic blood, using the multiple syringe technic and siliconized apparatus, were performed in 14 cases of "idiopathic" ("megakaryocytic") and 8 of "secondary" ("amegakaryocytic") thrombocytopenia. Effects on platelet number and function, and on hemorrhagic manifestations were studied.

2. Platelets disappeared very rapidly from the circulation (one-half to twelve hours) in the 12 patients with acute idiopathic thrombocytopenia and in 1 patient with chronic thrombocytopenia with a low platelet count. They disappeared more slowly (twenty-four hours) in the other case of chronic idiopathic thrombocytopenia in which there was a higher platelet count. Essentially similar platelet disappearance curves were obtained in 2 patients, before and after unsuccessful splenectomy. In the patients with secondary thrombocytopenia, the survival of platelets in the circulation was much longer (average forty-eight to ninety-six hours).

3. Behavior of clot retraction and prothrombin activity of serum paralleled closely that of the platelet count. The bleeding time and capillary fragility usually remained improved for twenty-four to forty-eight hours after the platelet count had fallen to the initial level. Likewise, the hemorrhagic manifestations were improved in every case for many hours after the platelet level had returned to its original value. Similar, but less marked beneficial effects on the hemorrhagic manifestations were also obtained with transfusions of fresh blood from donors with normal platelet counts, provided fresh blood was used and precautions taken to prevent excessive loss of platelets.

4. Remissions were observed in 2 patients with idiopathic thrombocytopenia following direct transfusion of polycythemic blood. In both cases, the injected platelets disappeared promptly from the circulation but within twenty-four hours the platelet count began to rise and a long-sustained remission became established. This finding suggests the possibility that polycythemic blood may contain a stimulatory principle which can determine or accelerate a spontaneous remission.

5. Conclusions. The prompt disappearance of injected platelets from the circulation of patients with "idiopathic" thrombocytopenia suggests a humoral destructive mechanism, which is not present in the "secondary" or "amegakaryocytic" cases. This destructive mechanism does not appear to originate in the spleen. Our observations do not exclude an inhibitory effect upon platelet production from the megakaryocytes of the bone marrow. Polycythemic blood may contain a platelet stimulatory factor.
Maintenance of Circulating Red Cell Volume in Rats after Removal of the Posterior and Intermediate Lobes of the Pituitary
Authors: VAN DYKE D. C. , GARCIA J. F. , SIMPSON M. E. , HUFF R. L. , CONTOPOULOS A. N. , EVANS H. M.
Published: October 2019

The anemia which follows hypophysectomy is apparently due to absence of the anterior lobe of the hypophysis as removal of the intermediate and posterior lobes did not change the hemoglobin concentration, hematocrit or volume of circulating red cells.
Correction for Plasma Trapped in the Red Cell Column of the Hematocrit
Authors: CHAPLIN H. , MOLLISON P. L.
Published: October 2019

1. In the hematocrit, the percentage of the red cell column occupied by trapped plasma increases directly with the packed cell volume; this is due to a corresponding reduction of the effective radius of centrifugation.

2. When P.C.V. is determined in Wintrobe hematocrit tubes spun at 3000 r.p.m. in a centrifuge of radius 15 cm., the variation in plasma trapping between one sample and another is much smaller after 55 minutes centrifugation than after 30 minutes. If a 55 minute period is used, variation is so small, even when pathologic bloods are tested, that it is satisfactory to use a calibration curve. Such a curve is presented.
Studies on the Radiosensitivity of Bone Marrow
Authors: PEARCE M. L. , GREENFIELD M. A. , VALENTINE W. N.
Published: October 2019

1. A method for the calculation of "birth rate" of erythrocytes is presented and the dynamics of erythrocyte regeneration are analyzed in terms of a two compartment system.

2. The regeneration of erythrocytes after hemorrhage in irradiated and nonirradiated cats is compared to predicted values.
Effect of Dextran on Blood Typing and Crossmatching
Authors: ROCHE PAT , DODELIN RICHARD A. , BLOOM WALTER L.
Published: October 2019

Partially hydrolyzed dextran used for the expansion of plasma volume has been shown to have no effect on typing, crossmatching, or Rh determination of blood from normal patients or patients in shock.
Erythremic Myelosis (Di Guglielmo's Disease)
Authors: SCHWARTZ STEVEN O. , CRITCHLOW JOAN
Published: October 2019

This article represents a critical attempt to survey the cumulative information of the rare syndrome ordinarily called Di Guglielmo’s disease. The historical background lies predominately in Europe, more particularly in Italy, where the recording of cases has been fairly extensive and where interpretation has been pioneered.

The present study does not presume to be definitive. It attempts to trace the thinking that has gone before, and to define the terms of reference so as not to compound nosologic confusion. It presents examples of cases in the literature acceptable diagnostically by definition. Criteria for the differential diagnosis of acute and chronic erythremic myelosis and erythroleukemia are offered. These three conditions are regarded as variants of reticulo-endotheliosis. This conclusion is based on what is known of the pathogenesis, and on what has been observed clinically and hematologically.

All evaluation is necessarily offered with the reservations that must be held regarding a rare disease of unknown etiology.
Adenine Nucleotide Content of Red Blood Cells of Normal Individuals and of Patients Suffering from Hodgkin’s Disease and Other Diseases
Authors: ROTTINO ANTONIO , HOFFMAN GEORGE T. , ALBAUM HARRY , KRISTAN JOSEPHINE
Published: October 2019

Levels of adenine nucleotides in the blood were determined, using an enzymatic spectrophotometric method.

The levels were determined for a large group of normal persons and a larger group of patients chronically ill from a variety of malignant and degenerative diseases.

The values for adenyl nucleotides were usually normal; a tendency to increased concentration occurred chiefly in anemia.

The varying degrees of increased concentration could not be related to the severity of the anemia or to physical alteration of the red cell. There was probably some relationship to an altered metabolism within the red cell—possibly glycolysis—which in turn might be linked to changes in concentration of electrolytes of the red blood cell, a fact observed in anemia.
Case Report: Possible Paroxysmal Nocturnal Hemoglobinuria with Pronounced Pancytopenia, Reticulocytopenia, and without Hemoglobinuria Simulating Aplastic Anemia
Authors: LETMAN HENNING
Published: October 2019

1. This paper describes a case of chronic severe anemia with very pronounced neutropenia and thrombocytopenia and a varying degree of reticulocytopenia. Crosby’s modification of the Ham test for paroxysmal nocturnal hemoglobinuria proved to be strongly positive. Despite a prolonged period of observation no unquestionable hemoglobinuria was found, but slight hemoglobinemia was found. As the serum bilirubin was normal and there was no reticulocytosis an ordinary hematologic examiation could not have revealed the hemolytic nature of the anemia.

Because of the pronounced pancytopenia and the lack of response to all therapy, the case would presumably have been classified as one of aplastic or "refractory" anemia.

2. The supposition suggests itself that other cases of aplastic or "refractory" anemia and of hemolytic anemia without hemoglobinuria are actually atypical forms of paroxysmal nocturnal hemoglobinuria. The hemoglobinuria in this disease is an immaterial symptom. It should be possible to reveal such atypical cases by means of Ham’s and Crosby’s tests.

3. Pancytopenia is seen not only in the so-called hypersplenic hemolytic anemia, in which the bone marrow is hyperplastic but in congenital hemolytic anemia during crisis and in certain cases of auto-immune acquired hemolytic anemia. It is therefore possible that other forms of hemolytic anemia may be cloaked by a picture of "aplastic anemia."

4. From the standpoint of therapy, it is naturally important to settle the question as to the true nature of the anemia. In congenital and in hypersplenic hemolytic anemia, splenectomy exerts a curative effective; in acquired hemolytic anemia caused by antibodies, treatment with ACTH and cortisone will be effective in many cases; and, finally, in PNH no therapy other than transfusions is presently available.
The Use of Thorium Dioxide Sol (Thorotrast) in the Roentgenologic Demonstration of Accessory Spleens
Authors: LOEB VIRGIL , SEAMAN WILLIAM B. , MOORE CARL V.
Published: October 2019

Experience with the use of thorium dioxide sol (Thorotrast) in the roentgen demonstration of accessory spleens is described. The importance of the application of this technic to the clinical management of patients with hematologic relapse following splenectomy is emphasized and illustrative case reports are presented.

Nine patients were given thorium dioxide in an attempt to demonstrate an accessory spleen. A remission was induced in 1 patient with acquired hemolytic anemia, who had failed to respond to splenectomy, by removal of an accessory spleen demonstrated with thorium dioxide. A second patient with hereditary spherocytosis who relapsed seven years after splenectomy was shown to have an accessory spleen. Seven patients with idiopathic thrombocytopenic purpura who had relapsed following splenectomy were given thorium dioxide and in no case was an accessory spleen found.
Methods: Blood Platelet Counts of the Golden Hamster, Cricetus auratus
Authors: OTTIS KENNETH , TAUBER OSCAR E.
Published: October 2019

Healthy, adult male and female golden hamsters, 3 months of age, showed blood platelet count means of 688,000 ± 141,000 per cu. mm. and 742,000 ± 120,000 per cu. mm., respectively, when direct counts were made with siliconized pipets and with Rees and Ecker fluid as a diluent.
Demonstration of a human epsilon-globin gene silencer with studies in transgenic mice
Authors: Raich N , Papayannopoulou T , Stamatoyannopoulos G , Enver T
Published: October 2019

The human epsilon-globin gene displays normal developmental regulation in transgenic mice; it is expressed only in embryonic and in definitive erythroid cells. We show here that deletion of a negative element located between -182 and -467 bp upstream of the epsilon-globin gene cap site results in continuation of epsilon gene expression in the definitive erythroblasts of the fetal liver and in the red blood cells of adult transgenic mice. These data provide direct in vivo evidence that cis acting silencing elements are involved in the developmental control of the epsilon-globin gene.
Platelet adhesion to laminin: role of Ca2+ and Mg2+ ions, shear rate, and platelet membrane glycoproteins
Authors: Hindriks G , Ijsseldijk MJ , Sonnenberg A , Sixma JJ , de Groot PG
Published: October 2019

The adhesion of platelets to purified laminin under flow conditions was investigated. Adhesion to laminin was strongly dependent on the presence of divalent cations. In the absence of cations platelet adhesion (8% coverage in 5 minutes) was maximal at a shear rate of 100/s and no adhesion could be detected at shear rates above 800/s. In the presence of 0.8 mmol/L Mg2+ and 2 mmol/L Ca2+ platelet adhesion reached its maximum (30% coverage) around 800/s. At 1,800/s platelets still adhered to purified laminin (coverage of 6%). Antibodies against the E8 domain of laminin and antibodies against the alpha 6 and beta 1 chains of platelet membrane glycoprotein very late activation antigen-6 (VLA-6), completely inhibited adhesion. No inhibition was found with antibodies against glycoprotein IIb:IIIa, against the alpha 2 chain of VLA-2, and against the alpha 5 chain of VLA-5. Fibronectin and von Willebrand factor were not involved in laminin-dependent adhesion. Anti- VLA-6 partly inhibited platelet adhesion to the extracellular matrix of endothelial cells at shear rates below 800/s. Preincubation of the matrices with antilaminin E8 antibodies did not influence the adhesion. These results show that purified laminin supports platelet adhesion and that the presence of VLA-6 is important for platelet adhesion under flow conditions. The protein in the matrix with which VLA-6 interacts is currently unknown.
Striking inverse association of IgG-anti-Fab gamma antibodies and CD4 cell counts in patients with acquired immunodeficiency syndrome (AIDS)/AIDS-related complex
Authors: Susal C , Daniel V , Oberg HH , Terness P , Huth-Kuhne A , Zimmerman R , Opelz G
Published: October 2019

The contribution of autoimmune phenomena to the pathogenesis of acquired immunodeficiency syndrome (AIDS) is poorly understood. We investigated the relationship between IgG-anti-Fab gamma autoantibodies and the main immunologic feature of AIDS, the decrease of CD4+ helper lymphocytes. Sera of 33 human immunodeficiency virus (HIV) infected (HIV+) hemophilia patients with AIDS/AIDS-related complex (ARC), 57 HIV+ patients without AIDS/ARC, 23 HIV-negative (HIV-) patients, and 76 healthy controls were tested for antibody activity against the Fab region of IgG. Patients with AIDS/ARC had significantly higher IgG-anti- Fab gamma activity than HIV+ patients without AIDS/ARC, HIV- patients, or controls (P less than .0001). A striking inverse association was found between IgG-anti-Fab gamma and CD4+ cell counts (r = -.69; P less than 10(-6)). Sequential testing in 16 AIDS/ARC patients showed that an increase in the IgG-anti-Fab gamma activity was invariably accompanied by a decrease in the CD4+ cell count. IgG-anti-Fab gamma antibodies may play an important role in the immunopathogenesis of AIDS.
The Function of the Eosinophile Leukocyte
Authors: VAUGHN JOHN
Published: October 2019

1. Eosinophilia has been produced experimentally in guinea pigs by the injection of extracts of Ascaris suum and histamine, the accompanying histologic changes, particularly the distribution of eosinophils, being noted.

2. The tissues of guinea pigs suffering from a natural eosinophilia have been studied and compared with those of experimental eosinophilia.

3. From these findings a path has been traced which, it is suggested, the eosinophil follows in its course through the body.

4. The function of the eosinophil has been discussed and the hypothesis advanced that its function is the carriage of histamine or a histamine-like toxic material for inactivation.

5. This viewpoint has been considered in relation to the occurrence in man of eosinophilia including eosinophilic infiltration of the lungs.
Demonstration of a Cytoplasmic Structure in Plasma Cells
Authors: BRAUNSTEINER H. , FELLINGER K. , PAKESCH F.
Published: October 2019

In normal and pathologic plasma cells of the bone marrow, a specific filamentous structure of the cytoplasm has been revealed by electron microscopy. The significance of this structure is briefly discussed.
Studies on Abnormal Hemoglobins
Authors: SINGER KARL , SINGER LILY
Published: October 2019

1. When sufficiently concentrated sickle cell hemoglobin containing solutions are exposed to a constant stream of CO2 gas, the hemolysates gel. This gelling phenomenon is indicative of the presence of S hemoglobin and cannot be obtained with any other type of human hemoglobin in the absence of S pigment. The lowest S hemoglobin concentration (Gm. per cent) of a hemolysate at which the gelling phenomenon can still be elicited is designated as its lowest gelling point.

2. A simple apparatus was developed to analyze the gelling phenomenon under standardized conditions. It could be shown that the lowest gelling points of hemolysates prepared from erythrocytes of the sickle cell trait (containing A + S hemoglobins), of the "C variant" (containing C + S hemoglobins), and from sickle cell anemia cells (containing S + F hemoglobins) differ distinctly. Further experiments suggest that the presence of A hemoglobin decreases the minimal amount of S pigment required for gel formation, and that type C hemoglobin reduces this amount even further. F hemoglobin seems to exert no significant influence on the gelling phenomenon. Serum albumin is also capable of decreasing the amount of S hemoglobin required for gelation.

3. A sickled erythrocyte is visualized as an S hemoglobin tactoid or gel, specifically influenced by the companion pigment which interacts with the S compound. Thus, in the sickle cell trait, a positive sickling test is not only caused by the presence of S hemoglobin, but also by its interaction with A hemoglobin. Only in the sickle cell anemia cells does sickling seem to depend solely upon the interaction of the S hemoglobin molecules.

4. The readily demonstrable differences of the lowest gelling points of hemolysates prepared from the various types of sickling red cells form the basis of the diagnostic gelling test which distinguishes sharply between sickle cell anemia and sickle cell trait erythrocytes. By this procedure atypical cases of sickle cell disease, for example, those whose erythrocytes contain C hemoglobin, may also be detected.
Studies in Rh Sensitization
Authors: WIENER ALEXANDER S. , NAPPI RAFFAELE , GORDON EVE B.
Published: October 2019

The distribution of the A-B-O blood groups among four hundred fifty-seven sensitized Rh negative mothers was compared with the distribution among eight hundred twenty-three nonsensitized Rh negative mothers. As expected, the frequencies of groups O and B were lower while that of group A was higher in the series of sensitized women. The difference in frequency of group A was 5 times its probable error and therefore statistically significant. Surprisingly, however, the difference proved to be due entirely to subgroup A1, while the frequency of subgroup A2 was the same in both series.

In a random series of four thousand nine hundred seventy-four Caucasoid individuals in New York City, the frequency of type rh’ was found to be 0.95 per cent. Statistical analysis of the Rh types of four hundred fifty-seven sensitized Rh negative mothers indicates, on the other hand, that the frequency of type rh’ is actually 0.41 per cent. The discrepancy is explained by postulating that in the control series, about half of the supposed type rh’ individuals were really type Rh1, since no special test had been made for Rh0 variants. In the series of sensitized Rh negative mothers, individuals of type Rh1 are automatically screened out since when factor Rh0 is present sensitization to the Rh0 factor is hardly likely to occur.
Hematopoietic Depression Induced by Chloromycetin
Authors: ERSLEV ALLAN
Published: October 2019

Two cases of salmonella infection receiving Chloromycetin therapy developed erythropoietic hypoplasia and myeloid maturation arrest. In both cases there was prompt hematologic recovery with well defined reticulocytosis after discontinuance of the drug.
Humoral Regulation of Red Cell Production
Authors: ERSLEV ALLAN
Published: October 2019

Large amounts of plasma from rabbits, rendered anemic by bleeding, were injected into normal rabbits. A significant rise in the number of reticulocytes was observed in these rabbits. Control rabbits receiving the same amount of plasma from normal donor rabbits failed to show any significant change in the reticulocyte count. A definite increase in red blood cell count, hematocrit and per cent nucleated red cells of the bone marrow was noticed in 2 rabbits receiving repeated injections of plasma from anemic rabbits. It is concluded that plasma from rabbits rendered anemic by bleeding contains a factor capable of stimulating red cell production.
Immunologic Studies of Hemoglobins
Authors: CHERNOFF AMOZ I.
Published: October 2019

1. A quantitative immunologic technic for the determination of fetal hemoglobin has been developed using a specific anti fetal hemoglobin serum.

2. Fetal hemoglobin occurs in many normal adult individuals in extremely small quantities: values of up to 1.0 per cent of the total hemoglobin may be in the form of the embryonic compound.

3. The alkali-resistant hemoglobin fractions detected by chemical procedures in a number of hematologic diseases are, in all probability, identical with the normally occurring fetal hemoglobin of the red cells of newborn infants. It is suggested, therefore, that these diseases are characterized by abnormal quantities of fetal hemoglobin rather than by an abnormal fetal-like component.
Some Observations on Anemia in Rheumatoid Arthritis
Authors: JEFFREY M. R.
Published: October 2019

1. The results of investigations upon some aspects of anemia in rheumatoid arthritis are reported and discussed.

2. The anemia was essentially normocytic and hypochromic, the mean corpuscular volume being usually normal and the mean corpuscular hemoglobin content and concentration usually reduced. Anemia was more pronounced in the presence of very active disease.

3. Undue hemolysis was not of importance.

4. The myelogram showed no gross abnormality but suggested impaired maturation of normoblasts and retarded hemoglobin formation.

5. The blood and plasma volumes were not essentially abnormal, though the frequent asthenic habitus and loss of weight might produce apparently exalted values in terms of body weight. In the more anemic cases, changes common to many anemias were found, viz., normal plasma volume with decreased corpuscle and whole blood volume.

6. The plasma iron concentration was usually reduced; subnormal values were invariable in cases with very active disease or marked anemia.

7. The iron-binding capacity of the serum might be raised by iron deficiency or slightly reduced but not enough to be a factor in producing or maintaining anemia.

8. As judged by changes in the plasma iron, intestinal absorption of iron after a single maximal therapeutic dose was variable. Some cases showed the excellent absorption and therapeutic response characteristic of simple iron deficiency. Others gave strong presumptive evidence of impaired absorption.
Control of the Postirradiation Hemorrhagic State by Platelet Transfusions
Authors: WOODS M. C. , GAMBLE FRANCES N. , FURTH J. , BIGELOW R. R.
Published: October 2019

Suspensions of concentrated, homologous blood platelets when introduced intravenously into lymph-cannulated dogs and rats (nine to twelve days after exposure to an LD50 or greater dose of x-rays) have been shown to eliminate the large scale diversion of erythrocytes into the lymph during the hemorrhagic phase of radiation injury. By means of injection of platelets preinjection levels of below 50,000 were raised to an average of 107,000 in 6 dogs, and to an average of 310,000 in five rats at 15 to 30 minutes after injection. All platelet-injected animals showed clearing of the bloody lymph within the first hour after injection, and within 5 to 6 hours the average erythrocyte count in the dogs dropped from an average of 270,000 to an average of 12,000 and in the rats from an average of 900,000 to an average of 30,000. No depression of lymph erythrocyte counts was found in control rats receiving plasma only. Lymph erythrocyte counts in the platelet recipients remained low for 24 to 36 hours after injection; bleeding into the lymph space reappeared in both rats and dogs when the platelet level dropped below approximately 50,000. Blood platelet and lymph erythrocyte levels are excellent indices to quantitate the severity of the hemorrhagic phase of radiation injury and to gauge the results of therapeutic procedures.

While the mechanism of platelet action remains to be demonstrated, the present studies support the thesis of earlier workers that platelets are essential to maintain vascular integrity and that a platelet deficiency is the main cause of the postirradiation hemorrhagic state.
Proteolytic and Fibrinolytic Activity of Serum
Authors: CLIFFTON EUGENE E. , CANNAMELA DOLLY A.
Published: October 2019

1. The fibrinolytic and proteolytic activity of serum plasminogen, activated by Streptokinase and Staphylokinase and measured by clot lysis and casein digestion, was studied in the following species of animals: human, cow, dog, horse, monkey, pig, rabbit, rat, and chicken.

2. A significant difference in fibrinolytic and proteolytic activity of serum plasminogen (activated by staphylokinase) was observed in three animal species: monkey, horse, and human. The significance of this observation is briefly discussed.
Hemolytic Disease of the Newborn Infant Caused by Maternal Sensitization to the Blood Factor hr' (c)
Authors: GRUNDORFER JOSEPH
Published: October 2019

1. Intragroup Rh incompatibility between an Rh-positive mother and an Rh-positive infant presents an unusual pattern of maternal isosensitization.

2. The antibody found in the sera of two Rh-positive mothers appeared to be of anti-hr' (anti-c) specificity.

3. Evidence of hemolytic disease in both infants was found and the findings directly correlated with this maternal antibody.

4. The welfare of the infant greatly depends on prompt recognition of this unusual pattern of maternal isosensitization and proper treatment of its disease.

5. An attempt has been made to evaluate the role of multiple transfusions of Rh-negative blood which had been given to an Rh-positive mother four years prior to gestation.

6. The wide spacing of several transfusions or transfusion and pregnancy respectively appears to be probably the most important factor in the development of isosensitization.

7. The possible danger in using Rh-negative blood as a universal safe donor is briefly discussed.
Paroxysmal Nocturnal Hemoglobinuria
Authors: CROSBY WILLIAM H.
Published: October 2019

1. Paroxysmal noctural hemoglobinuria is believed to be an acquired disease of the hematopoietic system in which abnormal red cells, white cells, and platelets are produced. The lesion of the cells probably involves the stromal proteins in such a fashion that they are susceptible to the proteolytic effect of a sytem of normal plasma enzymes. The severity of the disease depends upon the degree of sensitivity of a given patient’s blood cells to the plasma hemolytic system.

2. The plasma enzyme system that mediates the destruction of PNH cells consists of hemobytic factors and their inhibitors. The most important inhibitor is easily destroyed by thrombin, allowing increased hemolysis. As a consequence of this, any reaction in the patient that involves the blood coagulation system is apt to cause a hemolytic crisis.

3. PNH red cells are destroyed intravascularly causing hemoglobinemia. Hemoglobinuria, hemosiderinuria, and abnormalities of iron metabolism are sequels of this reaction.

4. Anemia in PNH is a result of hemolytic disease (short red cell life span) together with a relative bone marrow deficiency.

5. Hemolytic crises in PNH occur when plasma hemolytic activity increases. The intensity of activity depends upon a balance that exists between the hemolytic enzymes in the patient’s plasma and two inhibitory factors. Hemolytic crises take place when the balance is disturbed in favor of the hemolytic factors. Increased hemolysis at night may be due to changes in the balance of the inhibitor-hemolysin system in addition to the effect on pH that may be produced by retention of CO2 during sleep. Hemolytic crises have sometimes been due to the temporary appearance of an autoimmune reaction.

6. Aregenerative crises occur when the bone marrow temporarily suspends production of blood cells. This may result in severe anemia, agranulocytic angina, or both. Purpura is rare in PNH.

7. Chronic leukopenia in PNH is believed to be due to a leukocytic abnormality. The susceptibility to infection of patients with PNH may be related to the defective white cells.

8. Thrombocytopenia is believed to be due to a platelet abnormality. The platelets are prone to agglutinate, and their abnormality may be the basis of the susceptibility to thrombosis that these patients exhibit.

9. The sensitivity of the platelets to the PNH hemolytic enzymes and the sensitivity of the PNH inhibitor to the action of thrombin suggests that a vicious cycle of activity exists between the PNH hemolytic system and the coagulation system of the blood.

10. Dicumarol is able to impede this cycle. Its use has sometimes relieved the anemia. Dicumarol is of special value in protecting patients with PNH against thrombotic accidents. Heparin, on the other hand, by acting against the PNH inhibitor system actually causes increased hemolysis.

11. Etiologic factors are discussed, but the etiology of PNH is unknown.

12. As of July 1953, there had been published at least one hundred and sixty-two case reports of PNH.
The Immunologic Specificity of Antiserum for Trypsin-Treated Red Blood Cells and Its Reactions with Normal and Hemolytic Anemia Cells
Authors: Dodd Matthew C. , Wright Claude-Starr , Baxter J. Albert , Bouroncle Bertha A. , Bunner Alvinza E. , Winn Henry J.
Published: October 2019

The Addendum which appeared on page 647 of an article entitled "The Immunologic Specificity of Antiserum for Trypsin-Treated Red Blood Cells and Its Reactions with Normal and Hemolytic Anemia Cells" by Matthew C. Dodd, Claude-Starr Wright, J. Albert Baxter, Bertha A. Bouroncle, Alvinza E. Bunner, and Henry J. Winn (Blood 8: 640-647, 1953) belongs, instead, with the article entitled "Production of Nucleophagocytosis by Rabbit Antileukocytic Serum" by Hyman J. Zimmerman, John R. Walsh, and Paul Heller (Blood 8: 651-654, 1953).
A mutation of the common receptor subunit for interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor, and IL-5 that leads to ligand independence and tumorigenicity
Authors: D'Andrea R , Rayner J , Moretti P , Lopez A , Goodall GJ , Gonda TJ , Vadas M
Published: October 2019

The cytokines interleukin-3, interleukin-5, and granulocyte-macrophage colony-stimulating factor bind with high affinity to a receptor complex that contains a ligand-specific alpha-chain and a common beta-chain, h beta c. We report here the isolation of a mutant form of h beta c, from growth factor-independent cells, that arose spontaneously after infection of a murine factor-dependent hematopoietic cell line (FDC-P1) with a retroviral h beta c expression construct. Analysis of this h beta c mutation shows that a small (37 amino acid) duplication of extracellular sequence that includes two conserved sequence motifs is sufficient to confer ligand-independent growth on these cells and lead to tumourigenicity. Because this is a conserved region in the cytokine receptor superfamily, our results suggest that the large family of cytokine receptors has the capacity to become oncogenically active.
Roles for integrin very late activation antigen-4 in stroma-dependent erythropoiesis
Authors: Yanai N , Sekine C , Yagita H , Obinata M
Published: October 2019

Adhesion molecules are required for development of hematopoietic stem and progenitor cells in the respective hematopoietic microenvironments. We previously showed that development of the erythroid progenitor cells is dependent on their direct adhesion to the stroma cells established from the erythropoietic organs. In this stroma-dependent erythropoiesis, we examined the role of adhesion molecules in erythropoiesis by blocking antibodies. The development of the erythroid cells on stroma cells was inhibited by anti-very late activation antigen-4 (VLA-4 integrin) antibody, but not by anti-VLA-5 antibody, although the erythroid cells express both VLA-4 and VLA-5. Whereas high levels of expression of vascular cell adhesion molecule-1 (VCAM-1) and fibronectin, ligands for VLA-4, were detected in the stroma cells, the adhesion and development of the erythroid progenitor cells were partly inhibited by the blocking antibody against VCAM-1. VLA-5 and fibronectin could mediate adhesion of the erythroid progenitor cells to the stromal cells, but the adhesion itself may not be sufficient for the stroma-supported erythropoiesis. The stromal cells may support erythroid development by the adhesion through a new ligand molecule(s) for VLA-4 in addition to VCAM-1, and such collaborative interaction may provide adequate signaling for the erythroid progenitor cells in the erythropoietic microenvironment.
Effect of granulocyte-macrophage colony-stimulating factor on sepsis- induced organ injury in rats
Authors: Toda H , Murata A , Oka Y , Uda K , Tanaka N , Ohashi I , Mori T , Matsuura N
Published: October 2019

The administration of granulocyte-macrophage colony-stimulating factor (GM-CSF) to patients with severe active infections has been questioned because activation of neutrophils may cause tissue injury. To identify the effect of GM-CSF administration on severe sepsis, we examined the survival rate and pathologic changes in vital organs using the rat lethal sepsis model. Rats received 20 micrograms of recombinant murine GM-CSF (rmGM-CSF) 3 hours after the onset of peritonitis induced by cecal ligation and puncture. After 48 hours, the survival rate did not improve, and earlier deaths than in the control group were observed. In addition, the inhibition of early leuko-sequestration in the peritoneal cavity was seen in animals treated with GM-CSF. These results suggested that the administration of rmGM-CSF after the onset of sepsis was not beneficial; thus, we concluded that care should be taken in the clinical use of GM-CSF in severe infection.
A reverse transcriptase-polymerase chain reaction detects heterogeneous chimeric mRNAs in leukemias with 11q23 abnormalities
Authors: Yamamoto K , Seto M , Iida S , Komatsu H , Kamada N , Kojima S , Kodera Y , Nakazawa S , Saito H , Takahashi T
Published: October 2019

The MLL gene involved in 11q23 translocations found in the majority of infantile leukemias and some secondary leukemias makes fusion transcripts with genes such as LTG4 (chromosome 4), LTG9 (chromosome 9), and LTG19 (chromosome 19) as a result of reciprocal translocation. We have examined 25 cases of leukemias with 11q23 abnormalities by Southern blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). Using various primer pairs, chimeric mRNAs could be amplified in 6 of 7 leukemias with t(4;11), 6 of 8 leukemias with t(9;11) including secondary leukemia, 8 of 9 leukemias with t(11;19), and 1 with a deletion at 11q23. The chimeric mRNAs were heterogeneous and differential usage of the MLL exons was found, irrespective of the partner chromosomes. Sensitivity studies showed that a single clone with chimeric mRNA in 10(4) to 10(5) cells could be detected. These findings show that the present RT-PCR settings provide a rapid, accurate, and sensitive tool for diagnosing leukemias with 11q23 translocations and for monitoring response to therapy in these patients.
Interstitial insertion of retinoic acid receptor-alpha gene in acute promyelocytic leukemia with normal chromosomes 15 and 17
Authors: Hiorns LR , Min T , Swansbury GJ , Zelent A , Dyer MJ , Catovsky D
Published: October 2019

The translocation t(15;17)(q22;q21) is seen exclusively in patients with acute promyelocytic leukemia (APL) and in the promyelocytic blast crisis of chronic myeloid leukemia (CML). This translocation juxta- poses the promyelocytic leukemia (PML) gene on chromosome 15 and the retinoic acid receptor-alpha (RARA) gene on chromosome 17, resulting in the formation of a chimeric mRNA transcript. We describe a patient with the microgranular variant form of APL, with no detectable cytogenetic abnormality of either chromosomes 15 or 17, who nevertheless had juxtaposition of PML and RARA genes and expressed a chimeric transcript. Conventional cytogenetics showed the karyotype 46,XY,d- er(3)t(3;8)(p25;q12). Fluorescent in situ hybridization (FISH) with paints for chromosomes 8, 15, and 17 confirmed the presence of structurally intact chromosomes 15 and 17 and trisomy for chromosome 8q. Nevertheless, FISH using cosmid probes for PML and RARA showed their juxtaposition on one chromosome 15 homolog. Both genes were also present on their normal homologs; in addition, part of the RARA gene was still present on the remaining chromosome 17. DNA analysis by Southern blotting, performed with a variety of probes including PML, RARA and retinoic acid receptor-beta (RARB), showed a rearrangement in PML. Reverse transcriptase polymerase chain reaction (RT-PCR) confirmed the existence of hybrid transcripts of 276, 455 bp and 623 bp, from PML- RARA on the der(15) chromosome, consistent with alternate exon splicing of the long form of the transcript occurring in 50% to 60% of patients with APL. Our results show that APL patients with cytogenetically normal chromosomes 15 and 17 may, nevertheless, have involvement of both PML and RARA genes defining a subgroup of APL, t(15;17)- negative/PML-RARA-positive which is analogous to Philadelphia chromosome-negative/BCR-ABL-positive CML. In this case, the presence of chimeric transcripts suggests that treatment with all-trans RA may be warranted in APL, even in the absence of detectable cytogenetic change, showing the usefulness of RT-PCR or FISH to aid diagnosis.
The heterogeneity of azurophil granules in neutrophil promyelocytes: immunogold localization of myeloperoxidase, cathepsin G, elastase, proteinase 3, and bactericidal/permeability increasing protein
Authors: Egesten A , Breton-Gorius J , Guichard J , Gullberg U , Olsson I
Published: October 2019

Azurophil granules of myeloid cells form in promyelocytes. They store cytotoxic and digestive agents which when released are involved in the defense against infection. In order to characterize the intragranular distribution of these agents, ultrastructural methods using immunogold were used on promyelocytes. Azurophil granules were divided into nucleated, large spherical (large azurophil) and small electron-dense (small azurophil) granules. Myeloperoxidase showed a peripheral distribution of large azurophils and a uniform distribution of small and nucleated azurophils, consistent with previous findings. Likewise, the major neutral proteases of azurophils, cathepsin G, granulocyte elastase, and proteinase 3, displayed a similar distribution, with a peripheral localization in large azurophils and a uniform distribution in small and nucleated azurophils, except for proteinase 3, which was associated with the crystalloid structure in nucleated azurophils. In contrast, the bactericidal/permeability increasing protein, which is bacteristatic and bactericidal for Gram-negative bacteria, was localized to the membrane area in all types of azurophil granules, consistent with a suggested association of this protein with the granule membrane. The observed differences in intragranular distribution of the proteins investigated may reflect variations in binding to matrix structures and granule membranes.
Herpesvirus type 6 in patients undergoing bone marrow transplantation: serologic features and detection by polymerase chain reaction
Authors: Wilborn F , Brinkmann V , Schmidt CA , Neipel F , Gelderblom H , Siegert W
Published: October 2019

To evaluate the potential role of human herpesvirus type 6 (HHV-6) infection in patients after bone marrow transplantation (BMT) we sequentially analyzed buffy coat leukocytes, oral lavage fluid, and urine from 57 patients for the presence of HHV-6 DNA by polymerase chain reaction (PCR) before and after 60 BMTs. Twenty-four patients undergoing autologous BMT and 36 with allogeneic BMT were studied. Thirty-six patients (60%) were PCR positive in one or more tests. The majority of PCR-positive patients had positive results only sporadically, in 1 (n = 23) or 2 weeks (n = 5). Six patients were positive in 3 to 5 weeks. In 2 patients, we found a high frequency of positive tests, in 7 of 7 and 10 of 10 weeks analyzed. Twenty-four patients (40%) remained PCR negative throughout the post-BMT period. There was a significant correlation between the results of HHV-6 PCR and the occurrence of acute graft-versus-host disease (aGVHD). In grade II-IV, 6 of 8 (75%) patients had 2 or more positive PCR tests, compared with 5 of 25 (20%) patients without or with grade I aGVHD (P = .01). There was no difference in the outcome of PCR tests with respect to the type of BMT or pre-BMT HHV-6 enzyme-linked immunosorbent assay titers. Restriction enzyme analysis of PCR amplificates from 18 patients showed HHV-6 variant B in 16 (88.9%) and variant A in 2 cases (11.1%). We conclude that HHV-6 DNA can be detected in 60% of the patients after BMT. HHV-6 DNA can be detected more frequently in patients with moderate and severe aGVHD than in patients without aGVHD or with mild aGVHD.
The inv(11)(p15q22) Chromosome Translocation of De Novo and Therapy-Related Myeloid Malignancies Results in Fusion of the Nucleoporin Gene, NUP98, With the Putative RNA Helicase Gene, DDX10
Authors: Arai Yasuhito , Hosoda Fumie , Kobayashi Hirofumi , Arai Kyoko , Hayashi Yasuhide , Kamada Nanao , Kaneko Yasuhiko , Ohki Misao
Published: October 2019

The inv(11)(p15q22) is a recurrent chromosomal abnormality associated with de novo and therapy-related myeloid malignancies. Here we report the molecular definition of this chromosomal aberration in four patients. Positional cloning showed the consistent rearrangement of the DDX10 gene on chromosome 11q22, which encodes a putative RNA helicase. The translocation targets the NUP98 gene on 11p15, a member of the FG peptide repeat nucleoporin family. In DDX10 and NUP98, the inv(11) breakpoints occurred within two introns of each gene and the two genes merged in-frame to produce the chimeric transcripts characteristic of this translocation. Although two reciprocal chimeric products, NUP98-DDX10 and DDX10-NUP98, were predicted, only NUP98-DDX10 appears to be implicated in tumorigenesis. DDX10 is predicted to be involved in ribosome assembly. NUP98 has been identified as a nuclear pore complex protein and a target of chromosomal translocation in acute myeloid leukemia through the t(7; 11)(p15; p15) translocation. The predicted NUP98-DDX10 fusion protein may promote leukemogenesis through aberrant nucleoplasmic transport of mRNA or alterations in ribosome assembly.
The t(11;16)(q23;p13) Translocation in Myelodysplastic Syndrome Fuses the MLL Gene to the CBP Gene
Authors: Taki Tomohiko , Sako Masahiro , Tsuchida Masahiro , Hayashi Yasuhide
Published: October 2019

The recurrent translocation t(11; 16)(q23; p13) has been reported to be associated with therapy-related acute leukemia. The MLL gene involved in other 11q23 abnormalities was also rearranged by this translocation. We analyzed two patients with myelodysplastic syndrome with t(11; 16) and showed that the MLL gene on 11q23 was fused with CREB-binding protein (CBP) gene on 16p13 in these patients. The CBP gene encodes a transcriptional adaptor/coactivator protein and it is mutated in patients with Rubinstein-Taybi syndrome. The CBP gene is also involved in acute myeloid leukemia (AML) with t(8; 16)(p11; p13). In-frame MLL-CBP fusion transcripts combine the MLL AT-hook motifs and DNA methyltransferase homology region with a largely intact CBP. Our results combined with the finding of the MOZ-CBP fusion in t(8; 16)-AML suggest that the CBP gene may be associated with leukemogenesis through translocations.
Polyploidization and Functional Maturation Are Two Distinct Processes During Megakaryocytic Differentiation: Involvement of Cyclin-Dependent Kinase Inhibitor p21 in Polyploidization
Authors: Kikuchi Jiro , Furukawa Yusuke , Iwase Satsuki , Terui Yasuhito , Nakamura Mitsuru , Kitagawa Seiichi , Kitagawa Masatoshi , Komatsu Norio , Miura Yasusada
Published: October 2019

The mechanism of megakaryocytic differentiation was investigated using human megakaryocytic leukemia cell line UT-7. Polyploidization of UT-7 cells was induced by the microtubule-depolymerizing agent, nocodazole, and 12-O-tetradecanoylphorbol-13-acetate (TPA), but the effect was much more striking with nocodazole. By contrast, induction of cytoplasmic maturation, as judged by β-thromboglobulin production and platelet factor 4 expression, was more prominent in TPA-treated cells than in nocodazole-treated cells. Nocodazole and TPA could act synergistically to increase ploidy and to enhance the expression of mature phenotypes. Human thrombopoietin induced functional maturation but not polyploidization in UT-7 cells and also acts synergistically with nocodazole. Cyclin-dependent kinase inhibitor p21 was upregulated at the early stage of megakaryocytic differentiation, and overexpression of p21 resulted in an increase in ploidy of UT-7 cells. This suggests that p21 is implicated in polyploidization via suppression of CDC2 activity at mitosis. UT-7 but not HL-60 cells could incorporate [3H]thymidine in the presence of TPA, indicating the presence of megakaryocyte-specific licensing factor to allow DNA replication during differentiation. Taking these data together, we propose that megakaryocytic differentiation consists of two distinct processes, polyploidization and functional maturation, and that these two processes are independently regulated.
Genomic Sequence and Transcription Start Site for the Human γ-Glutamyl Carboxylase
Authors: Wu Sheue-Mei , Stafford Darrel W. , Frazier L. Dan , Fu Yuan-Yun , High Katherine A. , Chu Kirk , Sanchez-Vega Beatriz , Solera Jesús
Published: October 2019

The human gene for γ-glutamyl carboxylase is 13 kb in length and contains 15 exons. Transcription starts at a cytosine 217 base pair upstream of the first codon. There are two major transcripts in all tissues examined. They are distinguished by the presence of an Alu sequence in the 3′ nontranslated end of the longer species. Relative mRNA levels for 12 bovine tissues are presented.