bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of latent hematopoietic progenitor cells in cultures of newborn and adult baboon liver
Authors: Roodman GD , VandeBerg JL , Kuehl TJ
Published: October 2019

The anatomic site of hematopoiesis changes during fetal development from the yolk sac to the liver and finally to the marrow. Factors controlling this switch in the site of hematopoiesis are unknown. We assayed erythroid colony (CFU-E) and erythroid burst (BFU-E) formation in fetal, newborn, and adult baboon liver and marrow to determine the growth requirements of primate hematopoietic progenitor cells from different anatomic sites and developmental stages. We cocultured fetal, newborn, and adult liver and marrow nonadherent cells with adherent cells from these organs to assess the role adherent cells may play in determining the site of hematopoiesis. Fetal liver, fetal marrow, newborn marrow, and adult marrow cultures formed CFU-E and BFU-E colonies in vitro. In contrast, newborn and adult liver cell cultures very rarely formed colonies. However, when newborn or adult liver nonadherent cells were cocultured with marrow adherent cells, CFU-E and BFU-E colonies were detected. The colonies that formed in the newborn and adult liver cultures were derived from the liver and not from the marrow cells or peripheral blood trapped in the liver. These data suggest that in contrast to fetal liver, newborn and adult liver may not be hematopoietic organs in normal primates in vivo because of changes in the growth requirements of hematopoietic progenitor cells present in these organs.
Changes in intracellular Mg adenosine triphosphate and ionized Mg2+ during blood storage: detection by 31P nuclear magnetic resonance spectroscopy
Authors: Bock JL , Wenz B , Gupta RK
Published: October 2019

31P nuclear magnetic resonance (NMR) spectroscopy was used to measure changes in intra-erythrocyte Mg adenosine triphosphate (MgATP) and free Mg2+ during blood storage at 4 degrees C in standard citrate preservation media. The extent of Mg2+ complexation of ATP and the concentration of free Mg2+ were measured from the Mg2+-dependent chemical shift differences, at 22 degrees C, between the P beta and P alpha resonances of intracellular ATP. This difference changed from 721.0 +/- 1.4 Hz (mean +/- SE) on the day of collection to 741.0 +/- 3.4 Hz after three to seven days and 774.0 +/- 2.8 Hz after 11 to 40 days storage in either acid-citrate-dextrose (ACD) or citrate-phosphate- dextrose-adenine (CPDA-1). Changes in intracellular pH, detected from shifts in the intracellular Pi resonance, averaged 0.27 units after 11 to 40 days of storage. These data indicate a sizable decrease in the extent of Mg2+ complexation of ATP, and a decrease by a factor of 2.6 in free Mg2+, during the shelf-life of blood stored in ACD or CPDA-1.
LAV/HTLV III presence in peripheral blood lymphocytes of seropositive young hemophiliacs
Authors: Gomperts ED , Feorino P , Evatt BL , Warfield D , Miller R , McDougal JS
Published: October 2019

Recent studies indicate a high prevalence of seropositivity to the lymphadenopathy-associated virus/human T-lymphotropic virus (type III) among individuals with hemophilia exposed to clotting factor concentrates prepared from large donor pools. The peripheral blood lymphocytes of 19 young seropositive patients with inherited bleeding disorders were examined for the presence of this virus by coculture with phytohemagglutinin-stimulated lymphocytes. Viral isolates were obtained from six of 19 patients. While none of these patients have developed the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex, five of them had lymphadenopathy in two noncontiguous areas, and two showed clinically symptomatic enlarged tonsils and adenoids. Of the 13 patients in whom virus was not demonstrated, five were judged clinically normal and five had mild lymphadenopathy in one anatomical area. These results suggest that as many as 33% of hemophiliacs (six of 19 patients studied), who have circulating antibodies to mature viral proteins, have viral-infected peripheral blood lymphocytes capable of infecting other lymphocytes in vitro.
Hemolytic Disease and Polycythemia in Parabiosis Intoxication
Authors: CHUTE ROSANNA N. , SOMMERS SHELDON C.
Published: October 2019

Disharmoniouns parabiotic rats were studied, which had clinical, hematologic, pathologic and serologic evidences of blood incompatibility, isohemagglutinin hypersensitization and hemolytic anemia. Spherocytosis and increased incomplete isohemagglutinin antibodies, demonstrated by indirect Coombs-type agglutinations and with trypsinized rat erythrocytes, were found in both parabionts. One partner became anemic with reticulocytosis, erythroblastosis, bone marrow necrobiosis, reactive hematopoietic hyperplasia and enhanced extramedullary hematopoiesis. The other parabiont showed lymphoid and reticulo-endothelial hyperplasia, leukocytoid lymphocytosis and erythrophagocytosis. It was plethoric with polycythemia, increased blood volume and high blood pressure. The polycythemia was explained by mechanical trapping of sludged antibody-coated incompatible erythrocytes, which the reticulo-endothelial system was inadequate to process. Complicating hemoglobinemia and hemoglobinuric nephrosis developed. Other manifestations of parabiosis intoxication are briefly discussed.
Hemolytic Disease of the Newborn Due to Anti-S
Authors: LEVINE PHILIP , FERRARO LOUIS R. , KOCH ELIZABETH
Published: October 2019

The first case of hemolytic disease attributable to anti-S is reported. The serologic properties of this example of anti-S is fully described. The essential findings in 12 other anti-S sera reported in the literature are briefly summarized.
A Hemorrhagic Diathesis Due to a Qualitative Platelet Defect
Authors: SUSSMAN LEON N. , WALD NIEL , ROSENTHAL ROBERT L.
Published: October 2019

1. A patient with hemorrhagic diathesis associated with a prolonged clotting time is presented.

2. The clotting defect was corrected by normal and hemophiliac blood and normal platelet-containing plasma but was not corrected by idiopathic thrombocytopenic blood or by normal and leukemic platelet-poor plasmas as measured by the prothrombin consumption test.

3. The clotting defect appears most likely to be due to platelets which are defective in that area of platelet activity concerned with the activation of thromboplastinogen to form thromboplastin.
Effects of Cortisone and Hydrocortisone on the Numbers of Thoracic Duct Lymphocytes
Authors: HUNGERFORD GERALD F. , REINHARDT WILLIAM O. , LI CHOH HAO
Published: October 2019

The effects of cortisone and hydrocortisone on the levels of thoracic duct lymphocytes have been studied in normal and hypophysectomized rats under standardized conditions. It is shown that:

1. Four hours after its administration, hydrocortisone causes a marked thoracic duct lymphocytopenia, the effect being even more marked in hypophysectomized than in normal rats.

2. At comparable dose levels, hydrocortisone is demonstrably and significantly more effective in producing a thoracic duct lymphocytopenia than is cortisone.
The Metabolic Displacement of Folic Acid by Aminopterin
Authors: SWENDSEID MARIAN E. , SWANSON ANN LOUISE , MILLER STANLEY , BETHELL FRANK H.
Published: October 2019

The amount of Aminopterin excreted during treatment with this antagonist has been measured in 9 leukemia patients. It was found that considerable although variable amounts were retained in the body and that the smallest amounts were excreted by patients with chronic granulocytic leukemia. Urinary excretion of Aminopterin continued at a significant level for a period of at least six days after a course of therapy.

Following Aminopterin therapy, there was an increase in folic acid excretion as compared to the excretion of this vitamin prior to antagonist administration.

These data suggest that there is a metabolic displacement of folic acid by Aminopterin.
In Vitro Erythrophagocytosis in Acquired Hemolytic Anemia
Authors: ZINKHAM WILLIAM H. , DIAMOND LOUIS K.
Published: October 2019

It has been found that marked erythrophagocytosis occurred when the buffy coats of 3 patients with idiopathic acquired hemolytic anemia were incubated at 37 C. for one hour. It is suggested that this technic may be used as another in vitro method of demonstrating abnormalities of the red cells in patients with acquired hemolytic anemia, as well as other conditions in which there is damage or alteration of the red cells.
Transfer of Myelogenous Leukemia Induced by Gastric Instillation of Methylcholanthrene in Wistar Rats
Authors: SHAY HARRY , GRUENSTEIN MARGOT , HARRIS CHARLES , GLAZER LILLIAN
Published: October 2019

A third case of myelogenous leukemia that followed the gastric instillation of methylcholanthrene in Wistar rats is herein recorded. These 3 cases represent the first instances of myelogenous leukemia produced in rats by carcinogens that answer all the criteria for a true myelogenous leukemia; namely, positive peripheral blood picture, bone marrow, tissue infiltration and transferability.

A high percentage of transfer of the myelogenous leukemia was obtained with peripheral blood given intraperitoneally to very young animals of our random bred colony. Successful transfer was also accomplished to very young rats of the Long Evans and Sherman strains. While none of the three animals in which myelogenous leukemia was induced showed any green coloration, all transfers developed as chloroleukemia. Animals with leukemia will be made available to investigators.
Case Report: Hypersplenism
Authors: WIARD BARBARA A. , ROBBINS STANLEY L.
Published: October 2019

A case of hypersplenism giving rise to a panhematopenia is presented with complete postmortem study. Its association with rheumatoid arthritis permits it to be classified as a Felty’s syndrome. There was histologic evidence to support both of the now existing concepts of the mechanism by which splenic hyperactivity effects its cytopenias. Marked phagocytosis of both red and white cells was noted in the spleen. There was, in addition, similar phagocytic activity throughout the entire R-E system. Equal support for the hypothesis that splenic hyperfunction depresses or blocks bone marrow function was adduced from the finding of a complete maturation arrest of the white cell series of the bone marrow at the myelocytic level.

A third factor, to the best of our knowledge never previously identified in cases of hypersplenism, was also present. This was a circulating substance which produced active lysis of normal white cells and must have contributed to the leukopenia.

The anatomic observations in this case raise the possibility that generalized R-E hyperfunction and not simply splenic hyperfunction may be of significance in the production of cytopenias in some of the patients with so called hypersplenism.
Experiments on Plasma Clot Contraction
Authors: STILL BESSIE M.
Published: October 2019

1. Clot contraction was found to be associated with the action of whole, adhesive platelets. It was proportional to the number of adhesive platelets and not to the total number of platelets. It was inversely proportional to the fibrinogen concentration and a direct relationship was shown between the fibrinogen concentration and the number of platelets necessary to promote any fixed level of clot contraction.

2. Clot contraction could not be promoted solely by proteolytic agents and no association could be traced between clot contraction and proteolysis.

3. These findings are discussed with reference to the views concerning the mechanism of clot contraction.
The Immediate Hematologic Effects of Intravenous Saccharated Iron Oxide
Authors: WHITE L. P. , BIERMAN H. R. , KELLY K. H. , BYRON R. L. , CORDES F. L. , LITTMAN A. M.
Published: October 2019

1. Saccharated ferric oxide in colloidal suspension was administered intravenously on 17 occasions to 13 patients in doses of 100 to 1,000 mg. A prompt arterial leukopenia, followed within 30 to 90 seconds by a similar fall in the venous blood, was observed in all patients who received more than 200 mg. of iron. This leukopenia involved all white cell types.

2. The magnitude of the fall in leukocytes was directly proportional to the amount of material injected, and independent of the rate of injection under the conditions of the study.

3. There was no significant change in the platelet number in arterial or venous blood of 4 patients. In 2 patients an increased clotting power of venous blood containing large amounts of iron was observed only during the period of injection.

4. The colloidal nature of the material is probably the responsible factor in producing the leukopenia.
Case Report: Aplastic Anemia with Platelet Thrombi
Authors: FRUMIN ABRAHAM M. , CONSTON ALFRED S. , MERANZE DAVID R.
Published: October 2019

1. A case of aplastic anemia with platelet thrombi is presented.

2. The developmental features of this anemia are stressed and comparisons made with thrombotic thrombocytopenic purpura.

3. The role of platelet thrombi in other conditions are briefly reviewed.
Multiple bidirectional alterations of phenotype and changes in proliferative potential during the in vitro and in vivo passage of clonal mast cell populations derived from mouse peritoneal mast cells
Authors: Kanakura Y , Thompson H , Nakano T , Yamamura T , Asai H , Kitamura Y , Metcalfe DD , Galli SJ
Published: October 2019

Mouse peritoneal mast cells (PMC) express a connective tissue-type mast cell (CTMC) phenotype, including reactivity with the heparin-binding fluorescent dye berberine sulfate and incorporation of [35S] sulfate predominantly into heparin proteoglycans. When PMC purified to greater than 99% purity were cultured in methylcellulose with IL-3 and IL-4, approximately 25% of the PMC formed colonies, all of which contained both berberine sulfate-positive and berberine sulfate-negative mast cells. When these mast cells were transferred to suspension culture, they generated populations that were 100% berberine sulfate-negative, a characteristic similar to that of mucosal mast cells (MMC), and that synthesized predominantly chondroitin sulfate [35S] proteoglycans. When “MMC-like” cultured mast cells derived from WBB6F1-+/+ PMC were injected into the peritoneal cavities of mast cell-deficient WBB6F1- W/Wv mice, the adoptively transferred mast cell population became 100% berberine sulfate-positive. In methylcellulose culture, these “second generation PMC” formed clonal colonies containing both berberine sulfate-positive and berberine sulfate-negative cells, but exhibited significantly less proliferative ability than did normal +/+ PMC. Thus, clonal mast cell populations initially derived from single PMC exhibited multiple and bidirectional alterations between CTMC-like and MMC-like phenotypes. However, this process was associated with a progressive diminution of the mast cells' proliferative ability.
Significance of detection of occult non-Hodgkin's lymphoma in histologically uninvolved bone marrow by a culture technique
Authors: Sharp JG , Joshi SS , Armitage JO , Bierman P , Coccia PF , Harrington DS , Kessinger A , Crouse DA , Mann SL , Weisenburger DD
Published: October 2019

Prolonged disease-free survival of patients with recurrent or resistant non-Hodgkin's lymphoma (NHL) has been achieved with high-dose therapy followed by autologous bone marrow transplantation (ABMT). A concern with the use of ABMT is that the marrow that is reinfused may contain undetected NHL cells with the potential to reestablish metastatic disease in the recipient. Using a culture technique that is sensitive for detecting occult lymphoma cells in BM, we analyzed histologically normal marrow harvests from 59 consecutive patients with intermediate- or high-grade NHL who were candidates for high-dose therapy and ABMT. The culture results indicated that 22 of the patients had occult lymphoma in their marrow. Forty-three patients underwent high-dose therapy followed by ABMT. Twenty-four achieved a complete clinical remission. Those with occult lymphoma in their harvests (11 patients) continued to relapse for up to 3 years, whereas no relapses were observed beyond 8 months in 13 patients receiving marrow that did not contain detectable lymphoma cells using the culture technique. The relapses in the patients who achieved a complete remission occurred at sites of prior bulky disease rather than at new sites, suggesting that the ability to detect occult lymphoma cells in marrow is a marker of biologic aggressiveness and/or resistance to therapy, or that the reinfused cells could only grow in previously involved sites. The detection of lymphoma cells in marrow used for ABMT is an important adverse prognostic factor, and appears to be independent of other clinical predictors of outcome such as sensitivity or resistance of disease to prior chemotherapy.
Heterogeneity in filamentous actin content among individual human blood platelets
Authors: Oda A , Daley JF , Cabral C , Kang JH , Smith M , Salzman EW
Published: October 2019

The content of filamentous actin in individual platelets was measured by flow cytometry, using a fluorescent probe specific for filamentous actin (F-actin), 7-nitrobenz-2-oxa-1,3-phallacidin (NBD-phallacidin). NBD-phallacidin binding to fixed platelets was specific in that either pretreatment of platelets with unlabeled phallacidin or absorption of NBD-phallacidin by rabbit skeletal F-actin, but not globular actin (G- actin), resulted in a significant loss in the bound fluorescent probe. Mean NBD-phallacidin binding to fixed platelets varied with the agonist and paralleled the changes in F-actin reported with the DNAse I inhibition assay. (1) NBD-phallacidin binding increased with stimulation by ADP, U46619 (a prostaglandin H2 analogue), or collagen and paralleled shape change. (2) Epinephrine did not increase NBD- phallacidin binding. (3) Platelets treated at 4 degrees C contained more F-actin than did platelets kept at 37 degrees C. (4) Cytochalasin D (10 mumol/L) inhibited the increase of phallacidin binding to individual platelets stimulated by either ADP or U46619. In measurements of cytosolic free calcium concentration ([Ca2+]i) by flow cytometry in Indo-1-loaded platelets, ADP's dose-response for actin polymerization was similar to that for calcium mobilization. As shown by flow cytometry, a tail population that had a minimal increase in F- actin upon stimulation with ADP or U46619 also contained the platelets with the least forward and right angle light scattering, which are functions of platelet size and shape. When platelets treated with NBD- phallacidin were incubated with S12-murine monoclonal antibody (a marker of alpha-granule secretion detected by phycoerythrin-conjugated antimouse IgG second antibody), phallacidin fluorescence paralleled S12 binding. Thus, human blood platelets are heterogeneous in regard to actin polymerization at rest and in association with platelet activation; different degrees of phallacidin binding may identify functionally different platelet populations.
Adhesive protein expression on thrombin-stimulated platelets: time- dependent modulation of anti-fibrinogen, -fibronectin, and -von Willebrand factor antibody binding
Authors: Peerschke EI
Published: October 2019

Platelets contain a pool of endogenous adhesive proteins that can be released and may bind to surface membrane receptors under appropriate conditions. Because the binding of exogenous fibrinogen to platelets was shown previously to be accompanied by a time-dependent decrease in fibrinogen accessibility to antibody and enzymes, studies were performed to evaluate changes in the expression of endogenous fibrinogen released from thrombin-stimulated platelets using monospecific polyclonal and monoclonal antibody F(ab')2 fragments. Parallel studies were performed to compare the expression of released fibronectin and von Willebrand factor (vWF). Binding of polyclonal antibody F(ab')2 fragments directed against individual adhesive proteins was inhibited by EDTA or the 10E5 monoclonal antibody, suggesting that fibrinogen, fibronectin, and vWF expression was mediated, in large part, by divalent cation-dependent interactions with the glycoprotein IIb-IIIa complex. Interestingly, when polyclonal antibody F(ab')2 fragments were added to platelet suspensions at discrete times after thrombin stimulation, antifibrinogen F(ab')2 binding decreased by 72% +/- 15% (mean +/- SD, n = 22) over a 60-minute time course, whereas antifibronectin and anti-vWF antibody F(ab')2 fragment binding changed minimally (6% +/- 23%, n = 22 and 3% +/- 26%, n = 14, respectively). Similar observations were made with monoclonal antibodies. Parallel experiments using 125I-labeled fibrinogen as a marker indicated that the observed decrease in antifibrinogen F(ab')2 binding was not accompanied by fibrinogen dissociation. Moreover, antibody accessibility to platelet-bound fibrinogen could be restored after Triton X-100 platelet lysis. The data suggest that fibrinogen, fibronectin, and vWF are not coordinately expressed on thrombin- stimulated platelets. Rather, fibrinogen expression appears transient compared with the expression of fibronectin and vWF. The ability of platelets to secrete and organize adhesive proteins on their surface is likely to have important implications for hemostasis and thrombosis.
Antiproliferative effects of interleukin-4 on freshly isolated non- Hodgkin malignant B-lymphoma cells
Authors: Defrance T , Fluckiger AC , Rossi JF , Magaud JP , Sotto JJ , Banchereau J
Published: October 2019

The pattern of in vitro growth response of freshly isolated non-Hodgkin malignant lymphoma B cells (NHML) to cytokines was investigated. Ten tumor specimens of low- or intermediate-grade malignancy were selected for study. To assess their proliferative capacity in vitro, B-lymphoma cells were activated through ligation of their surface Ig receptor with insolubilized anti-IgM antibodies or Staphylococcus aureus strain Cowan I (SAC). In the great majority of cases, interleukin-2 (IL-2) was the sole factor that significantly and reproducibly stimulated DNA synthesis in NHML activated through their surface Igs. Other B-cell tropic factors, including IL-4, IL-5, IL-6, and tumor necrosis factor- alpha (TNF-alpha), failed to elicit a growth response in most of the IL- 2-responsive neoplastic samples. However, one specimen among 10 exhibited the opposite pattern of response and proliferated following culture with IL-4 and anti-Ig reagents, but not after IL-2 stimulation. Three specimens could also be induced for DNA synthesis on cross- linking of their surface Igs in the absence of exogenous growth factors. Although IL-4 could not support the in vitro growth of the majority of NHML cases, it strongly suppressed the proliferative signals delivered to these cells by anti-Ig reagents used alone or in combination with IL-2. Our data suggest that, in most cases, IL-4 essentially provides growth-inhibitory signals to NHML when they are activated through their surface Ig receptors and as such may be considered to be a valid candidate for future therapy of this type of mature B-cell malignancy.
Cyclic Neutropenia with Giant Follicular Lymphoblastoma and Lymphosarcoma
Authors: NATELSON ROBERT P.
Published: October 2019

1. A case of the rare disorder cyclic neutropenia with giant follicular lymphoblastoma and lymphosarcoma is reported.

2. Adrenocorticotropic hormone administration during a neutropenic phase resulted in a further depression of the total leukocyte count and no significant change in the already very low neutrophil count. Cortisone did not prevent or ameliorate a recurrence of neutropenia.

3. Splenectomy resulted in less depression of the total leukocyte and neutrophil values and a considerable amelioration of the symptoms during the continued cycles; however, because of the short follow-up, a definite statement on the long range value of splenectomy cannot be made.

4. The pathogenesis of cyclic neutropenia is discussed with particular reference to its relationship to lymphoma and hypersplenism.
Drowning Hemoglobinuria
Authors: RATH CHARLES E.
Published: October 2019

1. A case of hemoglobinemia and hemoglobinuria with recovery following incomplete drowning is presented.

2. The pathogenesis of the intravascular hemolysis is discussed.
The Significance of Stainable Iron in Sternal Marrow Sections
Authors: HUTCHISON H. E.
Published: October 2019

The amount of stainable iron in sternal aspirates has been estimated by the use of the prussian blue reaction. The superiority of histologic sections over smears for the purpose of iron staining has been stressed. The results of applying this simple histochemical test routinely to sections prepared from 141 marrow samples are described. It is shown that in disorders of the blood the procedure provides a simple means for the control of iron therapy. Not only does it indicate when iron is required but also when enough has been given and so can be used to obviate the danger of overdosage whets iron is being administered by the intravenous route. The method is put forward as a possible means of distinguishing the hypochromic anemias of chronic intoxication from those due to iron deficiency.
The Myeloproliferative Disorders With Special Reference to Myelofibrosis
Authors: HUTT M. S. R. , PINNIGER J. L. , WETHERLEY-MEIN G.
Published: October 2019

1. The clinical, hematologic and histologic findings in 10 patients with primary proliferative marrow disorders are presented.

2. It is considered that these findings indicate that myelofibrosis, myeloid leukemia, megakaryocytic myelosis and probably polycythemia vera are similar and related processes.

3. It is suggested that this relationship may be regarded as an example of a proliferative or neoplastic process involving multipotential primitive mesenchymal cells.

4. The significance of splenomegaly in myelofibrosis, the nature of extramedullary hemopoiesis and the relationship between this group of marrow disorders and the primary lymph node tumours are briefly discussed.
The Value of Routine Bone Marrow Culture for Histoplasma Capsulatum in Pediatric Hematology
Authors: ROHN ROBERT J. , BOND WILLIAM H. , HAYNES EDITH
Published: October 2019

1. Of 76 bone marrow cultures in pediatric patients with a variety of hematologic abnormalities, 5 (6.5 per cent) yielded H. capsulatum.

2. Methods used and brief case reports were described.

3. In 2 of the 5 cases with histoplasmosis, organisms were observed on direct examination of the marrow and cultures were only of confirmatory value.

4. In the remaining 3 patients with histoplasmosis, direct examination of marrow showed no features which distinguished them from the 71 negatives.

In these 3 bone marrow culture was the only means by which diagnosis could be established.

5. In no instance in which a negative bone marrow culture for H. capsulatum was obtained, was histoplasmosis diagnosed by other methods.

6. We are of the opinion that while routine marrow culture for H. capsulatum is of little value in adults, it is of considerable value for case finding in infants and children.
Immunologic Studies of Hemoglobins
Authors: CHERNOFF AMOZ I.
Published: October 2019

1. Antisera to hemoglobin solutions obtained from normal adult, cord blood and sickle cell anemia erythrocytes have been prepared by immunizing rabbits with adjuvant mixtures. Furthermore, anti F hemoglobin sera have been similarly prepared by using purified fetal hemoglobin. A species specific anti guinea pig hemoglobin serum was also developed.

2. Although anti F hemoglobin sera appear to be specific for fetal hemoglobin, no such immunologic specificity could be detected between anti A and anti SCA sera obtained from rabbits.

3. It is suggested that sera prepared by using hemoglobin solutions obtained from normal and sickle cell anemia erythrocytes may contain anti F antibodies in addition to anti A and anti S precipitins and that fetal hemoglobin may be present in many human adult blood specimens.
Concentration of Intrinsic Factor and Vitamin B12-Binding Activities of Fractions of Desiccated Hog Stomach
Authors: PRUSOFF W. H. , WELCH A. D. , HEINLE R. W. , MEACHAM G. C.
Published: October 2019

Fractionation of desiccated defatted hog stomach through the use of isoelectric and ammonium sulfate precipitation technics, and guided by assays in patients with pernicious anemia in relapse given daily oral doses of 5 µg. of vitamin B12, has led to the concentration of intrinsic factor activity in an amount of protein equivalent to 0.03 per cent of the original weight of crude material. The electrophoretic pattern of this active fraction indicated a lack of homogeneity.

The vitamin B12-binding activities of three fractions obtained by precipitation with ammonium sulfate were investigated by microbial inhibition and dialysis methods. There was no correlation between the ability of the fractions to bind vitamin B12 and to exert intrinsic factor activity. Among these fractions, that with the highest intrinsic factor activity in patients with pernicious anemia in relapse bound the least amount of vitamin B12.
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

1. Rabbits were immunized with both normal (Nrbc) and trypsinized (Trbc) human red blood cells and the antisera examined with normal, trypsinized, periodate-treated, and hemolytic anemia cells.

2. Absorption studies showed the presence of a fraction in both anti-Trbc and anti-Nrbc that was specific for trypsinized cells.

3. This T cell specific fraction from anti-Trbc serum (anti-TE) did not agglutinate or sensitize normal red blood cells, but agglutinated periodate-treated cells. This latter specificity was shown to be a part of the modification produced by trypsinization.

4. Anti-TE also agglutinated the cells of fifteen of nineteen patients with acquired hemolytic anemia and three of thirteen cases of hereditary spherocytosis.

5. Antibody for trypsinized and normal cells was also detected in antiserum to normal cells. Absorption data suggested the presence in this antiserum of antibody with a dual specificity for both types of cells.

6. The role of the antigenic modifications produced by trypsin in red cell immunization and in hemolytic anemia is discussed.
Studies on Platelets
Authors: PISCIOTTA ANTHONY V. , STEFANINI MARIO , DAMESHEK WILLIAM
Published: October 2019

1. The morphologic characteristics of the megakaryocytes were studied in fourteen normal or nonthrombocytopenic individuals, in nine patients with idiopathic thrombocytopenic purpura (I.T.P.), in two individuals with pernicious anemia, and in two normal individuals in whom thrombocytopenia was temporarily induced by the administration of plasma from a patient with a circulating high-titer platelet agglutinin. Phase contrast microscopy was used on unfixed, unstained materials.

2. The characteristics observed in normal megakaryocytes and in those from patients with I.T.P. confirmed data previously obtained with the use of fixed and stained material. In I.T.P. the bone marrow megakaryocytic series presented the following significant findings: (a) an increase in the number of the megakaryocytes, particularly of the younger forms. Young forms were particularly prominent in I.T.P. of the "acute" variety. (b) Production of platelets by promegakaryocytes, the platelets appearing large and bizarre. (c) Lack of platelet formation at the periphery of the cell and lack of granularity in the outer layer of the mature megakaryocyte. (d) Presensce of marked degenerative changes in the nucleus and cytoplasm of the megakaryocytes. The abnormalities observed suggested the possibility of a double mechanism: one in which the circulating platelets were injured, the other in which the megakaryocyte itself was attacked.

3. In a patient with a high-titer circulating platelet agglutinin the alterations of the megakaryocytes were fundamentally similar, although the granularity of the cytoplasm appeared to be less affected and the degenerative changes more pronounced. Similar abnormalities could be reproduced in the normal individuals receiving the patient's plasma. These results again suggested that, at least in those cases of I.T.P. in which the thrombocytopenia was due to a circulating antiplatelet factor, this acted by: (a) attacking the platelets being discharged from the megakaryocyte; (b) preventing the normal formation of platelets; (c) preventing the normal development of the megakaryocyte from younger to more mature forms.
Further Studies on Hemoglobin C
Authors: NEEL JAMES V. , KAPLAN EUGENE , ZUELZER WOLF W.
Published: October 2019

Three families are described in which there occur one or more children with sickle cell-hemoglobin C disease. Additional data are presented concerning two previously described families.
The balanced and the unbalanced chromosome aberrations of acute myeloid leukemia may develop in different ways and may contribute differently to malignant transformation
Authors: Pedersen-Bjergaard J , Rowley JD
Published: October 2019

Two general types of clonal chromosome abnormality are observed in de novo acute myeloid leukemia (AML): the unbalanced aberrations with visible gain or loss of chromosome material and the balanced aberrations without such visible gain or loss. AML can be induced by therapy with cytostatic drugs and radiation. The alkylating agents reacting directly with DNA induce AML which often presents as myelodysplasia with unbalanced aberrations, primarily loss of chromosome material. Cytostatic agents targeting DNA-topoisomerase II, frequently administered together with alkylating agents or cisplatin, induce the same type of leukemia. In addition, they often induce another type with a more rapid onset and with specific balanced chromosome aberrations rarely observed after therapy with alkylating agents alone. All of the most important chromosome aberrations found in de novo AML are now also found in therapy-related AML (t-AML); thus, t- AML may serve as a model in the search for mechanisms leading to the development of AML in general. Unbalanced chromosome aberrations with partial deletions or with loss of whole chromosomes may develop as a result of alkylation of DNA or other cellular targets. Balanced chromosome aberrations, on the other hand, may develop as illegitimate recombinations related to the activity of DNA-topoisomerase II. The balanced translocations contribute to malignant transformation by the formation of abnormal chimeric genes, whereas deletions may contribute by the loss of putative tumor suppressor genes. In either situation, the chromosome changes provide the altered cells with a proliferative advantage compared with normal cells.
Proteolysis of von Willebrand factor in therapeutic plasma concentrates
Authors: Mannuccio PM , Lattuada A , Ruggeri ZM
Published: October 2019

Therapeutic plasma concentrates containing vo Willebrand factor (vWF) lack the largest, most hemostatically active multimers. To evaluate whether this abnormality results from proteolysis during manufacturing, we have analyzed the subunit structure of vWF in several commercial products and found a marked reduction in the relative content of intact 225-kD subunit, paralleled by an increase in the proteolytic fragments normally present in plasma, particularly that of 176 kD. There was no heightened vWF fragmentation in blood-bank cryoprecipitate prepared from platelet-poor, single-donor plasma; in contrast, there was a marked degree of fragmentation in cryoprecipitate prepared from pooled plasmapheresis plasma representing the starting fraction for the production of commercial concentrates. In cryoprecipitate prepared experimentally from plasma containing varying numbers of platelets, the degradation of vWF was proportional to the platelet count, but was greatly diminished by adding protease inhibitors to the plasma. On the basis of these findings, we postulate that the loss of the largest vWF multimers in commercial products results from the use of poorly centrifuged plasmapheresis plasma containing an excessive number of residual platelets and leukocytes. These cells, lysing when plasma is frozen and thawed for the preparation of cryoprecipitate, may liberate proteolytic enzymes that cleave the vWF subunit and contribute to the degradation of the largest multimers. Our results should help devise new approaches for the preparation of more effective concentrates for the treatment of von Willebrand disease.
Bedside filtration of blood products in the prevention of HLA alloimmunization--a prospective randomized study. Alloimmunisation Study Group
Authors: Williamson LM , Wimperis JZ , Williamson P , Copplestone JA , Gooi HC , Morgenstern GR , Norfolk DR
Published: October 2019

To test the efficacy of poststorage bedside leucodepletion of blood products in the prevention of primary HLA alloimmunization and its clinical sequelae, 172 patients with hematologic malignancy requiring intensive red blood cell and platelet support were randomized to receive either standard or filtered red blood cells and platelets. Quality control of bedside filtration was explored by sequential sampling downstream of the filter, but this did not predict the total number of leucocytes transfused. After exclusions, 123 evaluable patients were assessed every two weeks until the end of therapy. HLA antibodies developed in 21 of 56 (37.5%) nonfilter (NF) and 15 of 67 (22%) filter (F) patients (risk ratio estimate, 0.60 [95% confidence interval, 0.34 to 1.05]; P = .07). Patients with acute myeloid leukemia (AML; n = 53) had higher alloimmunization rates in both arms of the study, with a greater effect of filtration (62.5% NF and 31.0% F; P = .025). Bedside filtration did not affect the overall incidence of febrile transfusion reactions (FTRs; 37% NF and 34% F; P = .71) or of platelet refractoriness assessed in 50 patients (30% NF and 26% F), despite an association between broad HLA reactivity and both FTRs and refractoriness. However, FTRs were also seen in 28 patients without HLA antibodies. Five alloimmunized refractory patients (2 F and 3 NF) required HLA-selected platelets. This report, the first prospective study of bedside filtration, has failed to show clear clinical benefit. Methodological limitations may account in part for this failure, notably the difficulties in accurately assessing the number of leucocytes transfused.
7E3 F(ab′)2 , a Monoclonal Antibody to the Platelet GPIIb/IIIa Receptor, Protects Against Microangiopathic Hemolytic Anemia and Microvascular Thrombotic Renal Failure in Baboons Treated With C4b Binding Protein and a Sublethal Infusion of Escherichia coli
Authors: Taylor F.B. , Coller B.S. , Chang A.C.K. , Peer G. , Jordan R. , Engellener W. , Esmon C.T.
Published: October 2019

We have used our previously described baboon model of infusion of both a sublethal dose of Escherichia coli and C4b binding protein to assess the impact of inhibiting platelet function with the F(ab′ )2 fragment of the monoclonal antibody 7E3, directed against the platelet glycoprotein (GP)IIb/IIIa receptor, on the characteristic microvascular changes. At a dose of 0.25 to 0.35 mg/kg bolus plus an infusion of 0.25 to 0.35 mg/kg over 6 hours, c7E3 F(ab′ )2 had only a minimal impact on fibrinogen consumption and delayed but did not prevent, the development of thrombocytopenia. Treatment with 7E3 F(ab′ )2 , however, produced significant protection from the development of microangiopathic hemolysis and renal insufficiency. Histologic examination supported these observations, with treated animals having fewer schistocytes on blood smear and less evidence of ischemic renal changes. Treated animals also had more rapid recovery of peripheral white blood counts, suggesting a possible protective effect of treatment on ischemic damage to the bone marrow. These data indicate that potent inhibition of platelet function via GPIIb/IIIa receptor blockade can decrease ischemic organ damage in this animal model that has features similar to those found in diffuse intravascular coagulation, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura.
Molecular Mechanisms of Monocyte Adhesion to Interleukin-1β–Stimulated Endothelial Cells Under Physiologic Flow Conditions
Authors: Kukreti Sharad , Konstantopoulos Konstantinos , Smith C. Wayne , McIntire Larry V.
Published: October 2019

This study identifies multiple pathways used by monocytes to adhere to 4-hour interleukin-1β stimulated human umbilical vein endothelial cells under flow conditions. Physiologic shear stresses were simulated in a flow chamber with parallel plate geometry; quantitation of primary adhesion, secondary adhesion, and transmigration was performed using phase contrast videomicroscopy. Neuraminidase treatment of monocytes reduced primary interaction by 50%, whereas blocking L-selectin or very late antigen-4 showed significant but smaller effects (∼30% inhibition). However, a combined treatment against all three pathways was able to reduce interaction by 80%. Blocking β2 and α4 integrin pathways together inhibited secondary/firm adhesion by 75%. Only 40% of firmly adherent monocytes transmigrated across the endothelial monolayer with significantly increased transmigration times when both β2 and α4 integrins were blocked. These results demonstrate that monocytes can use multiple receptors to interact with endothelial cells at both primary and secondary adhesion stages, and that these pathways have to be blocked simultaneously for maximum inhibition.
Differential Rates of Somatic Hypermutation in VH Genes Among Subsets of Chronic Lymphocytic Leukemia Defined by Chromosomal Abnormalities
Authors: Oscier David G. , Thompsett Andrew , Zhu Delin , Stevenson Freda K.
Published: October 2019

Chronic lymphocytic leukemia (CLL) is a B-cell tumor involving small lymphocytes that generally express the CD5 antigen and low levels of surface Ig. Within this definition, there is heterogeneity among cases in cell morphology, karyotypic abnormalities, and clinical course. Trisomy 12, the most frequent karyotypic abnormality, is commonly found in a subset of CLL with atypical morphology. It has also been associated with advanced disease, and possibly with a less favorable prognosis. A further subset of cases with abnormalities involving chromosome 13q14 have typical lymphocyte morphology. Occasionally, the two abnormalities are found together. To assess the clonal history of the cell of origin in disease subsets defined by these two chromosomal abnormalities, we investigated the usage of VH genes and the pattern of somatic mutation in 10 cases of trisomy 12 with atypical morphology and eight cases of 13q14 abnormality with typical morphology. In addition, four cases with both chromosomal abnormalities were analyzed. Results confirm a common usage of the VH1 family in all subsets. However, the patterns of somatic mutation were distinct, with cases of trisomy 12 showing a minimal level of mutation (mean ± SD, 0.34% ± 0.86%) and cases of 13q14 abnormality showing significant levels (6.5% ± 1.67%). The four cases with both abnormalities showed a mixed pattern. All mutated cases had intraclonal homogeneity, and three of 10 had a pattern indicative of antigen selection. These results suggest that the clonal history of the two subsets of CLL may differ.
Homozygous Deletion of the p16/MTS1 Gene in Pediatric Acute Lymphoblastic Leukemia Is Associated With Unfavorable Clinical Outcome
Authors: Kees Ursula R. , Burton Paul R. , Lü Changlong , Baker David L.
Published: October 2019

The p16 gene (MTS1, CDKN2, p16INK4A, CDKI) encoding an inhibitor of cyclin-dependent kinase 4 (cdk4) has been found to be deleted in various types of tumors, including leukemia, and is thought to code for a tumor suppressor gene. Our preliminary findings on eight pediatric patients with acute lymphoblastic leukemia (ALL) suggested that the survival of patients carrying a homozygous p16 gene deletion was significantly inferior to that of those without a deletion. The present study on 48 patients tested the hypothesis that the clinical outcome for pediatric ALL patients is correlated with the presence or absence of the p16 gene. Overall, nine of 48 children (18.3%) carried a homozygous p16 deletion. Such deletions were significantly more common (P = .003) among T-ALL patients (five of eight, 62.5%) than among precursor-B-ALL patients (four of 40, 10.0%). Of nine patients exhibiting p16 deletions, eight (88.9%) were classified as high-risk patients by the recognized prognostic factors of age, white blood cell count, and T-cell phenotype. The 4-year event-free survival in the study population as a whole was 72.7%. Without adjustment for other risk factors (univariate model), the presence of a homozygous p16 deletion was associated with a markedly increased probability of both relapse (P = .0003) and death (P = .002). These findings raise the question of whether the p16 deletion itself confers an increased risk of relapse after adjusting for the known risk factors. In this analysis, the estimated risk multiplier factor for relapse in patients carrying the p16 deletion was 14.0 (P = .0004) and for the risk of death 15.6 (P = .0008). We therefore conclude that the presence of a homozygous p16 deletion may well be an important risk factor for both relapse and death in childhood ALL, and that its prognostic effect is not a consequence of confounding by other factors already known to influence outcome in this disease.
Tissue Factor Pathway Inhibitor Blocks Cellular Effects of Endotoxin by Binding to Endotoxin and Interfering With Transfer to CD14
Authors: Park C. Thomas , Creasey Abla A. , Wright Samuel D.
Published: October 2019

Tissue factor pathway inhibitor (TFPI) is a Kunitz-type plasma protease inhibitor that inhibits factor Xa and the factor VIIa/tissue factor catalytic complex. It plays an important role in feedback inhibition of the coagulation cascade (Broze, Annu Rev Med 46:103, 1995). TFPI has also been used successfully to prevent lethality and attenuate coagulopathic responses in a baboon model of septic shock (Creasey et al, J Clin Invest 91:2850, 1993; and Carr et al, Circ Shock 44:126, 1995). However, the mechanism of reduced mortality in these animals could not be explained merely by the anticoagulant effect of TFPI, because TFPI-treated animals also had a significantly depressed interleukin-6 response. Moreover, inhibition of coagulopathic responses by other anticoagulants has failed to block the organ damage or lethal effect of endotoxic shock (Coalson et al, Circ Shock 5:423, 1978; Warr et al, Blood 75:1481, 1990; and Taylor et al, Blood 78:364, 1991). We show here that recombinant TFPI can bind to endotoxin in vitro. This binding prevents interaction of endotoxin with both lipopolysaccharide binding protein and CD14, thereby blocking cellular responses.