bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody-induced von Willebrand's disease: a newly defined inhibitor syndrome
Authors: Handin RI , Martin V , Moloney WC
Published: October 2019

A patient is described with clinical and laboratory evidence of von Willebrand's disease (VWD) in association with lymphosarcoma. He consistently had a bleeding time of over 20 min; factor VIIAHF, VIIAGN, and VIIVWF under 20%; and severe, diffuse hemorrhage, Following transfusion with cryoprecipitate, the patient had the expected rise in VIIAGN and VIIAHF. The patient's plasma contained an inhibitor which prevented aggregation of normal platelets by ristocetin, but which did not interfere with the measurement of VIIAGN or inactivate VIIAHF activity. The inhibitor was present in serum heated at 56 degrees C for 30 min, was present in a purified IgG fraction of serum, and was precipitated by anti-human IgG. The antibody did not interact directly with washed platelets, but appeared to interact with that portion of the factor VII protein that supports ristocetin aggregation (VIIIVWF). The data provide the first explanation for the pathophysiology of the acquired von Willebrand's syndrome.
Acquired storage pool disease in platelets during disseminated intravascular coagulation
Authors: Pareti FI , Capitanio A , Mannucci PM
Published: October 2019

A patient with clinical and laboratory evidence of disseminated intravascular coagulation associated with deep-vein thrombosis and pulmonary embolism developed a qualitative platelet abnormality characterized by a defective release reaction. Second-phase aggregation induced by ADP and adrenaline was impaired, and reduced collagen- induced aggregation was accompanied by defective release of ADP and ATP. The decrease in total platelet ATP and ADP, the high ATP:ADP ratio in the presence of normal amounts of metabolic adenine nucleotides, and the low content of serotonin associated with abnormal uptake and metabolism of the exogenous amine suggested that the defective platelet function was due to lack of the platelet organelles in which serotonin and nonmetabolic adenine nucleotides are normally stored. Acquired storage pool disease is likely to be related to exposure of circulating platelets to aggregating agents, with their degranulation occurring during disseminated intravascular coagulation.
A platelet abnormality in the Chediak-Higashi syndrome of man
Authors: Costa JL , Fauci AS , Wolff SM
Published: October 2019

Platelets from two probands homozygous for the Chediak-Higashi syndrome have approximately 10% of the normal number of serotonin-containing dense bodies as visualized electron microscopically in air-dried whole mounts. Since transport of serotonin across the platelet plasma membrane proceeds at a normal rate, and the few dense bodies present appear to store normal amounts of serotonin, the absence of dense bodies may account for the low platelet serotonin values found in these patients.
Fibrinolysis in normal plasma and blood: evidence for significant mechanisms independent of the plasminogen-plasmin system
Authors: Moroz LA , Gilmore NJ
Published: October 2019

Fibrinolytic activity of normal plasma and blood has been measured by 125l-fibrin solid phase assay. Activity of plasma is not affected by removal of plasminogenplasmin by affinity chromatography. Activities of euglobulin and pseudoglobulin fractions are approximately equal. epsilon-aminocaproic acid (EACA) (10 mM), tranexamic acid (10 mM), diisopropylfluorophosphate (DFP, 50 mM), and soybean and lima bean trypsin inhibitors (100 mug/ml) do not inhibit plasma activity at concentrations that inhibit pure plasmin and urokinase-activated plasma. Activity is not affected by glass contact and is not inhibited by inhibitors of contact or enzymatic activation of Hageman factor (hexadimethrine bromide, 100 mug/ml; cytochrome C, 250 mug/ml; spermidine, 2 mM; phenylmethylsulfonylfluoride, 1 mM). It is inhibited partially (30%-40%) by heating (56 degrees C, 30 min) and by zymosan (2.5 mg/ml; 40%-90% inhibition), and is increased by hydrazine (20 mM), salicylaldoxime (20 mM), DFP (50 mM), and tosyl-L-arginine methyl ester (TAMe, 10 mM)-the latter two at concentrations known to inhibit Cls of the classic, and factor D of the alternate complement pathways. Increase fibrinolytic activity with TAMe is associated with reciprocal decrease in classic and alternate complement pathway activity. It is concluded that normal plasma fibrinolytic activity is relatively independent of plasmin as the ultimate fibrinolytic enzyme, that Hageman factor-dependent pathways are of minor importance, and that significant heat-stable and heat-labile nonplasmin fibrinolytic activities are operative. These may include proteinases involved in complement activation, and in common control of classic and alternate complement pathways, as well as other nonplasmin proteinases.
Synthesis of cobalamin coenzymes by human lymphocytes in vitro and the effect of folates and metabolic inhibitors
Authors: Quadros EV , Matthews DM , Hoffbrand AV , Linnell JC
Published: October 2019

The uptake of 57Co-cyanocobalamin (CN-Cbl) and its conversion to 5- deoxyadenosylcobalamin (Ado-Cbl), methylcobalamin (Me-Cbl), and hydroxocobalamin (OH-Cbl) has been studied in phytohemagglutinin (PHA)- transformed lymphocytes from normal subjects and patients with patients with pernicious anemia. Uptake and conversion were much greater by PHA- stimulated lymphocytes than by mature non-transformed lymphocytes. In normal cells, uptake of 57Co-CN-Cbl and synthesis of the cobalamin coenzymes were approximately linear between 3 and 48 hr incubation. Ado- Cbl was the major cobalamin formed, and after 72 hr the cells contained about twice as much Ado-Cbl as Me-Cbl. Uptake by lymphocytes from patients with untreated pernicious anemia (PA) was greater than that by normal lymphocytes, but the proportions of Ado-Cbl and Me-Cbl synthesized by each were similar. Folic acid and methyltetrahydrofolate enhanced synthesis of Me-Cbl both in normal and in PA cells, while methotrexate and 5-fluorouracil depressed it. This depression was overcome by 5-formyltetrahydrofolate, suggesting that an uninterrupted folate cycle may play an important role in Me-Cbl synthesis.
Surface markers of human eosinophils
Authors: Gupta S , Ross GD , Good RA , Siegal FP
Published: October 2019

Peripheral blood eosinophils from patients with eosinophilia and from healthy subjects were studied for surface immunoglobulins, receptors for the Fc region of IgG, complement receptors, and spontaneous rosette formation with sheep and mouse erythrocytes. Eosinophils were found to have receptors for complement and for aggregated IgG, and to have the same two types of complement receptors as do lymphocytes and monocytes. Immune adherence type receptors were specific for C4 or C3b, while C3d receptors were specific for C3d but unreactive with C4. Eosinophils differed from fully mature neutrophils in that the former had C3d receptors and relatively weak immune adherence (C4 or C3b) receptors, while the later did not have the C3d receptors and had strong immune adherence receptors. Eosinophil phagocytosis of complement-receptor bound erythrocytes was dependent on the presence of IgG in the antibody coating the red blood cells; this requirement for IgG resembled that found in neutrophil phagocytosis. No surface Ig or spontaneous erythrocyte rosette formation was observed with eosinophils.
Mechanisms involved in enhancement of plasma fibrinolytic activity by chloroform
Authors: Moroz LA , Gilmore NJ
Published: October 2019

The effects of a single 1-min extraction with chloroform (CHCl3) on plasma fibrinolytic activity has been examined by 125I-fibrin solid phase assay, using normal plasma and plasma depleted of plasminogen (PLG) by lysine-Sepharose affinity chromatography. Fibrinolytic activity of normal plasma is increased (40%-175%), and more than 95% of antiplasmin activity is removed. The increase is demonstrable in PLG- depleted plasma, and is not inhibited by tranexamic acid (0.01 M). Purified PLG is not activated to plasmin by ChCl3 treatment. Bio-Gel A 0.5 m fractionation of CHCl3-extracted, PLG-depleted plasma reveals fractions with the following activities: (1) streptokinase-activatable, PLG-independent fibrinolytic activities; (2) PLG activator activities; and (3) plasmin-stimulated but PLG-independent fibrinolytic activities, which include activities inhibited by hexadimethrine bromide and which cofractionate in part with plasmin-stimulated procoagulant activities. In addition, similar fractionation of nonextracted plasma reveals two non-plasmin fibrinolytic activities (approximately 30,000 and 13,000 daltons) activated by streptokinase and plasmin, respectively. The findings indicate that the enhanced fibrinolytic activity resulting from CHCl3 treatment is independent of plasmin as the ultimate fibrinolytic enzyme, although activities stimulated by plasmin may contribute, and that such treatment is a useful maneuver for study of PLG-dependent and PLG-independent fibrinolytic mechanisms, and their interactions.
The effect of chelating agents on iron mobilization in Chang cell cultures
Authors: White GP , Jacobs A , Grady RW , Cerami A
Published: October 2019

The investigation of chelating agents with potential therapeutic value in patients with transfusional iron overload has been facilitated by the use of Chang cell cultures. These cells have been incubated with [59Fe]transferrin for 22 hr, following which most of the intracellular radioiron is found in the cytosol, distributed between a ferritin and a nonferritin form. Iron release from the cells depends on transferrin saturation in the medium, but when transferrin is 100% saturated, which normally does not allow iron release, desferrioxamine, 2,3- dihydroxybenzoic acid, rhodotorulic acid, cholythydroxamic acid, and tropolone all promote the mobilization of ferritin iron and its release from cells. They are effective to an approximately equal degree. The incubation of [59Fe]transferrin with tropolone in vitro at a molar ratio of 1:500 results in the transfer of most of the labeled iron to the chelator, reflecting the exceptionally high binding constant of this compound. How far these phenomena relate to therapeutic potentially remains to be seen.
Evidence suggesting the regulation of a coagulation factor levels in rabbits by a transferable plasma agent
Authors: Friedman EW , Karpatkin M , Karpatkin S
Published: October 2019

New Zealand white rabbits were given 30 ml of goat serum intravenously. This procedure resulted in an immediate decrease in platelet count, fibrinogen, and levels of coagulation factors II, V, VII, and X, due to consumption coagulopathy. These factors returned toward baseline levels approximately 12 hr after the injection. Plasma from rabbits who had received goat serum 48 hr previously (donor rabbits) was injected into recipient rabbits. This procedure resulted in a slight rise in the level of coagulation factor II (range, 20%-30%) and a significant rise in factors V (35%-75%), VII (35%-235%), and X (35%-75%) in the recipients. When plasma from control donor rabbits who had not received goat serum was injected into recipients, there was no change in these coagulation factors. It is postulated that the reduction in coagulation factor levels in donor rabbits induces a “coagulopoietin” for each factor or one “coagulopoietin” for all factors which stimulates increased synthesis and/or release of these factors in recipient rabbits.
Effect of induced fever on serum iron and ferritin concentrations in man
Authors: Elin RJ , Wolff SM , Finch CA
Published: October 2019

Previous reports have shown that endotoxin decreases serum iron in experimental animals. In this study fever was produced in nine female and nine male normal subjects in order to define the temporal and quantitative changes in serum iron and ferritin concentrations. Six volunteers were randomly given bacterial endotoxin (5 ng/kg) or saline intravenously and received the alternative compound a week later. Serial blood samples were drawn at 4-hr intervals for a 24-hr period, beginning when the compound was administered, for the determination of serum iron and ferritin concentrations. The same study was performed with intramuscular etiocholanolone (0.3 mg/kg) or the vehicle, propylene glycol, as a control, but the first blood sample was obtained 9 hr after the compound was given. In addition, blood samples were obtained at 12-hr intervals in six volunteers for 11 days after an intramuscular injection of etiocholanolone. The results showed a significant increase (p less than 0.005 for etiocholanolone, P less than 0.01 for endotoxin) in serum ferritin and a significant decrease (p less than 0.005 for etiocholanolone, p less than 0.001 for endotoxin) in serum iron for both pyrogenic compounds compared with the control compounds. However, the amount of fever and the changes in the iron parameters were greater with etiocholanolone. One episode of induced fever with etiocholanolone effected changes in serum ferritin and iron concentrations that lasted 10 days. Thus this study demonstrated that a single episode of fever in man produced rapid and prolonged changes in serum iron and ferritin concentrations.
Heterogeneity of human whole blood platelet subpopulations. I. Relationship between buoyant density, cell volume, and ultrastructure
Authors: Corash L , Tan H , Gralnick HR
Published: October 2019

A quantitative high yield method utilizing isosmotic arabino-galactan (Stractan) solutions and isopycnic centrifugation was developed to isolate and to fractionate total human platelet populations into density-dependent subpopulations. Isolated platelets were free of factor VIII/von Willebrand factor and other plasma proteins. They responded to ADP, epinephrine, and collagen with a sensitivity equal to platelet-rich plasma platelets. The correlation of platelet density with volume and ultrastructure was examined for normal subjects. Recovery of total platelet populations averaged 92.76%+/-3.64% (SD). Normal individuals exhibited a narrow range of platelet buoyant density distribution. Computerized probability plot analysis of platelet volume distribution for 15 normal subjects' total platelet populations showed a mean volume of 5.17+/-0.46 cu mum (SD). Platelets with buoyant density less than or equal to 1.062 g/ml had a mean volume of 4.50+/- 0.48 cu mum, while platelets with buoyant density greater than 1.071 g/ml, but less than or equal to 1.084 g/ml, had a mean volume of 5.32+/- 0.63 cu mum (SD). The volume difference by paired t test was significant, p less than 0.001. Thin-section electron microscopy demonstrated a significant reduction of granule content in light platelets, as compared to heavy platelets, but an equal number of mitochondria for both groups. These differences of platelet volume and structure between light and heavy platelets suggested that aging may be a determinant of platelet heterogeneity.
Demonstration of granulocytic proteases in plasma of patients with acute leukemia and septicemia with coagulation defects
Authors: Egbring R , Schmidt W , Fuchs G , Havemann K
Published: October 2019

To show whether direct proteolysis of coagulation factors may play a role in patients with so-called consumption coagulopathy, granulocytic neutral proteases in the plasma of patients with acute myelocytic leukemia and septicemia were assayed by one- and two-dimensional Laurell electrophoresis. Complexes between serum alpha1-antitrypsin and elastase-like granulocytic protease could be demonstrated in those patients with acute myelocytic leukemia and septicemia who also had moderate or severe coagulation defects. Despite the presence of a high antiprotease potential, addition of the elastase-like enzyme to normal plasma resulted in coagulation defects in vitro comparable to those seen in the patients. These results and the ability of the elastase- like protease to destroy isolated clotting factors suggested that in certain types of coagulation factor deficiencies direct proteolysis rather than consumption of clotting factors due to disseminated intravascular coagulation may be operational.
Ph1-positive acute lymphocytic leukemia with chromosome 7 abnormalities
Authors: Mandel EM , Shabtai F , Gafter U , Klein B , Halbrecht I , Djaldetti M
Published: October 2019

A 56-yr-old woman with acute lymphocytic leukemia showed the presence of a Philadelphia chromosome in 90% of the bone marrow metaphases and in 10% of the peripheral blood metaphases. Part of the long arm of a G22 chromsome was translocated to the long arm of a C9 chromosome. A second cytogenetic abnormality was found in chromosome 7. Monosomy 7 was found in 60% of the marrow and in 20% of the peripheral blood metaphases. Chromosome 7q- was also found in a small percentage of the metaphases examined. Three months later, when the patient was in partial remission, only 10% of the marrow cells showed chromosome 7 monosomy and a Ph1 chromosome. During complete remission, no chromosomal abnormalities were found, except for a high breakage rate. The finding of a Ph1 chromosome in acute lymphocytic leukemia indicates that different precursors, both granulocytic and lymphocytic, may be involved in the Ph1 process.
Combination chemotherapy ("CHOP-Bleo") in advanced (non-Hodgkin) malignant lymphoma
Authors: Rodriguez V , Cabanillas F , Burgess MA , McKelvey EM , Valdivieso M , Bodey GP , Freireich EJ
Published: October 2019

Forty-seven adults with advanced malignant lymphoma (the majority in stage IV) were treated with a combination of cyclophosphamide, hydroxyldaunorubicin (Adriamycin), vincristine (Oncovin), prednisone, and bleomycin (CHOP-Bleo). The complete remission (CR) rate was 66%. The overall response (complete + partial remission) was 92%. The CR rate in patients with diffuse histiocytic lymphoma (DHL) was 69%. Only 3 of the 18 patients with DHL in CR have relapsed; the projected median duration of response was calculated to be greater than 2 yr. In patients with nodular poorly differentiated lymphocytic lymphoma (NPDL), the CR rate was 62%. One of the eight patients with NPDL in CR has relapsed; the projected median duration of complete response will be greater than 4 yr. The median survival for all patients entered in this study has not been reached; however, it was estimated that it will be greater than 3 yr. The survival curves became flat at 70 wk for the patients with DHL and at 1 yr for the patients with NPDL. Major complications during chemotherapy with CHOP-Bleo were myelosuppression and alopecia. Only six severe infections occurred during myelosuppression. No hemorrhagic problems were observed. This study indicates that combination chemotherapy with these agents is effective in increasing the CR rate and survival in patients with diffuse histiocytic lymphoma. In patients with NPDL, further observation will be needed to assess the effect of this combination on survival.
Electrolyte and acid-base disturbances in the management of leukemia
Authors: O'Regan S , Carson S , Chesney RW , Drummond KN
Published: October 2019

Electrolyte disturbances in leukemia can be the result of the disease process or drug therapy. One group of electrolyte abnormalities is related to the stage of the leukemic process. Included in this group are newly diagnosed patients who may show elevated serum potassium, phosphorus, and magnesium--a result of their release from malignant cells after cytotoxic therapy or their accumulation due to urate nephropathy. Patients in remission usually have normal serum electrolyte concentrations, but acute leukemia patients during relapse may have hypokalemia, hypophosphatemia, and hypomagnesemia. This imbalance may be related to cellular uptake of these electrolytes in the presence of inadequate dietary intake. Other factors contributing to electrolyte derangements, and related to the leukemic process, include hyponatremia and hypochloremia secondary to the SIADH, hypokalemia in acute monocytic or acute myelomonocytic leukemia due to lysozyme-induced tubular damage, hypercalcemia possibly secondary to leukemic infiltration of bone or parathyroid glands (with PTH release), or production of a PTH-like substance by leukemic cells. Nonspecific factors related to the disease process which may aggravate the electrolyte imbalance include gastrointestinal loss through nausea, vomiting, and malnutrition. The drug-related electrolyte abnormalities include cyclophosphamide- and vincristine-induced SIADH; decreased serum sodium, chloride, potassium, and calcium concentrations as a result of polymyxin B nephrotoxicity; hypokalemia and hypomagnesemia secondary to amphotericin B; hypocalcemia, hypophosphatemia, and hyperphosphaturia due to L-asparaginase-induced hypoparathyroidism; hypokalemia due to a nonreabsorbable anion effect of antibiotics in the distal tubule or changes in membrane ionic transport of all cells by large doses of antibiotics. Electrolyte disturbance in leukemia thus have a multifactorial pathogenesis which can best be delineated according to the stage of the leukemic process and the drugs being used. Recognition of the cause or causes in a particular patient is essential for an effective approach to management. This review emphasizes the need for routine measurement of serum electrolytes during all phases of the leukemic process.
Free erythrocyte porphyrin: hemoglobin ratios, serum ferritin, and transferrin saturation levels during treatment of infants with iron- deficiency anemia
Authors: Thomas WJ , Koenig HM , Lightsey AL Jr , Green R
Published: October 2019

Free erythrocyte porphyrin:hemoglobin (FEP:Hb) ratios were determined on 20 infants with iron-deficiency anemia. FEB:Hb ratios were compared with simultaneously drawn serum ferritin and serum transferrin saturation levels. FEP:Hb ratios dropped steadily during treatment of the anemic infants, but remained elevated compared to age-matched nonanemic infants, until the anemia was corrected. FEP:Hb ratios detected iron deficiency when acute inflammatory disease was present. Serum ferritin levels and transferrin saturation failed to detect iron deficiency after iron therapy was started or when acute inflammatory disease was present. Measurement of FEP:Hb ratios is a reliable and practical way to make a prompt diagnosis of iron-deficiency anemia in infants.
Glucose-6-phosphate dehydrogenase deficiency and red cell glutathione peroxidase
Authors: Beutler E
Published: October 2019

The activity of glutathione peroxidase (GSH Px), glucose-6-phosphate dehydrogenase (G-6-PD), hexokinase, and glutamic oxaloacetic transaminase (EGOT) was measured in 78 blood samples. GSH Px activity was not found to correlate with hexokinase or EGOT activity, indicating that it was not a strongly age-dependent enzyme. Although modest elevations of GSH Px activity were observed in the red cells of patients with a variety of hematologic disorders, the most consistent and striking increases in activity were observed in G-6-PD-deficient subjects.
Significance of the intact polypeptide chains of human fibrinogen in ADP-induced platelet aggregation
Authors: Niewiarowski S , Budzynski AZ , Lipinski B
Published: October 2019

The presence of human fibrinogen in suspensions of washed human platelets is a requirement for ADP-induced platelet aggregation. Digestion of fibrinogen with plasmin destroys this function of the protein. The high solubility fraction of Kabi fibrinogen, fragment X (stage 1) and framgent X (stage 2), are two, eight, and ten times, respectively, less potent in promoting ADP-induced platelet aggregation, as compared with intact fibrinogen. Fragments Y and D and the mixture of reduced and carboxymethylated chains of human fibrinogen do not support ADP-induced platelet aggregation at all. SDS polyacrylamide gel electrophoresis of nonreduced and reduced fibrinogen and its derivatives indicates that the intact fibrinogen molecule is essential for ADP-induced platelet aggregation. It is suggested that the carboxy-terminal part of the Aalpha chain and possibly also the amino-terminal part of the Bbeta chain are required for the platelet aggregation-promoting function of fibrinogen.
Current status of bone marrow transplantation for aplastic anemia and acute leukemia
Authors: Thomas ED , Fefer A , Buckner CD , Storb R
Published: October 2019

Clinical results of marrow transplantation in the treatment of aplastic anemia and acute leukemia are reviewed. The principal problem areas in this field at this time are discussed.
Circulating anticoagulant in a family with prolonged bleeding time and factor VIII deficiency
Authors: Diez-Ewald M , Lian EC , Nunez R , Deykin D , Harkness DR
Published: October 2019

A circulating anticoagulant against factor VIII activity was demonstrated in the plasma of a boy from a family with both factor VIII deficiency and prolonged bleeding time. However, the factor VIII- related antigen, ristocetin-induced platelet aggregation activity, platelet retention in glass bead columns, platelet aggregation with adenosine 5′-diphosphate, collagen and epinephrine, and clot retraction among affected members were normal. The electrophoretic mobility of factor VIII-related antigen on crossed immunoelectrophoresis was normal. The inactivation of factor VIII activity by the inhibitor was time dependent and was nonlinear as the concentration of the inhibitor was increased. Immunotyping showed that the inhibitor was IgG with k light chains.
Heterogeneity of factor VIII antibodies: further immunochemical and biologic studies
Authors: Hultin MB , London FS , Shapiro SS , Yount WJ
Published: October 2019

Previous studies using immunoneutralization techniques have shown that many factor VIII inhibitors are IgG antibodies of a single light chain type. We have investigated this apparent homogeneity by immunoneutralization assay and liquid isoelectric focusing of inhibitor fractions from five hemophiliacs and two nonhemophiliacs. By immunoneutralization assay, inhibitors from four hemophiliacs and one nonhemophiliac were exclusively k light chain type: the fifth hemophilic inhibitor was predominantly k1 and the second nonhemophilic inhibitor was a mixture of k and gamma. However, heavy chain subtyping of the six predominantly or exclusively k inhibitors showed all to be mixtures of IgG4 and IgG1. By isoelectric focusing, two inhibitors showed multiple peaks of activity between pH 5 and 9. The remaining five showed predominant peaks of activity, which were solely IgGk1 between pH 5.8 and 7, with smaller peaks between pH 7 and 9. The most acidic major peak, focusing at pH 6, was IgG4 in the three cases tested. Two of these acidic peaks neutralized factor VIII more efficiently than other peaks in the same focusing profiles, suggesting greater affinity for factor VIII. These studies demonstrate that factor VIII inhibitors are composed of heterogenous subpopulations of immunoglobulins which can be separated by isoelectric focusing.
Human marrow cells capable of erythropoietic differentiation in vitro: definition of three erythroid colony responses
Authors: Gregory CJ , Eaves AC
Published: October 2019

A systematic study has been undertaken to analyze the spectrum of erythropoietic colonies obtained in cultures of human marrow cells plated in methyl cellulose. Colonies were identified as erythropoietic on the basis of the appearance in them of hemoglobin-containing erythroblasts. As found previously in mouse marrow cultures, three sequentially appearing types of colonies which differed in their ultimate cluster content could be readily distinguished. Small erythroid colonies containing 1–2 clusters reached a peak after 7–8 days; small bursts containing 3–8 clusters reached a peak after 10–12 days; and large bursts containing greater than 16 clusters reached a peak after 17–20 days. The previously reported enhancing effect of human leukocyte conditioned medium on burst formation seen in cultures of human nonadherent cells was found to be due largely to an effect on the formation of the largest, late appearing type of burst. By analogy with the mouse, the progenitors of such bursts would represent a primitive cell type which has a close relationship with pluripotent stem cells, as well as a second and independent close relationship to the progenitors of granulopoietic colonies.
Prognostic classification of Hodgkin disease in pathologic stage III, based on anatomic considerations
Authors: Desser RK , Golomb HM , Ultmann JE , Ferguson DJ , Moran EM , Griem ML , Vardiman J , Miller B , Oetzel N , Sweet D , Lester EP , Kinzie JJ , Blough R
Published: October 2019

Fifty-two patients with pathologic stage III Hodgkin disease were studied in an effort to determine whether location of involved abdominal nodes influenced survival. Treatment consisted of total nodal radiotherapy with or without subsequent combination chemotherapy. Th initial radiation field was the “extended mantle,” which included supradiaphragmatic nodes, the splenic hilar area, and paraaortic nodes to the level of L2-L4. Subsequently, lower paraaortic and iliac regions were treated (“lower inverted Y”). Patients with disease limited to the spleen and/or splenic, celiac, or portal nodes (“anatomic substage” III1) had a more favorable 5-yr survival than did patients with involvement of paraaortic, iliac, or mesenteric nodes (“anatomic substage” III2): 93% versus 57%, respectively (p less than 0.05). The addition of combination chemotherapy to total nodal irradiation was associated with improved survival of patients in stage III2, but not of those in stage III1.
Effect of procaine HCLl on ATP: calcium-dependent alterations in red cell shape and deformability
Authors: Palek J , Liu A , Liu D , Snyder LM , Fortier NL , Njoku G , Kiernan F , Funk D , Crusberg T
Published: October 2019

Procaine hydrochloric acid, a cationic anesthetic, although unable to prevent the effect of calcium ionophore A23187 on erythrocytes, inhibited the discocyte--echinocyte transformation, increased viscosity, and decreased filterability of red cells undergoing ATP depletion. The effects were abolished by washing ATP-depleted, procaine HCl-treated red cells prior to these determinations. Procaine HCl had no effects on volume, incubated osmotic fragility, or monovalent cation composition of ATP-depleted red cells. The drug increased 45Ca uptake by ATP-depleted red cells but did not change the fraction of membrane- bound calcium. Sodium dodecyl sulfate acrylamide gel electrophoresis of membrane proteins from ATP-depleted red cells revealed formation of high molecular weight protein complexes, which were not formed when biconcave shape and ATP content were maintained by incubation with adenine (0.54 mM) and inosine (12.7 mM); Formation of these complexes was not prevented when the biconcave shape was maintained by procaine HCl. It was concluded that the maintenance of the biconcave shape and normal deformability during ATP depletion by procaine HCl was not related to a displacement of membrane-bound calcium and inhibition of ATP-dependent rearrangement of red cell membrane proteins.
Metabolic activity of phagocytosing granulocytes in chronic granulocytic leukemia: ultrastructural observation of a degranulation defect
Authors: Cramer E , Auclair C , Hakim J , Feliu E , Boucherot J , Troube H , Bernard JF , Bergogne E , Boivin P
Published: October 2019

The functional capacities of granulocytes in patients with chronic granulocytic leukemia are still a subject of controversy, probably due to the heterogeneity of the abnormalities observed from patient to patient. For a better definition of these abnormalities, 14 patients with untreated chronic granulocytic leukemia were studied. The patients were divided into three groups on the basis of the functional activities of their phagocytosing granulocytes. In four patients (group I), the granulocytes were normal in respect to particle ingestion, nitroblue tetrazolium (NBT)-stimulated reduction, cyanide-insensitive oxygen (O2) consumption, superoxide anion (O2-)-stimulated production, hydrogen peroxide (H2O2) production, and iodination. They also had a normal myeloperoxidase (MPO) content. In four patients (group III), the granulocytes were significantly defective in all of these activities. In the six remaining patients (group II), all the initial metabolic steps of the phagocytosing granulocytes (ingestion, NBT reduction, O2 consumption, O2-production, H2O2 production) were normal, as were the MPO content of the granulocytes, while iodination was strikingly decreased. These metabolic features suggested a degranulation defect which was observed ultrastructurally in the only patient studied among these six. The phagocytosing granulocytes of this patient did not degranulate and no deposits of MPO activity were seen in the phagosomes.
Cold agglutinins in infectious mononucleosis and heterophil-antibody- negative mononucleosis-like syndromes
Authors: Horwitz CA , Moulds J , Henle W , Henle G , Polesky H , Balfour HH Jr , Schwartz B , Hoff T
Published: October 2019

Cold agglutinins (CA) were evaluated prospectively in patients with various mononucleosis syndromes and in a large control group. Cold agglutinins with anti-i specificity were seen mainly in heterophil- positive or -negative Epstein-Barr virus (EBV)-induced infectious mononucleosis (31.8% of cases). Unclassified CA with equal reactivity against cord and adult erythrocytes were seen in 56 of 150 (37.3%) cases of heterophil-antibody-positive infectious mononucleosis (IM), in 1 of 7 (14.3%) cases of heterophil-negative EBV-induced IM, and in 12 of 31 (38.7%) cases of the heterophil-negative mononucleosis-like syndrome due to cytomegalovirus or other unspecified agents. One patient with heterophil-positive IM had a persistent, partially papain sensitive CA with anti-Pr-like activity. Anti-i CA were seen in less than 1.0% of healthy young adults (500) or patients without mononucleosis (500) submitted for heterophil studies. Unclassified CA were noted in 3.2% of the latter 1000 samples.
Effect of heparin on complement activation and lysis of paroxysmal nocturnal hemoglobinuria (PNH) red cells
Authors: Logue GL
Published: October 2019

The effect of heparin upon the binding of the third component of complement (C3) to PNH red cells in vitro and their subsequent hemolysis is described. Heparin, in increasing concentrations, progressively inhibits membrane C3 fixation and hemolysis when the classic complement pathway is activated by anti-red cell antibodies. Heparin has a biphasic effect upon membrane C3 fixation and hemolysis when complement is activated in serum at decreased ionic strength (sucrose lysis) or in serum at decreased pH (Ham test). Heparin in concentrations above 2 U/ml inhibits C3 binding and hemolysis while lower concentrations of heparin enhance the consequences of complement activation by these two procedures. This enhanced complement activation may explain the increased hemolysis sometimes reported in PNH patients treated with heparin, and suggests that heparin may aggravate the consequences of pathologic alternative pathway complement activation in other diseases.
Slide chamber culture system for the in vitro study of humoral regulation of granulocyte and monocyte-macrophage proliferation and differentiation
Authors: Winton EF , Vogler WR , Kellar KL , Parker MB , Kinkade JM Jr
Published: October 2019

A liquid culture system employing slide chambers was developed to facilitate the study of proliferation and differentiation of mouse neutrophilic granulocytes and cells of the monocyte-macrophage series. Cultures were initiated with 1–2.5 X 10(4) light density marrow cells which had been fractionated on Ficoll-Hypaque. In the presence of colony-stimulating activity (CSAHU), two types of clusters were observed. One was tight, spheroidal, and composed of neutrophilic granulocytes, while the other was a loose grouping of flattened cells of the monocyte-macrophage series. The slide chamber culture is adaptable to microscopic assay techniques (e.g., cluster size, autoradiography, immunofluorescence) as well as quantitative biochemical methods (e.g., rate of 3H-thymidine incorporation, DNA quantitation). We have demonstrated both a shortening of the generation time of granulocytes in tight clusters and increasing rates of 3H- thymidine incorporation into DNA, with a corresponding increase in total culture DNA as a function of CSAHU concentration. Granulocytic differentiation in the tight spheroidal clusters has been demonstrated by histochemical stains and immunofluorescence of an antibody to a marker protein specific for the secondary granule of the neutrophil (lactoferrin).
Tissue-factor coagulant activity of cultured human endothelial and smooth muscle cells and fibroblasts
Authors: Maynard JR , Dreyer BE , Stemerman MB , Pitlick FA
Published: October 2019

The tissue-factor (thromboplastic) activity of cultured human endothelial cells and fibroblasts is low at time of transfer into fresh medium but increases 3–10 fold. Endothelial cells reach peak activity (400 U/10(5) cells) 5–8 hr after subculture. Activity in fibroblast cultures peaks (3000–12,000 U/10(5) cells) 7–12 hr after subculture. After attaining maximum activity, endothelial and fibroblast tissue- factor content decreases in a time course similar to other cells studied in this laboratory, approaching basal levels by 24–50 hr after subculture. If medium over fibroblasts is changed every 12 hr, activity can be sustained at the peak level for an additional day but cannot be maintained at a high level indefinitely. The kinetics of expression of smooth muscle cell tissue factor are markedly different from other cell types. There is always a pronounced lag (30 hr or more) before the activity increases, and then, in most cases, there is no subsequent decline in activity even though the cells are not refed or restimulated. The activity of each of these cell types is cryptic but becomes available after freeze-thaw disruption of cells.
Effect of blood donation on iron stores as evaluated by serum ferritin
Authors: Finch CA , Cook JD , Labbe RF , Culala M
Published: October 2019

Serum ferritin was measured in 2982 blood donors. First-time male donors had a geometric mean of 127 microgram/liter and female donors 46 microgram/liter. While values were essentially constant in the women between the ages of 18 and 45, there was a rapid increase in the men between 18 and 30 years of age consistent with the establishment of iron stores during that time. Blood donation was associated with a decrease in serum ferritin. One unit per year, equivalent to an increased requirement of 0.65 mg/day, halved the serum ferritin level in the male. More frequent donations were associated with further decreases. From the data obtained it would appear that male donors, while depleting their iron stores, were able to donate 2–3 U/yr without an appreciable incidence of iron deficiency. Women could donate only about half that amount, and more frequent donations were associated with a high incidence of iron deficiency and donor dropout. These data have provided information on the effect of graded amounts of iron loss through bleeding on iron balance.
Effects of partial hepatectomy on extrarenal erythropoietin production in rats
Authors: Anagnostou A , Schade S , Barone J , Fried W
Published: October 2019

Studies were performed to determine the effects of partial hepatectomy on extra-renal erythropoietin production. Rats were either partially hepatectomized or sham operated. At intervals of from 5 min to 7 days afterward, both kidneys were removed from cohorts of the above two groups of rats and the animals were then exposed to hypoxia for 7.5 hr. Immediately afterward, their plasma was collected and its erythropoietin titer was assayed. Rats which were partially hepatectomized 2–4 days prior to nephrectomy and hypoxia had significantly higher plasma erythropoietin levels than did sham- operated controls, whereas rats hepatectomized 5 min, 1 day, or 7 days prior to nephrectomy and hypoxia did not. These data are consistent with the conclusion that extrarenal erythropoietin production is enhanced in association with rapid regeneration of hepatic cells.
Evidence for intracellular amyloid formation in myeloma
Authors: Kjeldsberg CR , Eyre HJ , Totzke H
Published: October 2019

Evidence for intracellular formation of amyloid fibrils in a patient with kappa light-chain myeloma is described. Amyloid fibrils were seen as intracytoplasmic inclusions within plasma cells, histiocytes, renal tubule cells, and possibly in hepatocytes. Extracellular amyloid was also present. The electron microscopic studies suggest that amyloid fibrils may form in the Golgi apparatus and within lysosomes.
Culture in vitro of isolated guinea pig megakaryocytes: recovery, survival, morphologic changes, and maturation
Authors: Levine RF
Published: October 2019

Isolated guinea pig megakaryocytes were maintained in liquid cultures for up to 4 days. Megakaryocytes were incubated in siliconized glass vials in Dulbecco's Modified Eagle Medium with 5%-10% guinea pig serum and 2.3% bovine serum albumin. Cultured megakaryocytes did not adhere to glass vials and were almost entirely recovered by aspiration. No reproduction or cell division of megakaryocytes occurred. A small decline in viability occurred promptly on placing the freshly isolated cells in culture medium and could be attributed to reexposure to calcium. On incubation there was little further cell death. Up to 2 days in culture the megakaryocytes remained morphologically intact and appeared similar to megakaryocytes in situ. Megakaryocytes matured in culture with a loss of cytoplasmic basophilia, an increase in granule content, and progressive changes in nuclear configuration. The most mature megakaryocytes developed pseudopod formation but large-scale platelet liberation was not seen. The ability to culture megakaryocytes in vitro will allow more extensive biochemical and physiologic studies of this cell than previously possible.
Characterization of colony-stimulating activity produced by human monocytes and phytohemagglutinin-stimulated lymphocytes
Authors: Shah RG , Caporale LH , Moore MA
Published: October 2019

Human colony-stimulating activity (CSA) may support the proliferation of both human and murine granulocyte-macrophage progenitor cells (CFU- C) or, in the case of human urinary CSA, may only stimulate murine bone marrow CFU-C. CSA produced in the culture media of monocytes and macrophages and phytohemagglutinin-stimulated lymphocytes from human peripheral blood was characterized for both human and mouse marrow CFU- C stimulating activities. During the initial phase of a long-term cultures of monocytes, both human- and mouse-active CSA (MnCM-HM) were produced. In later phases of culture, however, only mouse-active CSA (MnCM-M) was produced. Fractionation on Sephadex G-150 revealed two functionally distinct species from MnCM-HM and lymphocytes conditioned medium, a high molecular weight factor (MW greater than 150,000) which stimulated mouse but not human colony formation, and a low molecular weight species (MW 25,000–35,000) which was active against both mouse and human target cells. However, MnCM-M revealed only one high molecular weight species (greater than 150,000), active only on mouse marrow. The possible biologic significance of such an activity is discussed.
Heterogeneity in human neutrophil, macrophage and eosinophil progenitor cells demonstrated by velocity sedimentation separation
Authors: Johnson GR , Dresch C , Metcalf D
Published: October 2019

Progenitor cells of neutrophils, monocyte-macrophages, and eosinophils in human marrow were enumerated in agar cultures stimulated by placental conditioned medium or white cell underlayers. Fractionation of marrow populations by velocity sedimentation showed that the profiles of neutrophil and macrophage colony-forming cells shifted from a peak of 8–9 mm/hr in 7-day cultures to a peak of 6–7 mm/hr in 14-day cultures. This shift was due to degeneration of some early colonies formed by rapidly sedimenting cells and the delayed formation of colonies by slowly sedimenting cells. Eosinophil colony formation was delayed until the second week of incubation. Further evidence of heterogeneity was the observation that rapidly sedimenting colony forming cells were more responsive to stimulation than more slowly sedimenting cells. In the macrophage and eosinophil populations, cluster-forming cells were partially segregatable form colony-forming cells. The observed heterogeneity was similar to the described previously in the mouse and suggests that separate subpopulations of progenitor cells may exist within each hemopoietic family that could possibly give rise to functionally different progeny.
Erythroid cell growth from normal and W/WV murine bone marrow on macrophage-coated membranes
Authors: Torok-Starb BJ , Wolf NS , Boggs DR
Published: October 2019

Cellulose acetate membranes (CAM) placed in the peritoneal cavity of mice develop a macrophage layer capable of supporting in vivo hematopoietic colonies from intraperitoneally injected bone marrow cells. Modifications allowing for routine morphologic identification of colonies showed that both erythrocytic (E) and granulocytic (G) colonies occur with a consistent E:G ratio of 0.19 +/- 0.037. Stimulating recipients by bleeding or phenylhydrazine injection did not produce a significant change in the total number of colonies and a reduction in granulocytic colonies so that the E:G ratio significnatly increased. Hypertransfusion of donor animals had no effect on the number of erythroid colonies that grew on CAM of average recipients. The total colony-forming ability of bone marrow cells from genetically anemic W/WV mice was found not to differ from that of normal +/+ littermates; however, the E:G ratio of W/WV marrow in bled recipients was significantly lower (p less than 0.01) then that of +/+ marrow. These studies suggest that a CAM system supports an erythroid progenitor which is not affected by hypotransfusion of the donor animal, yet is dependent upon erythropoietin for colony formation, and that it is defective in the W/WV mouse.
Platelet-bound complement (C3) in immune thrombocytopenia
Authors: Hauch TW , Rosse WF
Published: October 2019

The fixation of complement to the circulating platelet in immune thrombocytopenia was detected by measurement of one of the complement components, C3, on the surface of platelets from patients with idiopathic thrombocytopenic purpura (ITP) and systemic lupus erythematosus (SLE) using the anti-C3 consumption assay. The surface IgG was determined simultaneously using the previously described anti- IgG consumption assay. Washed platelets from normal controls had 3.5 fg (10(-15) g) of C3, or about 11,000 molecules, per platelet, an amount comparable to the IgG (4.1 FG, or 15,000 molecules, per platelet). For most patients with ITP both C3 and IgG were increased on the platelet surface, although for 5 of 16 patients only IgG was increased. Two patients with SLE and thrombocytopenia had an increase in both C3 and Ig, six patients with SLE who were not thrombocytopenic had normal amounts of membrane-bound C3 and IgG. In 5 patients, 3 with ITP and 2 with collagen vascular disease, both surface immunoproteins decreased with successful treatment of the thrombocytopenia.
Purification and characterization of granulocytic progenitor cells (CFU- C) from human peripheral blood using immunologic surface markers
Authors: Richman CM , Chess L , Yankee RA
Published: October 2019

The concentration of committed granulocytic progenitor cells (CFU-C) in functionally unique subpopulations of human peripheral blood mononuclear cells has been determined by the in vitro methyl-cellulose assay. Using immunoabsorbent column chromatography and rosette-depletion techniques, we have demonstrated that CFU-C, although not present in either purified T or B lymphocyte populations, are highly concentrated in the “null” cell population, which lacks sheep erythrocyte receptors and surface immunoglobulin. Further fractionation of this null subset has demonstrated that CFU-C do not bear complement receptors, but require the presence of peripheral blood mononuclear cell feeder layers for maximum proliferation.