bluuebunny/crossref_metadata_2025 huggingface dataset:
Reticulocyte motility and form: studies on maturation and classification
Authors: Mel HC , Prenant M , Mohandas N
Published: October 2019

A new two-stage living cell cytological classification is presented for reticulocytes in the rat and shown on an overall population basis to be the equivalent of the classical new methylene blue staining characterization. Class 1 reticulocytes (R1) are motile and multilobular. They comprise about 25% of normal bone marrow reticulocytes, but none of the reticulocytes in normal blood. Class 2 reticulocytes (R2) are asymmetrically “deep dished” in appearance and contain visible refractile granules, viewed in phase-contrast microscopy. In animals “stimulated” by bleeding, the percentage of motile marrow reticulocytes is found to be a linear function of hematocrit over the range of about 25%-70% total reticulocytes. In the blood, the associated maximum rise of class R1 is only to about 8%. These and related cellular properties are discussed with respect to the maturation and release of normal reticulocytes. Applicability of this work to studies on humans and to pathology is also indicated.
Factors controlling stem cell recirculation. III. Effect of the thymus on the migration and differentiation of hemopoietic stem cells
Authors: Petrov RV , Khaitov RM , Aleinikova NV , Gulak LV
Published: October 2019

Experiments were carried out to investigate the effect of thymectomy on the migration and differentiation of hemopoietic stem cells released from shielded parts of bone marrow after irradiation of mice with lethal doses of x-rays. In 2–3 wk after thymectomy, the rate of migration declined and the differentiation of stem cells into granulocytic colonies was inhibited. Transplantation of syngeneic thymus or lymph node cells into thymectomized mice enhanced the migration of stem cells from the bone marrow and restored the usual pathways of their differentiation.
Cell marker analysis in acute monocytic leukemias
Authors: Koziner B , McKenzie S , Straus D , Clarkson B , Good RA , Siegal FP
Published: October 2019

Leukemic cells from nine cases of acute monocytic leukemia (AMoL) were characterized by multiple differentiation markers. Cells in most cases were phagocytic, carried an Fc receptor, and stained positively for alpha-naphthyl acetate esterase but negatively for naphthol AS-D chloroacetate esterase. However, subtle differences in marker expression were observed which suggested different degrees of leukemic cellular maturation or activation. Cell marker analysis proved to be a useful adjunct to conventional morphology in confirming the diagnosis and the recognition of the neoplastic cells in AMoL, and may ultimately provide insight into the functional state of these cells.
Marrow culture studies in adult acute leukemia at presentation and during remission
Authors: Vincent PC , Sutherland R , Bradley M , Lind D , Gunz FW
Published: October 2019

Culture of bone marrow and/or blood cells in a semisolid agar system from 43 adults with acute nonlymphoblastic leukemia at first presentation showed two distinct growth patterns at 14 days. In 53% of patients cells failed to grow (type O), while in the remainder an abnormal growth pattern (type B) with small numbers of diffuse colonies and excessive numbers of cell clusters was seen. The response following chemotherapy was significantly better in the patients whose cells failed to grow. Serial culture studies, performed in 9 patients throughout remissions of 100–1112 days, which had been maintained by intermittent chemotherapy, showed wide fluctuations in proliferative activity. These ranged from no growth to marked proliferation with predominance of clusters and small numbers of diffuse colonies, indistinguishable from the type B pattern seen in 47% of patients at first presentation. The possibility is discussed that the periods of failure to grow, and/or those in which a type B pattern emerged, represented sporadic reactivation of leukemic cells.
Red cells in sickle cell crisis: observations on the pathophysiology of crisis
Authors: Rieber EE , Veliz G , Pollack S
Published: October 2019

The pathophysiology of the occurrence and resolution of sickle cell crisis is unknown. The molecular abnormality is constant, while crisis is episodic. In the present study, red cell filterability and sickling with deoxygenation have been measured during sickle cell crises. Recovery from sickle crisis is associated with an increased filterability of the circulating red cell and a decreased susceptibility of the red cell to sickle with deoxygenation (p less than 0.05). The possibility that these changes are responsible for the resolution of crisis is suggested.
Circulating antibody to transcobalamin II causing retention of vitamin B12 in the blood
Authors: Carmel R , Tatsis B , Baril L
Published: October 2019

A patient with recurrent pulmonary abscess, weight loss, and alcoholism was found to have extremely high serum vitamin B12 and unsaturated vitamin B12-binding capacity (UBBC) levels. While transcobalamin (TC) II was also increased, most of his UBBC was due to an abnormal binding protein which carried greater than 80% of the endogenous vitamin B12 and was not found in his saliva, granulocytes, or urine. This protein was shown to be a complex of TC II and a circulating immunoglobulin (IgGkappa and IgGlambda). Each IgG molecule appeared to bind two TC II molecules. The reacting site did not interfere with the ability of TC II to bind vitamin B12, but did interfere with its ability to transfer the vitamin to cells in vitro. The site was not identical to that reacting with anti-human TC II antibody produced in rabbits. Because of this abnormal complex, 57Co-vitamin B12 injected intravenously was cleared slowly by the patient. However, no metabolic evidence for vitamin B12 deficiency was demonstrable, although the patient initially had megaloblastic anemia apparently due to folate deficiency. The course of the vitamin B12-binding abnormalities was followed over 4 yr and appeared to fluctuate with the status of the patient's illness. The IgG-TC II complex resembled one induced in some patients with pernicious anemia by intensive treatment with long-acting vitamin B12 preparations. The mechanism of induction of the antibody formation in our patient is unknown.
Inhibition of ristocetin-induced platelet agglutination by vancomycin
Authors: Moake JL , Cimo PL , Peterson DM , Roper P , Natelson EA
Published: October 2019

Ristocetin and vancomycin are structurally similar glycopeptide antibiotics. Both vancomycin and ristocetin in high concentrations (3.0 mg/ml) cause the precipitation of fibrinogen, plasminogen, and IgG from platelet-poor plasma (PPP). In contrast to ristocetin, vanomycin (0.5–1.5 mg/ml) does not agglutinate platelets in normal platelet-rich plasma (PRP) or formalin-treated platelets in the presence of normal PPP. Preincubation of vancomycin (0.5–1.25 mg/ml) with normal PRP, von Willebrand platelets in normal PPP, or formalinized platelets results in inhibition of platelet agglutination induced by ristocetin (0.7–1.25 mg/ml) or ristocetin and normal PPP. This inhibition can be overcome by increasing the final concentration of ristocetin in the platelet suspension. Preincubation of formalin-treated platelets with the major fraction obtained by carboxymethyl-Sephadex C-50 chromatography of commercial vancomycin also results in inhibition of agglutination induced by ristocetin and normal PPP. Incubation with vancomycin (1.25 mg/ml) does not interfere with von Willebrand factor (vWF) or factor VIII coagulant activities in normal PPP or in Sepharose 4B void volume fractions of PPP. These results indicate that vancomycin interacts with normal, von Willebrand, and formalin-treated platelets and inhibits the binding of ristocetin (or ristocetin-vWF complexes).
Prognostic significance of thrombocytosis in idiopathic sideroblastic anemia
Authors: Streeter RR , Presant CA , Reinhard E
Published: October 2019

In order to determine the prognostic significance of thrombocytosis in idiopathic sideroblastic anemia, the clinical courses of 17 patients were reviewed. Six patients (36%) had thrombocytosis, and none developed acute leukemia. Nine patients (53%) had normal platelet counts, and one developed acute leukemia. Two patients (12%) were thrombocytopenic, and one died of acute leukemia. There was little correlation between survival and platelet count. Sixty-three additional case reports of idiopathic sideroblastic anemia were collected from the literature. Analysis of those patients and the patients in the present study documented transformation to acute leukemia in 5 of 9 (56%) thrombocytopenic patients, 4 of 54 (7.4%) patients with normal platelet counts, and 0 of 17 patients with thrombocytosis (p less than 0.05). Therefore patients with idiopathic sideroblastic anemia and thrombocytosis appear to have a decreased likelihood of leukemic transformation.
Superoxide-forming enzyme from human neutrophils: evidence for a flavin requirement
Authors: Babior BM , Kipnes RS
Published: October 2019

The superoxide-forming activity of 27,000-g particles prepared from homogenates of zymosan-treated human neutrophils is lost if the assay is conducted in the presence of 0.045% Triton X-100. This loss in activity in the presence of detergent is prevented by 40 micron flavin adenine dinucleotide (FAD), but not by flavin mononucleotide, riboflavin, adenosine 5′-diphosphate, or adenosine 5′-monophosphate. With resting particles or particles from zymosan-treated chronic granulomatous disease neutrophils, no superoxide-forming activity is detectable even in the presence of FAD; this is true whether or not detergent is present in the assay. Particles extracted with detergent prior to assay are fully active if assayed in the presence of FAD, but show little activity if FAD is omitted from the assay mixture. These results suggest that the superoxide-forming enzyme from human neutrophils is a FAD-requiring enzyme.
Oxidative metabolism of the human eosinophil
Authors: DeChatelet LR , Shirley PS , McPhail LC , Huntley CC , Muss HB , Bass DA
Published: October 2019

We have compared the oxidative metabolism of human eosinophils (80%-90% purity) to that of neutrophils. Hexose monophosphate (HMP) shunt activity of eosinophils was higher than that of neutrophils under either resting or phagocytizing conditions. Eosinophil HMP shunt activity also was stimulated by phorbol myristate acetate, a membrane- active agent. Eosinophils showed a marked incorporation of 125I into trichloroacetic acid-insoluble material under resting conditions, which increased markedly during phagocytosis. Eosinophils likewise showed a greater reduction of nitroblue tetrazolium dye during phagocytosis than did neutrophils. Measurement of other parameters of oxidative metabolism indicated that eosinophils generated superoxide anion following phagocytosis and also elicited a burst of chemiluminescence similar to that observed during phagocytosis by neutrophils. Measurement of NADPH oxidase activity demonstrated that this enzyme was 3–6 times more active in fractions isolated from eosinophils than in corresponding fractions isolated from neutrophils; this was observed over a range of substrate concentrations. The eosinophil enzyme sedimented differently than the neutrophil enzyme with differential centrifugation; neither showed sedimentation characteristics of peroxidase. These data indicate that eosinophils possess a similar, although in some ways more potent, oxidative burst than neutrophils and are consistent with a role for NADPH oxidase in the initiation of that burst.
Initial prognostic factors and lymphoblast-erythrocyte rosette formation in 109 children with acute lymphoblastic leukemia
Authors: Dow LW , Borella L , Sen L , Aur RJ , George SL , Mauer AM , Simone JV
Published: October 2019

Bone marrow lymphoblasts from 109 children admitted with untreated acute lymphoblastic leukemia (ALL) were tested for spontaneous rosette formation with sheep erythrocytes. Twenty-six children (24%) had lymphoblasts that formed rosettes (E+). Of 13 initial clinical characteristics, 8 were significantly associated with E+ lymphoblasts: mediastinal enlargement (86% of patients E+), leukocyte counts over 100 X 10(9)/liter (65% E+), nodes greater than 2 cm in any diameter (65% E+), age over 5 yr (46% E+), hemoglobin over 8 g/dl (44% E+), hepatomegaly greater than 5 cm (38% E+), boys (35% E+), and lymph node enlargement outside of the cervical area (28% E+). Spleen size, initial platelet counts, and periodic acid-Schiff scores did not distinguish E+ from E- patients. Since few patients were black and few presented with central nervous system leukemia, the association of these two characteristics with E+ blasts could not be determined. A hierarchical classification scheme and a linear logistic regression model were used to define the patterns of characteristics associated with E+ lymphoblasts. The initial clinical characteristics and the poorer course of E+ patients suggest that ALL comprises at least two biologically and clinically distinct types. The E+ ALL may result from a leukemic transformation of a non-Hodgkin lymphoma.
Discrete clusters of hematopoietic cells in the marrow cavity of man after bone marrow transplantation
Authors: Cline MJ , Gale RP , Golde DW
Published: October 2019

Discrete aggregates of hematopoietic cells were observed in the bone marrows of patients with aplastic anemia or acute leukemia 14 days after marrow transplantation. The great majority of such colonies were of a single cell type, and less than 3% contained two or more cell types. Erythroid, myeloid, and undifferentiated hematopoietic colonies were approximately equal in frequency.
Globin synthesis in mouse erythroleukemia cells in vitro: a switch in beta chains due to inducing agent
Authors: Alter BP , Goff SC
Published: October 2019

Friend erythroleukemia cells, originally derived from DBA/2 mice, differentiate when cultured with inducing agents. Studies of the different effects of inducing agents on clone 745 have revealed that both dimethyl sulfoxide (DMSO) and hemin produce benzidine-positive cells. Butyric acid produced mature but benzidine-negative cells in this clone. All agents induced globin synthesis above the 0.1% of protein synthesis found in uninduced cells. DMSO induction stimulated globin synthesis 9%, hemin 2%, and butyric acid 3%. Total beta/alpha ratios were approximately unity with all agents. Although the inducing agents all stimulated total globin synthesis in Friend cells, the relative rates of synthesis of the two mouse beta chains were affected differently by the various agents. Hemin markedly increased the proportion of beta minor. For example, DBA/2 mouse reticulocytes synthesized 20% beta minor and 80% beta major. DMSO induction of clone 745 caused 20%-33% synthesis of beta minor. In contrast, hemin increased the proportion of beta minor to 64%-69%. Thus the Friend erythroleukemia cell system provides an in vitro approach to the study of the regulation of globin-chain switching.
Cellular fluxes of vitamin B12
Authors: Ostray F , Gams RA
Published: October 2019

Incorporation of vitamin B12 into L1210 cells requires the protein binder transcobalamin II (TCII). The process is saturable, follows Michaelis-Menten kinetics (Km = 2.5 X 10(-9) M at 37 degrees C), is both temperature and calcium (K50 - 1 X 10(-6) M) dependent, and is inhibited by apo-TCII, indicating the presence of a TCII specific receptor on the cell membrane. B12 also leaves the cell by a calcium- independent pathway bound to either TCII or to a protein with chromatographic properties similar to those of TCIII. Since intact TCII- B12 can be found in the cytosol and can promote B12 uptake by mitochondria, it is proposed that the B12 released from the cell bound to the TCIII-like protein is derived by mitochondrial processing of incorporated TCII-B12. The slower time course of release of the latter B12 is consistent with this postulate.
Filtration leukapheresis: effects of donor stimulation with dexamethasone
Authors: Higby DJ , Henderson ES , Burnett D , Cohen E
Published: October 2019

Dexamethasone was administered to 51 donors prior to filtration leukapheresis. The results of this maneuver, including the consequences of transfusion, were contrasted with results in 52 donors who did not receive the steroid. Yields of polymorphonuclear leukocytes, the posttransfusion increments in recipients, themorphologic polymorphonuclear leukocytes obtained, and the incidence of donor and recipient reactions were all beneficially influenced by this manipulation. Possible mechanisms responsible for these observations are discussed. It is recommended that dexamethasone stimulation be used in filtration leukapheresis when circumstances do not otherwise contraindicate such a maneuver.
Comparison of daily versus intermittent chlorambucil and prednisone therapy in the treatment of patients with chronic lymphocytic leukemia
Authors: Sawitsky A , Rai KR , Glidewell O , Silver RT
Published: October 2019

Ninety-six patients with stage III and stage IV chronic lymphocytic leukemia (CLL) were randomized into one of three treatment schedules. Prednisone was common to all three schedules and was given daily in an initial dosage of 0.8 mg/kg for the first 14 days, with successive halving of the daily dose on days 15 and 29 for a total 6-wk course. Prednisone was then given once a month at 0.8 mg/kg once a day for each of 7 consecutive days. Schedule I was prednisone plus chlorambucil (CLB) given as a once-a-month dose of 0.4–0.8 mg/kg; schedule II was both drugs, but the CLB was given as a daily dose of 0.08 mg/kg; schedule III was prednisone alone. Complete and partial remission (CR + PR) was 47% for schedule I, 38% for schedule II, and 11% for schedule III. Patients who responded (CR + PR) in each of the treatment schedules survived longer than the nonresponders. Complete remission was obtained in both CLB treatment schedules, but not with the prednisone alone regimen. Although overall survival was best in the intermittent CLB arm, there was no significant difference in survival time between the three treatment schedules. Toxicity was minimal in all three regimens. Augmentation of the intermittent monthly CLB, even to 1.5 and 2.0 mg/kg, was tolerated without undue marrow toxicity. About 22% of these patients either had diabetes mellitus at the time of entry on the study or manifested hyperglycemia during the course of treatment and observation.
Congenital nonspherocytic hemolytic anemia with an unstable hexokinase variant
Authors: Board PG , Trueworthy R , Smith JE , Moore K
Published: October 2019

We report a family with a new hexokinase variant that gives rise to nonspherocytic hemolytic anemia in one apparently homozygous family member. The variant enzyme has a normal pH optimum, normal reaction kinetics, and normal electrophoretic properties, but has reduced activity and is apparently inactivated rapidly as the affected erythrocytes age.
Heterogeneity of human colony-forming cells
Authors: Inoue S , Ottenbreit MJ
Published: October 2019

Culture supernatants from established human fibroblast cell lines (FCM) and from phytahemagglutinin-stimulated peripheral blood leukocytes (LCM) were compared with respect to their stimulatory effects on the colony formation by human marrow leukocyte precursors in vitro using the methylcellulose culture system. The cultures were performed using specimens from children with a variety of disorders. LCM stimulation consistently produced a significantly higher proportion of macrophage colonies than did FCM stimulation, whereas FCM stimulation resulted in a significantly higher number of number of neutrophil colonies. Based on the sequential examination of colony numbers in 35 FCM-and 11 LCM- stimulated cultures, the colonies in LCM-stimulated cultures (predominantly macrophage colonies) survived much longer than those in FCM-stimulated cultures (predominantly neutrophil colonies). Cell fractionation experiments by velocity sedimentation on four different specimens revealed that the majority of FCM-responsive cells were large and formed pure neutrophil colonies, while the majority of LCM- responsive cells were smaller and formed either pure macrophage or mixed neutrophil-macrophage colonies. These observations suggest that human colony-forming cells consist of at least two distinct cell populations different in (1) cell size, (2) response to two different conditioned media, LCM and FCM, and (3) type of colonies each population forms.
Lack of protective effect of autotransplanted splenic tissue to pneumococcal challenge
Authors: Schwartz AD , Goldthorn JF , Winkelstein JA , Swift AJ
Published: October 2019

Studies in animals and clinical experience in patients have demonstrated that splenectomy may lead to an increased susceptibility to infection. The infections are usually caused by encapsulated bacteria such as penumococcus. It has been shown in a variety of experimental animals that autotransplanted splenic tissue is capable of regenerating into implants that are microscopically indistinguishable from normal spleen and of restoring a number of normal splenic functions. The response to intravenous challenge with Streptococcus pneumoniae, type 25, was therefore studied in control, asplenic, and autotransplanted Sprague-Dawley rats. Despite previous observations that a number of immune functions can be restored in this animal model by autotransplanted splenic tissue, the present study indicates that splenic tissue autotransplants do not restore the ability to resist intravenous pneumococcal challenge.
Endotoxin-induced changes in human platelet membranes: morphologic evidence
Authors: Ausprunk DH , Das J
Published: October 2019

Interaction between human platelets and bacterial endotoxin was studied in vitro with transmission and scanning electron microscopy. Washed human platelets, whose aggregation was blocked with apyrase, were incubated in a plasma-free medium containing crude endotoxin that had previously been complexed with copper. Thirty minutes of incubation resulted in adherence of endotoxin particles to the platelet surface, breaks in the platelet plasma membrane with apparent attempts at repair, pseudoped formation, and centralization of platelet organelles. Copper appeared to potentiate these phenomena, since neither Cu2+ at low concentrations nor endotoxin alone altered the morphology of the platelet membrane. This platelet-endotoxin interaction may be an intermediary step in the detoxification and clearance of endotoxin from the plasma.
Effect of protein deprivation on extrarenal erythropoietin production
Authors: Anagnostou A , Schade S , Barone J , Fried W
Published: October 2019

The production of erythropoietin by rats fed a protein-deficient diet is markedly decreased. This defect can be rapidly reversed by feeding protein. In the reported experiments we investigated the effect of protein deficiency on the extra-renal production of erythropoietin. The results indicate that the production of erythropoietin in nephrectomized animals is insensitive to protein depletion.
Discrimination between human T and B lymphocytes and monocytes by computer analysis of digitized data from scanning microphotometry. I. Chromatin distribution patterns
Authors: Durie BG , Vaught L , Chen YP , Olson GB , Salmon SE , Bartels H
Published: October 2019

Digitized images of Feulgen-stained normal human B and T lymphocytes and monocytes were analyzed by computer and microphotometry. The algorithms assessed the chromatin distribution patterns. A substantial number of parameters descriptive of the chromatin distributions were found to show statistically highly significant differences between B and T cells and monocytes. Although the effects of a number of biologic variables, such as cell cycle phase and immune stimulation, need to be carefully evaluated, the preliminary results indicate potential for automatic classification and subclassification of human B and T lymphocytes in normal and disease states.
Centrifugal method of determining red cell deformability
Authors: Corry WD , Meiselman HJ
Published: October 2019

A recently developed technique for deforming red blood cells (RBC) in which they are centrifuged through buffer and into a glutaraldehyde solution was evaluated as a method of assessing cellular deformability (i.e., the ability of the entire RBC to form a new configuration). To accomplish this, RBC populations of differing cellular deformability were tested, using three generally accepted techniques to obtain these differences: partial fixation with low concentrations of glutaraldehyde, density fractionation, and suspension of RBC in nonisotonic media. Our results indicate that at a constant deforming force the mean deformed length of the RBC decreased under conditions where cellular deformability is known to decrease, thus suggesting the usefulness of this centrifugal method for the estimation of this cellular property.
Water distribution in blood during sickling of erythrocytes
Authors: Fales FW
Published: October 2019

Plasma urea and protein determinations proved suitable for measuring changes in total diffusible water and plasma volume in whole blood. Deoxygenation by saturation with carbon dioxide at 25 degrees C caused no change in plasma urea, but a significant increase in plasma protein concentration was induced with both normal and sickle-cell (HbSS) blood. Thus in HbSS blood there was no binding or trapping of water as a result of sickling and there was a normal influx of water into the cells (Bohr effect) despite the polymerization of the hemoglobin molecules with sickling. Consistent with this observation was the finding that the deoxygenation induced a similar increase in concentration of the plasma cations, sodium plus potassium. HbSS erythrocytes neither lost nor gained water under the more physiologic conditions of deoxygenation with a 95% nitrogen, 5% carbon dioxide gas mixture.
Inhibition in vivo of mouse granulopoiesis by cell-free activity derived from human polymorphonuclear neutrophils
Authors: Broxmeyer HE
Published: October 2019

Cell-free extracts from human polymorphonuclear neutrophils (PMN) inhibited rebound granulopoiesis in the bone marrow and spleen of mice pretreated with cyclophosphamide to remove endogenous PMN. Absolute numbers of granulocytemonocyte progenitor cells (CFU-C) and net endogenous colony-stimulating activity (CSA) production were found to be increased 3 days after cyclophosphamide in the bone marrow and 6 days after in the spleen. Administration of PMN extract to the drug- treated mice prior to rebound granulopoiesis substantially decreased CSA production and CFU-C but not spleen B lymphocyte colony-forming cells. In addition, mice treated with PMN extract had decreased levels of CSA in serum and in conditioned medium of marrow-free bone, heart, and lung cultures. Inhibition was reversed by injection of bacterial lipopolysaccharide. Extracts from PMN of patients with chronic myelogenous leukemia, inactive in vitro, had no effect in vivo. These results demonstrate that inhibitory activity derived from PMN can control granulopoiesis in vivo.
Colony-stimulating factors active on human bone marrow cells from a Yoshida sarcoma cell line
Authors: Ohno T , Seki M , Shikita M
Published: October 2019

Colony-stimulating factors (CSF) active on both human and murine bone marrow colony-forming units in culture (CFU-C) were found in the conditioned medium of Yoshida sarcoma cells (line YSSF-212T), although the cells originated from rats. The CSF were inactivated by digestion with trypsin, alpha-chymotrypsin, and pronase. By chromatography on DEAE-cellulose, the CSF were separated into five subgroups with different capacities to stimulate mouse granulocyte and macrophage progenitors. CSF in these five peaks were eluted from an Ultrogel AcA 44 gel filtration column with apparent molecular weights of 22,000– 25,000 daltons. CSF of nonhuman origin stimulating human CFU-C would be useful in hematologic studies of bone marrow cells.
Hereditary nonspherocytic hemolytic anemia and hexokinase deficiency
Authors: Beutler E , Dyment PG , Matsumoto F
Published: October 2019

An 11-yr-old child with mild chronic hemolytic anemia was found to have decreased red cell hexokinase activity in spite of the reduced mean age of her red cell population. Similar decreases in red cell hexokinase activity were documented in the patient's parents and in one sib. The red cells were morphologically normal. Red cell 2,3-DPG levels were normal and ATP and glucose-6-phosphate levels were diminished. The kinetic properties, electrophoretic mobility, and thermal stability of the residual red cell hexokinase were normal or nearly so. Glucose consumption of the hexokinase-deficient cells was not appreciably decreased, probably because less of the potent inhibitor glucose-6- phosphate was present in the erythrocytes. It is likely, although not certain, that in this patient nonspherocytic hemolytic anemia resulted from hexokinase deficiency.
Polyamine synthesis in bone marrow granulocytes: effect of cell maturity and early changes following an inflammatory stimulus
Authors: Evans WH , Grieshaber CK , Miller WC , Wilson SM , Hoffman HA
Published: October 2019

Enriched fractions of mature and immature neutrophil granulocytes, isolated from guinea pig bone marrow, were assayed for ornithine decarboxylase activity and polyamine content. The results show that immature granulocytes contain at least ten times more ornithine decarboxylase activity and two times more spermidine than mature granulocytes. The incorporation of 14C-ornithine into putrescine and spermidine of intact immature granulocytes was three to four times and ten times, respectively, that of mature granulocyte preparations. Six hours after an inflammatory stimulus, transient increases of 14-fold and 3-fold in the activities of ornithine decarboxylase and S-adenosyl- L-methionine decarboxylase, respectively, were observed in immature bone marrow granulocytes. At this time the incorporation of 14C- ornithine into putrescine and spermidine in bone marrow granulocytes from stimulated animals was 14 times that of cells from controls. A maximum increase in DNA synthesis in these cells during the inflammatory response occurred 6 hr after the maximum increase in the polyamine synthetic activity. Together these data suggest that polyamine synthesis in the granulocyte compartment of the bone marrow is associated chiefly with immature proliferating cells and that increased polyamine synthesis precedes increased granulocyte proliferation in the bone marrow following an inflammatory stimulus.
Evidence for distinct lymphocytic and monocytic populations in a patient with terminal transferase--positive acute leukemia
Authors: Mertelsmann R , Koziner B , Ralph R , Filippa D , McKenzie S , Arlin ZA , Gee TS , Moore MA , Clarkson BD
Published: October 2019

Two distinct cell populations with lymphoblastic and monocytic characteristics were separated and characterized by multiple cell markers in a patient with terminal transferase-positive acute acute leukemia. The clinical course and sequential cell marker studies were consistent with the interpretation of a defect at the level of a common stem cell giving rise to a terminal transferase--positive lymphoblastic cell population at diagnosis and, following initial therapy, a terminal transferase--negative monocytic population.
Hormone-stimulated heme synthesis by isolated mitochondria
Authors: Agam G , Djaldetti M
Published: October 2019

Erythropoietin (Ep), insulin, and prostaglandins E1 and A1 (PGE1, PGA1) enhanced heme synthesis by mitochondria isolated from embryonic and adult mouse liver cells. The combination of Ep with PGE1 had an antagonistic effect, while that of Ep with PGA1 gave an additive result. The capability of isolated human platelet mitochondria to synthesize heme was also shown. This activity was stimulated by Ep.
Failure of the alpha-thalassemia gene to decrease the severity of sickle cell anemia
Authors: Natta C
Published: October 2019

A 15-yr-old black male with homozygous sickle cell disease was severely growth retarded and had a chronic hemolytic anemia requiring transfusions. Globin chain synthetic studies of both peripheral blood reticulocytes and bone marrow cells revealed a ratio of alpha to betas globin synthesis (alpha/betas ratio) of 0.5, indicating the presence of an alpha-thalassemia gene. Messenger RNA isolated from the bone marrow of the patient was translated in the wheat germ cell-free system, and the globin synthesized had an alpha/betas ratio of 0.7. The hemolysate prepared from incubated bone marrow cells was fractionated on a Sephadex G100 column. The results showed that there was a peak of radioactivity that eluted after the hemoglobin peak. When this pooled peak was analyzed by CMC chromatography, the alpha/betas ratio was 0.9. These globin intermediates, probably dimers, may have contributed to the hemolysis in this patient.
Clonal evolution in atypical chronic granulocytic leukemia: a non- Philadelphia translocation
Authors: Bagby GC Jr , Kaiser-McCaw B , Hecht F , Koler RD , Linman JW
Published: October 2019

Hemopoietic cells in chronic granulocytic leukemia (CGL) frequently contain a chromosome translocation involving chromosome 22 and another autosome, usually number 9. The translocated chromosome 22 is known as the Philadelphia (Ph) chromosome. The appearance of a second Ph chromosome is the most common cytogenetic abnormality in CGL signaling the blastic phase. For 6 yr we serially studied a man with atypical CGL whose marrow cells were marked by a translocation from chromosome 18 to chromosome 11 [46XY,t(11;18)(q23;q12)]. Three months prior to blast transformation there appeared an extra copy of the marker chromosome 18: 47XY,t(11;18)(q23;q12),+(18p11 leads to 18q12). This man presents a new cytogenetic pattern of clonal evolution in CGL. The pattern is analogous to that of the Ph chromosome and is characterized by a balanced chromosomal rearrangement and the subsequent acquisition of an extra copy of the small translocation chromosome immediately prior to blast transformation.
Inhibition of platelet aggregation by protease inhibitors. Possible involvement of proteases in platelet aggregation
Authors: Aoki N , Naito K , Yoshida N
Published: October 2019

The possible participation of proteases in human platelet aggregation was explored using various protease inhibitors and substrates. Protease inhibitors used included naturally occurring inhibitors of serine proteases and synthetic inhibitors that modify the active site of protease. Substrates used were synthetic substrates for the trypsin type as well as for the chymotrypsin type of protease. All these inhibitors and substrates inhibited platelet aggregation and serotonin release induced by ADP, collagen, epinephrine, or thrombin. In ADP- and epinephrine-induced platelet aggregation the second phase of aggregation was most efficiently inhibited. The inhibitors suppressed the formation of malondialdehyde during platelet aggregation. Release by aggregating agents of arachidonate and its metabolites from indomethacin-treated platelets as well as nontreated platelets was also inhibited. The inhibitors apperar to interact with stimulated platelets but not with unstimulated platelets. These observations suggest that the interaction of an aggregating agent with its platelet receptor activates a unique precursor serine protease that in turn activates platelet phospholipase to liberate arachidonic acid (the precursor of the potent platelet aggregating agent thromboxane A2) from platelet phospholipids.
Erythropoiesis-stimulating factor production by rabbit kidney cultures from "programmed" rabbits
Authors: Ogle JW , Lange RD , Dunn CD
Published: October 2019

Rabbit kidney tissue either from normal rabbits or from rabbits “programmed” by phlebotomy or varying periods of hypoxia was cultured and shown to release into the culture medium an erythropoietically active substance. The cultures established from rabbits programmed by 6- 12 hr of hypoxia yielded the highest levels of erythropoiesis- stimulating activity.
Hypergranular promyelocytic leukemia (APL): cytogenetic and ultrastructural specificity
Authors: Testa JR , Golomb HM , Rowley JD , Vardiman JW , Sweet DL Jr
Published: October 2019

Cytogenetic and ultrastructural findings were important diagnostic indicators of hypergranular promyelocytic leukemia (APL) in a patient whose bone marrow morphology appeared, by light microscopy, to be similar to that in acute myeloblastic leukemia (AML) with maturation. Peripheral blood smears and bone marrow specimens examined by light microscopy showed few cells with the numerous coarse, azurophilic granules typical of APL. Cytogenetic analyses, with several banding techniques, of cells from bone marrow and unstimulated peripheral blood revealed the 15;17 translocation, which has been observed only in APL. A reinterpretation of the reciprocal translocation, based on R banding, suggests that the breakpoints are distal to q24 in No. 15 and at or near the junction of q21 and q22 in No. 17. In addition, the patient had disseminated intravascular coagulation. The characteristic morphology of granules seen in APL was observed in this case only when transmission electron microscopy was used, since the granules were quite small. Since treatment for AML differs from that for APL, identification of the 15;17 translocation and ultrastructural evidence of granules represent valuable diagnostic aids for APL.
Beta O-thalassemia intermedia
Authors: Cividalli G , Kerem H , Ezeckiel E , Rachmilewitz EA
Published: October 2019

Three patients with a relatively mild form of beta O-thalassemia who did not require regular blood transfusions are described. Globin synthesis was studied by gel filtration and urea-carboxymethylcellulose chromatography of stroma-free hemolysates prepared from peripheral blood and bone marrow cells labeled in vitro with 14C-leucine. gamma/alpha Synthetic ratios in peripheral blood were in the same range as in patients with the severe clinical form of beta O-thalassemia, while gamma/alpha synthetic ratios in bone marrow cells were higher than in that group of patients. The size of the free alpha-chain pool measured in one case was smaller than in other patients with “classical” Cooley anemia. It is concluded that the severity of the clinical course in beta O-thalassemia does not correlate with the imbalance in alpha verus gamma chain synthesis in peripheral blood and is determined by the synthetic ratio in bone marrow cells, where the bulk of hemoglobin synthesis takes place.
Modified method of erythropoietin (ESF) bioassay in vitro using mouse fetal liver cells. I. Effect of serum iron on 59Fe incorporation into heme
Authors: de Klerk G , Kruiswijk C , Hart AA , Goudsmit R
Published: October 2019

Investigations on the mouse fetal liver cell bioassay for erythropoietin (ESF) have revealed that iron present in test sera significantly dilutes the radiolabel (59Fe) and thus decreases 59Fe incorporation into heme. A method of correction for the influence of iron on the dose-response relationship of human sera is presented. Application of this method made it possible to assay human sera up to culture concentrations of 150 microliter/ml. It was shown that a corrected serum dose-response curve showed parallelism to the curve of an ESF standard preparation. This suggests similarity of the active principles and allows valid estimation of a potency ratio.
Modified method of erythropoietin (ESF) bioassay in vitro using mouse fetal liver cells. II. measurement of ESF in human serum
Authors: de Klerk G , Hart AA , Kruiswijk C , Goudsmit R
Published: October 2019

A modification of the mouse fetal liver cell bioassay for erythropoietin (ESF) that allows quantitative detection of ESF in human serum is described. The modification consists of (1) correction for the effect of serum iron on 59Fe incorporation into heme and (2) the use of an “internal standard,” i.e., a standard ESF preparation dissolved in the assayed test serum. As a result of this modification the statistical method of bioassay analysis had to be changed fundamentally. The mean serum concentration of ESF measured in 20 normal males was 48 microunit/ml, as compared to 29 microunit/ml in 18 females. The difference was significant at p = 0.017. The stimulatory activity of a human serum on 59Fe incorporation into heme could be neutralized by an anti-human ESF immune serum.
Proliferative state of blast cell progenitors in acute myeloblastic leukemia (AML)
Authors: Minden MD , Till JE , McCulloch EA
Published: October 2019

Peripheral blood from patients with acute myeloblastic leukemia (AML) contains cells capable of giving rise to colonies in culture when stimulated by media conditioned by leukocytes (LCM) in the presence of phytohemagglutinin (PHA). Two types of colonies are recognized with high frequency: The first grows in the presence of low concentrations of PHA LCM, have a blast-like morphology, and are numerically correlated with morphologically identified blast cells. The second requires either high PHA LCM concentrations or PHA alone with or without 2-mercaptoethanol and consists of cells capable of forming rossettes with sheep erythrocytes and resembles. T-lymphocyte colonies from normal blood. Precursors of blast cell colonies from 15 leukemic patients were tested for cycle state, using either the 3H-thymidine or hydroxyurea techniques. All were found to have a high proportion of cells in the S phase of the cycle. In contrast, T lymphocyte precursors from three normal individual were quiescent. The data are consistent with the maintenance of the leukemic blast cell populations by the proliferative activity of a small subpopulation of blasts.
Fibrin formation: effect of calcium ions
Authors: Brass EP , Forman WB , Edwards RV , Lindan O
Published: October 2019

Using laser fluctuation spectroscopy, a technique that measures particle size change in solution, the kinetics of fibrin clot formation from fibrinogen can be studied. With this technique the effect of calcium on the three distinguishable phases of clot formation, (1) proteolysis of fibrinogen, (2) fibrinogen-fibrin monomer complex formation, and (3) fibrin monomer polymerization, were investigated. Only a small change in the length of the induction period that results from the fibrinogen-fibrin monomer interactions was observed. However, there was a marked increase in the rate of fibrin monomer polymerization in the presence of calcium ions. These data show that calcium decreases the time required for fibrin formation from fibrinogen by markedly accelerating the phase of fibrin monomer polymerization.
Colony formation in vitro by leukemic cells in acute lymphoblastic leukemia (ALL)
Authors: Smith SD , Uyeki EM , Lowman JT
Published: October 2019

An assay system in vitro for the growth of malignant lymphoblastic colony-forming cells (CFC) was established. Growth of malignant myeloblastic CFC has been previously reported, but this is the first report of growth of malignant lymphoblastic CFC. Established assay systems in vitro have been very helpful in elucidating the control of growth and differentiation of both normal and malignant bone marrow cells. Lymphoblastic CFC were grown from the bone marrow aspirates of 20 children with acute lymphoblastic leukemia. Growth of these colonies was established on an agar assay system and maintained in the relative hypoxia (7% oxygen) of a Stulberg chamber. The criteria for malignancy of these colonies was based upon cellular cytochemical staining characteristics, the presence of specific cell surface markers, and the ability of these lymphoid cells to grow without the addition of a lymphoid mitogen. With this technique, specific nutritional requirements and drug sensitivities can be established in vitro, and these data may permit tailoring of individual antileukemic therapy.
Absence of beta mRNA in beta0-thalassemia in Kurdish Jews
Authors: Ramirez F , Starkman D , Bank A , Kerem H , Cividalli G , Rachmilewitz EA
Published: October 2019

We report the characterization of the amount of beta mRNA in a Kurdish Jewish population with beta0-thalassemia using the same methods employed for characterization of the Catania and Ferrara beta0 patients. We found very low amounts of beta mRNA sequences, consistent with the presence of beta0-thalassemia of the beta mRNA-negative population type. In addition, no globin gene deletion was detected that could account for the absence of beta mRNA.
Plasma cell leukemia (PCL): A report on 15 patients
Authors: Woodruff RK , Malpas JS , Paxton AM , Lister TA
Published: October 2019

Fifteen patients presenting with plasma cell leukemia (PCL) are reported in detail. The clinicopathologic features of PCL differ from typical myeloma and resemble those of acute leukemia: patients with PCL have less bone disease but a much higher incidence of organomegaly and tissue infiltration as well as diffuse marrow involvement and more pronounced pancytopenia. One of the reported patients developed meningeal plasma cell leukemia and is reported in detail. Cytomorphologic assessment of PCL cells showed nuclear immaturity and obvious nuclear/cytoplasmic asynchrony. Despite the use of cytotoxic agents known to be effective in myeloma, the prognosis in PCL is poor, and the median survival of the reported patients was only 2 mo.
Atypical lymphoid leukemia in ataxia telangiectasia
Authors: Levitt R , Pierre RV , White WL , Siekert RG
Published: October 2019

We observed two sisters with ataxia telangiectasia, one of whom developed an atypical subacute lymphocytic leukemia characterized by atypical lymphocytes and absence of palpable lymphadenopathy or hepatosplenomegaly. The lack of organomegaly in this patient may have been due to the underlying ataxia telangiectasia, which was associated with lymphoid hypoplasia. Cytogenetic studies showed a marker chromosome 14 [t(14q11:14q34)] in both patients. The sister with leukemia had other complex chromosomal aberrations in addition to the marker chromosome 14 that were stable for more than 14 mo before the patient's death from complicating infection. The development of atypical T cell leukemia has not been previously described in ataxia telangiectasia. This case further illustrates the interesting interrelationships amoung immunosuppressed states, development of lymphoid malignancy, and an emerging pattern of a propensity to chromosome 14 abnormalities in various lymphoid malignancies.
Deletion of the long arm of chromosome 20 [del(20)(q11)] in myeloid disorders
Authors: Testa JR , Kinnealey A , Rowley JD , Golde DW , Potter D
Published: October 2019

Detailed clinical and cytogenetic studies were performed in five patients who had abnormal hematopoiesis and an acquired deletion of an F-group chromosome. Cytogenetic analyses, with banding techniques, of cells from bone marrow, spleen, or unstimulated peripheral blood showed a partial deletion of the long arm of one chromosome 20 [del(20)(q11)] in all five patients. Three patients had myeloproliferative disorders of uncertain classification, the fourth had possible preleukemia, and the fifth had acute myelomonocytic leukemia. Although the five cases showed certain similarities, the clinical and hematologic findings seen with the 20q- abnormality were not specific. None of the patients showed evidence of polycythemia vera or idiopathic acquired refractory sideroblastic anemia, two diseases previously associated with the 20q-. Our studies indicate that the 20q- abnormality is not limited to diseases primarily affecting erythropoiesis but that it can be found in the broader spectrum of myeloid disorders. In polycythemia vera, the 20q- has sometimes been regarded as a possible result of previous therapy with cytotoxic agents; however, four of our patients were untreated when the deletion was first noted.