bluuebunny/crossref_metadata_2025 huggingface dataset:
Hand-mirror lymphocytes in infectious mononucleosis
Authors: Thomas WJ , Yasaka K , Strong DM , Woodruff CM , Stass SA , Schumacher HR
Published: October 2019

The identification of a morphologically unique lymphocyte in the peripheral blood of patients with infectious mononucleosis (IM) led to additional studies. Examination of Wright-stained smears of EDTA anticoagulated blood obtained from 25 patients with IM at presentation revealed significantly increased (p less than 0.0001) percentages of hand-mirror lymphocytes (HML) (mean 9.2%) compared with normal controls (mean 2.7%) or controls with nonspecific viral syndromes (mean 2.2%). Follow-up blood samples obtained on 10 of these patients demonstrated a marked increase in the HML count (mean 12.1%) that coincided with the onset of recovery. E-rosette separation of Ficoll-Hypaque-derived peripheral blood lymphocytes from 5 patients with early recovery IM showed the HML to be present almost exclusively in the T-cell population, representing about 25% of the T cells. Identical procedures on 5 controls showed less than 5% HML in the T-cell sample. Cytochemistries supported a T-cell derivation for HML. Electron microscopic examination of HML in IM demonstrated that these cells have unique ultrastructural features that may be related to functions of cellular attachment and cytotoxicity.
The regulation of hemopoiesis in long-term bone marrow cultures. II. Stimulation and inhibition of stem cell proliferation
Authors: Toksoz D , Dexter TM , Lord BI , Wright EG , Lajtha LG
Published: October 2019

The isolation of a DNA synthesis inhibitor (NBME fraction IV) and stimulator (RBME fraction III) specific for the hemopoietic stem cell (CFU-s) from freshly isolated normal adult and regenerating murine bone marrow, respectively, has been well documented. We have utilized long- term liquid bone marrow cultures in a further analysis of the role of these factors in the regulation of CFU-s proliferation. Our results show that shortly after feeding, at a time when the cultured CFU-s are actively proliferating, high levels of the hemopoietic stem cell proliferation stimulator fraction III can be isolated from the culture medium. In contrast, the presence of essentially noncycling CFU-s found in cultures fed 8–10 days previously correlates with high levels of the hemopoietic stem cell inhibitor fraction IV. These results suggest that a certain balance between these factors determines CFU-s proliferation in the long-term cultures. In support of this, DNA synthesis in actively cycling CFU-s in the long-term cultures is inhibited for at least 3 days by the addition of excess NBME fraction IV (inhibitor). Furthermore, DNA synthesis in noncycling cultured CFU-s is stimulated for at least 5 days by the addition of RBME fraction III (stimulator).
Effect of peripheral blood mononuclear cells from aplastic anemia patients on the granulocyte-macrophage and erythroid colony formation in samples from normal human bone marrow in vitro--a cooperative work
Authors: Takaku F , Suda T , Mizoguchi H , Miura Y , Uchino H , Nagai K , Kariyone S , Shibata A , Akabane T , Nomura T , Maekawa T
Published: October 2019

Thirty-six patients with aplastic anemia and three with aplastic anemia- PNH syndrome in eight institutes were studied for the presence of peripheral blood mononuclear cells having a suppressive effect on granulocyte-macrophage (G-M) and erythroid colony formation using a uniform protocol. In 11 cases of 29 (38%) and 9 cases of 29 (31%), the presence of mononuclear cells with a suppressive effect on G-M and erythroid colony formation, respectively, was demonstrated. However, the presence of mononuclear cells suppressive both to G-M and erythroid colony formation was demonstrated only in 3 of 19 cases (16%).
Histiocytic lymphoma fifteen years following remission of acute lymphoblastic leukemia
Authors: Dosik H , Anon R , Lee SL , Allen AC , Krishnamurthy M
Published: October 2019

A 19-yr-old female presented with diffuse histiocytic lymphoma after a 15-yr remission of acute lymphoblastic leukemia. All treatment had been discontinued 7 yr before the onset of the lymphoma. The possible relationships of these two neoplasms are discussed.
A plaque assay to enumerate circulating Ig-secreting cells of each type of the different Ig classes
Authors: Shimizu K , Hirano A , Kunii A
Published: October 2019

A new method to quantitate circulating cells secreting immunoglobulin (Ig) of each type of the different classes by hemolytic plaque technique is reported. The ratio of kappa (k)-secreting cells and lambda(lambda)-secreting cells of the different lg classes in normal individuals is almost equal to that of kappa:lambda light chains in corresponding serum lg class, whereas the ratio in plasma cell myeloma patients is extraordinarily high or low, indicative of the type and class specificity of this assay system as well as the monoclonal nature of circulating Ig-secreting cells in such patients.
Cryopreservation of human granulocytes: study of granulocyte function and ultrastructure
Authors: Boonlayangoor P , Telischi M , Boonlayangoor S , Sinclair TF , Millhouse EW
Published: October 2019

Human granulocytes can be cryopreserved with dimethyl sulfoxide (DMSO) at--80 degrees C. However, the percent recovery of functional cells has been unsatisfactory to date. Throughout this study we have been able to prepare relatively pure granulocytes approximately equal to 85%), cryopreserve them with 5%--10% DMSO with and without serum, and store them at--80 degrees C for up to 4 mo. The parameters studied were absolute cell counts and viability determination, myeloperoxidase activity, phagocytosis, candidacidal activity, bactericidal activity, nitroblue tetrazolium reduction, chemiluminescence, and cell morphology by transmission and scanning electron microscopy. Based on our investigation, granulocytes cryopreserved without serum showed an intact membrane of superior integrity as compared with those preserved with serum. At least 50% of the cells recovered were functional after 2 mo of storage, but there was a progressive loss of viability and function on prolonged storage. The property of phagocytosis was the best preserved after storage for 4 mo, whereas myeloperoxidase activity, killing activity, nitroblue tetrazolium reduction, and chemiluminescence were maintained less efficiently. Morphological studies of cryopreserved granulocytes revealed that the nuclear, cytoplasmic, and cell surface architectures were altered by storage. Depletion of nuclear and cytoplasmic material, as well as changes in configuration, were also noted.
Chronic myelodysplastic syndrome (preleukemia) with the Philadelphia chromosome
Authors: Roth DG , Richman CM , Rowley JD
Published: October 2019

A patient with severe anemia, reticulocytopenia, and erythroid hyperplasia of the bone marrow developed fatal acute nonlymphocytic leukemia after 3 yr. A Philadelphia chromosome with the typical 9/22 translocation t(9q +;22q-) was identified by banding techniques in a small number of bone marrow cells throughout the preleukemic phase of the illness (14%--38% of metaphases) and during the acute transformation (50%). Granulocytic colony formation in vitro was abnormal in the preleukemic phase. The diagnosis of chronic granulocytic leukemia was excluded on the basis of clinical and laboratory findings. The identification of the Ph1 chromosome in this form of chronic myelodysplastic syndrome (preleukemia) provides a new example of a hematologic disorder predisposing to acute leukemia in which this chromosomal abnormality occurs.
Acute lymphoblastic leukemia with Burkitt cell morphology and cytoplasmic immunoglobulin
Authors: Ganick DJ , Finlay JL
Published: October 2019

A case of acute lymphoblastic leukemia with morphological characteristics of Burkitt's leukemia (L3 morphology) is presented. This patient's lymphoblasts were lacking in surface immunoglobulin, but were found to contain cytoplasmic IgM. This is the first report of a morphologically B-cell leukemia showing pre-B-cell characteristics immunologically.
Serum lactic dehydrogenase (LDH) levels in acute leukemia: marked elevations in lymphoblastic leukemia
Authors: Kornberg A , Polliack A
Published: October 2019

Serum total lactic dehydrogenase (LDH) levels were examined in 42 patients with acute leukemia, 9 patients with chronic myeloid leukemia, 6 of them in blastic crisis, and 53 patients with lymphoma and other lymphoproliferative disorders. The mean range of serum LDH leveles in Hodgkin's and non-Hodgkin's lymphoma was 402 +/- 210 IU/liter and 313 +/- 113 IU/liter, while that of patients with nonmalignant disorders was 308 +/- 74 IU/liter. In acute nonlymphoblastic leukemia (ANLL), the range was 126–684 IU/liter (mean value 413 +/- 146 IU/liter). In 6 of the patients (11.3%) with lymphoma and in 6 cases (26.8%) with ANLL, the LDH levels were above 500 IU/liter. None of these patients had levels over 900 IU/liter. Patients with acute lymphoblastic leukemia (ALL) had a range of 402–3582 IU/liter (mean value of 1669 + 1038 IU/liter). In 15 of the 19 patients (78.9%) with ALL, serum LDH values were above 900 IU/liter. In addition, 3 patients with chronic myeloid leukemia (CLM) in blastic crisis had levels of 970–1940 IU/liter. One of these 3 patients had lymphoblastic crisis, while the second case responded clinically to vincristine and prednisone, but was not regarded as ALL. The differences in serum LDH levels between ALL and ANLL are statisticaly significant (p < 0.001). It appears that markedly elevated serum LDH levels in acute leukemia are suggestive of ALL, and that in individual patients, the LDH levels were correlated with the number of blasts during remission and relapse.
Purine nucleoside phosphorylase activity in acute lymphoblastic leukemia
Authors: Blatt J , Reaman GH , Levin N , Poplack DG
Published: October 2019

Purine nucleoside phosphorylase (PNP) activity has been measured in the lymphoblasts of 22 patients with acute lymphoblastic leukemia (ALL) and correlated with routine immunologic cell surface markers. Fourteen of the 22 patients were considered to have non-T, non-B-cell ALL; 8 patients had T-cell disease. The median PNP activity in 21 control samples of normal peripheral blood mononuclear cells was 83 U. The median PNP activity of the non-T, non-B lymphoblasts was 79 U. No statistical difference in PNP activity between these two groups could be discerned (p < 0.37). In contrast, T-cell lymphoblasts demonstrated diminished PNP activity with a median of 38 U. The differences in activity between T lymphoblasts and both non-T, non-B leukemic cells and normal peripheral blood mononuclear cells were significant (p < 0.001 and p < 0.003, respectively). Evaluation of PNP activity provides further evidence of biochemical heterogeneity among immunologic subclasses of ALL.
Cytotoxicity as an indicator for transport mechanism: evidence that murine bone marrow progenitor cells lack a high-affinity leucine carrier that transports melphalan in murine L1210 leukemia cells
Authors: Vistica DT
Published: October 2019

Cytotoxicity of melphalan to murine L1210 leukemia cells was reduced to a limiting maximum value of 50% by 2-aminobicyclo[2,2,1]heptane-2- carboxylic acid (BCH), indicating that cytotoxicity is partially dependent on drug uptake by system L. L-Leucine, but not alpha- aminoisobutyric acid (AIB), completely reduced the remaining 50% of drug cytotoxicity. These results contrast with those obtained with a sensitive host tissue, the bone marow progenitor cells of the white cell series (CFU-C), in that a high-affinity leucine transport system corresponding to system L was not identified.
Release of human platelet factor V activity is induced by both collagen and ADP and is inhibited by aspirin
Authors: Vicic WJ , Lages B , Weiss HJ
Published: October 2019

Factor V activity in suspensions of human platelets washed by albumin density gradient separation increased in response to stimulation by both collagen and adenosine diphosphate (ADP). The appearance of factor V activity extracellularly had the characteristics of platelet secretion and was partially inhibited by aspirin and by the antimetabolites 2-deoxyglucose and antimycin A. Some increase in factor V activity was also observed in platelet suspensions during the initial response to ADP; this activity was not detected extracellularly, but remained associated with the platelets. Patients with storage pool deficiency (SPD) whose platelets are deficient only in dense granule substances released normal amounts of factor V activity, whereas decreased amounts were released in a patient whose platelets have both dense and alpha granule deficiencies. These findings suggest that a portion of platelet factor V is associated with, and released from, alpha granules.
Ultrastructural evidence for the common origin of human mast cells and basophils
Authors: Zucker-Franklin D
Published: October 2019

Although the functional similarity of basophils and mast cells is widely accepted, their distinctive morphological features have been taken to indicate the existence of two different, albeit functionally complementary, cell systems. The recent demonstration that mast cells as well as basophils originate from the bone marrow raises the possibility that these cells derive from the same precursor. This report provides evidence for this theory by describing a distinctive “intermediate” cell possessing the ultrastructural features typical of both basophils and mast cells. These cells were encountered in three patients with myeloproliferative diseases and may thus be more readily found in states of disturbed myelopoiesis. These observations have given impetus for the first comparative description of the ultrastructure of human basophils, human mast cells, and the newly recognized intermediate cell within a single report.
Factor V deficiency in Philadelphia-positive chronic myelogenous leukemia
Authors: Hasegawa DK , Bennett AJ , Coccia PF , Ramsay NK , Nesbit ME , Krivit W , Edson JR
Published: October 2019

Factor V deficiency has been identified in 8 of 8 patients 7--20 yr of age, with Philadelphia-positive (Ph1+) chronic myelogenous leukemia (CML). In these 8 patients, factor V deficiency was not due to hepatic dysfunction, factor V inhibitors, or disseminated intravascular coagulation. In 3 patients, factor V activity rose 10%--12% (0.10--0.12 U/ml) after the infusion of 28--31 ml/kg body weight of fresh frozen plasma (FFP). The rise persisted less than 14 hr. The mean measured postinfusion rise in factor V was 18% of the expected rise calculated from the volume of FFP infused in the patients' plasma volume. In 4 patients, a small transient rise in factor V activity occurred after splenectomy or plateletpheresis. Factor V deficiency was completely corrected after a marked reduction in bone marrow cellularity in 2 patients with Ph1+ CML treated with extensive chemotherapy, total body irradiation, and bone marrow transplantation. Factor V deficiency was retrospectively observed in 6 of 20 patients, ages 20--80 yr, with Ph1+ CML and 3 of 6 patients with other myeloproliferative disorders. The factor V deficiency appears to be associated with the large myeloid- megakaryocytic cell mass characteristic of CML and other myeloproliferative disorders.
Autoimmune and alloimmune phenomena in patients with aplastic anemia: cytotoxicity against autologous lymphocytes and lymphocytes from HLA identical siblings
Authors: Warren RP , Storb R , Thomas ED , Su PJ , Mickelson EM , Weiden PL
Published: October 2019

We have studied peripheral blood lymphocytes of 117 patients with severe aplastic anemia and 237 healthy individuals for reactivity against autologous lymphocytes and/or lymphocytes from HLA-identical siblings using a 51Cr release assay. Lymphocytes from 29% of the patients exhibited reactivity against their own lymphocytes, while only 3% of lymphocytes from normal individuals showed such reactivity. Lymphocytes from 49% of the patients showed reactivity against lymphocytes from their HLA-identical siblings compared to 4% of normal individuals. Correlation existed between allogeneic and autologous reactivities (p < 0.001), suggesting a common pathway for cytotoxicity. Both reactivities showed an association with extremely low granulocyte counts (p < 0.01) and an inability of the patient's mononulcear cells to stimulate in allogeneic mixed leukocyte culture (p < 0.05) as well as an inverse correlation with time from diagnosis (p < 0.001). At least two explanations exist for the transfusion-independent autologous and allogeneic cytotoxicities: (1) they could be epiphenomena secondary to the stem cell defect, such as the loss of a cell that suppresses (or regulates) naturally occurring cytotoxic cells, or (2) they may be involved in the cause of the disease.
Modification of hemoglobin by ninhydrin
Authors: Kokkini G , Stevens VJ , Peterson CM , Cerami A
Published: October 2019

The Strecker degradation reaction was evaluated as a means of modifying hemoglobin in vitro, utilizing ninhydrin as a model compound. Ninhydrin led to modification of hemoglobin (when incubated with hemoglobin or red cells) at physiologic temperature and pH. Isoelectric focusing documented the formation of new hemoglobin bands, all with decreased (more negative) isoelectric points that hemoglobin A. Both alpha and beta chains were modified to an equal degree, although electrophoretic studies documented two modified species of alpha-chains and three modified species of beta-chains. Amino acid analysis of modified hemolysate following NaB3H4 reduction revealed peaks that coeluted with deaminated valine, epsilon-deaminated lysine, and a product with the guanidino group of arginine. The oxygen affinity of hemoglobin increased following its incubation with increasing concentrations of ninhydrin. These studies suggest that ninhydrin is representative of a class of carbonyl compounds that could be utilized to specifically modify that structure and function of hemoglobin variants.
Morphological classification, response to therapy, and survival in 263 adult patients with acute nonlymphoblastic leukemia
Authors: Mertelsmann R , Tzvi Thaler H , To L , Gee TS , McKenzie S , Schauer P , Friedman A , Arlin Z , Cirrincione C , Clarkson B
Published: October 2019

Bone marrow smears of 263 protocol patients with acute nonlymphoblastic leukemia (ANLL) and related disorders treated between 1970 and 1978 at MSKCC were reviewed blindly by two pairs of hematomorphologists and classified according to the FAB system. It was found necessary to add one category (MO) for acute undifferentiated leukemia and to define more precise quantitative criteria for the categories M1-M6 based on bone marrow differential counts. Using this modified FAB classification, agreement between the two observer groups based on morphology alone was 69%. Cytochemical stains were essential in establishing the diagnosis in 9%, led to a change of diagnosis by one observer team in 14%, and helped to confirm the diagnosis in 32% of cases. Complete remission rates, remission duration, and survival were not significantly different among diagnostic categories. Myelodysplastic syndromes (MDS: M6, RAEB, CMML; CR rate 48%) and ANLL without differentiation (M0, M1, M5a; CR rate 50%) appeared to do less well than ANLL with partial differentiation (M2, M3, M4, M5b; CR rate 59%) on all three protocols studied. Auer rods were present in 53% of all cases with 63% in the myeloid categories (M1-M4). Auer rods were found to be the single most important prognostic parameter in this study, with a complete remission (CR) rate of 68% in the Auer-rod- positive and of 40% in the Auer-rod-negative group (p < 0.0001). Survival was significantly longer for patients exhibiting Auer rods (p < 0.0002). Median survival for the total group was 13.5 mo and median remission duration for responders was 11.5 mo in the Auer-rod-positive group compared to 6.2 mo median survival and 9.2 mo median remission duration for the Auer-rod-negative group.
Membrane abnormality in red blood cells with weak type B expression
Authors: Yoshida A , Fujii H , Dave V , Cozant MJ , Morel PA
Published: October 2019

The mechanisms of unusually weak blood group (A and B) expressions are not yet well understood. We examined properties of blood group galactosyltransferase (B-enzyme) and characteristics of red cell membrane components obtained from family members with A2Bm character. B- enzyme activity of the A1Bm plasma is in normal range, and kinetic properties (i.e., Km for UDP-Gal, Km for 2′-fucosyllactose, and pH optima) of B-enzyme from the A1Bm subjects are identical to that of normal B-enzyme. When A1Bm red cell were incubated with UDP-Gal and B- enzyme, the cells became strongly agglutinable with anti-B. When A1Bm membranes were incubated with B-enzyme or A-enzyme (i.e., blood group N- acetylgalactosaminyltransferase) and the appropriately labeled nucleotide sugar (UDP-Gal3H for B-enzyme and UDP-GalNAc3H for A- enzyme), significant incorporation of the sugar was observed. The amounts of the sugar incorporated into A1Bm membranes were about 40%- 50% of that incorporated into O membranes at saturation, indicating that about one-half of H-sites remained unglycosylated in A1Bm red cells. Examination of radioactive components by isoelectric focussing revealed that the labeled components of A1Bm membranes were distinctively different from that of O membranes. Therefore, one can conclude that the weak B expression is not due to direct mutation of ABO locus, but due to a secondary consequence of genetic abnormality of a membrane component (or components) associated with blood group substances.
Hormonal effects on cell proliferation in a human erythroleukemia cell line (K562)
Authors: Gauwerky C , Golde DW
Published: October 2019

We have investigated the hormonal responsiveness of K562 cells using a serum-substituted in vitro clonogenic assay. Dexamethasone inhibited colony formation by the K562 cells, and the inhibitory effect could be reversed by progesterone (10(-6) M). Fluoxymesterone caused a prominent enhancement of K562 colony growth, whereas estriol had no effect. Stimulation by triiodothyronine was maximal at 10(-7) M, and the thyroid effect could be abrogated by the beta 2-adrenergic antagonist butoxamine in equimolar concentrations. Using standard tissue culture conditions, the beta-adrenergic agent isoproterenol, but not the alpha catecholamine phenylephrine, enhanced the proliferation of K562 cells. When K562 cells were grown under hormone-depleted conditions, they developed responsiveness to phenylephrine and were no longer stimulated by isoproterenol. DbcAMP and prostaglandins of the E series also caused K562 colony enhancement. Prostaglandin F2 alpha had no effect on cell proliferation. Insulin was an effective stimulant of colony formation of K562 cells, as were human growth hormone and ovine prolacin. Bovine growth hormone had no effect. Our results are consistent with the identificaiton of K562 as an erythroid line, and they indicate that K562 cells respond to endocrine hormones in a manner analogous to normal erythroid progenitors.
Human T-cell antigens defined by monoclonal antibodies. Absence of T65 on committed myeloid and erythroid progenitors
Authors: Taetle R , Royston I
Published: October 2019

The complement-dependent cytotoxicity of monoclonal T-cell antibody (T101) for normal and abnormal hemopoietic progenitors was assessed. T101 demonstrated toxicity for normal T-colony-forming cells from peripheral blood and bone marrow. Cytotoxicity was absent for normal peripheral blood and bone marrow granulocytes/macrophage (CFU-C) and erythroid (BFU-E) progenitors. The antibody was also not toxic for peripheral blood blast progenitors from patients with acute myelogenous leukemia (AML). These studies indicate the absence of the antigen defined by T101 (T65) from normal progenitor cells and from blast progenitors in patients with AML. T101 may be used in the treatment of T-cell malignancies and in the prevention of graft-versus-host disease (GVHD) without damage to normal progenitor cells.
Clinical implications of glucocorticoid receptor studies in childhood acute lymphoblastic leukemia
Authors: Mastrangelo R , Malandrino R , Riccardi R , Longo P , Ranelletti FO , Iacobelli S
Published: October 2019

We have performed in parallel, in 19 children with acute lymphoblastic leukemia, a quantitative determination of glucocorticoid levels, in vitro steroid induced inhibition of nucleic acid precursors, and a short-term clinical trial of corticosteroids alone, before the treatment was given, which included corticosteroids and other drugs. From our results it appears that high glucocorticoid receptor levels in acute lymphoblastic leukemia of children do not guarantee a clinical response to corticosteroids. On the other hand, glucocorticoid receptors may turn out to be of value in predicting a poor response to corticosteroids only if their levels are considerably low.
Cord blood screening for hemoglobin abnormalities by thin layer isoelectric focusing
Authors: Galacteros F , Kleman K , Caburi-Martin J , Beuzard Y , Rosa J , Lubin B
Published: October 2019

Hemoglobin variants can be successfully identified in cord blood samples. The methods most commonly used include cellulose acetate (CAC) and citrate agar (CAG) electrophoresis. Recently thin layer isoelectric focusing (TLIF) has been shown to be an excellent method for identifying hemoglobin variants. To determine the applicability of TLIF for cord blood screening, we compared the results of 835 samples obtained by TLIF with that obtained by CAC, CAG, and the combination of both CAC and CAG. In 100 of these samples we detected an abnormal hemoglobin pattern using TLIF. In contrast, we detected only 80 abnormal samples by CAC, 70 by CAG, and 80 by using the combination of CAC and CAG. Due to the increased resolution provided by TLIF, we correctly diagnosed two sickle cell trait samples by TLIF that were incorrectly suspected to be homozygous for sickle cell disease by CAC and CAG. We identified 41 samples containing Bart's hemoglobin by TLIF in contrast to only 21 using CAC and 14 using CAG. The time and cost of TLIF was comparable to that using the combination of both methods. We, therefore, conclude that TLIF is the method of choice for cord blood screening.
Significance of Ph1-negative marrow cells in Ph1-positive chronic granulocytic leukemia
Authors: Sokal JE
Published: October 2019

Fifty-six of 195 Ph1-positive patients with chronic granulocytic leukemia were found to have Ph1-negative metaphases in marrow aspirates on one or more occasions. In 22 cases, Ph1-negative cells were found prior to initiation of antileukemic therapy. Five patients were in the blastic stage of the disease when Ph1-negative mitoses were seen. The finding of Ph1-negative cells appeared to be related principally to short duration of CGL and to administration of antileukemic therapy (conventional agents and doses, in most cases). Ph1-negative cells were usually not found more than 2 yr after the diagnosis of leukemia, but in a few cases, they were seen as long as 5–10 yr after diagnosis. Only a minority of metaphases analyzed were Ph1-negative, except in the case of 6 patients who transiently had 50% or more Ph1-negative cells after antileukemic therapy. The presence of Ph1-negative cells in marrow was not associated with any survival advantage in this series.
The in vitro sensitivity of leukemic and normal leukocytes to hydrocortisone induced cytolysis
Authors: Galili U , Prokocimer M , Izak G
Published: October 2019

A marked sensitivity of CLL lymphocytes to hydrocortisone in vitro was demonstrated in each of the 25 patients tested. The sensitivity was manifested by the eventual lysis of the affected cells. Malignant lymphocytes from 8 out of 14 ALL patients were found also to be in vitro sensitive, whereas CML cells, AML cells, normal BM cells, thymocytes, peripheral blood lymphocytes, and polymorphonuclear cells were resistant. Within a tested CLL lymphoid suspension it is proven that the hydrocortisone causes the specific lysis of the malignant cells leaving the normal lymphocytes undamaged. The cytolysis is not an immediate action, but is expressed within 7–8 hr of incubation. However, 30 min incubation with the hormone is sufficient for the cytolytic effect to occur 20 hr later. The possible mechanisms involved in the specific glucocorticoid induced cytolysis are discussed.
Prenatal diagnosis of hemoglobinopathies: comparison of the results obtained by isoelectric focusing of hemoglobins and by chromatography of radioactive globin chains
Authors: Dubart A , Goossens M , Beuzard Y , Monplaisir N , Testa U , Basset P , Rosa J
Published: October 2019

Isoelectric focusing (IEF) of hemoglobin was compared to the classical chromatography of labeled globin chains for 22 antenatal diagnoses of hemoglobinopathies: 11 for beta thalassemia, and 11 for sickle cell disease. In all cases, the two methods gave identical results. The diagnosis was confirmed after birth or abortion. Three fetuses homozygous for beta thalassemia and one homozygous for sickle cell disease exhibited no Hb A by IEF, in contrast to normal fetuses or those heterozygous for one of the two hemoglobinopathies. In addition, blood samples obtained in other centers after abortion of 22 fetuses homozygous for beta + or beta 0 thalassemia exhibited no Hb A when analyzed by IEF. When Hb A was present, the respective proportions of Hb A and acetylated Hb F were determined by densitometry of the IEF gel. The Hb A/acetylated Hb F ratio obtained by IEF correlated well with the beta A/gamma ratio of globin chain synthesis, IEF requires 0.1 mg of unlabeled hemoglobin. It is performed in 90 min and several samples can be analyzed simultaneously. If present, maternal contamination of fetal blood must be eliminated by selective lysis of maternal (RBC) using the Orskov reaction. Improvements in this method to obtain suitable samples for IEF analysis are described.
Improved exercise performance after exchange transfusion in subjects with sickle cell anemia
Authors: Miller DM , Winslow RM , Klein HG , Wilson KC , Brown FL , Statham NJ
Published: October 2019

Ten patients with sickle cell anemia underwent partial exchange transfusion with hemoglobin-A-containing cells using a technique that allowed hemoglobin concentration and blood volume to remain constant. The mean fraction of hemoglobin-A in these patients increased from 9% to 55%, but the mean hemoglobin concentration increased by only 1.44 g/dl. The exchange resulted in a large improvement in submaximal exercise capacity: the mean of the anaerobic threshold (the work at which lactic acid begins to accumulate in the blood) increased from 68 to 114 W. The mean work performed at a heart rate of 170/min, an estimation of maximal work capacity, increased from 128 to 187 W. Improved exercise performance after partial exchange transfusion may result from the superior flow properties of hemoglobin-A-containing red cells. Furthermore, we believe that exercise testing in sickle cell anemia has great potential utility as a means to monitor therapy and to evaluate the benefits of exchange transfusion.
The nature and action of granulocyte-macrophage colony stimulating factors
Authors: Burgess AW , Metcalf D
Published: October 2019

Granulocyte-macrophage colony stimulating factor (GM-CSF) stimulates the in vitro proliferation and differentiation of granulocytic and macrophage cells. This regulator is now known to act at other levels of hemopoietic regulation. The heterogeneity of GM-CSFs is not only related to the tissue of origin and the in vitro production method, but also to functional subclasses of the molecule that have distinct biologic specificities. Most adult mouse organs produce GM-CSF (mol wt 23,000), but a macrophage (M)-CSF has been detected in fetal conditioned medium (CM) and isolated from L-cell CM. Murine endotoxin serum appears to contain a M-CSF, GM-CSF, and G-CSF, the last of which cofractionates with a differentiation factor active on leukemic cells. Human GM-CSFs, G-CSF, and EO-CSFs active on human cells have been detected in a variety of CM, but as yet none have been purified. Again, there are subclasses of progenitor cells that respond to particular forms of human active CSFs. GM-CSF isolated from mouse lung CM stimulates multipotential progenitor cells, the initial proliferatin of progenitors in the erythroid, eosinophil, and megakaryocyte series, as well as mature cells in the GM series. While GM-CSF is also able to stimulate the differentiation of myeloid leukemic cells, other factors appear to be more potent in this respect. Information on the regulation of GM-CSF production, on the modulators of its action on specific target cells, and on its role in vivo will be required before the physiologic function of this molecule can be properly assessed.
Human red cell membrane adenylate cyclase in normal subjects and patients with hereditary spherocytosis, sickle cell disease and unidentified hemolytic anemias
Authors: Piau JP , Delaunay J , Fischer S , Tortolero M , Schapira G
Published: October 2019

We have investigated adenylate cyclase in ghosts from normal and pathologic human red blood cells. Basic parameters such as specific activity, apparent Michaelis constant (KMapp), and response to effectors: sodium fluoride (NaF), 5′-guanylyl imidodiphosphate (Gpp (NH)p), isoproterenol, and PGE1 were investigated. Basal and NaF- stimulated activities were measured in ghosts from patients with hereditary spherocytosis, sickle cell disease, and various unidentified hemolytic anemias. Both activities were increased in any of these pathologic conditions as compared with those of normal red blood cells. Normal values were found in patients with hereditary spherocytosis after splenectomy and in patients with heterozygous sickle cell disease. There was a good correlation between the reticulocyte count and adenylate cyclase activity in hereditary spherocytosis and in sickle cell disease with reticulocyte count lower than 10%. The enzyme activity of the first group was about three times that of the second group. There was no correlation at all in sickle cell disease with higher reticulocytosis and in the group of unidentified hemolytic anemias. These results suggest that increased adenylate cyclase activities are not specific of any of these diseases. In the patients with hereditary spherocytosis, the adenylate cyclase activity seems to be essentially related to younger mean age of red blood cell population while in the patients with sickle cell disease and in others with unidentified hemolytic anemias some additional factors might interfere directly with the enzyme and alter its activity.
The mechanism of platelet release
Authors: Radley JM , Scurfield G
Published: October 2019

The mechanism of platelet release has been studied in mouse bone marrow, using scanning and transmission electron microscopy. Cytoplasmic processes extended into sinusoids by megakaryocytes were found to undergo attenuation, and at the distal end develop constrictions between segments of platelet size. Rupture at sites of constriction is thought to release platelets. Microtubules were present in the processes, orientated longitudinally, and became concentrated in the constrictions. The narrowest constrictions observed were about 0.2 micrometer in diameter and several microns in length. The structural similarity of the narrowest constrictions with those found in bridges separating midbody and cell in the terminal phase of cytokinesis suggests that platelets and cells share a common mechanism for release. The presence of a centriole in each cytoplasmic process was revealed by serial sectioning. It is suggested that the microtubules in each cytoplasmic process originate from an organizing center associated with the centriole, and that the number of cytoplasmic processes that emerge from a megakaryocyte is governed by its ploidy.
5-Azacytidine and renal tubular dysfunction
Authors: Peterson BA , Collins AJ , Vogelzang NJ , Bloomfield CD
Published: October 2019

During initial trials of 5-azacytidine in adults with advanced acute leukemia, we unexpectedly observed acid-base, fluid, and electrolyte abnormalities that contributed directly to the deaths of two early patients. To evaluate this toxicity further, we studied 22 patients who received a total of 33 courses of combination chemotherapy that included 5-azacytidine. During 29 courses (88%) of treatment, polyuria, glucosuria, and/or transient changes in the serum concentrations of bicarbonate or phosphorus were detected. Spontaneous polyuria with demonstrable salt wasting and orthostatic hypotension occurred during seven courses (21%) of treatment. Inappropriate glucosuria was observed in nine courses (27%). In 24 courses (73%) the serum bicarbonate fell below the normal range. The urine became alkaline during 12 of these instances; the anion gap was not increased during the acidosis. Hypophosphatemia with serum phosphorus concentrations as low as 0.3 mg/dl occurred in 21 of 32 evaluable courses (66%). In the three patients studied the tubular reabsorption of phosphorus was 10%-18%. The renal abnormalities that were observed suggest both proximal and distal tubular damage from 5-azacytidine. Patients receiving 5- azacytidine should be monitored closely for manifestations of renal toxicity.
Absence of platelet-specific alloantigens in Glanzmann's thrombasthenia
Authors: van Leeuwen EF , von dem Borne AE , von Riesz LE , Nijenhuis LE , Engelfriet CP
Published: October 2019

The platelets of 11 patients with Glanzmann's thrombasthenia and their nearest family members were studied for the expression of the platelet- specific alloantigens of the Zw-, Ko- and Bak systems. The strength of the expression of the Zwa antigen was diminished on the platelets of 3 patients, and the antigen was absent from the platelets of the other 8. The platelets of none of the patients reacted with anti-Zwb serum. Therefore, Glanzmann's thrombasthenia is probably a “Zw-null disease.” The expression of the Zwa antigen on the platelets of all the relatives was normal, as indicated on the cytoflurograph. Investigations on the expression of the Koa antigen were complicated by agglutinations of the platelets from genetically Koa-negative thrombasthenic patients with the anti-Koa serum. The Kob antigen was normally expressed. The Baka antigen was absent from the platelets of all thrombasthenic patients and a relatively high percentage of the relatives. A close association between the Glanzmann gene and the Bak(a-) gene is assumed on statistical grounds. Thrombasthenic platelets showed no reaction with EDTA-dependent antibodies, which are reactive with all normal platelets. Owing to immunization by multiple blood and platelet transfusions, serum samples of most patients studied contained HLA antibodies and platelet-specific alloantibodies. However, antibodies directed against the Zw-antigen-bearing glycoproteins were detected in the serum of only one patient and, therefore, seem to be rare.
Sucrose density gradient analysis of erythrocyte membranes in hemolytic anemias
Authors: Flynn TP , Johnson GJ , Allen DW
Published: October 2019

To investigate the membrane abnormalities that may play a pathophysiologic role in several hemolytic anemias we determined the density distribution on sucrose density gradients of human red blood cell (RBC) membranes from patients with these disorders, from normal controls, and from incubated normal RBC. We analyzed the fractions for membrane-adsorbed hemoglobin (Hb), globin, and nonglobin cytoplasmic proteins. The relationship between the cytoplasmic proteins adsorbed on the membranes and the specific gravity (SG) of the membranes was linear. An increase in SG of the entire membrane population was seen in Hb C disease due to adsorbed Hb. Subpopulations of membranes with increased SG due to adsorption of nonglobin protein were evident in the membranes from two splenectomized patients with hemolytic glucose-6- phosphate dehydrogenase (G6PD) variants. Dense membrane subpopulations found in RBC membranes from three splenectomized patients with Hb Koln were associated with adsorbed globin, while similar subpopulations in RBC membranes from three splenectomized patients with hereditary spherocytosis demonstrated increased SG due to adsorbed Hb. Splenectomized normals had no such abnormality in membrane density. Sucrose density gradients demonstrate that membrane bound cytoplasmic protein is characteristic of the RBC membranes in several hemolytic disorders. Additionally, gradients are useful for the isolation and further analysis of those subpopulations of RBC membranes with abnormal SG and exaggerated membrane protein abnormalities.
Genetic variation in the quantitative levels of an NADP (H)-binding protein (FX) in human erythrocytes
Authors: Lenzerini L , Benatti U , Morelli A , Pontremoli S , De Flora A , Piazza A , Rinaldi A , Filippi G , Siniscalco M
Published: October 2019

FX is a red cell NADP(H)-binding protein that has been well defined biochemically and immunologically but whose function is still unknown. Preliminary data indicated that the levels of this protein are significantly increased in hemizygotes, heterozygotes, and homozygotes for the G6PD Mediterranean mutant, thus raising the question of whether or not the individual variation in FX levels is more or less directly influenced by X-linked genes. The present study, based on a large series of population and family data collected in Sardinia, confirms unequivocally the above mentioned interaction, but shows at the same time that the variances in FX levels “between sibships” are 2–3 times larger than those “within sibships,” when the analysis is done separately for the G6PD-normal or the G6PD-deficient sibs. From the comparison of the interclass and intraclass correlation coefficients, it appears that about 60% of the total variation of FX is of genetic origin. Moreover, the FX levels of children, analyzed in a pairwise manner, were found to be more positively correlated with those of their fathers (r = 0.39) than with those of their maternal grandfathers (0.20). This latter finding obviously favors the conclusion that “autosomal”; rather than “X-linked” genes are involved in the determination of the FX levels.
The relationship between plasma iron and plasma iron turnover in the rat
Authors: Bauer W , Stray S , Huebers H , Finch C
Published: October 2019

Plasma iron turnover has been evaluated in the growing rat. Consistent data were obtained with the intravenous injection of radioiron in the form of ferrous sulfate or ferric citrate. Plasma iron turnover changed as a function of plasma iron concentration. Only part of this effect in the rat was due to the different rates of clearance of mono-and differic transferrin, the latter having a higher iron delivery rate in vivo. An additional effect was shown to relate to the rate of red cell production. With decreased production, the effect of plasma iron on plasma iron turnover was reduced, whereas with increased erythropoiesis there was an additional increment in plasma iron turnover for any increase in plasma iron. Since this effect was observed when increased iron demands were due to an increase in erythroid precursors in the marrow but not in the circulating blood, it is attributed to limitations in iron flow to the marrow. This suggests that erythroid marrow activity and the adequacy of iron supply when studied by ferrokinetic techniques can best be defined by the response curve relating plasma iron concentration to plasma iron turnover.
Human myelogenous leukemia: enhanced clonal proliferation in the presence of phorbol diesters
Authors: Koeffler HP
Published: October 2019

Phorbol diesters, including 12–0-tetradecanoyl-phorbol-13-acetate (TPA) at low concentrations, enhanced the clonal proliferation of a human myelogenous leukemia cell line (KG-1). TPA (1.0 X 10(-9) -5.0 X 10(-11) M) increased KG-1 clonal growth by four to tenfold in the presence of suboptimal concentrations of colony stimulating factor (CSF) and by two to fourfold in the presence of maximally stimulating concentrations of CSF. The ability of TPA analogs to enhance KG-1 clonal growth paralleled their reported ability to promote tumors in mice. Only slight CSF activity was detected in the conditioned medium from KG-1 cells cultured in liquid medium with TPA. TPA was unable to increase myeloid clonal growth of CSF-stimulated normal human model to study TPA enhancement of CSF-stimulated growth.
Measurement of ploidy distribution in megakaryocyte colonies obtained from culture: with studies of the effects of thrombocytopenia
Authors: Levin J , Levin FC , Penington DG , Metcalf D
Published: October 2019

Microdensitometric measurement of the DNA content of individual megakaryocytes was performed using megakaryocyte colonies obtained following culture, in soft agar, of hematopoietic cells from C57BL/6J mice. Two types of colonies were detected. After 7 days of culture, the big cell type contained 16 /+- 2.3 acetylcholinesterase (AChE) positive cells/colony, with a mean ploidy level of 16.8 /+- 0.8/cell and the ploidy distribution characteristic of recognizable megakaryocytes in bone marrow. The heterogeneous type contained 44 /+- 9.6 cells/colony (some of which were AChE negative), with a mean ploidy level of 6.8 /+- 0.7/cell. The ploidy distribution of heterogeneous colonies differed markedly from big cell colonies, with preponderance of 2N and 4N cells. Colony-forming cells, obtained 4–5 days after induction of acute thrombocytopenia, gave big cell colonies with a marked increase in DNA content. Mean ploidy level increased to 21.5 /%- 1.8/cell; the frequency of 32N cells increased from 17% to 30% and 64N cells from 0% to 6%. This is the pattern of change observed in bone marrow, in vivo, 24 to 48 hr after induction of acute thrombocytopenia. The number of cells/colony did not increase. In contrast, acute thrombocytopenia did not alter the ploidy of heterogeneous colonies. The different responses to the stimulus of acute thrombocytopenia suggest that there are at least two types of Meg-CFC. The delayed appearance of altered Meg-CFC that produced big cell colonies indicates that the pool of stem cells, from which committed megakaryocyte precursors are derived, may respond indirectly to the stimulus of platelet depletion.
Further evidence of the in vivo role of erythropoietin or companion molecules induced by hypoxia on proliferation and continuing differentiation of BFU-e in PCDC
Authors: Harigaya K , Cronkite EP , Miller ME , Moccia G
Published: October 2019

Normal and plethoric bone marrow cells were grown in plasma clot diffusion chambers (PCDC) implanted into the peritoneum of normal mice or mice submitted to 7 her of hypoxia (23,000 ft) daily, on a single day or on 2 consecutive days at different times after implantation of the PCDC's. Daily discontinuous hypoxia (DDH) produced more 6-day bursts than other treatments. Hypoxia on days 1 and 2 after implantation was nearly as effective as DDH on day-6 bursts. Later bouts of hypoxia or a singly hypoxic exposure on day 1 or 2 was less effective. Erythropoietin (Ep) levels were measured by bioassay on both diffusion chamber (DC) contents and serum. Serum Ep levels peaked at 160 mU/ml after a 7-hr hypoxic exposure while the DC content Ep levels were in the nondetectable range (less than 50 mU/ml). The data implies that either higher than normal Ep levels or a companion molecules (s) produced by hypoxia are required for 1–2 days early in the culture period of force an increasing number of BFU-d-e down the erythrocytic pathway and thus increase red cell production at times of need in vivo.