bluuebunny/crossref_metadata_2025 huggingface dataset:
Impaired granulocyte superoxide production and prolongation of the respiratory burst due to a low-affinity NADPH-dependent oxidase
Authors: Shurin SB , Cohen HJ , Whitin JC , Newburger PE
Published: October 2019

A 7-yr-old girl with self-limited pulmonary aspergillosis was found to have a defect in granulocyte superoxide production. Her cells produced superoxide at 3% of control rates in response to phorbol myristate acetate (PMA) and opsonized zymosan. Lag times for O-2 production were normal with PMA, opsonized zymosan, and concanavalin-A stimulation. Her granulocyte membranes depolarized in response to all of these stimuli. Superoxide produced by podosomes and a particulate fraction demonstrated an enzyme activity with a normal maximal velocity but a decreased affinity for NADPH. NADH-dependent superoxide production by particles was similar with patient and control material. The duration of superoxide production was prolonged in the patient's intact granulocytes and in the particulate fractions from her cells. Bacterial killing by the patient's granulocytes was initially low, but approached control values after 90 min of incubation. These results are explained by an enzyme activity that has a decreased affinity for its substrate and a decreased rate of inactivation. Family studies indicate an autosomal recessive mode of inheritance.
Prednisone pulse therapy for refractory myeloma
Authors: Alexanian R , Yap BS , Bodey GP
Published: October 2019

The utility of vindesine and a frequent prednisone schedule was evaluated in 70 patients with refractory myeloma. No patient responded to vindesine alone, but about one-fourth achieved significant tumor reductions from intermittent high-dose prednisone, either alone or in combination with vindesine. Forty-seven percent responded when prednisone pulses were combined with vincristine and doxorubicin, providing the best results yet achieved in our patients with refractory myeloma. In responding patients, remissions were of excellent quality and survival was prolonged significantly. These results supported the utility of a more frequent corticosteroid schedule with increased doxorubicin dose in patients with advanced and resistant multiple myeloma.
Differences among myeloproliferative disorders in the behavior of their restricted progenitor cells in culture
Authors: Croizat H , Amato D , McLeod DL , Eskinazi D , Axelrad AA
Published: October 2019

We have studied the behavior in culture of circulating restricted hemopoietic progenitor cells from patients with idiopathic myelofibrosis (IMF), polycythemia vera (PV), and essential thrombocytopenia (ET). We have found differences in circulating granulocyte-macrophage, erythroid, and megakaryocytic progenitors that appear to be specific for these chronic myeloproliferative disorders. In IMF, most affected were granulocyte-macrophage progenitor cells (CFU- C), which circulated in increased numbers and were heterogeneous in their sensitivity to the regulatory factor(s) present in phytohemagglutinin (PHA) stimulated T-lymphocyte conditioned medium (CM). Most CFU-C were either highly sensitive to, or independent from, stimulatory factors, while others showed normal sensitivity. In some IMF patients, circulating megakaryocytic progenitors (CFU-M) were present that were capable of giving rise to colonies in the absence of added CM or erythropoietin (EPO). In PV, we confirmed the presence of circulating erythroid progenitor cells that give rise to colonies in culture without the addition of EPO. The number of circulating CFU-C was normal and they responded normally to CM. In ET, failure to detect 7-day circulating restricted progenitor cells was a common observation; the level of other circulating restricted progenitors was in the low normal range. Thus, despite certain common features, including a primary lesion at the level of the pluripotential hemopoietic stem cell, the myeloproliferative disorders differ with respect to the behavior in culture of their circulating restricted progenitor cells. These results have led us to postulate a second regulatory lesion in the pluripotential stem cell that differs in these disorders and is expressed at the level of the respective restricted progenitor cells.
A monokine regulates colony-stimulating activity production by vascular endothelial cells
Authors: Bagby GC Jr , McCall E , Bergstrom KA , Burger D
Published: October 2019

Human umbilical vein endothelial cells were cultured in supernatants of peripheral blood monocytes that had been cultured for 3 days with and without lactoferrin. Colony-stimulating activity (CSA) was measured in supernatants of the endothelial cell cultures and appropriate control cultures using normal, T-lymphocyte-depleted, phagocyte-depleted, low- density bone marrow cells in colony growth (CFU-GM) assays. Monocyte- conditioned medium contained a nondialyzable, heat labile factor that enhanced 4–15--fold the production of CSA by endothelial cells. The addition of lactoferrin to monocyte cultures reduced the activity of this monokine by 69%. Lactoferrin did not inhibit CSA production by monokine-stimulated endothelial cells. Therefore, vascular endothelial cells are potent sources of CSA, the production of CSA by these cells is regulated by a stimulatory monokine, and the production and/or release of the monokine is inhibited by lactoferrin, a neutrophil- derived putative feedback inhibitor of granulopoiesis. Inasmuch as a similar monokine is known to stimulate CSA production by fibroblasts and T lymphocytes, we suggest that mononuclear phagocytes play a pivotal role in the regulation of granulopoiesis by recruiting a variety of cell types to produce CSA.
High serum colony-stimulating activity of leukocytopenic patients after intravenous infusions of human urinary colony-stimulating factor
Authors: Motoyoshi K , Takaku F , Miura Y
Published: October 2019

Eight adult patients suffering from leukocytopenia have been injected by droplet intravenous infusions with partially purified human urinary colony-stimulating factor (CSFHU), which stimulated human monocytes to produce colony-stimulating activity (CSA) in vitro. Three of eight patients were injected with low-dose CSFHU and five with high-dose CSFHU. The infusion of high-dose CSFHU led to a slightly earlier rise in absolute neutrophil counts and a higher CSA level in the serum, as compared to noninjected leukocytopenic patients. There was little toxicity associated with it. Human urinary colony-stimulating factor used in the clinical studies did not have any CSA in vitro in the presence or absence of the patients' sera. These data may suggest that the intravenous infusion of CSFHU increased the serum CSA level, probably through the stimulation of CSA-producing cells in vivo.
Leukocyte interferon-induced myeloid cytoreduction in chronic myelogenous leukemia
Authors: Talpaz M , McCredie KB , Mavligit GM , Gutterman JU
Published: October 2019

We investigated the antiproliferative effect of partially purified human leukocyte interferon (HuIFN-alpha) given in a dose of 9–15 X 10(6) U daily by intramuscular injection to 7 patients with chronic myelogenous leukemia (CML). Hematologic remission of the disease was obtained in 5 patients. Among the responding patients, the mean white blood cell count decreased from 97.4 X 10(3)/cu mm (range from 35 X 10(3)/cu mm to 239 X 10(3)/cu mm) to 4.2 X 10(2)/cu mm (range from 3.0 X 10(3) to 7.9 X 10(3) cu/mm). Parallel reduction occurred in serum B12, from a mean of 1,435 pg/ml to a mean of 726 pg/ml, and lactate dehydrogenase levels, from a mean of 325 mU/ml to 112 mU/ml. Enlarged spleens decreased in 3 of 3 patients. The 5 responding patients have been maintained on HuIFN-alpha, 3 X 10(6) U daily or every other day, for 6+-35+ wk.
Induction of differentiation of human acute myelogenous leukemia cells: therapeutic implications
Authors: Koeffler HP
Published: October 2019

A prominent phenotypic abnormality of human acute myelogenous leukemia cells is the inability of the cells to differentiate to functional mature cells; instead, the cells are blocked at an early stage of development and remain in the proliferative pool and rapidly accumulate. Investigation of the induction of myeloid leukemic cell differentiation has made recent advances with the development of several human myelogenous leukemia cell lines. The lines provide models to study the biology of myeloid differentiation and to identify inducers of differentiation of myeloid leukemic blood cells. This review critically examines the inducers of leukemic cell differentiation and their potential therapeutic importance.
Hemolytic anemia, recurrent metabolic acidosis, and incomplete albinism associated with glutathione synthetase deficiency
Authors: Prchal JT , Crist WM , Roper M , Wellner VP
Published: October 2019

The clinical and laboratory features of a 3-mo-old black male infant with glutathione (GSH) synthetase deficiency of the generalized type was evaluated. Partial albinism, brisk hemolytic anemia, recurrent febrile episodes, and mental retardation were noted. Also, severe recurrent metabolic acidosis and marked oxoprolinemia and oxoprolinuria were found in the proband but not in his first-degree relatives. The relationship of these disease manifestations to the underlying metabolic defect is discussed.
A proposal for smoldering adult T-cell leukemia: a clinicopathologic study of five cases
Authors: Yamaguchi K , Nishimura H , Kohrogi H , Jono M , Miyamoto Y , Takatsuki K
Published: October 2019

We have observed five patients with smoldering adult T-cell leukemia (ATL) who had skin lesions as premonitory symptoms. The illness developed slowly, but flared up after several years. Skin lesions appeared in the form of erythema, papules, or nodules. Infiltration of the skin by ATL cells was slight, and the proportion of ATL cells in the peripheral blood was 0%-2%. The serum lactate dehydrogenase (LDH) value was within normal range and was not associated with hypercalcemia; lymphadenopathy, hepatosplenomegaly, and bone marrow infiltration were very slight. In most cases, hypergammaglobulinemia was seen, and in one case, monoclonal hypergammaglobulinemia was observed. All five patients had lived in an area in which ATL was endemic, and their anti-ATLA antibodies were positive; none had ever received a blood transfusion. One patient developed typical ATL after more than 13 yr of illness and died of renal insufficiency. Another patient developed typical ATL after 5 yr of illness and died of cryptococcus meningitis. Based on clinical and pathologic differences, we believe that these cases should be distinguished from typical ATL cases for the purposes of prognosis and treatment.
Additional glycoprotein defects in Bernard-Soulier's syndrome: confirmation of genetic basis by parental analysis
Authors: Berndt MC , Gregory C , Chong BH , Zola H , Castaldi PA
Published: October 2019

The glycoprotein profile of Bernard-Soulier platelets was examined by labeling washed platelets with periodate 3H-sodium borohydride, a procedure that labels greater than 30 glycoproteins on the membrane surface of normal platelets. Three Bernard-Soulier patients were studied; two were siblings and the third was unrelated. The platelet protein and glycoprotein profiles were evaluated under nonreduced and reduced conditions using 5%-15% exponential SDS-polyacrylamide gel electrophoresis. The two siblings completely lacked glycoprotein Ib (GPIb). The unrelated patient had congruent to 7% of the normal level. This was confirmed by two-dimensional nonreduced-reduced SDS- polyacrylamide gel electrophoresis, a procedure that allows clear separation of the disulfide-linked subunits of GPIb, GPIb alpha (mol wt 145,000), and GPIb beta (mol wt 25,000) from other membrane glycoproteins. On one-dimensional analysis, Bernard-Soulier's syndrome (BSS) platelets also lacked the peripheral membrane glycoprotein, GPV (mol wt 82,000) and a low molecular weight glycoprotein, GPIX, (nonreduced or reduced, mol wt congruent to 22,000). The two- dimensional gel system also revealed the absence of a minor glycoprotein with a molecular weight of congruent to 100,000 (GP 100). Quantitation of these proteins solubilized from electrophoretograms showed that the siblings' parents had congruent to 50% levels of GPIb, GPIX, and GP 100. A monoclonal antibody against glycoprotein Ib, FMC 25, was negative by immunofluorescence against Bernard-Soulier platelets and immuneprecipitated both GP Ib and GPIX from Triton X100 solubilized, labeled platelets. The combined results suggest that the apparent genetic absence of multiple proteins in Bernard-Soulier platelets is due, in part, to the presence in normal platelets of a tight membrane complex between glycoprotein Ib and at least one of the other absent glycoproteins.
Studies on mitochondrial and cytoplasmic malate dehydrogenase in childhood myelodysplastic syndrome
Authors: Muchi H , Yamamoto Y
Published: October 2019

Three cases of uncommon childhood hematologic disorders are reported. At presentation, one patient had refractory anemia with an excess of blasts (RAEB) with partial 7-monosomy and was reclassified into RAEB “in transformation” thereafter. Another case was diagnosed as acute myelogenous leukemia with complete 7-monosomy. The other case was diagnosed as RAEB “in transformation” without chromosome aberrations. The cytogenetic studies of the patients with 7-monosomy revealed abnormal karyotypes on bone marrow cells, but normal karyotypes on peripheral blood cells. Polymorphonuclear cells from the two patients with 7-monosomy revealed reduced mitochondrial malate dehydrogenase activity, but those from the patient with RAEB “in transformation” without chromosome aberrations did not. Cytoplasmic malate dehydrogenase activity, having been defined as located on chromosome 2, was within the normal range in those three patients. The decreased mitochondrial enzyme activity in the two patients with 7-monosomy would be a dosage effect of the chromosome aberration, but not caused by their hematologic disorders. The level of mitochondrial enzyme activity in the patients with 7-monosomy was reduced in polymorphonuclear cells, but not in mononuclear cells in peripheral blood. This fact would indicate that such chromosome evolution had involved myeloid cells only, but not lymphoid cells. Both enzymes from leukemic cells of four patients with active disease revealed much higher activities than controls, an expression of partially enhanced oxidative phosphorylation.
Inhibition of human lymphocyte proliferation by monoclonal antibody to transferrin receptor
Authors: Mendelsohn J , Trowbridge I , Castagnola J
Published: October 2019

A monoclonal antibody, 42/6, which blocks the binding of transferrin to its receptor on the cell membrane, inhibits proliferation of human lymphocytes stimulated by phytohemagglutinin. Anti-receptor antibody B3/25, which does not block transferrin binding, does not alter the mitogenic response. Addition of soluble iron, in the form of ferric nitrilotriacetic acid, results in partial reversal of inhibition. Lymphocytes in the quiescent phase of the cell cycle at the time of 42/6 antibody addition are unable to traverse S phase, whereas cells actively proliferating when antibody is added are sensitive to its inhibitory effects throughout all phases of the cell cycle. Inhibition is static rather than cidal, since it can be reversed by removal of antibody after up to 48 hr of exposure.
The effect of X chromosome inactivation on the inhibition of Plasmodium falciparum malaria growth by glucose-6-phosphate-dehydrogenase- deficient red cells
Authors: Roth EF Jr , Raventos Suarez C , Rinaldi A , Nagel RL
Published: October 2019

Previous data on in vitro culture of Plasmodium falciparum malaria demonstrated that red cell glucose-6-phosphate dehydrogenase deficiency (G6PD-) inhibited parasite growth in deficient hemizygous males. This study investigated the effect of heterozygosity for G6PD- on parasite growth. Blood was obtained from 8 female Sardinian G6PD- heterozygotes with G6PD normal cells ranging from 13% to 60%. For comparison, blood from a G6PD- hemizygous male, containing 100% deficient red cells, was mixed in different proportions with compatible normal blood. In both experiments, parasite growth was inhibited by the presence of deficient cells. In both cases, it was found that the inhibition could be explained by a simple dilution of normal cells by G6PD- cells. Thus, the typical female heterozygote is also protected to a significant extent. When considering the “malaria hypothesis” as it relates to G6PD, protection of the female heterozygote as well as the male hemizygote must be taken into account.
Removal of T cells from bone marrow for transplantation: a monoclonal antilymphocyte antibody that fixes human complement
Authors: Hale G , Bright S , Chumbley G , Hoang T , Metcalf D , Munro AJ , Waldmann H
Published: October 2019

Graft-versus-host disease is one of the major problems in clinical bone marrow transplantation. Many experiments in animals have shown that it could be greatly reduced if mature T lymphocytes were removed from the donor marrow. Here we describe a new rat monoclonal antibody, CAMPATH 1, which is suitable for depleting lymphocytes from human marrow grafts. CAMPATH 1 is an IgM that fixes human complement. It binds to both T and B lymphocytes and some monocytes but not to other hemopoietic cells. When peripheral blood mononuclear cells were treated with CAMPATH 1 and complement, more than 99% of lymphocytes were killed and viable T cells could no longer be detected. Under these conditions, in vitro multipotential erythroid and myeloid colony-forming cells were unaffected. As well as being used for in vitro treatment of bone marrow to remove T cells, CAMPATH 1 could potentially be applied to other experimental and clinical situations where depletion of lymphoid cells is required, including serotherapy to achieve immunosuppression for organ transplants or to treat lymphocytic leukemias.
Antithoracic duct lymphocyte globulin therapy of severe aplastic anemia
Authors: Camitta B , O'Reilly RJ , Sensenbrenner L , Rappeport J , Champlin R , Doney K , August C , Hoffmann RG , Kirkpatrick D , Stuart R , Santos G , Parkman R , Gale RP , Storb R , Nathan D
Published: October 2019

We performed a prospective randomized trial of antithoracic duct lymphocyte globulin (ATDLG), HLA-haploidentical marrow, and androgen (regimen ABA) versus androgen alone (concurrent STANDARD care controls) in 42 newly diagnosed individuals with severe aplastic anemia. ABA patients also were matched with patients from our preceding study (historical STANDARD care controls). Supportive care and pretreatment patient characteristics were the same in all groups. By life table analysis, 76% of patients receiving ABA are alive at 2 yr compared to 31% of the concurrent control group (p less than 0.002 versus ABA) and 19% of the historical controls (p less than 0.0001 versus ABA) given STANDARD care. ABA patients had greater hematologic improvement than either control group (p less than 0.001). However, improvement with ABA was often incomplete. Toxicity of ATDLG was considerable but manageable. Further studies to determine the mechanism of action and active component(s) of ABA are indicated.
Iron storage in ferritin following intracellular hemoglobin denaturation in erythroleukemic cells
Authors: Fibach E , Bauminger ER , Konijn AM , Ofer S , Rachmilewitz EA
Published: October 2019

Murine erythroleukemia (MEL) and human K-562 cell lines were cultured in the presence of 57Fe, and the quantities of cellular iron-containing compounds were determined with the aid of Mossbauer spectroscopy. Upon induction of differentiation, both ferritin-iron and hemoglobin (Hb) iron could be detected. Treatment of the cells with 0.01%-0.02% acetylphenylhydrazine (APH) resulted in gradual denaturation of Hb and incorporation of the released Hb-iron into ferritin. Following treatment with APH, the ratio of Hb-57Fe to ferritin-57Fe decreased from 2.6 to 0.2 in MEL cells and from 0.56 to 0.12 in K-562 cells. No change was observed in the total intracellular iron. Using fluorescence ELISA, an increased level of immunologically detectable ferritin was found in hemoglobinized K-562 cells treated with APH, as compared to the amount of ferritin found in untreated cells. Ferritin may thus function not only as an intermediate during Hb synthesis, but also as storage protein for iron released during Hb denaturation.
A family with segregating triplicated alpha globin loci and beta thalassemia
Authors: Galanello R , Ruggeri R , Paglietti E , Addis M , Melis MA , Cao A
Published: October 2019

In this article we report a Sardinian family, in which a beta- thalassemia gene and a triple alpha-globin loci, counterpart of the rightward deletion type alpha-thalassemia-2, were segregating. The analysis of the genotype-phenotype correlations in the different family members allowed us to give an outline of the manifestations associated with different genotype combinations. The heterozygote for the triple alpha-loci showed no consistent abnormal clinical or hematologic characteristics and presented balanced alpha/beta-globin chain synthesis. In the homozygous state for this lesion, the only phenotypic expression was a slightly imbalanced globin chain synthesis. The combination of heterozygous beta-thalassemia with the heterozygous state for the triple alpha-globin loci produced no clinical manifestations and showed a hematologic phenotype indistinguishable from that of heterozygous beta-thalassemia. On the other hand, the combination of the homozygous state for the triple alpha-globin gene loci and the heterozygous state for beta-thalassemia produced a clinical picture of thalassemia intermedia with a very mild clinical course, minor increase of fetal hemoglobin (HbF) levels, and a pronounced imbalance of globin chain synthesis.
Biologic and clinical significance of red cell ferritin
Authors: Cazzola M , Dezza L , Bergamaschi G , Barosi G , Bellotti V , Caldera D , Ciriello MM , Quaglini S , Arosio P , Ascari E
Published: October 2019

Red cell ferritin was measured in normal subjects and patients with disorders of iron metabolism, inflammation, liver dysfunction, impaired hemoglobin synthesis, and increased red cell turnover by means of radioimmunoassays with antibodies to liver (basic) and heart (acidic) ferritins. The normal mean values for basic and acidic ferritin were 8.9 and 22.7 altogram (ag)/cell, respectively. The red cell ferritin content reflected changes occurring in tissues both in iron deficiency and iron overload. Basic ferritin was more closely related to the body iron status than acidic ferritin, and the acidic/basic ferritin ratio was increased in iron deficiency and decreased in iron overload. The major factor determining the red cell ferritin content appeared to be the transferrin saturation, that is, the distribution of iron between monoferric and diferric transferrin. This is in keeping with recent data indicating a competitive advantage of diferric transferrin in delivering iron to erythroid cells. In addition, the red cell ferritin content was increased in thalassemic patients with normal iron status, appearing to be inversely related to the rate of hemoglobin synthesis. The determination of red cell ferritin, based on a commercially available basic ferritin assay, can have clinical application. It can be used for evaluating the adequacy of the iron supply to the erythroid marrow, particularly in patients with increased red cell turnover. Moreover, it may be useful in evaluating the body iron status in patients with hemochromatosis and liver disease.
Pyrimidine nucleotidase deficiency with active dephosphorylation of dTMP: evidence for existence of thymidine nucleotidase in human erythrocytes
Authors: Paglia DE , Valentine WN , Keitt AS , Brockway RA , Nakatani M
Published: October 2019

Erythrocytes from a patient with classical pyrimidine nucleotidase (PyN) deficiency had less than 10% residual PyN activity with uridine 5′-monophosphate (UMP) or cytidine 5′-monophosphate (CMP) as substrate, but exhibited brisk nucleotidase activity with thymidine 5′- monophosphate (dTMP). This strongly suggests the existence of separate enzymes or isozymes of PyN in normal human erythrocytes--an hypothesis that should be tested by similar studies in other cases of severe PyN deficiency, whether induced by genetic defects or lead toxicity.
Monoclonal antibody therapeutic trials in seven patients with T-cell lymphoma
Authors: Miller RA , Oseroff AR , Stratte PT , Levy R
Published: October 2019

We have studied the clinical effects of a murine monoclonal anti-human T-cell antibody in seven patients with T-cell lymphoma. Four to 17 treatments with anti-Leu-1 were given to each patient over periods of 14–75 days. Doses of antibody ranged from 250 micrograms to 100 mg. Antibody treatments usually caused a rapid fall in circulating T cells, with return to baseline levels within 24–48 hr. The optimum dose appeared to vary for each patient. Clearance of circulating tumor cells correlated with the amount of antibody bound to cells. Other than dyspnea in one patient, no serious toxicity was noted. Five patients had definite tumor responses, but these were of short duration (1.5–4 mo). Four patients developed anti-mouse immunoglobulin (Ig) antibodies, and in three patients, this was responsible for tumor escape from therapy. Although 95% of the host anti-mouse Ig response was directed against mouse Ig constant region determinants, a small but significant component was found to be antiidiotype.
Estrogen receptor analysis in chronic lymphocytic leukemia
Authors: Rosen ST , Maciorowski Z , Wittlin F , Epstein AL , Gordon LI , Kies MS , Kucuk O , Kwaan HC , Vriesendorp H , Winter JN
Published: October 2019

Estrogen receptor (ER) determinations were performed on cytosol preparations of Ficoll-Hypaque density separated mononuclear cells from 11 patients with chronic lymphocytic leukemia (CLL). The presence of ER was noted in 8 of 11 specimens (73%). ER ranged from 431 fmole/mg to 4.3 fmole/mg cytosol protein. Two types of receptor subunits were observed at the 8S and 4S region of the sucrose gradient. In addition, 1 of 3 Epstein-Barr virus (EBV) transformed B-lymphoblastoid cell lines from healthy donors had a measurable amount of ER. Patient R.L., who was refractory to standard chemotherapy and radiation and was ER positive, experienced a minor response to Tamoxifen therapy, with subsequent loss of ER. The demonstration of ER in CLL suggests that this malignancy may have a hormone-dependent subpopulation of cells.
The effect of bloodletting on exercise performance in a subject with a high-affinity hemoglobin variant
Authors: Winslow RM , Butler WM , Kark JA , Klein HG , Moo-Penn W
Published: October 2019

We studied two young army recruits with erythrocytosis. One had a variant hemoglobin with high affinity for oxygen (hemoglobin Osler, also known as Fort Gordon and Nancy, beta 145 Tyr leads to Asp). The other had normal oxygen affinity and erythrocytosis of undetermined etiology. Both were asymptomatic. We studied exercise capacity on a cycle ergometer before and after hemodilution. In the subject with high oxygen affinity, hemodilution resulted in reduced maximal work and increased heart rate at every work level. In addition, minute ventilation and arterial lactic acid increased, while anaerobic threshold decreased, indicating diminished oxygen supply to tissues. In contrast, the subject with normal oxygen affinity had no significant changes in exercise performance after hemodilution. These results suggest that when blood oxygen affinity is high, loss of efficiency in tissue oxygenation can be expected after phlebotomy or hemodilution. Therefore, it may be useful to measure blood oxygen affinity and exercise performance in polycythemic subjects in whom such procedures are intended to ameliorate symptoms of hyperviscosity.
Intracellular ferriprotoporphyrin IX is a lytic agent
Authors: Fitch CD , Chevli R , Kanjananggulpan P , Dutta P , Chevli K , Chou AC
Published: October 2019

Human erythrocytes were treated with menadione to oxidatively denature hemoglobin and release ferriprotoporphyrin IX (ferriheme, FP) intracellularly. The high affinity of FP for chloroquine was used to detect its release. After incubation for 1 hr at 37 degrees C and pH 7.4 with 0.5 mM menadione, erythrocytes bound 14C-chloroquine with an apparent dissociation constant of 10(-6)M. Untreated erythrocytes did not bind chloroquine with high affinity. At a chloroquine concentration in the medium of 2 microM, for example, menadione-treated erythrocytes bound 70 mumole chloroquine/kg and untreated erythrocytes bound 13.4 mumole/kg. The intracellular location of FP released by menadione was verified by finding that Tween 80 did not prevent chloroquine binding. By contrast, Tween 80 inhibited the binding of chloroquine to erythrocytes treated with extracellular FP. The hemolytic response to menadione was characteristic of the hemolytic response to FP. Thus, 5 microM chloroquine caused hemolysis to increase to 60% from baseline values of 5% in experiments using erythrocytes treated either with 0.5 mM menadione or with 5 microM FP; and, in both cases, the potentiating effect of chloroquine was inhibited by 1 microM mefloquine or 10 microM quinine. Higher concentrations of menadione caused hemolysis in the absence of chloroquine. We conclude that FP released by menadione exists intracellularly in a form that is accessible to bind chloroquine and to express its lytic activity.
Coincidence of B-cell chronic lymphocytic leukemia and cutaneous T-cell lymphoma (mycosis fungoides): immunologic characterization by monoclonal antibodies
Authors: Sheibani K , Forman SJ , Winberg CD , Rappaport H
Published: October 2019

Although rare cases of chronic lymphocytic leukemia (CLL) of the T-cell type have been reported, CLL is more commonly found to be a neoplastic lymphoproliferative disease of B-cell origin. In this article, we describe a patient with long-standing CLL that was immunologically shown to be of the B-cell type, who, during the course of his disease, developed cutaneous T-cell lymphoma (CTCL), which was shown to be of the helper/inducer subtype. The neoplastic lymphoid cells in the skin infiltrate differed morphologically and immunologically from those in the peripheral blood. The occurrence of CTCL during this patient's clinical course represents a second neoplasm arising from a different cell line, rather than a tissue manifestation of the patient's CLL. To our knowledge, this is the first report in which the occurrence of CTCL is documented in a patient with immunologically known B-cell CLL. In addition to establishing the presence of B-cell CLL and CTCL of the helper/inducer T-cell type in the same patient, this case report demonstrates the usefulness and necessity of evaluating lymphoproliferative disorders by means of a multidisciplinary approach.
Sulfhydryl reagents induce altered spectrin self-association, skeletal instability, and increased thermal sensitivity of red cells
Authors: Smith DK , Palek J
Published: October 2019

Incubation of erythrocytes with the sulfhydryl reagent N-ethyl- maleimide (NEM) results in altered spectrin self-association and formation of dimers on the membrane. Skeletons isolated from these cells exhibit marked skeletal instability. In addition, NEM treatment induces increased thermal sensitivity of both cells and purified spectrin. These effects were not produced in aerobically incubated glucose-6-phosphate dehydrogenase deficient cells and were therefore presumably not due to depletion of intracellular reduced glutathione. These effects were produced by another permeant sulfhydryl reagent, monobromobimane, but not by its membrane-impermeant derivative. We conclude that spectrin sulfhydryl groups play an important role in spectrin self-association and thermal stability.
Carbohydrate antigen profiles of human erythroleukemia cell lines HEL and K562
Authors: Kannagi R , Papayannopoulou T , Nakamoto B , Cochran NA , Yokochi T , Stamatoyannopoulos G , Hakomori S
Published: October 2019

The expression of major carbohydrate antigens carried by polylactosaminyl chains in human erythroleukemia cell lines, K562 and HEL, was investigated by applying monoclonal antibodies recognizing specific carbohydrate determinants. The two cell lines showed common differences in their glycolipid compositions: (1) the presence of significant amounts of ganglio-series glycolipids, which are absent in normal erythrocytes; and (2) a remarkable reduction in the amount of globo-series glycolipids, which are the major glycolipids in normal human erythrocytes. A variety of differences were also detected in the carbohydrate antigens carried by lacto-series glycolipids and glycoproteins having related carbohydrate chains. K562 cells were i+H- X+, with a minor population of I+ cells. HEL cells were I-i+H+X-. The presence of the I+ population in K562 cells is particularly noteworthy, since I-antigen is characteristic of adult mature erythrocytes and is absent in most human leukemic cell lines. Several clones showing I+, I+/-i+/-, or I-i+ specificities were isolated from K562 cells by cloning in either methylcellulose media or limiting dilution, and I+ and I- cells were sorted by FACS fluorometer. HEL cells and these K562 clones provide a useful experimental model for studying the biologic significance and enzymatic control in expression of cell surface polylactosamines.
Antithrombin "Chicago": a functionally abnormal molecule with increased heparin affinity causing familial thrombophilia
Authors: Bauer KA , Ashenhurst JB , Chediak J , Rosenberg RD
Published: October 2019

A family with a high incidence of spontaneous thromboembolism over four generations has been investigated. The propositus is a 21-yr-old male with a history of thrombophlebitis. Medical histories of 46 family members were obtained. Twelve of these individuals have experienced deep venous thromboses and/or pulmonary emboli. Seven members of the kindred, with a prior history of thrombotic phenomena, were investigated in detail. These subjects were found to have normal plasma concentrations of immunoreactive antithrombin (mean 96%), decreased plasma levels of progressive antithrombin activity (mean 50%), and greatly reduced amounts of plasma heparin cofactor activity (mean 42%). The abnormal antithrombin (“Chicago”) was found to elute from heparin- Sepharose at a higher ionic strength than normal inhibitor. The functionally defective antithrombin molecules exhibit a reduced ability to neutralize thrombin in the presence or absence of heparin (approximately 10%-20% of normal). The molecular defect of this protease inhibitor thus appears to be distinct from those of previously described abnormal antithrombins.
Inhibition of human bone marrow fibroblast colony formation by leukemic cells
Authors: Nagao T , Yamauchi K , Komatsuda M , Noguchi K , Shimizu M , Yonekura S , Nozaki H
Published: October 2019

The number of bone marrow fibroblast colony formations decreases in most cases of acute leukemia before the initiation of chemotherapy. This study was undertaken to clarify the mechanism of suppression of fibroblast colony formation in leukemic patients. Titration of the number of bone marrow cells did not indicate a linear relationship between the number of bone marrow cells cultured and the number of fibroblast colony formations. The number of fibroblast colony formations recovered by removal of nonadherent leukemic cells following one day of incubation increased. The cloning efficiency of patient bone marrow still showed increases in colony formation at higher plating concentrations after the nonadherent cells were removed. When leukemic and normal bone marrow cells were cocultured, the suppressive effect of leukemic cells on normal marrow fibroblast colony formation was clearly observed. The suppressive effect disappeared at complete remission, and then reappeared at relapse. Heat-inactivated serum and bone marrow culture media from leukemic patients whose fibroblast colony formations were small in number suppressed fibroblast colony formation from normal bone marrow. From these results, it was concluded that the suppression of fibroblast colony formation in leukemic patients was through humoral factors produced by leukemic cells.
Antigenic heterogeneity of human mononuclear phagocytes: immunohistologic analysis using monoclonal antibodies
Authors: Hancock WW , Zola H , Atkins RC
Published: October 2019

Eight monoclonal antibodies to cell surface antigens of human monocytes were evaluated as immunologic markers for recognition of macrophages in sections of normal and diseased tissues, using immunoperoxidase and enzyme histochemical techniques. Monoclonal antibodies assessed were PHM2, PHM3, FMC17, FMC32, FMC33, FMC34, OKM1, and 63D3. Sites studied were human bone marrow, blood, lymph node, spleen, thymus, liver, kidney, lung, and peritoneal lavages, rejecting renal allografts containing inflammatory macrophages, and granulomata showing epithelioid and multinucleate giant cell formation. All antibodies bound to at least some tissue macrophages and, except for FMC32 and FMC33 antibodies, which were identically distributed, each antibody had a distinctive tissue distribution. Some antigens were shared by other bone-marrow-derived cells (megakaryocytes and cortical thymocytes), endothelium, epithelium, and dendritic cells. Antigenic differences were also detected between mononuclear phagocytes present at different sites, different stages of differentiation, and likely different states of activation. These studies provide evidence of major antigenic differences between various populations of human mononuclear phagocytes. They therefore indicate the need for careful evaluation of experiments involving the recognition of macrophages in tissue sections and smears based solely on the use of antimonocyte monoclonal antibodies.
Effect of cell cycle position on dexamethasone binding by mouse and human lymphoid cell lines: correlation between an increase in dexamethasone binding during S phase and dexamethasone sensitivity
Authors: Distelhorst CW , Benutto BM , Bergamini RA
Published: October 2019

We determined the effect of cell cycle position on the amount of dexamethasone that was specifically bound by mouse and human lymphoid cell lines. Cell lines that were either sensitive or resistant to growth inhibition by dexamethasone were compared. Exponentially growing cells were separated by centrifugal elutriation into fractions that corresponded to different positions in the cell cycle. The cell cycle phase distribution of each fraction was estimated by flow cytometry and autoradiography. The amount of dexamethasone bound per cell in each fraction was measured by a whole cell binding assay. In three dexamethasone-sensitive cell lines (two mouse and one human), we found that the amount of dexamethasone bound per cell increased 2–4-fold between G1 phase and S phase, and then decreased during G2/M phase. Results were the same when the amount of dexamethasone bound per milligram of cell protein was measured. Binding affinity was the same during G1 phase and S phase, but the proportion of bound dexamethasone that translocated to the nucleus was greater during S phase. In contrast, we found that the amount of dexamethasone bound per cell by three dexamethasone-resistant cell lines (two mouse and one human) did not increase during S phase. Our results indicate that cell cycle changes in dexamethasone binding are not simply related to changes in cell protein or cell volume during the cell cycle. An increase in dexamethasone binding during S phase may be required for dexamethasone to inhibit cell growth, and a failure of dexamethasone binding to increase during S phase might represent a new mechanism of dexamethasone resistance in lymphoid cells.
Is activation of the granulocyte by concanavalin-A a reversible process?
Authors: Cohen HJ , Whitin JC , Chovaniec ME , Tape EH , Simons ER
Published: October 2019

The stimulation of granulocyte O2- production by concanavalin-A can be reversed with alpha-methylmannoside. Such cells can be reactivated to generate O2- by adding phorbol myristate acetate or N-formyl-methionyl- leucyl-phenylalanine. Opsonized zymosan, however, is not an effective stimulant to these cells. alpha-Methylmannoside prevents, but does not reverse, depolarization of granulocytes by concanavalin-A. Previously activated cells have a shorter lag time for reactivation by phorbol myristate acetate. Incubation in 2-deoxyglucose of cells previously treated with concanavalin-A and alpha-methylmannoside prevents reactivation. EGTA prevents concanavalin-A-stimulated O2- production only when added prior to the stimulant. EGTA has only a slight effect on reactivation. alpha-Methylmannoside prevents concanavalin-A- stimulated release of lysozyme only when added prior to the stimulant. Prior treatment of cells with concanavalin-A and alpha-methylmannoside inhibits subsequent ingestion of complement-coated particles. We conclude that although the O2--generating system can be reversibly activated with concanavalin-A followed by alpha-methylmannoside, these cells are different from untreated cells. Cells treated in such a way do not respond to all stimuli, remain depolarized, have shortened lag times, no longer require calcium for activation, continue to degranulate, and do not ingest well. Thus, although some changes that accompany the interaction of stimuli with granulocytes are reversible, some are not, and the previously activated cell does not return to a true resting state.
Adhesion of platelets to human artery subendothelium: effect of factor VIII-von Willebrand factor of various multimeric composition
Authors: Sixma JJ , Sakariassen KS , Beeser-Visser NH , Ottenhof-Rovers M , Bolhuis PA
Published: October 2019

The relationship between the multimeric size of factor VIII-von Willebrand factor (FVIII-vWF) and the support of platelet adhesion to subendothelium was studied in an annular perfusion chamber, employing human renal and umbilical arteries. Commercial factor VIII concentrates containing multimers of low molecular weight that had been shown not to correct the bleeding time upon infusion into patients with von Willebrand's disease did not support platelet adhesion in the perfusion chamber. Cryoprecipitate and two experimental FVIII-vWF concentrates containing multimers of high molecular weight supported platelet adhesion. Factor VIII-vWF purified from cryoprecipitate was subdivided into three fractions of different molecular weights (6.0–14.0, 4.0–9.0, and 3.0–7.5 X 10(6) dalton). These fractions appeared to bind equally well and to be equally effective in supporting platelet adhesion. Factor VIII-vWF with multimers of low molecular weight (0.5–1.5 X 10(6) dalton) were prepared by partial reduction. Binding of FVIII-vWF to subendothelium was not impaired, and the support of platelet adhesion appeared to be more resistant to the effect of reduction than the ristocetin cofactor activity. At high shear rate (2,500 sec-1), increased platelet adhesion was observed with partially reduced FVIII- vWF. These data indicate that the ability of FVIII-vWF preparations to correct the bleeding time is reflected in enhanced platelet adhesion to subendothelium in a perfusion chamber. These data also emphasize that multimeric size is not the only factor determining whether FVIII-vWF will support platelet adhesion.
Two distinct categories of warm autoantibody reactivity with age- fractionated red cells
Authors: Branch DR , Shulman IA , Sy Siok Hian AL , Petz LD
Published: October 2019

Using age-fractionated erythrocytes, warm autoantibodies can be classified into two distinct categories, depending on their reactivity with reticulocyte-enriched (younger) or reticulocyte-poor (older) red cell fractions. The strength of the direct antiglobulin test (DAT) on the age-fractionated red cells of 24 patients indicated that 19 (79%) had an IgG warm autoantibody that reacted preferentially with older red blood cells. In 7 of these 19 patients (37%), the DAT was negative using reticulocyte-enriched red cell fractions. We have termed this preferential reactivity of warm autoantibodies with older red cells as type I. Five of the 24 patients studied (21%) had an IgG warm autoantibody that demonstrated no preference for young or older red cells. We have termed this pattern of warm autoantibody reactivity as type II. All 5 patients having type II warm autoantibodies had severe anemia. In contrast, 6 of 19 patients having type I warm autoantibody did not have clinical evidence of anemia when tested, and 11 of the 19 had only slight to moderate anemia. Additionally, our results using type I warm autoantibody raise questions regarding the blood group specificity of warm autoantibodies. The antigen recognized by type I warm autoantibody may be a cryptantigen. Rh specificity or relative Rh specificity, often associated with warm autoantibodies, may simply be a coincidental finding.
Time course of reversibility of accelerated fibrinogen disappearance in diabetes mellitus: association with intravascular volume shifts
Authors: Jones RL , Jovanovic L , Forman S , Peterson CM
Published: October 2019

Accelerated fibrinogen disappearance in diabetic patients is reversible with normalization of blood glucose. To define the time course of this reversal, we measured 125I-fibrinogen disappearance in 19 diabetic patients experiencing acute changes in blood glucose, as monitored and controlled by a microprocessor-controlled closed loop insulin infusion system (artificial beta cell). The data were corrected for blood volume dilutional changes and fit to a model describing two sequential exponential functions and a single exponential function. The sequential model provided the best fit for all but one patient. This indicates that there were two distinct rates of fibrinogen disappearance and suggests that the time course of reversal of accelerated fibrinogen disappearance in diabetic patients is very rapid, if not immediate. Rapid fibrinogen turnover during hyperglycemia was temporally associated with vascular volume changes, reflected as dilutional changes of 51Cr-RBC concentrations. These findings were also associated with an increase in pulse pressure during hyperglycemia, suggesting blood volume expansion due to an osmotic mechanism. The results of this study suggest a picture of vascular volume expansion and contraction, perhaps secondary to the osmotic effects of hyperglycemia. Accelerated fibrinogen turnover associated with these events may be related to increased vascular permeability and/or increased fibrin formation. These events, in concert, may contribute to the initiation and/or propagation of diabetic vascular sequelae.
Increased HbF in sickle cell anemia is determined by a factor linked to the beta S gene from one parent
Authors: Milner PF , Leibfarth JD , Ford J , Barton BP , Grenett HE , Garver FA
Published: October 2019

Members of 7 large families, containing 20 patients with sickle cell anemia (SS) characterized by high levels of fetal hemoglobin (HbF), were studied using immunofluorescence to count F cells and a radioimmunoassay to measure small amounts of HbF. In five of these families, one of the sickle cell trait (AS) parents had a much higher HbF and F-cell count than the other; in one family, both parents had a marked increase in HbF and F cells; in the remaining family, HbF and F cells were at borderline values in both parents. Seven of 14 AS siblings, but only 1 of 8 normal hemoglobin (AA) siblings, also had HbF and F-cell counts above the “normal” range. It seems that a factor for increased F cells, linked to the beta S gene of one parent, is segregating in these families and is responsible for the greatly increased HbF and F cells in the SS subjects. HbF per F cell in AS parents and siblings was the same as that of normal AA subjects, whereas in the SS offspring it was greatly increased, suggesting that it was the result of marrow hyperplasia associated with their hemolytic anemia. The similarity of this “increased F-cell gene” to heterocellular hereditary persistence of fetal hemoglobin (HPFH). Swiss type, is discussed, and it is suggested that it may control the persistent synthesis of HbF in sickle cell anemia by its presence in early infancy.