bluuebunny/crossref_metadata_2025 huggingface dataset:
Antithrombin Pittsburgh: an alpha1-antitrypsin variant causing hemorrhagic disease
Authors: Lewis JH , Iammarino RM , Spero JA , Hasiba U
Published: October 2019

A 10-year-old boy had a severe lifelong hemorrhagic disorder that had necessitated more than 50 hospitalizations. Laboratory examination showed prolonged bleeding, clotting, partial thromboplastin, prothrombin, and thrombin times. These findings were due to a potent inhibitor of the thrombin-fibrinogen reaction. This inhibitor was similar to heparin in that it acted immediately and did not interfere with the coagulant activities of certain venoms. It differed from heparin in not being adsorbed to barium citrate or neutralized by protamine sulfate. The inhibitory effect was found in the alpha1-globulin fraction. It was identified immunologically and functionally as a double-banded alpha1-antitrypsin of a previously unreported phenotype. The inhibitory effects were depressed by trypsin and heterologous anti-alpha1-antitrypsin.
Agnogenic myeloid metaplasia: a clonal proliferation of hematopoietic stem cells with secondary myelofibrosis
Authors: Jacobson RJ , Salo A , Fialkow PJ
Published: October 2019

The glucose-6-phosphate dehydrogenase (G-6-PD) types and chromosomes of hematopoietic and other tissues were determined in a woman with agnogenic myeloid metaplasia. The patient was heterozygous at the X- linked G-6-PD locus so that both B and A isoenzymes were found in nonhematopoietic cells. In contrast, only one G-6-PD type was found in granulocytes, red cells, and platelets. She also had a distinctive chromosome abnormality in blood cells but not in other tissues. These results indicate that agnogenic myeloid metaplasia is a disorder of a pluripotent stem cell and provide strong evidence that it is of clonal origin. In contrast to blood cells, the patient's cultured marrow “fibroblasts” had normal chromosomes and both B and A G-6-PD types, suggesting that the marrow fibrosis is a secondary abnormality. Thus, at least in this case of agnogenic myeloid metaplasia, the hematopoietic cell proliferation appears to be clonal, and, by inference, possibly neoplastic, whereas the marrow fibrosis is probably not clonal, and therefore appears to be secondary.
Interaction between Hb Hasharon and alpha-thalassemia: an approach to the problem of the number of human alpha loci
Authors: Pich P , Saglio G , Camaschella C , David O , Vasino MA , Ricco G , Mazza U
Published: October 2019

We report the case of an Italian infant girl from Polesine (Po delta region in northern Italy) who was heterozygous for Hb Hasharon and alpha-thalassemia, did not synthesize any normal HbA, and had 3% HbH on electrophoresis. Hematologic and biosynthetic studies on Hb Hasharon carriers of the propositus' family suggest the possibility that the Hb Hasharon gene is linked to an alpha-thalassemia gene. On the other hand, in the Askenazy carriers of Hb Hasharon, Hb Harsharon is probably linked to a normal alpha gene. In comparing Hb Hasharon's behavior with that of other alpha variants, particularly HbG Philadelphia, frequent recombinations between alpha structural genes were suggested. The possible identity between the single alpha locus and the alpha2- thalassemia genotype is discussed.
Immunologic abnormalities in an animal model of chronic hypoplastic marrow failure induced by busulfan
Authors: Pugsley CA , Forbes IJ , Morley AA
Published: October 2019

The immunology of chronic hypoplastic marrow failure (CHMF, aplastic anemia) was studied in an experimental murine model of the disease induced by busulfan. B lymphocytes of peripheral blood, spleen, and bone marrow were reduced to 30%–40% and T lymphocytes of thymus, spleen, marrow, and blood were decreased to 20%–70% of control values. IgG and IgM antibody titer to sheep red blood cells were reduced to one- third of control levels, and splenic IgG, but not IgM, plaque-forming cells were fewer on day 7 after antigen stimulation. The proliferative responses to phytohemagglutinin or concanavalin A were reduced in cultures of peripheral blood lymphocytes, splenic lymphocytes, and thymocytes, and cutaneous delayed-type hypersensitivity induced by dinitrofluorobenze was not detected in mice with CHMF. The results demonstrate disturbance of a variety of cellular and humoral functions and suggest that the disturbance was due to quantitative and possibly qualitative abnormalities of the cell types subserving these functions. The results suggest that residual cell injury, the lesion underlying experimental CHMF, is not confined to the myeloid stem cell but also involved cells of the lymphoid series.
Aplastic anemia with fetallike erythropoiesis following androgen therapy
Authors: Rao AN , Brown AK , Rieder RF , Clegg JB , Marsh WL
Published: October 2019

A 43/4-yr-old black girl with acquired aplastic anemia had an increase in total hemoglobin (Hb) from 4.5 to 16.8 g/dl and fetal hemoglobin (HbF) from 0.8 g/dl (18.8%) to 9.6 g/dl (60.2%) following combined androgen-adrenal steroid therapy. Discontinuation of the drugs was followed by a decline in both HbF and total Hb. Reinstitution of the combined steroids prompted a second rise in total and fetal hemoglobin. During these responses the subject's erythrocytes exhibited an increased i antigen score and a low level of red cell carbonic anhydrase. The glycine:alanine ratio at position 136 of the gamma chains of HbF was of the fetal type (proportion of chains with glycine residues, 0.74). Hemoglobin A2 was low (0.4%). The synthesis of alpha and non-alpha chains was balanced. These results indicate that the stimulation of red cell proliferation in this subject, in response to androgen therapy, resulted in the production of cells with several characteristics of “fetal” erythrocytes.
Platelet transfusion therapy. Optimal donor selection with a combination of lymphocytotoxicity and platelet fluorescence tests
Authors: Brand A , van Leeuwen A , Eernisse JG , van Rood JJ
Published: October 2019

Although the value of HLA matching for the selection of platelet donors for patients refractory to random platelets is beyond doubt, even perfectly matched combinations sometimes fail to give a satisfactory transfusion response. With HLA typing and negative lymphocytotoxicity crossmatches, 35% of the platelet transfusions administered to 15 patients gave disappointing results (29 of 82). Additional crossmatching with the newly developed platelet fluorescence test described in this paper reduced the unexpected transfusion failures to 7% (6 of 82). Five of these failures were observed in one patient. The target of the antibodies detected with this platelet fluorescence test is not yet fully specified. It seems probable that both HLA and platelet-specific non-HLA antibodies were detected. No correlation of the results of platelet transfusions with the presence or absence of leukoagglutinating antibodies was found.
Cation permeability alterations during sickling: relationship to cation composition and cellular hydration of irreversibly sickled cells
Authors: Glader BE , Nathan DG
Published: October 2019

Sickle erythrocytes (RBC) incubated under 100% nitrogen for 4 hr manifested marked Na gain with an equivalent K loss. There were no changes in cell total cation or water content under these conditions, and no irreversible sickle cells (ISC) were formed. In contrast, sickle RBC incubated for 24 hr under 100% nitrogen in a glucose-free Na medium containing calcium manifested marked ISC formation. ISC formed under these conditions also had elevated Na content, although K content was much more reduced, and consequently ISC were cation depleted and dehydrated. When sickle RBC were incubated 24 hr under 100% nitrogen in a glucose-free K medium containing calcium no ISC formed and there were no major changes in cation or water content. These studies indicate that the Na+K content and dehydration of ISC was not directly related to the increased cation permeability associated with sickling. Rather, the ISC changes appear to reflect the well-known Gardos effect (K and water loss occurring in ATP-depleted RBC incubated with calcium). In addition, these studies suggest that ISC formation per se is related to K and water loss, since no ISC were formed when ATP-depleted sickle RBC were deoxygenated in calcium-containing high-K media that prevented K loss and dehydration.
Visualization of tubulin in lymphocytes. I. Comparison of normal and chronic lymphocytic leukemia (CLL) lymphocytes
Authors: Dighiero G , Karsenti E , Follezou JY , Bornens M
Published: October 2019

Cell structures containing tubulin were studied in peripheral blood lymphocytes from 8 normal donors and 11 patients with CLL using specific antitubulin antibodies revealed by immunoperoxidase assay. The centriole and microtubules were clearly visible in both groups. A “nucleus-associated tubulin-containing structure” was revealed by antitubulin antibodies and was found in virtually all lymphocytes of normal subjects but in a considerably lower number of CLL lymphocytes. The nature of this structure and its relationship to other cell structures are discussed.
Cytogentics of fibroblastic colonies in Ph1-positive chronic myelogenous leukemia
Authors: Greenberg BR , Wilson FD , Woo L , Jenks HM
Published: October 2019

The cytogenetic status of bone marrow stromal elements obtained from six patients with Ph1-positive chronic myelogenous leukemia (CML), two in blast crisis, was studied in vitro utilizing the potential of marrow to form surface-adherent colonies morphologically compatible with mesenchymal elements. We demonstrated the absence of both the marker chromosome and other chromosomal abnormalities in all the fibroblastic colonies studied, indicating that the progenitors of such colonies (plaque-forming units in culture, PFU-C) are not closely related to hematopoietic elements including macrophages. This supports previous reports suggesting that the stromal elements in myelofibrosis associated with CML are not derived from the primary Ph1-positive malignant clone but represent a stromal reactive component of benign or independent malignant potential.
Acquired IgG antibody occurring in a thrombasthenic patient: its effect on human platelet function
Authors: Levy-Toledano S , Tobelem G , Legrand C , Bredoux R , Degos L , Nurden A , Caen JP
Published: October 2019

In subagglutinating amounts, an IgG antibody isolated from the plasma of a polytransfused thrombasthenic patient (L) inhibited ADP-, epinephrine-, collagen-, and thrombin-induced aggregation of normal human platelets. The inhibition of ADP-induced aggregation was strongly diminished following the prior incubation of the antibody with control human platelet stroma but not with the stroma prepared from the platelets of two different thrombasthenic patients. The IgG(L) did not affect the binding of 14C-ADP to control human platelet membranes and did not inhibit the ADP-induced shape change. Bovine factor VIIIVWF- induced agglutination and ristocetin-induced aggregation of control human platelets were not inhibited in the presence of the antibody. The IgG(L) strongly inhibited ADP-induced retraction of reptilase clot and thrombin-induced clot retraction. This antibody therefore induced a thrombasthenialike state in normal human platelets, suggesting that the antigenic site recognized by the antibody plays a central role in the later stages of the mechanism of platelet aggregation induced by physiologic aggregation-inducing agents.
Abnormalities of iron metabolism and erythropoiesis in vitamin E- deficient rabbits
Authors: Chou AC , Broun GO Jr , Fitch CD
Published: October 2019

Rabbits fed a vitamin E-deficient diet developed severe muscular dystrophy in 3–4 wk, but they did not become anemic. Nevertheless, reticulocyte counts increased in deficient rabbits (3.2%) compared to control rabbits (0.9%), and erythroid hyperplasia was evident in the bone marrow. Comparing deficient rabbits to controls, the plasma iron concentration was lower (134.4 versus 206.6 microgram/dl); the TIBC was higher (335.9 versus 228.3 microgram/dl); the whole blood protoporphyrin concentration was higher (131.6 versus 81.7 microgram/dl); and the total iron content was lower in spleen (71 versus 153 microgram), higher in skeletal muscle (4956 versus 3054 microgram), and unchanged in bone marrow, liver, and heart. Studies of iron absorption and excretion using 59Fe showed no abnormalities in deficient rabbits. There were abnormalities of ferrokinetics, however. The half-time of disappearance of 59Fe was shorter (100.6 versus 169.4 min), the plasma iron turnover was greater (1.25 versus 0.95 mg/dl blood/day), and the reappearance of 59Fe in circulating erythrocytes at day 9 was greater (77.2% versus 57.2%) in deficient rabbits. Anemia induced by phlebotomy accentuated the abnormal iron metabolism of deficient rabbits, and the animals were unable to correct the anemia. These findings show that vitamin E deficiency in rabbits causes abnormal erythropoiesis associated with abnormal iron metabolism and sequestration of iron in skeletal muscle.
Development of a practical oral dexamethasone premedication schedule leading to improved granulocyte yields with the continuous-flow centrifugal blood cell separator
Authors: Winton EF , Vogler WR
Published: October 2019

Studies were conducted to improve the yield of granulocytes collected for transfusion from normal donors by means of the continuous-flow centrifugal blood cell separator. Nonleukapheresed donors were medicated with varying schedules of corticosteroids to learn the magnitude and duration of granulocytosis. Normal donors were medicated with varying schedules of corticosteroids prior to a 4-hr leukapheresis and the granulocyte yields determined. It was found that maximum yields (32.2 x 10(9) granulocytes) were obtained by use of dexamethasone given orally 12 and 3 hr prior to leukapheresis. There was a good correlation between the yields and the circulating granulocyte count at the start and during the procedure.
T cell chronic lymphocytic leukemia: presence in bone marrow and peripheral blood of cells that suppress erythropoiesis in vitro
Authors: Hoffman R , Kopel S , Hsu SD , Dainiak N , Zanjani ED
Published: October 2019

The pathogenesis of the anemia associated with malignancy was investigated in a patient with T cell chronic lymphocytic leukemia. The plasma clot culture system was used as a measure in vitro of erythropoiesis. The patient's peripheral blood and marrow T lymphocytes obtained both before and after transfusion therapy suppressed erythroid colony formation by normal human bone marrow cells. Pretreatment of the patient's bone marrow T cells by antithymocyte globulin (ATG) and complement reversed this suppression. In addition, pretreatment of the patient's marrow cells with ATG and complement markedly augmented erythropoiesis in vitro. The expression of erythroid activity caused by the selective destruction of the suppressor T lymphocytes in the patient's bone marrow with ATG and the suppression of normal erythropoiesis by the patient's bone marrow and peripheral blood lymphocytes suggest that interaction between the malignant T cell and the erythropoietin-responsive stem cell is important in production of anemia in this patient.
Successful engraftment of cryopreserved autologous bone marrow in patients with malignant lymphoma
Authors: Appelbaum FR , Herzig GP , Ziegler JL , Graw RG , Levine AS , Deisseroth AB
Published: October 2019

In an effort to evaluate the possible utility of cryopreserved autologous bone marrow infusions in man, 22 patients with malignant lymphoma resistant to conventional chemotherapy were treated with high- dose chemotherapy. This was followed in 12 patients by an infusion of their cryopreserved autologous bone marrow; 10 patients received chemotherapy alone and serve as controls. Following chemotherapy, severe leukopenia (less than 100 leukocytes/mm3) lasted 6–10 (median 8) days in patients receiving cryopreserved marrow, compared to 10–29 (median 16) days in controls (p less than 0.001). Recovery to 1000 leukocytes/mm3 occurred 10–18 (median 13) days after chemotherapy in autograft recipients but was delayed until 12–38 (median 23) days after chemotherapy in controls (p less than 0.001). Autografted patients also recovered granulocyte and platelet function significantly faster and had significantly fewer febrile days after chemotherapy than did controls. Cryopreserved autologous bone marrow infusions can hasten hemopoietic recovery in man after high-dose chemotherapy; this earlier reconstitution may be of clinical benefit to the patient.
Ultrastructural study of acute lymphocytic leukemia: comparison with immunologic studies
Authors: Glick AD , Vestal BK , Flexner JM , Collins RD
Published: October 2019

Leukemic cells from 29 cases of acute lymphocytic leukemia (ALL), studied for T and B cell markers by the use of sheep erythrocyte rosetting and surface immunoglobulin determinations, were examined by electron microscopy. The majority of patients (76%) were found to have non-T, non-B neoplasms composed predominantly of relatively small, inactive-appearing cells with frequent nuclear folds. T cell cases (21%) were associated with mediastinal masses and were predominantly composed of large, active-appearing cells with nuclear irregularity and little rough endoplasmic reticulum. One case of B cell origin was not morphologically distinct from the non-T, non-B cell cases.
Hereditary abnormality of platelet aggregation attributable to nucleotide storage pool deficiency
Authors: Ingerman CM , Smith JB , Shapiro S , Sedar A , Silver MJ
Published: October 2019

An abnormality of platelet aggregation has been detected in six family members with mild bleeding tendencies. In citrated platelet-rich plasma, primary aggregation induced by ADP or epinephrine and agglutination in response to ristocetin were present but second wave aggregation and aggregation in response to collagen suspension were absent or greatly reduced. Sodium arachidonate-induced aggregation was normal although aggregation in response to prostaglandin G2 was reduced and depended entirely on the presence of plasma or ADP. Further tests indicated that the platelets produced prostaglandins but did not release ATP in response to thrombin or sodium arachidonate. Platelets from the patients were found to contain reduced amounts of ADP and 5- hydroxytryptamine and to be unable to retain radioactivity during prolonged incubation at 37 degree C with radiolabeled 5- hydroxytryptamine. Although electron microscopy revealed an absence of very dense bodies, the platelets appeared otherwise normal. The findings are discussed in relation to previous studies of nucleotide storage pool deficiency and the light they shed on platelet physiology in general.
Efficacy of granulocyte transfusions in the control of systemic candidiasis in the leukopenic host
Authors: Ruthe RC , Andersen BR , Cunningham BL , Epstein RB
Published: October 2019

An experimental canine model was designed to evaluate the effect of granulocyte transfusions on systemic infection with Candida albicans in the granulocytopenic host. Each of a pair of dogs was rendered granulocytopenic with a single intravenous (i.v.) dose of cyclophosphamide (50 mg/kg body weight) and challenged with 10(6) Candida albicans organisms administered i.v. when granulocyte counts were less than or equal to 500/mm3. Granulocytes procured by leukofiltration were infused into six experimental dogs 1, 24, 48, and 72 hr after challenge with Candida. An average of 13 +/- 1.3 X 10(9) granulocytes were administered per infusion, producing an average 1-hr increment of 588 +/- 146 granulocytes/mm3 over the pretransfusion granulocyte count. Experimental and control dogs were killed 96 hr after challenge and organs examined grossly and by quantitative culture techniques to measure the extent of infection. All animals receiving granulocyte transfusions had significantly less tissue infection than nontransfused controls (p less than 0.05). It was concluded that granulocyte transfusions are effective in reducing the severity of infection by Candida albicans during periods of leukopenia.
Safety and efficacy of jet anesthesia for bone marrow aspirations
Authors: Dyment PG , Doering EJ , McHugh MJ
Published: October 2019

Aspirations or Jamshidi needle biopsies (n = 287) of bone marrow were performed on children and adolescents with acute leukemia or other malignant disease following the use of a spring-loaded instrument that delivered local anesthetic in a jet spray; 89% of the patients were receiving chemotherapy, 12% were thrombocytopenic, and 23% of the 269 patients who were afebrile at the time of the procedure were severely neutropenic. None of these patients had an infection or a hemorrhage as a complication of the procedure. We conclude that not only is this procedure safe, but it is also much less painful than the traditional method of local anesthetic infiltration using a syringe and needle.
Heterogeneity of human whole blood platelet subpopulations. II. Use of a subhuman primate model to analyze the relationship between density and platelet age
Authors: Corash L , Shafer B , Perlow M
Published: October 2019

A subhuman primate model was developed to ascertain whether or not platelet heterogeneity could be explained by aging in the peripheral circulation. Density-dependent platelet cohorts, postulated to represent cells of different ages, were isolated on isosmolar arabinogalactan gradients and labeled with radiochromium. Mean platelet lifespan was measured for the different density cohorts, and simultaneous sequential density distribution analysis was performed to follow changes in cell density during aging. The average mean lifespan of light platelets was 74.6 hr, compared to 313.6 hr for heavy platelets. After injection, labeled light platelets were recovered only in the gradient light region, in contrast to labeled heavy platelets, which were initially restricted to the dense region and progressively migrated to the light region during their lifespan. This study supports the hypothesis that platelet age in unstressed primates correlates with cell density and provides a rationale for the use of “age-dependent” markers to estimate platelet turnover rates.
Fetal hemoglobin in paroxysmal nocturnal hemoglobinuria (PNH): evidence for derivation of HbF-containing erythrocytes (F cells) from the PNH clone as well as from normal hemopoietic stem cell lines
Authors: Papayannopoulou T , Rosse W , Stamatoyannopoulos G
Published: October 2019

The cellular distribution of HbF was studied in nine patients with paroxysmal nocturnal hemoglobinuria (PNH) by measuring the level of HbF and determining the number of HbF-containing red cells (F cells) in whole blood and in the population of normal cells obtained after immune lysis of the abnormal erythrocytes. The amounts of HbF and the F cell frequencies found in the normal red cells were strikingly similar to the values seen in whole blood. The observed frequencies of F cells in normal cells best fitted those expected under the assumption that the F cells arise equally from normal hemopoietic stem cells and from the stem cells with the PNH defect. Since PNH appears to be a clonal hemopoietic stem cell disorder, this evidence argues against a derivation of F cells from distinct pluripotent stem cell lines.
Primary systemic amyloidosis: comparison of melphalan and prednisone versus placebo
Authors: Kyle RA , Greipp PR
Published: October 2019

Satisfactory treatment for primary amyloidosis does not exist. Because the amyloid fibrils consist of a portion of a monoclonal light chain, it appears reasonable to treat amyloidosis with alkylating agents that are effective against the plasma cells that synthesize monoclonal light chains. Fifty-five patients with primary systemic amyloidosis were randomized (double blind) to melphalan-prednisone or placebo. In comparison with the placebo group, patients given melphalan-prednisone were able to continue on treatment for a longer time and to receive larger doses before the code was broken. Among this group, the nephrotic syndrome disappeared in two patients and urinary excretion of protein was reduced by more than 50% in eight others. Of 13 patients who received melphalan-prednisone for more than 12 mo, 6 improved, 3 were stable, and 4 had progression of disease. Survival did not differ significantly between the groups.
Sex differences in the antithrombotic effects of aspirin
Authors: Kelton JG , Hirsh J , Carter CJ , Buchanan MR
Published: October 2019

Aspirin inhibits platelet function by acetylating platelet cyclooxygenase. Recent clinical trials indicate that aspirin is a promising antithrombotic agent against both venous and arterial thrombosis, but somewhat surprisingly this protective effect appears to be limited to males. To examine the potential sex-related differences in response to aspirin, we developed an animal model for quantitating fibrin accretion into an injury-induced thrombus and used it to study the effects of aspirin on thrombus size in male and female rabbits. Platelet prostaglandin synthesis was estimated by assay of platelet malondialdehyde and was significantly decreased in both male and female rabbits following treatment with 10 mg/kg aspirin (p less than 0.001). This inhibitory effect was not different for platelets from male and female rabbits. Thrombus size was significantly decreased in aspirin- treated male rabbits when compared to controls (p less than 0.05), but this aspirin effect was not apparent in female rabbits or rabbits of either sex treated with 10 mg/kg sodium salicylate. These findings support the results of clinical trials that were obtained by retrospective subgroup analysis. The reason for the sex difference is not known, but the findings raise an important issue in relationship to this mechanism of the antithrombotic effect of aspirin.
Linkage and gene localization of hereditary spherocytosis (HS)
Authors: Kimberling WJ , Taylor RA , Chapman RG , Lubs HA
Published: October 2019

Fifteen kindreds with dominant hereditary spherocytosis (HS) were studied. Expansion of the data from a family with an 8/12 translocation provided further evidence that at least one locus for HS is located near the breakpoint of the translocation. Linkage analysis of all families showed a lack of linkage with all marker loci studied except for Gm (IgG). Linkage between Gm and HS was shown to be significant with a maximum lod score of 3.42 at a recombination fraction of 22%. No heterogeneity of the recombination fraction was observed either between sexes or between families. These results are compatible with the hypothesis that HS is not a heterogeneous disorder.
Effect of C3b inactivator on monocyte-bound C3-coated human erythrocytes
Authors: Schreiber AD , McDermott PB
Published: October 2019

As a model of IgM-induced hemolytic anemia in man, human erythrocytes were sensitized with IgM antibody and coated with complement components, including C3 and C4, using human serum as a source of complement. These coated red cells were then interacted with monolayers of human mononuclear phagocytic cells (monocytes). Complement-coated red cells so bound could be displaced from their monocyte attachment site in a dose- and time-dependent manner by serum factors, including C3b inactivator (C3bINA). These factors were more efficient in inactivating red cell-bound complement components prior to interaction of the coated cells with monocytes. With large amounts of complement per erythrocyte, measured as membrane-bound C3, the ability of the serum inactivating factor(s) to remove the complement-coated red cells from the monocyte surface was compromised and persistently bound red cells were progressively phagocytosed. These studies implicate C3bINA in the displacement of complement-coated erythrocytes, formed from the interaction of IgM antibody and serum complement, from the hepatic macrophage in IgM-induced immune hemolysis. They suggest that both the concentration of complement components, especially on the erythrocyte surface, and the level of C3bINA and perhaps other inactivators may be important features regulating hemolysis in this disorder.
Expression and specificity of FcIgG receptor sites on neoplastic lymphocytes
Authors: Huber C , Flad HD , Nilsson K , Wigzell H , Michlmayr G , Frischauf H , Braunsteiner H
Published: October 2019

We investigated the ability of an FcIgG receptor marker to discriminate between subtypes of malignant lymphoproliferative diseases that differed in their clinical presentations. A quantitative radioimmuno assay was established that enabled us to evaluate average receptor densities on a population basis. Surface receptors were first saturated with IgG complexes. The number of membrane associated IgG molecules was subsequently determined with 125I-staphylococcal protein A. Results obtained with this assay on a battery of malignant lymphocytes suggested that the range of receptor densities of malignant B and T cells might overlap each other but would correlate with the degree of tumor cell differentiation and the clinical stage of the underlying disease. This behavior limits the use of this marker in the characterization of the derivation of malignant lymphocytes; these findings, however, may be useful in the prognostic classification of lymphomas of known origin.
Local development of mast cells from bone marrow-derived precursors in the skin of mice
Authors: Hatanaka K , Kitamura Y , Nishimune Y
Published: October 2019

A mechanism to control development of mast cells was investigated in mice. Although mast cells in the skin of normal C57BL/6 mice were still of host type 290 days after irradiation and injection of bone marrow cells from beige (Chediak-Higashi syndrome, C57BL/6 bgJ/bgJ) mice, donor-type mast cells with giant granules appeared after painting of methylcholanthrene on the dorsal skin. Since donor-type mast cells appeared only at the painted portion of the skin, with an increase in the labeling index of such donor-type mast cells with 3H-thymidine, proliferation and differentiation of bone marrow-derived precursors of mast cells seem to be controlled locally. Although the morphologic feature of marrow-derived precursors was not identified, the finding that all fibroblasts cultured from the methylcholanthrene-treated skin were of host type may exclude the possibility that fibroblasts are the precursors of mast cells.
Fetal hemoglobin (HbF) synthesis in baboons, Papio cynocephalus. Analysis of fetal and adult hemoglobin synthesis during fetal development
Authors: De Simone J , Mueller AL
Published: October 2019

Fetal hemoglobin (HbF) and adult hemoglobin (HbA) synthesis was studied in fetal baboons, Papio cynocephalus, to determine the normal pattern of hemoglobin production during fetal development. Fetuses ranging from 53 to 180 days gestation (term gestation 184 days) were used. Erythroid cells were incubated with 3H-L-leucine, and the rates of globin chain synthesis and the distribution of radioactivity into hemoglobin intermediates and completed hemoglobin molecules were determined. Gamma chain synthesis accounted for approximately 97% of the total nonalpha chain synthesis up to 140 days gestation; beta chain synthesis accounted for the remainder. After 140 days gestation, approximately equal quantities of gamma and beta chain were synthesized in the bone marrow. Prior to 140 days gestation, total alpha chain synthesis was 30% greater than total non-alpha chain synthesis, while there was balanced chain synthesis after 140 days gestation. During the period of excess alpha chain synthesis, fetal erythrocytes contained a large pool of alpha-hemoglobin (alpha chain with heme attached) molecules uncombined with beta or gamma chains. In view of the possibility that alpha chains may have a lower affinity for gamma chains than beta chains, excess alpha chain synthesis may be required to maintain low levels of free gamma chains.
Acquired coagulation inhibitor delaying fibrinopeptide release
Authors: Marciniak E , Greenwood MF
Published: October 2019

A 14-yr-old girl Down syndrome developed a unique type of circulating inhibitor causing a mild bleeding tendency and interfering strongly with coagulation tests, including reptilase time, and with the reaction of purified fibrinogen and thrombin. The concentration of all coagulation factors was normal. The inhibitor had no direct effect on thrombin activity or on the aggregation of fibrin monomer in plasma. Chromatography on DEAE-cellulose and neutralization by immune sera revealed that the inhibitor was an immunoglobulin of IgG class with both kappa and lambda determinants. Isolated inhibitor delayed the release of fibrinopeptide A from a normal fibrinogen reacting with thrombin and retarded the onset of visible clotting, but had no effect on the the final degree of clottability. The clinical and laboratory features of this patient resemble those of patients with congenital dysfibrinogenemia associated with abnormal fibrinopeptide release.
Normal iron absorption and decreased red cell iron uptake in the aged
Authors: Marx JJ
Published: October 2019

Absorption of iron was studied with a double-isotope technique that allowed differentiation between “mucosal uptake,” “mucosal transfer,” and ultimate “retention” of iron. A physiologic dose of ferrous sulfate was administered to 25 healthy young adults, 40 active aged persons, and 20 patients with uncomplicated iron deficiency. Radioactivity was measured with a whole-body scanner. Iron absorption values were not decreased in aged subjects compared to young adults. Mucosal uptake, mucosal transfer, and retention of iron were equally increased in both young and old patients with iron deficiency. In 12 young adults and 33 aged persons red cell iron uptake was studied in addition to iron absorption. Young adults utilized 91% of the retained, orally administered iron and the aged only 66%. An increase in ineffective erythropoiesis in old age is suggested.
Quantitative anisocytosis as a discriminant between iron deficiency and thalassemia minor
Authors: Bessman JD , Feinstein DI
Published: October 2019

The coefficient of variation (CV) of red cell size, as measured by electronic red cell sizing (erythrography), was less than 14.0% in 20 normal subjects. In 22 of 25 patients with beta-thalassemia minor and microcytosis (mean corpuscular volume [MCV] less than 70 fl), CV was less than 14.0%; in the other 3, CV was 14.0%--14.9%. In 53 patients with iron deficiency anemia and MCV less than 70 fl, CV always was greater than 14.0%. In 7 patients with alpha-thalassemia minor and MCV less than 70 fl, CV was less than 14.0% in all 7. Among patients with microcytosis, erythrography appears to be an excellent technique for rapidly distinguishing between iron deficiency and alpha or beta thalassemia minor.
Presence of a platelet aggregating factor in the plasma of patients with thrombotic thrombocytopenic purpura (TTP) and its inhibition by normal plasma
Authors: Lian EC , Harkness DR , Byrnes JJ , Wallach H , Nunez R
Published: October 2019

Three patients with thrombotic thrombocytopenic purpura (TTP) were treated by infusion of normal plasma with dramatic responses. The plasmas collected from these patients during relapse induced in vitro aggregation of washed platelets from both normal donors and the patients during remission. The platelet aggregating factor was not dialyzable or adsorbable by Al(OH)3 and was not inactivated by diisopropylfluorophosphate, hirudin, or heparin in the presence of normal amounts of antithrombin. In contrast to the platelet aggregation induced by platelet isoantibody, the platelet aggregating activity of TTP plasma diminished as a function of time when it was incubated with normal plasma at 37 degrees C. These observations suggest that at least some instances of TTP appear to be due to deficiency of a plasma inhibitor to counteract a platelet aggregating factor demonstrated to be present in the plasma of these patients.
Protective effect of prophylactic penicillin on splenectomized mice exposed to an aerosolized suspension of type III Streptococcus pneumoniae
Authors: Dickerman JD , Bolton E , Coil JA , Chalmer BJ , Jakab GJ
Published: October 2019

Prophylactic penicillin has been recommended for use in asplenic patients and postsplenectomy patients. A laboratory model using aerosolized pneumococci has been devised to test the effectiveness of prophylactic penicillin in a manner analogous to human experience. There is increased mortality, over time, in asplenic mice exposed to aerosolized type III Streptococcus pneumoniae. One hundred twenty-one male Swiss mice (mean weight 26 g) were divided into four groups: splenectomized, sham-operated, splenectomized + penicillin, and sham- operated + penicillin. After 2 wk the four groups were exposed for 30 min to an aerosolized atmosphere of 2.4 x 10(9) colony-forming units of type III S. pneumoniae using a Tri-R model A-42 airborne infection apparatus. Penicillin was given at a daily intramuscular dosage of 40,000 units procaine penicillin G beginning 2 days prior to exposure and continuing through the third day after exposure. The splenectomized and sham-operated mice given penicillin showed significantly lower mortality (p less than 0.001) than mice not given penicillin.
Immune thrombocytopenia in lymphoproliferative diseases
Authors: Kaden BR , Rosse WF , Hauch TW
Published: October 2019

We have studied the thrombocytopenia of lymphoproliferative disorders using a measurement of membrane-bound IgG by an antiglobulin consumption assay. Nine patients with chronic lymphocytic leukemia (CLL) and thrombocytopenia had increased membrane-bound IgG. Two patients with non-Hodgkins lymphoma and 1 patient with Hodgkins disease also had thrombocytopenia and increased membrane-bound IgG. Five of the patients with CLL had positive direct antiglobulin (Coombs) tests on red cells; of these, 3 patients had hemolytic anemia. In eight of the 9 patients with CLL, thrombocytopenia, and increased platelet-bound-IgG, the platelet count increased with the administration of prednisone or an alkylating agent, with splenectomy, or with a combination of these.
Correlation of total body potassium and leukemic cell mass in patients with chronic lymphocytic leukemia
Authors: Chandra P , Sawitsky A , Chanana AD , Chikkappa G , Cohn SH , Rai KR , Cronkite EP
Published: October 2019

Total body leukemic mass in patients with chronic lymphocytic leukemia (CLL) was measured by quantitation of total body potassium (TBK) with a whole-body counter. In addition, the predicted normal total body potassium (Kp) for each patient was calculated from an empirically derived relationship involving height, weight, age, and sex. Both the absolute TBK and the relative excess of total body potassium (TBK/Kp) were related to the stage of disease. Patients in the early stages of CLL were found to have lower TBK and TBK Kp than patients in the late stages of disease. Both of these parameters increased with the successively advanced stages of the disease. The clinically monitored reduction of leukemic cell mass following therapy was accompanied by reductions in TBK and TBK/Kp. Data presented support the notion that TBK/Kp is a useful indicator of the total body leukemic mass. Furthermore, the results of these studies quantitatively validate the proposed clinical staging system for CLL. Quantitation of TBK by a whole-body counter is an accurate and noninvasive procedure and does not require administration of isotopes.
Application of flow cytometry and cell sorting to megakaryocytopoiesis
Authors: Nakeff A , Valeriote F , Gray JW , Grabske RJ
Published: October 2019

We have employed flow cytometry (FCM) and cell sorting to quantitate and study megakaryocytes in mouse and rat femoral marrow following their 20- to 30-fold concentration by centrifugal elutriation (CE). This enrichment of megakaryocytes permitted the first determination of their DNA-related fluorescence by FCM analysis following DNA staining. Fluorescence distributions of CE-enriched cell fractions following supravital staining with Hoechst 33342 were similar to those following chromomycin A3 staining of ethanol-fixed cells. Microscopic examination of cells sorted onto glass slides on the basis of their DNA-related fluorescence following supravital staining together with specific acetylcholinesterase staining for megakaryocytes indicated that megakaryocytes generally increased in cell size with increasing DNA content. This technologic application represents a significant advance in the study of megakaryocytopoiesis, since the kinetics of either the normal or perturbed population can now be studied rapidly and quantitatively.
An assay for intrinsic factor based on blocking of the R binder of gastric juice by cobinamide
Authors: Begley JA , Trachtenberg A
Published: October 2019

An in vitro assay for measurement of gastric juice intrinsic factor (IF) was developed based on the ability of the cobinamide (Cbi) [(CN, OH) Cbi] to bind to the gastric juice R-type binders of cobalamin (Cbl) and not to the IF binder. Subsequently added radioactive Cbl, CN-[57Co] Cbl, binds only to the IF binders and allows for direct measurement of this Cbl binding protein. This Cbi blocking assay was found to function as well as the more conventional methods of IF measurement, G-100 column chromatography, and IF blocking antibody assay. The present assay has the advantage of eliminating the need for elaborate forms of protein separation and does not rely on a source of antibody.
Comparison of sizes and shapes of tumor cells in plasma cell leukemia and plasma cell myeloma
Authors: Isobe T , Ikeda Y , Ohta H
Published: October 2019

Thirteen cases of plasma cell leukemia and 13 cases of nonleukemia plasma cell myeloma were compared with respect to the sizes and shapes of tumor cells in marrow smears. It was demonstrated that the leukemia cells were elongated or ovoid and significantly smaller in size, whereas the myeloma cells were generally more nearly round and larger.
Arsenic-induced bone marrow toxicity: ultrastructural and electron- probe analysis
Authors: Feussner JR , Shelburne JD , Bredehoeft S , Cohen HJ
Published: October 2019

A patient with severe arsenic poisoning that resulted in marked peripheral blood and bone marrow abnormalities, including megaloblastic erythropoiesis experienced many of the previously reported hematologic complications of arsenic poisoning: leukopenia, granulocytopenia, absolute eosinophilia, and profound anemia. In this study we report an ultrastructural and electron-probe analysis of the bone marrow. Although megaloblastic anemia associated with arsenic poisoning has been described rarely, the presence of arsenic in the local bone marrow milieu has not been demonstrated previously. The ultrastructural features of arsenic-induced bone marrow toxicity are similar to those described in other dyserythropoietic states and include marked nuclear aberrations involving shape, chromatin distribution, and nuclear envelope. Using the technique of energy-dispersive x-ray analysis (electron probe) we demonstrated arsenic in bone marrow spicules; this supports the contention that arsenic can cause megaloblastic anemia. We suggest that this technique may be a useful tool in further studies that attempt to explore the mechanism of arsenic-induced hematologic toxicity. Finally, we suggest that arsenic has a direct toxic effect on DNA synthesis that results in marked disturbances of nuclear division. We recommend that the most appropriate screening procedure to evaluate possible arsenic poisoning is tissue arsenic measurements (hair and nails) rather than 24-hr urinary measurements.
Glucocorticoid effects on peripheral blood lymphocytes in cows infected with bovine leukemia virus
Authors: Bloom JC , Kenyon SJ , Gabuzda TG
Published: October 2019

We examined the effects of glucocorticoids on peripheral blood lymphocytes (PBL) in lymphoproliferative conditions associated with bovine leukemia virus (BLV): persistent lymphocytosis (PL) and lymphosarcoma cell leukemia (BLSL). The effects of hydrocortisone 21- sodium succinate (HSS) on spontaneous incorporation (SI) and mitogen- stimulated incorporation of radiolabeled-thymidine and the effects of intramuscular administration of prednisolone acetate were studied. An expanded population of B lymphocytes in cows with PL was remarkable sensitive to glucocorticoids in vitro and in vivo. SI was markedly inhibited by concentrations of HSS as low as 10(-7) M. These results correlated well with in vivo observations, where an 80%-90% decrease in PBL occurred during the course of glucocorticoid administration. The decrease in total lymphocytes was accounted for almost entirely by a decrease in the expanded B lymphocyte population. Steroid-sensitive lymphocytes together with steroid-resistant cells were observed in cows with BLSL. The reduction in the steroid-sensitive lymphocytes was associated with rapid disease progression in cows with lymphosarcoma. Steroid-sensitive lymphocyte populations in cows with BLSL may include the same reactive B-cell population found in cows with PL. Glucocorticoids may prove to be a useful tool for study of the immune response to the oncogenic virus and lymphoma in BLV-infected cattle.
Comparison of red cell creatine level and reticulocyte count in appraising the severity of hemolytic processes
Authors: Fehr J , Knob M
Published: October 2019

In seeking a sensitive indicator for quantitative assessment of hemolytic disease, we found a close dependence of red cell creatine level on cell age. Studies in 21 patients with steady-state hemolysis showed high correlation (r=0.89, p less than 0.001) between reticulocyte counts and red cell creatine levels. Excluding elevation of the creatine level as a variable epiphenomenon of increased erythropoietic activity, density separation of normal red cells revealed distinctly higher creatine levels in younger cells. The reticulocyte counts and creatine levels as quantitative predictors of hemolytic processes were compared: in severe hemolytic anemias (T50Cr less than 11 days), erythrocyte survival (T50Cr) correlated well with creatine levels (r = -0.86, p less than 0.01) and, to a lesser degree, with reticulocyte counts (r = -0.72, p less than 0.05). In milder disease (T50Cr greater than 11 days), however, no correlation existed between reticulocyte counts and T50Cr, whereas the creating levels correlated closely with T50Cr (r = -0.84, p less than 0.001). Thus, on the basis of our regression equations, useful estimation of red cell survival may be obtained from single measurements of erythrocyte creatine.
Distribution of iron between the binding sites of transferrin in serum: methods and results in normal human subjects
Authors: Leibman A , Aisen P
Published: October 2019

When it is incompletely saturated with iron, transferrin may exist in four molecular forms: apotransferrin, monoferric (A) transferrin (with iron occupying only the A site of the protein), monoferric (B) transferrin, and diferric transferrin. By combining electrophoresis in urea-polyacrylamide gels with crossed immunoelectrophoresis using specific antihuman transferin antiserum, it is possible to display and estimate the concentration of each of these four forms in normal human serum. The distribution of iron between the binding sites of transferrin is neither random nor determined by the relative binding strengths of transferrin's two sites. Rather, the more weakly binding and acid-labile B site of the protein is predominantly occupied.