bluuebunny/crossref_metadata_2025 huggingface dataset:
Metabolic dependence of protein arrangement in human erythrocyte membranes. I. Analysis of spectrin-rich complexes in ATP-depleted red cells
Authors: Palek J , Liu SC , Snyder LM
Published: October 2019

The discocyte-echinocyte transformation and the decrease in deformability associated with red cell ATP depletion have been attributed to changes in the physical properties of spectrin and actin, membrane proteins located at the membrane-cytosol interface. We investigated the spontaneous formation of spectrin-rich complexes in human erythrocyte membranes, employing two-dimensional SDS- polyacrylamide gel electrophoresis. Membranes of red cells depleted in ATP under aerobic conditions exhibited (1) an increase in components 4.5 and 8 and globin subunits, (2) a spontaneous formation of heterodimers of spectrin 1 + 2 and spectrin 2 + component 4.9, and (3) a large molecular weight (greater than 10(6) daltons) protein complex with a high spectrin to band 3 ratio. These complexes were dissociated with dithiothreitol and were prevented by anaerobic incubation or the maintenance of red cell ATP and GSH levels with glucose, adenine, and inosine. The complexes 1 + 2 and 2 + 4.9 were also seen in acetylphenylhydrazine-treated, glucose-6-phosphate dehydrogenase- deficient fresh erythrocytes that showed marked GSH depletion but preserved greater than 70% of the original ATP level. However, membranes of these cells did not contain the greater 10(6) dalton aggregate with a high spectrin to band 3 ratio. We concluded that the formation of the latter complex results from rearrangement of spectrin and other polypeptides in membranes of ATP-depleted red cells. Under aerobic conditions, the rearranged proteins undergo spontaneous intermolecular crosslinkings through disulfide couplings.
Microscopic platelet size and morphology in various hematologic disorders
Authors: Zeigler Z , Murphy S , Gardner FH
Published: October 2019

Microscopic evaluation of apparent platelet size and morphology was examined in a variety of hematologic disorders. The time of preparation of the blood smear was important. An artifactual increase in platelet size was noted on blood films from 20 normal individuals that were prepared either immediately or 180 min after venipuncture. The clearest differentiation of patient categories was obtained with smears prepared 60 min after venipuncture using blood anticoagulated with K3EDTA. Under these conditions, normal size and morphology values were found in thrombocytopenic patients with aplasia or with increased splenic pooling. In contrast, large size values were a reliable finding in idiopathic thrombocytopenic purpura patients, whose platelet counts were less than 50,000/microleter. Large size values were also noted in patients with infiltrated bone marrows or myeloproliferative syndromes regardless of the platelet count. The last two groups usually showed abnormal platelet morphology with greater than 10% hypogranular platelets. Normal platelet size and morphology were observed in patients with iron-deficiency and megaloblastic anemias and in patients with idiopathic thrombocytopenic purpura and systemic lupus erythematosus who had normal platelet counts.
Concentration gradient of blood stem cells in mouse bone marrow--an open question
Authors: Maloney MA , Lamela RA , Dorie MJ , Patt HM
Published: October 2019

The distribution of pluripotential blood stem cells (CFU-S) in bone marrow was studied in four strains of mice. Analyses of paired samples containing various fractions of axial and marginal bone marrow cells, as well as longitudinal 200-micrometer sections, revealed a rather uniform spatial distribution of CFU-S across the diameter of the femoral medullary cavity, in contrast to the finding by others of a CFU- S concentration gradient extending from the endosteum to the central longitudinal axis of bone marrow. The existence of a stem cell gradient is therefore open to question.
Fast-acting plasmin inhibitor in human plasma
Authors: Collen D , Wiman B
Published: October 2019

The fast-acting and physiologically most important inhibitor of plasmin in human plasma is a recently discovered and purified alpha 2-glycoprotein with a molecular weight of 65,000–70,000 daltons occurring at a concentration of 1 muM. The inhibitor rapidly forms a completely inactive 1:1 stoichometric complex with plasmin through reaction with the B chain (light chain) of the enzyme, which contains the active center. It also reacts with trypsin and very slowly with urokinase and with some other enzymes in purified systems, but its role in vivo as an inhibitor of proteases other than plasmin seems negligible. Antiplasmin is the only plasma protein that can inhibit the fibrinolysis associated with transformed or malignant cells. The plasmin-antiplasmin complex contains neoantigenic structures not present in the parent molecules that may form the basis of immunochemical methods for detecting activation of the fibrinolvtic system in blood.
Asynchronous globin chain synthesis in rabbit reticulocyte lystates induced by hemin-controlled repressor
Authors: Franco RS , Hogg JW , Martelo OJ
Published: October 2019

To define further the role of hemin-controlled repressor (HCR) in globin synthesis, we studied its effect on the synthesis of individual globin chains in a rabbit reticulocyte lysate cell-free system. In the presence of HCR there was a marked globin chain imbalance, resulting in a lowered alpha/beta ratio. These findings in vitro may have relevance to certain clinical heme deficiency states in which a similar globin chain imbalance has been observed.
Surface membrane characteristics and cytochemistry of the abnormal cells in adult acute leukemia
Authors: Gordon DS , Hubbard M
Published: October 2019

Membrane marker and cytochemical analyses were carried out on the abnormal cells from 70 adult acute leukemia patients. Such information may (1) supplement standard morphology and serve as a basis for a new classification scheme for acute leukemia, and (2) characterize the surface membranes of granulocyte, lymphocyte, and monocyte “progenitors.” Classification of acute lymphoid leukemias solely on the basis of morphology was unsatisfactory. The presence or absence of T-or B-cell markers was helful in classifying lymphoid leukemias. Monocyte progenitors were characteristically nonspecific esterase positive and Fc-receptor and membrane-IgG positive, but poorly phagocytic. Promyelocytes and myelocytes were frequently Fc-receptor positive and consequently positive for surface immunoglobulin. Myeloblasts were characteristically Fc-receptor negative. We conclude that surface membrane markers are essential in diagnosing lymphoid leukemias and helpful in nonlymphoid acute leukemias, and that cytochemistry is essential in delineating lymphoid from nonlymphoid leukemias and in subclassifying the latter.
Congenital hypoplastic anemia (Diamond-Blackfan syndrome) terminating in acute myelogenous leukemia
Authors: Wasser JS , Yolken R , Miller DR , Diamond L
Published: October 2019

A 31-yr-old female with congenital hypoplastic anemia (Diamond-Blackfan syndrome) whose long course terminated in acute myelogenous leukemia is described. In contrast to Fanconi anemia, malignant transformation rarely occurs in congenital hypoplastic anemia. This patient's diagnosis of congenital hypoplastic anemia is supported by her clinical course, absence of renal abnormalities, a negative family history for hematologic disorders, normal chromosome studies, failure of her skin fibroblasts to transform in culture with SV-40 virus, macrocytic erythrocyte indices, erythrocyte enzyme studies, and bone marrow findings. Only two other cases of malignancy have been reported in patients with congenital hypoplastic anemia. The development of malignancy in these patients suggests that malignant transformation may be a concern in the long-term progression of congenital hypoplastic anemia.
Role of microtubules in granulocyte adherence
Authors: Boxer LA , Allen JM , Watanabe AM , Besch HR Jr , Baehner RL
Published: October 2019

The adherence of human polymorphonuclear leukocytes (PMN) to nylon fibers is inhibited in a dose-dependent fashion by exposure in vitro of these cells to either colchicine or VM-26, both of which agents prevent microtubule assembly. Mean adherence of human PMN was 48% +/- 2%, following treatment with 10(-5) M colchicine or 10(-4) M VM-26 it was reduced to 31% +/- 2% and 7%, respectively. Peritoneal PMN obtained from mice and mink with Chediak-Higashi syndrome (CHS) thought to have a microtubule-membrane disorder affecting the PMN had a mean adherence of 29% +/- 3% and 40% +/- 8% compared to control values of 46% +/- 5% and 73% +/- 8%, respectively, from the mice and mink. Both ascorbic acid and bethanechol, shown previously to enhance microtubule assembly in humans with CHS, normalized granulocyte adherence in PMN obtained from mice with CHS. Cyclic nucleotide levels were not altered by treatment of human PMN with colchicine, nor did they differ between normal and CHS animals. Thus it appears that the state of microtubule assembly may directly affect the properties of the PMN plasma membrane without requiring alterations of cyclic nucleotides as an intermediary.
Role of the hypophysis in erythropoietin production during hypoxia
Authors: Peschle C , Rappaport IA , Magli MC , Marone G , Lettieri F , Cillo C , Gordon AS
Published: October 2019

Hypophysectomized or sham-operated male rats were exposed to hypoxia (0.42--0.40 or 0.37--0.35 atm for 6, 12, or 24 hr) applied 2 wk to 7 mo after surgery. Erythropoietin (Ep) levels in rat serum were evaluated on the basis of the exhypoxic polycythemic mouse assay. Ep activity evoked by hypoxia was significantly lower in hypophysectomized rats than in sham-operated controls. Progressive increase of the EP response to hypoxia correlated with extension of the time interval between hypophysectomy and hypoxia from 2 wk to 2--4 mo apparently mediated by the simultaneous inverse decline of red cell mass (RCM) values, i.e., of the “relative plethora” induced by a low O2 demand associated with relatively high RCM values. However, after 3--7 mo hypoxic Ep activity was still lower than in sham-operated controls. In these ablated animals the relative plethora became negligible or absent; accordingly, the Ep response apparently had reached plateau levels. These studies indicate that hypophysis (hypophyseal and target hormones, with the exception of estrogens) modulates Ep production under hypoxic conditions, possibly via a permissive enhancement of renal Ep activity.
Marked reduction of spectrinin hereditary spherocytosis in the common house mouse
Authors: Greenquist AC , Shohet SB , Bernstein SE
Published: October 2019

In contrast to the disease in humans, hereditary spherocytosis in the common house mouse produces an extreme spherocytosis. The cells show a broad distribution in size ranging from microcytic to macrocytic. Of particular interest is the finding of a substantial reduction in the major membrane polypeptide called spectrin, supporting a critical role for this protein in the control of erythrocyte shape and membrane stability.
Intermediate syndrome of platelet dysfunction
Authors: Czapek EE , Deykin D , Salzman E , Lian EC , Hellerstein LJ , Rosoff CB
Published: October 2019

We studied eight patients with intermittent bleeding episodes usually following trauma and associated with the ingestion of medicine known to interfere with platelet function. All patients had a normal or minimally prolonged baseline bleeding time. All had a normal platelet count, glass bead retention test, and platelet serotonin content and a variable pattern of abnormalities in prothrombin consumption and platelet factor 3 availability. However, all showed abnormal platelet aggregation reactions using epinephrine, adenosine diphosphate, and collagen. Following the administration of 975 mg aspirin, our patients' bleeding times became prolonged to a greater extent than the bleeding times of normal controls (range 13 to greater than 20 min). Review of the literature showed approximately 5% of “normal” controls had findings similar to those we report. We believe we are describing a group of individuals with an intermediate form of platelet dysfunction. Although their bleeding diathesis is not as severe as that of patients with platelet dysfunction syndromes previoulsy described, they do bleed significantly when subjected to trauma following the ingestion of drugs such as aspirin. We propose that this defect is common and should be screened for. The aspirin tolerance test is a simple test for detecting these patients.
Clinical and cell surface marker characterization of the early phase of chronic lymphocytic leukemia
Authors: Rudders RA , Howard JP
Published: October 2019

The immunologic surface markers on lymphocytes and clinical characteristics of 35 patients with established (stages 0–4) CLL with absolute lymphocyte counts greater than 15,000/cu mm were compared to those of a group of 25 patients with CLL in an early or preleukemic phase (counts of less than 15,000/cu mm). We found a monoclonal B cell proliferation in most cases in the latter group, in spite of the paucity of clinical and laboratory findings. Furthermore, early CLL can readily be distinguished from benign lymphocytosis by surface marker criteria. In untreated CLL, surface marker characteristics are stable with time and predominantly reflect expansion of clones expressing only B cell markers; however, small increase of blood T cells are often seen. Surface markers are a simple and clinically useful tool for definding and characterizing the preleukemic phase of CLL and its ultimate progression to established CLL.
Cellular maturation in human preleukemia
Authors: Koeffler HP , Golde DW
Published: October 2019

Bone marrow cells from three preleukemic patients with prominent marrow karyotypic abnormalities were studied in liquid culture to determine if the neoplastic clones were capable of maturation. Parallel cytogenetic and cytologic studies were performed in sequentially harvested bone marrow cultures. Maturation, albeit delayed, occurred in cultures from all three patients. By 14 days of culture in vitro, morphologic, cytochemical, and functional evidence of maturation was observed in about 70% of the cells. By day 21, 85% of the cells were mature by these criteria. All but 2 of 249 metaphases from the cultured cells contained the cytogenetic abnormality of the neoplastic clone. We conclude that some preleukemic cells identified by a chromosomal abnormality can mature in vitro. Preleukemia may be viewed as a syndrome of “early leukemia” in which the neoplastic clone is established and manifested functionally as ineffective hematopoiesis. Hematopoietic cell differentiation becomes progressively abnormal with termination in the nearly complete maturational block characteristic of acute myelogenous leukemia.
Expression of the major sialoglycoprotein (glycophorin) on erythroid cells in human bone marrow
Authors: Gahmberg CG , Jokinen M , Andersson LC
Published: October 2019

The major sialoglycoprotein of human erythrocyte membranes (glycophorin) is one of the most-studied membrane proteins. Although the structure is relatively well known, almost nothing is known about its expression in erythroid cells. To study this we raised an antiserum that reacted specifically with this protein. This was accomplished by immunization of rabbits with a preparation of glycophorin followed by absorption with En(a-) erythrocyte membranes, which lack glycophorin. By use of this antiserum and a staphylococcus protein A technique we could establish that only bone marrow cells of erythrocyte lineage express glycophorin at the cell surface. This occurs in basophilic normoblasts and later stages of erythrocyte differentiation, whereas pronormoblasts do not seem to contain glycophorin.
Decrease of mast cells in W/Wv mice and their increase by bone marrow transplantation
Authors: Kitamura Y , Go S , Hatanaka K
Published: October 2019

Production of tissue mast cells was evaluated in genetically anemic mice of W/Wv genotype and was found to be abnormal. In the skin of adult W/Wv mice the number of mast cells/cm was less than 1% of the number observed in the congeneic +/+ mice. No mast cells were detectable in other tissues of the W/Wv mice. After transplantation of bone marrow cells from +/+ mice the number of mast cells in the skin, stomach, caecum, and mesentery of the W/Wv mice increased to levels similar to those of the +/+ mice. These results show that the W/Wv mouse is a useful tool for the investigations concerning the physiologic roles and the origin of mast cells.
Membrane mechanical properties of ATP-depleted human erythrocytes
Authors: Meiselman HJ , Evans EA , Hochmuth RM
Published: October 2019

Although the relations between the metabolic state and the mechanical properties of human red blood cells (RBC) continue to be of current interest, literature reports in this area are not in agreement. The present investigation was designed to determine several intrinsic mechanical properties of human RBC membranes before and after metabolic depletion via incubation at 37 degrees C for 24 hr. Using micropipette and flow channel techniques, three properties were measured: (1) mu, surface shear modulus of elasticity; (2) K, elastic area compressibility modulus; (3) etap, shear viscosity in the plastic domain. Our results indicate no significant differences in these parameters between fresh and ATP-depleted human RBC membranes. These present data are thus in disagreement with other literature reports indicating large changes in membrane mechanical properties consequent to metabolic depletion. A brief discussion of the possible reasons for this disagreement is presented.
Chronic lymphocytic leukemia (CLL) terminating in multiple myeloma: report of two cases
Authors: Kough RH , Makary AZ
Published: October 2019

Two cases of multiple myeloma (MM) developed late in the course of chronic lymphocytic leukemia (CLL). An 81-yr-old white female developed, after 6 yr of CLL, IgAk MM with sheets of plasma cells abutting sheets of lymphocytes in the bone marrow, multiple pathologic fractures, and 0.26 g/24 free k light chains in the urine. A 74-yr-old white male developed, after 16 yr of CLL, k light chain MM with 20% plasma cells in the bone marrow, multiple panthologic fractures, and 3.7 g/24 hr free k light chains in the urine. In both cases the CLL had responded well to intermittent low-dose chlorambucil therapy, but the MM failed to respond to cyclic melphalanprednisone therapy. A review of 105 cases of CLL seen at the Geisinger Medical Center failed to turn up any other cases of MM developing during the course of CLL. The suggestion that there is an increased prevalence of MM in CLL is an attractive one because both diseases are B cell neoplasms and because of the increased frequency of asymptomatic monoclonal gammopathies in CLL found by others.
Synthesis of a peroxidase activity by the cells of hairy cell leukemia: a study by ultrastructural cytochemistry
Authors: Reyes F , Gourdin MF , Farcet JP , Dreyfus B , Breton-Gorius J
Published: October 2019

The nature of cells present in the blood, marrow, and spleen of patients with hairy cell leukemia is largely debated. These cells have been tentatively categorized on the basis of either monocytic or lymphocytic markers, and the accumulating data points to the fact that they share some characteristics of both cell types. Although hairy cells are known to lack myeloperoxidase-positive granules, present in normal human monocytes, we investigated the possible presence of other peroxidase activities differing from the granule-bound myeloperoxidase. The study was carried out with several methods based on the incubation of fixed and unfixed cells in the presence of diaminobenzidine and hydrogen peroxide. A peroxidase activity was found in hairy cells, located always in the endoplasmic reticulum but not in the Golgi apparatus or in any granule. By its cytochemical characteristics it appears to be closely related to that of tissue macrophages, activated blood monocytes, and other nonlymphocytic hematopoietic cells. This peroxidase is not found in lymphocytes with B or T phenotypes.
Detection of factor XIIIa (active fibrin-stabilizing factor) in normal plasma
Authors: Nelson JC , Lerner RG
Published: October 2019

Factor XIIIa (active fibrin-stabilizing factor) generated in heat- defibrinated plasma by the addition of thrombin can be measured by 14C- putrescine incorporation into casein. Modification of this assay be substituting 3H-putrescine of high specific activity as the donor amine permits measurement of amine incorporation by plasma even in the absence of added thrombin. Incorporation is calcium dependent, inhibited by iodoacetamide, and absent from congenital factor XIII- deficient plasma and from normal platelets. The transamidating activity detected by radioenzymatic assay catalyzed the formation of gamma-gamma dimers and alpha polymers of fibrin and was thus biologically functional. This fibrin cross-linking activity was absent from factor XIII-deficient plasma. These experiments show (1) some factor XIII is present in plasma as factor XIIIa; (2) this factor XIIIa can cross-link fibrin and thus has biologic activity as well; and (3) this activity is not present in factor XIII-deficient plasma. Factor XIIIa in normal plasma is possibly activated in vivo, perhaps by circulating thrombin, factor Xa, or other proteolytic enzymes.
Microscopic method for assaying F cell production: illustrative changes during infancy and in aplastic anemia
Authors: Dover GJ , Boyer SH , Bell WR
Published: October 2019

Fetal hemoglobin (HbF)-bearing reticulocytes (F reticulocytes) can be detected in peripheral blood by a modification of the microscopic single-cell radial immunodiffusion method. Thereby otherwise inappreciable changes in HbF production can readily be recognized. F reticulocyte frequencies are reporducibly measurable whenever the product of whole blood HbF-bearing red cell (F cell) frequency and reticulocyte frequency is approximately 5 X 10(-4) or greater. Serial analyses of F reticulocytes and nonreticulocyte F cells (F erythrocytes) illustrate that (1) levels of F reticulocytes and F erythrocytes are persistently similar in normal adults with more than 6% F cells and thus cell survival times of F and non-F cells must be essentially the same, (2) changing levels of F reticulocytes can be sensitive predictors of later changes in mature F cell frequencies during infancy and in adults recovering from aplastic anemia, and (3) alterations in F reticulocyte frequency and the amount of HbF per F reticulocyte are discordant in some settings but concordant in others.
Monoclonal IgG anticoagulants delaying fibrin aggregation in two patients with systemic lupus erythematosus (SLE)
Authors: Galanakis DK , Ginzler EM , Fikrig SM
Published: October 2019

There is paucity of information regarding the prolonged plasma thrombin time known to occur in some patients with systemic lupus erythematosus. Detailed investigations of plasma from two such patients disclosed that IgG accounted for this defect in each case. IgG isolated from plasma of either patient possessed the property of delaying fibrin aggregation and prolonging the clotting times of fibrinogen. Preincubation of IgG from either patient with anti-IgG or anti-Fab (rabbit) serum abolished this anticoagulant property. Moreover, the anticoagulant IgG from the first patient was neutralized with anit-k chain and anti-IgG3, that from the second patient with anti-lambda chain and anti-IgG1 serum. These anticoagulants were also dissimilar with respect to their interactions with fibrin(ogen). IgG from the first patient had no anticoagulant activity against fibrin(ogen) species lacking intact Aalpha chains. IgG from the second patient displayed undiminished anticoagulant effect on such fibrin(ogen) species. We conclude that each anticoagulant interacted with a distinct region(s) on the fibrinogen molecule and that these interactions affect or involve sites that participate in the fibrin self-assembly process.
Human T lymphocyte cell line producing colony-stimulating activity
Authors: Golde DW , Quan SG , Cline MJ
Published: October 2019

We derived a permanent human T lymphocyte cell line that elaborates a potent colony-stimulating activity (CSA). The line was established with spleen cells from a patient with a T lymphocyte variant of hairy-cell leukemia. These cells form rosettes with sheep erythrocytes, show a proliferative response to phytohemagglutinin, and are lysed by antithymocyte globulin. They do not synthesize immunoglobulin, nor do they contain Epstein-Barr virus. CSA is regularly detected in the supernatant medium after 3 days culture. In the presence of PHA there is augmented elaboration of CSA; maximal activity is reached by 2 days and is 20% greater than that produced by a feeder layer of 1 X 10(6) peripheral blood leukocytes. One microliter of the supernatant material stimulated colony formation from the light-density nonadherent fraction of human bone marrow; there was maximal activity between 10 and 50 microliter/ml. Conditioned medium from these cells has little effect in stimulating CFU-C from murine bone marrow. The availability of a human T lymphocyte line producing CSA will provide a source for large quantities of the lymphocyte-derived hormone and permit a definition of factors modulating the interaction of T lymphocytes with granulocyte and monocyte stem cells.
Adriamycin and daunomycin generate reactive oxygen compounds in erythrocytes
Authors: Henderson CA , Metz EN , Balcerzak SP , Sagone AL Jr
Published: October 2019

Adriamycin and daunomycin produce dose-related cardiac toxicity that may be related to oxygen radicals. Addition of these compounds to human erythrocyte suspensions resulted in stimulation of hexose monophosphate shunt activity that was markedly impaired in the absence of oxyhemoglobin. Evidence for generation of hydrogen peroxide by these compounds was provided by oxidation of reduced glutathione, by 14C- formate oxidation, and by the catalase-aminotriazole trapping technique. These experiments indicate that Adriamycin and daunomycin interact with oxyhemoglobin to generate reactive oxygen metabolites. A similar interaction with oxymyoglobin may occur in the heart and produce oxygen radicals that injure cardiac myocytes.
Activation of factor X by factors IXa and VIII; a specific assay for factor IXa in the presence of thrombin-activated factor VIII
Authors: Hultin MB , Nemerson Y
Published: October 2019

We studied the activation of factor X by the intrinsic pathway of blood coagulation using a new assay of factor X activation. When factor X tritiated in its sialic acid residues is activated, activation can be measured by the release of tritiated activation peptide, and the initial rate of activation can be determined under varying conditions. In the presence of phospholipid and calcium ions, factor IXa activated factor X slowly without factor VIII, and this activation was blocked by a specific factor IX inhibitor. These data provide strong evidence that factor IXa is the enzyme responsible for factor X activation by the intrinsic pathway. The role of factor VIII was also investigated. Factor VIII could be reproducibly thrombin activated and then stabilized by the addition of 2 mM benzamidine hydrochloride; this suggests that inactivation is due to proteolysis. Neither unactivated nor thrombin-activated factor VIII produced factor X activation without factor IXa. With a constant level of factor IXa, factor X activation was directly proportional to the level of activated factor VIII. With a constant level of activated factor VIII, factor X activation was proportional to the factor IXa concentration. This observation was exploited to develop a specific, sensitive assay for factor IXa.
Release of arachidonic acid from human platelets. A key role for the potentiation of platelet aggregability in normal subjects as well as in those with nephrotic syndrome
Authors: Yoshida N , Aoki N
Published: October 2019

Low (nonaggregating) concentrations of collagen that potentiate platelet aggregation did not induce the formation of measurable amount of malondialdehyde (MDA) but released small but significant amounts of radioactivity from 14C-arachidonic acid-labeled platelets. A major portion of the radioactive compounds released by nonaggregating concentrations of collagen existed as arachidonic acid and a minor part as thromboxane B2. The nephrotic syndrome enhances platelet aggregability, and this effect is abolished by correcting hypoalbuminemia in vitro and in vivo by the addition of albumin, which is the main carrier for free fatty acids, including arachidonic acid. Human albumin (fatty acid free) inhibited collagen-induced aggregation, MDA formation, and release of the radioactivity from 14C-arachidonic acid-labeled platelets in normals as well as in those with nephrotic syndrome. These data support our hypothesis that the main mechanism responsible for the potentiation of platelet aggregation is the release of arachidonic acid from platelet membrane phospholipids via the activation of phospholipase A2. Furthermore, enhanced platelet aggregation in the nephrotic syndrome was at least partly attributable to an increased availability of arachidonic acid released secondary to hypoalbuminemia. Albumin inhibits aggregation probably by binding to released arachidonic acid preventing arachidonic acid from being metabolized to potent aggregating substances, endoperoxides and thromboxane A2. The mechanism of release of arachidonic acid may play a key role in the potentiation of platelet aggregability in normals as well as in pathologic conditions such as the nephrotic syndrome.
Adenosine deaminase, terminal deoxynucleotidyl transferase (TdT), and cell surface markers in childhood acute leukemia
Authors: Coleman MS , Greenwood MF , Hutton JJ , Holland P , Lampkin B , Krill C , Kastelic JE
Published: October 2019

The purpose of this report is to compare measurements of enzymatic activities and cell surface markers as methods of distinguishing subtypes of lymphoid leukemias of childhood. Twenty-six children ages 2- 15 yr were studied. Terminal deoxynucleotidyl transferase (TdT) activity was high in blasts from all 20 children with either null or T cell acute lymphoblastic leukemia. The activity of adenosine deaminase per cell was higher (P less than 0.005) and that of TdT lower (p less than 0.05) in T than in null cell lymphoblasts, although there was some overlap in values. Blasts from 3 children with acute lymphoid leukemia were positive for surface-associated immunoglobulins. The neoplastic lymphoid cells from these children differed from T and null cell leukemic lymphoblasts by having very low levels of TdT and adenosine deaminase activity. Measurements of adenosine deaminase and TdT may complement measurements of cell surface markers and distinguish biochemical subtypes of acute lymphoid leukemia.
Hematologic changes during spleen colony development in nonirradiated mice
Authors: Tamai M , Kitamura Y
Published: October 2019

The mechanism of spleen colony formation in nonirradiated mice was investigated. When the spleen cells of C57BL mice immunized against CBA- T6T6 mice were injected into the nonirradiated BT6F1 (C57BL X CBA-T6T6) hybrid mice, the number of hematopoietic stem cells (CFU-S of C57BL mouse origin that settled in the spleen of the BT6F1 mice continued to decrease in the first 9 days and then started to increase, with a doubling time of about 36 hr. Colonies were detected on the surface of the spleen 16–22 days after the cell injection. The slower appearance of spleen colonies in nonirradiated mice (compared with 6–10 days in the irridiated animals) appears to be due to retarded start of differentiation and to the prolonged doubling time of CFU-S in non- irradiated mice.
Granuloerythropoietic colonies in human bone marrow, peripheral blood, and cord blood
Authors: Fauser AA , Messner HA
Published: October 2019

Colonies that contain granulopoietic and erythropoietic cells can be grown in specimens of human bone marrow, peripheral blood, and cord blood. Growth of these colonies is promoted by media conditioned by leukocytes in the presence of phytohemagglutinin (PHA-LCM) and the addition of erythropoietin on days 4 or 5 to the cultures. Sedimentation velocity profiles for these granuloerythrocytic colonies suggest their origin from single cells (CFU-G/E) rather than from doublets or clumps. This hypothesis is supported by cocultivation of male and female specimens. Cells in granuloerythrocytic colonies that developed in such mixing experiments were either uniformly female by Y- chromatin analysis or contained Y-chromatin bodies in both the granulocytic and erythroid cells.
Parallel decrease of erythrocyte membrane deformability and spectrin solubility at low pH
Authors: Smith BD , La Celle PL
Published: October 2019

Reduction of pH over the range 6.0–4.5 results in a decrease of erythrocyte deformability in parallel with the induced progressive sphericity of cells. At low deformation rates employed, increase of hemoglobin viscosity was not significant. A decrease of membrane elasticity was detected in cells when sphering, the major determinant of cellular deformability, was prevented at pH 5.0 by hyperosmotic medium. The pronounced change of deformability and the reduced elasticity occurred at calculated intracellular pH values at which solubility in vitro of extracted erythrocyte spectrin is also markedly reduced. The parallel decrease of deformability and spectrin solubility supports the hypothesis that spectrin aggregation may contribute to regulation of erythrocyte deformability through effects on shape and membrane elastic characteristics.
B cell acute lymphoblastic leukemia (ALL) with a 14q+ chromosome abnormality
Authors: Roth DG , Cimino MC , Variakojis D , Golomb HM , Rowley JD
Published: October 2019

An adult patient with acute lymphoblastic leukemia associated with a 14q+ marker chromosome is presented. The abnormality resulted from a translocation of material from the long arm of chromosome 11. The leukemic cells were found to be B cells on the basis of surface immunoglobulins, lack of receptors for sheep erythrocytes, and a characteristically low level of adenosine deaminase activity. In other patients with ALL studied by us or reported by others in whom chromosome banding was done, a 14q+ chromosome was present in only one instance, also a case of B cell ALL. These two cases are the only examples of B cell ALL studied with chromosome banding reported to date. The frequent occurrence of a 14q+ chromosome in other malignant lymphoproliferative diseases of B cell origin suggests that a general association may exist between the 14q+ abnormality and B cell neoplasms. Cytogenetic analysis may therefore be useful in defining subtypes of ALL and in relating specific chromosomal abnormalities to lymphoproliferative disorders.
Turnover of antiplasmin, the fast-acting plasmin inhibitor of plasma
Authors: Collen D , Wiman B
Published: October 2019

Human antiplasmin, the fast-acting plasmin inhibitor in plasma, was purified to homogeneity and labeled with 125I. This material, which was indistinguishable from antiplasmin in plasma with respect to several physicochemical and functional properties, was injected intravenously and its turnover measured in control subjects and in patients undergoing thrombolytic therapy. In eight control subjects (four healthy persons and four patients with atherosclerosis), the following turnover characteristics were obtained: plasma radioactivity half-life 2.64 +/- 0.32 days, fractional catabolic rate 0.53 +/- 0.09 of the plasma pool per day, intravascular fraction 0.51 +/- 0.05, and synthetic (catabolic) rate 1.4 +/- 0.27 mg/kg/day. The half-life of the plasmin-antiplasmin complex in plasma, measured from the disappearance rate of labeled antiplasmin, plasmin or plasmin-antiplasmin complex during thrombolytic therapy was approximately 0.5 days.
Reduction of salivary tissue factor (thromboplastin) activity by warfarin therapy
Authors: Zacharski LR , Rosenstein R
Published: October 2019

The coagulant of normal human saliva has been identified as tissue factor (thromboplastin, TF) by virtue of its ability to cause rapid coagulation in plasmas deficient in first-stage coagulation factors and to activate factor x in the presence of factor VII and by virtue of the fact that its activity is expressed only in the presence of factor VII and is inhibited by an antibody to TF. The TF is related to cells and cell fragments in saliva. Salivary TF activity has been found to be significantly reduced in patients taking warfarin. The decline in TF activity during induction of warfarin anticoagulation occurs during the warfarin-induced decline in vitamin-K-dependent clotting factor activity, as judged by the prothrombin time. The decrease in TF activity is not related to a reduction in salivary cell count or total protein content or to a direct effect of warfarin on the assay. It is hypothesized that the mechanism by which warfarin inhibits TF activity may be related to the mechanism by which it inhibits expression of the activity of the vitamin-K-dependent clotting factors. Inhibition of the TF activity may be involved in the antithrombotic effect of warfarin.
Hereditary myeloperoxidase deficiency
Authors: Cech P , Papathanassiou A , Boreux G , Roth P , Miescher PA
Published: October 2019

The functional properties of granulocytes in a diabetic patient deficient in myeloperoxidase (MPO) were compared with those of granulocytes in healthy subjects. The granulocytes of this patient had normal phagocytic activity. The microbicidal activity of the granulocytes was partially diminished with regard to Staphylococcus aureus and was almost nil with regard to Candida albicans. Fungicidal activity of normal granulocytes was shown to be impaired during the in vitro artificial hyperglycemic condition. The relationship among diabetes mellitus, MPO deficiency, and serious C. albicans infection was examined. Genetic investigation was carried out in 28 members of the proband's family. In close relatives of the patient, MPO values were found to be diminshed to a greater or lesser degree, thus suggesting variable expressivity of the heterozygote state of MPO deficiency.
Treatment of acute nonlymphocytic leukemia: use of anthracycline- cytosine arabinoside induction therapy and comparison of two maintenance regimens
Authors: Preisler HD , Rustum Y , Henderson ES , Bjornsson S , Creaven PJ , Higby DJ , Freeman A , Gailani S , Naeher C
Published: October 2019

Patients with acute nonlymphocytic leukemia were given remission induction therapy consisting of cytosine arabinoside and an anthracycline. Those patients who experienced complete remission received two courses of consolidation therapy and were randomized to receive maintenance therapy consisting of either daily chemotherapy with reinforcements every 3 mo or reinforcement therapy only every 6 wk. The overall complete remission rate was 66%, with 80% complete remission for previously untreated patients less than 60 yr of age who did not have a prior history of malignancy. Remission durations were the same for patients treated with both maintenance regimens. The major determinant for successful remission induction therapy was patient age, with older patients frequently succumbing to intercurrent infection. Documented leukemic cell resistance to the therapy employed was only rarely encountered. Once remission was achieved, age was no longer a determinant of patient survival, since duration of remission was independent of age. Remission durations were directly related to leukemic cell retention of cytosine arabinoside triphosphate. Hence therapy for acute nonlymphocytic leukemia can be divided into two separate areas: remission induction and remission maintenance.
Hemoglobin biosynthesis in individual bursts in culture: studies of human umbilical cord blood
Authors: Kidoguchi K , Ogawa M , Karam JD , McNeil JS , Fitch MS
Published: October 2019

We cultured human umbilical cord blood erythropoietic precursors in methyl cellulose clonal assay and analyzed the synthetic rates of Hb A and Hb F in individual erythropoietic bursts. Hemoglobin was labeled with 14C-amino acids in culture, separated by slab gel isoelectric focusing techniques, and quantitated by fluorographic methods. Almost all bursts exhibited both Hb A and Hb F in varying ratios. Frequencies of the individual bursts differing in percentage Hb F biosynthesis had normal distributions. Natal erythropoietic precursors appeared to be randomly committed to Hb F synthesis.
Chromosome analysis in two unusual malignant blood disorders presumably induced by benzene
Authors: Van den Berghe H , Louwagie A , Broeckaert-Van Orshoven A , David G , Verwilghen R
Published: October 2019

Two patients with presumably benzene-induced malignant blood disorders with preleukemic phases were cytogenetically monitored through the courses of their diseases. Patient 1, in addition to a familial chromosome translocation [t(3;16)], developed karyotypic abnormalities in 100% of the marrow cells, including two translocation: t(9;16)] and t(4;15). Monosomy of chromosome 7 characterized the cells of patient 2. Cytogenetic monitoring of the patients at various phases of their diseases served as an important indicator of the transformation or progression of the preleukemia into frank leukemia and of the unusual behavior of such leukemic cells.
Effect of anti-P1A1 antibody on human platelets. I. The role of complement
Authors: Cines DB , Schreiber AD
Published: October 2019

We studied the interaction of complement with human platelets. Complement was activated by IgG anti-P1A1 antibody obtained from 3 patients with the post-transfusion purpura syndrome. We used a heparin- plasma buffer system that permits complement activation and also preserves platelet function. With this system complement activation was efficient, and platelet immune alteration was extensive. Anti-P1A1 antibody was effective only in the presence of complement, in which case both platelet lysis and serotonin release (release reaction) in the absence of lysis were observed. Platelet lysis, as assessed by 51Cr loss, required 10-fold more antibody than was necessary to induce platelet aggregation and release of 14C-serotonin. This platelet release reaction required an intact classic complement sequence through C6. The extent of platelet serotonin release parallelled the depletion of C1 and C4 from platelet-rich plasma. Concentrations of antibody insufficient to induce platelet aggregation and serotonin release could still activate C1 and deposit increased C3 on the platelet surface. These studies demonstrated that complement activation by anti-P1A1 antibody can alter human platelets in a nonlytic system. Several phases of complement-mediated human platelet alteration are possible, depending on the concentration of anti-P1A1 antibody.
Congenital deficiency of blood clotting factors II, VII, IX, and X
Authors: Chung KS , Bezeaud A , Goldsmith JC , McMillan CW , Menache D , Roberts HR
Published: October 2019

A patient congenitally deficient in factors II, VII, IX, and X has been further investigated after a follow-up of 15 yr. At birth, these factors, when determined by clotting assays, were undetectable. Following therapy with vitamin K1, the clotting activity of these factors rose but never exceeded 18% of normal. Immunologic assays revealed much higher levels of these factors than did clotting assays, thus suggesting that the vitamin-K-dependent factors were present in abnormal forms. Two-dimensional crossed immunoelectrophoresis showed that at least two forms of prothrombin were present in the patient's plasma. One form was similar to normal prothrombin; the other had the same mobility as acarboxyprothrombin. In addition, the majority of this fast-migrating peak was not adsorbable onto insoluble barium salts. These observations suggested that some molecules of the patient's prothrombin lacked the normal complement of gamma-carboxyglutamic acid residues. This observation was confirmed by a specific assay for gamma- carboxyglutamate. Since malabsorption of vitamin K, warfarin intoxication, and hepatic dysfunction were excluded as causes of this patient's syndrome, this rare congenital abnormality could represent either a defective gamma-carboxylation mechanism within the hepatocyte or faulty vitamin K transport.
Hematin administration to an adult with lead intoxication
Authors: Lamon JM , Frykholm BC , Tschudy DP
Published: October 2019

Lead poisoning and acute intermittent porphyria (AIP) may exhibit similar neurologic manifestations, and they have in common elevated excretion of urinary aminolevulinic acid (ALA). Despite their similarities, the possible pathophysiologic connection between AIP and lead poisoning in not known. Because intravenous hematin administration has produced biochemical improvement in AIP, a hematin trial in lead intoxication was of interest with respect to some of the heme metabolism abnormalities observed in the condition. Significant diminution of urinary ALA and coproporphyrin excretion occurred in association with intravenous hematin administration.
Clonal identification in acute lymphoblastic leukemia
Authors: Goldstone AH , McVerry BA , Janossy G , Walker H
Published: October 2019

In a case of acute lymphoblastic leukemia, two distinct types of leukemia blast cells could be identified throughout the course of the disease. The initially dominant type of blast cell was sensitive to chemotherapy; the other was drug-resistant, gradually becoming dominant as the disease progressed. The cell types could be clearly separated by their morphologic and surface membrane marker characteristics. The same chromosomal constitution was present in both types of blast cells, indicating a common clonal origin. Additional chromosomal abnormalities were present in the later stages of the disease, demonstrating that a distinct subclone had proliferated. This study illustrates that in some cases of acute leukemia, disease relapse is caused by growth of drug- resistant subclones that may be clearly identified by changes in morphology and surface membrane marker characteristics.
Genetics of classic von Willebrand's disease. II. Optimal assignment of the heterozygous genotype (diagnosis) by discriminant analysis
Authors: Miller CH , Graham JB , Goldin LR , Elston RC
Published: October 2019

In classic von Willebrand's disease (vWd), assignment of the heterozygous genotype for genetic studies and diagnosis for clinical purposes (which are not exactly the same) are formidable problems. We have pointed out in the first report in this series that almost 50% of the members of two large kindred who transmitted this disease, and were therefore heterozygous, were scored as normal by the usual tests of hemostasis. This report describes how this large proportion can be significantly reduced by application of discriminant analysis. Using linear discriminants in three variables--coagulation factor VIII (VIII:C), factor-VII-related antigen (VIIIR:Ag), and the ristocetin cofactor related to factor VIII (VIIIR:WF)--we were able to classify as heterozygous more than 80% of the transmitters in the two large kindred. It was of particular interest that the four parents of two related vWd homozygotes could be scored as heterozygous by discriminant analysis even though all their laboratory tests were within the normal ranges.