bluuebunny/crossref_metadata_2025 huggingface dataset:
Determination of glycosylated hemoglobins in neonatal blood by isoelectric focusing
Authors: Fitzgerald M. D. , Cauchi M. N.
Published: February 2007

Isoelectric focusing (IEF) with suitable „spacers”︁ can be used to distinguish glycosylated hemoglobins from HbA and HbF in adults and infants, respectively. Using this technique we have shown that there is a good correlation between glycosylated hemoglobins measured by IEF and column chromatography (r = 0.778, P < 0.001). There was no significant difference in the rate of glycosylation between adult and cord blood hemolysates following incubation with glucose in vitro: In both instances there was an increase from the normal levels of about 5% to 26% through 31% after 18 hours of incubation. The levels of glycosylated hemoglobins in infants of varying gestation, neonates, and normal adults was remarkably constant, varying from 4.7 ± 1.1% to 6.1 ± 1.6%.

These results show that IEF can be useful in the quantitation of glycosylated hemoglobins, particularly in the presence of hemoglobins (eg, HbF) where standard chromatographic techniques cannot be used. IEF has the added advantage of using small quantities of blood, and allows a number of samples to be tested at the same time, thus facilitating the identification of abnormal hemoglobins.
Childhood CML in blastic stage: An analysis of cell markers and cell kinetics
Authors: Beck J. D. , Andreeff M. , Mertelsmann R. , Haghbin M. , Tan C. , Miller D. R. , Good R. A. , Gupta S.
Published: February 2007

The clinical course of Ph+ CML and their terminal blastic stage is described in the following case histories. During blastic phase, cell surface markers, terminal deoxynucleotidyl transferase (TdT) activity, and flow cytometric measurements were used for determination of the blast cell phenotype which was undifferentiated by morphological and cytochemical criteria. A high proportion of blast cells expressed Fc receptors for IgG, TdT activity was normal in both children, and RNA measurements of single cells by flow cytometry showed a high RNA content in the majority of blasts. These findings are compatible with the phenotype seen in myelomonocytic leukemias. Flow cytometry appears to be a useful adjunct to immunological methods and TdT activity for the rapid characterization of CML in blastic phase.
A clinicopathologic analysis of chronic lymphocytic leukemia
Authors: van Scoy‐Mosher Michael B. , Bick Miriam , Capostagno Vince , Walford Roy L. , Gatti Richard A.
Published: February 2007

Sixty consecutive patients with chronic lymphocytic leukemia (CLL) were studied from both a clinical and laboratory standpoint. Hypoimmunoglobulinemia was found in 45% of patients; many of these patients suffered from severe bacterial infections. Second primary malignancies were diagnosed in 20% of patient's; most of these predated the diagnosis of CLL. HLA‐B17 typing was found in unexpectedly high frequency in a small group of Coombs'‐positive patients. The Rai‐staging system was found to be generally useful for determining prognosis of groups of patients, although less useful in any individual patient. Findings relating to surface membrane immunoglobulin‐positive (B) and E‐rosetting (T) lymphocytes are described. A patient with null‐cell CLL is described. All patients with proliferation of gamma heavy‐chain‐bearing cells were diagnosed in early Rai stages suggesting that this marker may identify a subset of patients who present early and have a good prognosis. These data suggest that lymphocyte marker studies augment the Rai criteria in evaluating prognosis and may eventually be of value to the clinician in evaluating stage of disease and response to treatment.
The immunological and cytochemical characterization of T‐cell lymphoproliferative diseases
Authors: Spira T. J. , Ades E. W. , Gordon D. S.
Published: February 2007

Surface markers, terminal transferase, cytochemical staining, and function of peripheral blood lymphocytes were examined in nine patients with T‐cell lymphoproliferative disease. Anti‐T‐cell antiserum proved useful in characterizing the phenotype of the abnormal cells when only a minority of the abnormal cells formed E‐rosettes. Anti‐B‐cell antiserum was consistently unreactive with the abnormal cells. Terminal transferase was elevated in T‐cell ALL but low in T‐cell CLL, lymphoma‐leukemia, or Sézary syndrome. Lymphocyte proliferative response to nonspecific mitogens was generally depressed as was the response to allogeneic cells. The cells also failed to stimulate in mixed lymphocyte culture, implying a lack of Ia antigens on their surface. Acid phosphatase stain was positive in the three patients tested. Peroxidase and esterase stains were negative, whereas PAS was positive in five of eight patients tested. Prognosis in general, regardless of the clinical picture, (ie, acute, chronic, or lymphoma‐leukemia), was poor with seven of the nine patients dying within 14 months of diagnosis. Therefore, the use of anti‐T‐cell antisera and the E‐rosette surface marker should be used to characterize the phenotype of the malignant cells in lymphoproliferative disease to enable more vigorous treatment of those that are of T‐cell origin and that therefore carry a poor prognosis.
Late relapses in hodgkin disease
Authors: Kanofsky Jeffrey R. , Golomb Harvey M. , Vardiman James E. , Sweet Donald L. , Ultmann John E.
Published: February 2007

Among a cohort of 142 patients with Hodgkin disease (HD), pathologic stages (PS) IA through IVB, 84 remained in complete remission (CR) at least 3 years after the completion of initial protocol therapy. Eight of these patients subsequently developed a recurrence of HD and were defined as having a late relapse. The disease‐free interval ranged from 37 to 76 months (median 53 months). Six of the relapse patterns were those of dissemination. All patients were salvaged and are in CR from 14 to 50 months after relapse. The actuarial survival, measured from the time of relapse, of the eight late‐relapsing patients was superior to the actuarial survival of 38 patients with early relapse (disease‐free interval less than 36 months), but as yet this difference is not statistically significant (P = 0.07, Gehan).
Effects of dexamethasone on fetal hemoglobin synthesis in peripheral blood erythroid burst‐forming units
Authors: Steinberg Martin H. , Coleman Mary B. , Garver Fred A. , Grenett Hernan E. , Pressley Annette , Adams Junius G.
Published: February 2007

Colonies derived from erythroid burst‐forming units (BFU‐E) synthesize fetal hemoglobin (HbF) in amounts that far exceed in vivo levels. There is some evidence that HbF synthesis is controlled at the level of a primitive erythroid precursor cell. Dexamethasone may potentiate the development of BFU‐E. Since a means of augmenting HbF production in sickle cell anemia or severe β‐thalassemia would be of great therapeutic value, we studied the effects of dexamethasone on HbF and γ‐globin chain synthesis in BFU‐E from patients with sickle cell anemia and controls. HbF was measured by radioimmunoassay of BFU‐E lysate and γ‐chain synthesis by the incorporation of 3H‐leucine into globin, which was then purified by gel filtration and column chromatography. Dexamethasone (10−9 M) produced an increase in the number of BFU‐E in 16 of 19 subjects when compared with numbers of BFU‐E cultured with only erythropoietin. The individual BFU‐E were larger and contained more subcolonies. Dexamethasone did not increase HbF or γ‐chain synthesis, and there was no relationship between increased proliferation of BFU‐E and augmented HbF production. Thus, although dexamethasone augmented the development of erythroid bursts, there was no increment in HbF.
Human thymus antigen: Characterization and its expression on human leukemias
Authors: Fujimoto Junichiro , Ishii Yoshifumi , Koshiba Hirofumi , Matsuura Akihiro , Ogasawara Minoru , Uede Toshimitsu , Kikuchi Kokichi
Published: February 2007

Specific anti‐human thymus xenoantiserum (ATS) was utilized for characterizing a human thymus antigen (HTA) preferentially expressed on human thymocytes. Binding of ATS with different cell types was studied by immunofluorescence and immunoperoxidase techniques, as well as by radioimmunoprecipitation (RIP) followed by SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE). ATS reacted selectively with the majority (87%) of human thymocytes among normal lymphoid cell populations tested. In the thymus, HTA was expressed largely on cortical thymocytes but not on those cells located in the thymic medulla. When 20 cases of different types of lymphatic leukemias were studied with ATS, HTA was found on acute lymphoblastic leukemia cells rosetting with sheep erythrocytes (T‐ALL) but not on other lymphatic leukemia cases, including chronic lymphocytic leukemia cells rosetting with sheep red blood cells (T‐CLL). SDS‐PAGE analysis of immunoprecipitates formed between ATS and radiolabeled cell‐surface components of human thymocytes demonstrated that ATS bound a 48,000‐molecular‐weight component that could be adsorbed to Sepharose 4B coupled with Lens culinaris hemagglutinin and could be labeled by periodate‐tritiated borohydride, indicating that HTA is a sialoglycoprotein. Similar antigen molecules were also precipitated by ATS from T‐ALL cells but not from other lymphatic leukemia cells. Five T‐ALL‐derived cell lines were tested with ATS by immunofluorescence and by RIP and SDS‐PAGE, but only two cell lines (MOLT‐3 and P12/Ich) possessed HTA molecules on their cell membrane.
A defect in the formation of uropod‐bearing lymphocytes (hand‐mirror cells) in patients with the wiskott‐aldrich syndrome
Authors: McGraw Thomas P. , Folds James D. , Whisnant John K. , Phillips Terry M. , Stass Sanford A.
Published: February 2007

The formation of a uropod by lymphocytes (hand‐mirror cells) represents a morphologic stage of immune activation and motility in lymphocytes. Immune complexes have been previously shown to induce hand‐mirror‐cell (HMC) formation in human lymphocytes and have been associated with increased number of HMC in acute leukemia. We studied the ability of immune complexes to induce HMC in patients with Wiskott‐Aldrich syndrome (WAS), a disease of cellular immune deficiency. Our findings indicate that lymphocytes from patients with WAS adhere to immune complexes but have a defect in their ability to form HMC when compared with normal human lymphocytes. These results support the concept that immune complexes induce HMC formation and that the HMC is related to the normal cellular immune response.
In vitro synthesis of hemoglobin and hemoglobin chains in the BFUe‐derived colonies from persons with α‐ or β‐thalassemia
Authors: Huisman T. H. J. , Reese A. L. , Webber B. , Okonjo K. , Altay C. , Felice A. E.
Published: February 2007

The synthesis of α and non‐α chains of human hemoglobin (Hb) was studied in reticulocytes and in BFUe‐derived cell colonies from patients with α chain or β chain deficiencies. The subjects included normal adults (αα/αα) with or without a β chain variant (Hb S, Hb Leslie) or an α chain variant (Hb G‐Georgia); α‐thalassemia‐2 heterozygotes (α0α/αα) with an α chain variant (G‐Georgia or G‐Philadelphia); an α‐thalassemia‐1 heterozygote (α0α0/αα); α‐thalassemia‐2 homozygotes (α0α/α0α) with a β chain variant (Hb S), an α chain variant (G‐Philadelphia), a Hb S homozygosity with Hb G‐Philadelphia, or a Hb G‐Philadelphia homozygosity; and three black β+ ‐thalassemia homozygotes. Data from reticulocyte in vitro synthesis analyses showed the expected deficiencies. However, similar analyses of the Hb synthesized in cell colonies (even from the black β‐thalassemia homozygotes) gave (nearly) balanced ∑α/∑non‐α ratios. It is speculated that this balanced synthesis is due to a most effective proteolysis in the immature erythroblast which rapidly removes free α or β chains. The levels of Hb F and Hb A2 were considerably increased in these proerythroblasts; a two‐ to threefold increase in the synthesis of Hb A2 was observed over that seen in the reticulocytes.
Aspects of platelet formation and release
Authors: Scurfield G. , Radley J. M.
Published: February 2007

The surface architecture and location of megakaryocytes in the extravascular compartment of mouse bone marrow are considered. The finger‐like cytoplasmic processes that extend into sinuses pass through apertures, approximately 3.0 mμ in diameter, occurring singly or in groups. The cytoplasmic processes enlarge on first entering a sinus and form villi that seem to anchor them to the endothelium. The villi consist largely of microfilaments. The cytoplasmic processes extend, attenuate, and undergo irregular constriction along their lengths. The constrictions demarcate modicums of cytoplasm of platelet size one from another that are released when they break. Extravascular platelet release may also occur, and evidence is presented that points to the likelihood of the platelet's being engulfed and phagocytosed by macrophages before reaching the microcirculation.
Priapism in a non‐black with sickle cell anemia associated with α‐thalassemia
Authors: Fich Alexander , Rachmilewitz Eliezer A.
Published: February 2007

Sickle cell anemia, the most prevalent type of hemoglobinopathy, appears almost exclusively in blacks. Since the first report of sickle cell anemia in a Caucasian by Cooley and Lee in 1929 [1], additional cases have been reported, mainly from Mediterranean countries, the Persian Gulf, and India [2]. Several cases were also found among Arabs in Israel with a relatively benign clinical course [3, 4].

The present report describes a unique case of sickle cell anemia associated with α‐thalassemia in a non‐black young adult. The diagnosis was made at the age of 23 years. Severe priapism was almost the first manifestation of the disease and subsided following transfusion of five units of packed red blood cells.
Cellular kinetics in a patient with sezary syndrome
Authors: Chandra P. , Chanana A. D. , Chikkappa G. , Cronkite E. P.
Published: February 2007

Cellular kinetics and proliferation of Sezary cells (SC) were studied in a 48‐year‐old woman with Sezary syndrome (SS). In vitro flash labeling indices of peripheral blood (PB) SC were studied by labeling with tritiated thymidine (3HTdR) and tritiated cytidine (3HCdR). Intradermal SC were labeled in vivo by local injection of 3HTdR followed by skin biopsies of the injected sites. Traffic patterns of DNA labeled PB SC were studied by intravenous 3HTdR followed by sampling of PB, skin, lymph node (LN), and bone marrow (BM). Fluxes of PB SC in various tissues were investigated by autotransfusion of 3HCdR labeled PB SC followed by serial sampling of PB, LN, BM, and skin. In vitro response of PB SC to phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed (PWM) were also investigated. The results from these studies in this patient indicate that 1) proliferation of SC was primarily in the skin, 2) there was a negligible flux of SC from blood into skin, LN, and BM, and 3) the mitogenic response of PB mononuclear cells (mostly SC) to PHA, PWM, and Con A was poor.
Nonimmune interaction of leukocytes with platelets and megakaryocytes
Authors: Poon Man‐Chiu , Parmley Richard T. , Chang‐Poon Vivien Y‐H , Embry Joseph H. , Austin Ronald L.
Published: February 2007

Platelet–leukocyte interaction was observed in an asymptomatic woman. After incubation in the patient's EDTA‐plasma, autologous and allogeneic platelets adhered to the surfaces of neutrophils, monocytes, macrophages and, rarely, eosinophils. Monocytes, macrophages, and occasionally neutrophils phagocytosed platelets. Degranulation of peroxidase‐positive lysosomes into the platelet‐containing phagosome was demonstrated ultrastructurally. Bone marrow studies indicated that bands and earlier neutrophilic precursors did not participate in the reaction, and that neutrophils adhered to, and were rarely engulfed by megakaryocytes.

Sequential exposure of the patient's EDTA‐plasma to platelets and leukocytes indicated that a nondialyzable factor(s) was first absorbed by platelets which then interacted with leukocytes. The reaction proceeded best in the presence of EDTA at 21°C, and was inhibited or dissociated by divalent cations or at 37°C. Metabolic integrity of both platelets and leukocytes was also essential for the reaction since each was inhibited by formalin fixation and partially inhibited by the metabolic inhibitor 2‐deoxyglucose. Formalin‐treated platelets continued to absorb the plasma factor(s). The plasma and the cell fractions were inactivated by periodate and nonspecific protease. Treatment of the platelets with trypsin or the leukocytes with neuraminidase diminished the interaction by 50%. The reaction was also interfered with by concanavalin A. Immunofluorescent and immunoabsorption studies failed to identify an immune component of this interaction. These findings indicate that the plasma factor(s) and the cell surface receptors are nonimmune glycoconjugates and consequently differ from previously documented cases of platelet‐leukocyte interaction.
Clinical use of β‐thromboglobulin levels in diagnosing and treating consumptive and immune thrombocytopenia
Authors: Rubenstein Martin D. , Wall Robert T.
Published: February 2007

Plasma β‐thromboglobulin (β‐TG), a platelet‐specific protein, is a marker of intravascular platelet degranulation. We measured plasma β‐thromboglobulin by radioimmunoassay in 13 patients with thrombocytopenia of various etiologies to determine whether or not the test is clinically useful in the differential diagnosis of thrombocytopenia. Four patients with intravascular platelet consumption (three with thrombotic thrombocytopenic purpura and one with vasculitis) had significantly higher plasma β‐thromboglobulin levels than four patients with extravascular platelet destruction due to idiopathic thrombocytopenic purpura. Five patients with thrombocytopenia and decreased numbers of megakaryocytes in the bone marrow also had β‐thromboglobulin levels that were not elevated. Two patients with thrombotic thrombocytopenic purpura achieved clinical remission associated with a decrease in β‐TG level to the normal range. Plasma β‐thromboglobulin determination can be useful in determining the mechanism of thrombocytopenia when bone marrow examination demonstrates adequate megakaryocyte numbers.
Acute lymphocytic leukemic transformation of chronic lymphocytic leukemia: Substantiation by flow cytometry
Authors: Frenkel Eugene P. , Ligler Frances S. , Graham Mateel S. , Hernandez Jose A. , Kettman John R. , Smith R. Graham
Published: February 2007

Acute leukemia, during the course of chronic lymphocytic leukemia, is a rare event and only a small fraction of such cases have been shown to be true transformations to acute lymphocytic leukemia. In one previous case, evidence suggested this was transformation of a single neoplastic clone. The present report describes the clinical and laboratory findings in a case of chronic lymphocytic leukemia in which an acute leukemic pattern evolved. The fluorescence‐activated cell sorter provided the resolution needed to distinguish low numbers of surface immunoglobulins on the acute leukemic cells, verifying their identity as B lymphoblasts. Furthermore, a small population of large cells bearing the same light chain type as the acute leukemic cells were detected in the spleen 3½ months before clinical or laboratory evidence of the onset of the acute leukemia.
A case of lead intoxication: Clinical and biochemical studies
Authors: Miwa Shiro , Ishida Yoji , Takegawa Susumu , Urata Gumpei , Toyoda Toshio
Published: February 2007

A 23‐year‐old Japanese male with severe lead intoxication accompanied by hemolytic anemia was studied. The patient had taken 12 g of lead in about a month. He had moderate hemolytic anemia (Hb 8.9 g/100 ml) with reticulo‐cytosis ranging from 2.5 to 11.7%. Peripheral blood smear showed nucleated red cells (42/200 white blood cells) and marked basophilic stippling in the red cells. Activities of erythrocyte enzymes were either normal or increased except for pyrimidine 5′‐nucleotidase (P5N) and δ‐aminolevulinic acid dehydratase (ALA‐D) both of which were found to be decreased, being 48.8% and 4.1% of the normal controls respectively. Erythrocyte reduced glutathione (GSH) was high (145.6 mg/100ml RBC). Erythrocyte pyrimidine nucleotides were accumulated up to 10.2% of total nucleotides. The level of lead in peripheral blood was 112 μg per 100 ml blood.

Ca2+‐Na2‐EDTA was given to the patient as treatment. The level of lead in the blood decreased gradually and hemolytic anemia improved. Basophilic stippling in the red cells disappeared.

These results confirmed the findings of Valentine et al [1] that lead‐induced deficiency of P5N resulted in basophilic stippling and hemolytic anemia just like hereditary hemolytic anemia due to P5N deficiency.
Effect of chemical modification of sulfhydryl groups of human erythrocyte enzymes
Authors: Valentine W. N. , Paglia D. E.
Published: February 2007

Erythrocyte lysate proteins prepared by five methods were chemically modified by methanethiolation with methyl methanethiolsulfonate (MMTS). This relatively newly studied agent delivers the small uncharged, non‐hydrogen‐bonding CH3S‐group to accessible ‐SH groups under mild conditions. Glycolytic and nonglycolytic red cell enzymes studied could be grouped into four categories based on degree of catalytic inhibition by MMTS, which ranged from 100% to nil. NADH (0.06 mM) protected against lactate dehydrogenase (LDH) inhibition nearly completely, and partial protection was evident at 1 μM concentrations. NAD was virtually ineffective at comparable molarities, and pyruvate failed to protect significantly. Glucose‐6‐phosphate dehydrogenase (G‐6‐PD) (but not 6‐PGD) was protected by NADP (but minimally if at all by NADPH) at low molarities. Substrate G‐6‐P failed to protect. Mg++ alone protected enolase to a major degree against MMTS inhibition. Residual activities of several enzymes were rendered highly unstable to thermal stress, and in the case of selected enzymes, alteration of pH activity curves and catalytic activity at diminished substrate concentration was demonstrable. The highly specific modification of sulfhydryl groups of cysteinyl residues under mild conditions preserving other aspects of enzyme structure provides insights into the role of these most reactive of all amino side‐chains in the enzyme proteins derived from human red cell lysates.
Characteristics of enzymes of erythrocytes from newborn infants and adults: Activity, thermostability, and electrophoretic profile as a function of cell age
Authors: Mohrenweiser H. W. , Fielek S. , Wurzinger K. H.
Published: February 2007

The level of enzyme activity, the enzyme thermostability profile, and the isozyme electrophoretic pattern were determined in young and old erythrocytes from newborn infants and adults and in samples from adult individuals with increased reticulocyte counts. Cord blood samples had higher levels of enzymatic activity for 12 of the 14 enzymes measured, adenylate kinase and phos‐phoglucomutase being the exceptions. The largest differences in activity between newborns and adults were for glutamic oxaloacetic transaminase, hexo‐kinase, glucose 6‐phosphate dehydrogenase, and glutathione reductase, while glutamic oxaloacetic transaminase and pyruvate kinase showed the largest differences between young and old cells. The levels of activity of glutathione reductase, adenylate kinase, phosphoglucomutase, lactate dehydrogenase, phos‐phoglycerokinase, and glucose phosphate isomerase in cord blood samples suggest the regulation of expression of these enzymes is different in fetal erythrocytes than in erythrocytes from an adult. Differences in the thermostability profile of enzymes from cells from different sources and/or of different ages were noted for 5 of 9 enzymes. No unique electrophoretically identifiable fetal isozymes were observed, although differences in isozyme distribution and staining intensity associated with cell source and/or cell age were noted for many of the 23 enzymes examined. Many of these differences in enzyme characteristics have the potential to be confused with genetic alterations in enzyme structure and function.
Immune complexes in hemophilia
Authors: Poskitt Thomas R. , Poskitt Paula K. F. , Bean Cheryl A. , Arkel Yale S.
Published: February 2007

Circulating immune complexes (ICs), assayed by the L1210 enzyme‐linked immunoassay, were detected in 48% of patients with hemophilia A, 50% of patients with von Willebrand's disease, and in none of our patients with hemophilia B. Eighty‐five % of the hemophilia A and B patients had mild to moderate disease with only one patient demonstrating a circulating inhibitor. No correlation was found between IC levels and hepatitis B infection, SGOT, disease severity, total quantity of factor VIII or IX infused, time interval from last infusion, or rheumatoid factor positivity. Although the nature of the ICs is not known, the similarity of IC levels between hemophilia A and von‐Willebrand's disease is discussed with regard to antibodies generated to non‐procoagulant portions of the factor VIII molecule.
Effect of endotoxin on arachidonic acid release and thromboxane B2 production by human platelets
Authors: Stuart Marie J.
Published: February 2007

Plasma thromboxane A2, which is elevated during endotoxemia, has previously been shown to be a major factor contributing to the mortality and morbidity that occurs in endotoxin shock in the experimental animal. Using a minimal dose of Escherichia coli endotoxin (1 μg/ml), we have demonstrated that the preincubation of human platelets with endotoxin induces changes in platelet arachidonic acid release and the subsequent conversion of the released arachidonic acid to thromboxane B2, the stable end product of thromboxane A2. In paired experiments, in the presence of endotoxin, the addition of the aggregating agent thrombin (0.5 U/ml) caused human platelets to release 29.1 ± 3.4% of 14C‐arachidonic acid from prelabeled platelet phospholipids. This value was significantly elevated (P < 0.02) when compared with the release of 14C‐arachidonic acid from platelets in the absence of endotoxin (21.9 ± 3.6%). Similarly, comparison of the results of the conversion of the released arachidonic acid to platelet thromboxane B2 (TxB2) revealed that TxB2 production was significantly increased (P < 0.01) when human platelets were preincubated with endotoxin prior to the addition of thrombin (6.1 ± 0.6%) when compared with TxB2 formation observed in the absence of endotoxin (3.4 ± 0.5%). The absolute amount of released arachidonic acid that was converted to TxB2 in the presence of endotoxin (1.8 ± 0.3%) was also significantly higher (P < 0.01) than the value observed in its absence (0.8 ± 0.2%). This study suggests that one of the tissue sources of the proaggregatory vasoconstrictor thromboxane A2 during endotoxemia is the platelet.
Morphologic and functional heterogeneity of chronic neutropenia of childhood with normal neutrophil colony formation in vitro
Authors: Komiyama A. , Yamazaki M. , Yoda S. , Saitoh H. , Morosawa H. , Akabane T.
Published: February 2007

In order to obtain further knowledge of chronic neutropenia of childhood, we studied nine neutropenic infants six to ten months of age by in vitro techniques, including bone marrow culture, electron microscopy, and chemotaxis assay. Eight of the nine patients had a benign clinical course and the bone marrow aspirates showed a reduced number of segmented neutrophils. The ninth patient had a moderately severe course and the bone marrow showed maturation arrest at the promyelocyte stage. Bone marrow cultures demonstrated that the in vitro neutrophil colony formation and production of colony‐stimulating activity were normal in all of the eight patients studied. Neutrophils from one of the nine patients had ultrastructural abnormalities such as a decrease in number of primary and secondary granules and the presence of myelin figures in primary granules. Neutrophil chemotaxis was defective in three of the nine patients. All of the six patients in whom the neutrophil colony formation in agar, the ultrastructure of neutrophils, and neutrophil chemotaxis were normal recovered from the neutropenia between 11 and 30 months of age. These in vitro parameters appear to be useful for evaluating chronic neutropenia of childhood.
Standardization of platelet function tests
Authors: Roper‐Drewinko Pamela R. , Drewinko Benjamin , Corrigan Gail , Johnston Dennis , McCredie Kenneth B. , Freireich Emil J.
Published: February 2007

Methods for the standardization of several currently used platelet function tests (bleeding time, capillary fragility, platelet retention, platelet aggregation, platelet factor 3, and platelet volume profiles) are presented. Different variables that may interfere with the reproducibility of the results of each assay were identified and standardized. Using the standardized techniques, the range of normal values for each test was determined in a large population of normal volunteers and used to identify disease states by comparing patient results with those of the normal population. A format for presenting the entire profile of platelet function parameters is proposed.
Evidence for the generation of hydroxyl radical during arachidonic acid metabolism by human platelets
Authors: Singh Daljeet , Greenwald James E. , Bianchine Joseph , Metz Earl N. , Sagone Arthur L.
Published: February 2007

Reactive oxygen species, probably hydroxyl radicals (OH·), have been suggested to be generated during arachidonic acid (AA) metabolism and, once released, these species can modify the rate and extent of various reactions involved in AA metabolism. We have studied this phenomenon in washed human platelets. OH· generation was quantitated using 14C‐benzoic acid as a specific trap in a continuous ionization chamber system. Resting platelets did not produce any detectable signal, whereas addition of AA resulted in gradual OH· production with peak values detected at approximately 20 min. Similar studies conducted under nitrogen or after boiling the platelets almost abolished OH· generation. Aspirin had no significant effect, whereas 5,8,11,14‐eicosatetraynoic acid decreased the signal by > 90%, thus suggesting that OH· is produced primarily through the lipoxygenase pathway. Superoxide dismutase (SOD) and catalase had no effect and, as expected, phenol and mannitol decreased OH· production considerably, by > 50% and 90%, respectively. Azide and cyanide also reduced the OH· generation by about two‐thirds. We conclude that OH· is generated during AA metabolism by human platelets. It is primarily produced via the lipoxygenase pathway and may require a heme‐dependent peroxidase. This highly reactive oxidant may play an important role in normal and abnormal hemostasis.
Evidence that circulating immune complexes remove transfused platelets from the circulation
Authors: Kutti Jack , Zaroulis Charles G. , Safai‐Kutti Soodabeh , Dinsmore Robert E. , Day Noorbibi K. , Good Robert A.
Published: February 2007

Fifteen recently diagnosed patients with acute leukemias admitted for induction chemotherapy were selected for study. When thrombocytopenic (venous platelet count <20 × 109/1) these patients received prophylactic platelet transfusions. A total of 67 platelet transfusion therapies were administered and evaluated. Using the Raji cell radioimmunoassay, the serum concentrations of circulating immune complexes (CIC) were measured immediately before and 10–12 hr after each platelet transfusion. In 36 instances, elevated values of CIC were present in the recipient's pretransfusion samples, the corresponding posttrans‐fusion values being significantly lower (P < 0.05). Furthermore, in those 36 instances the mean percentage for the posttransfusion platelet increment was significantly lower (P < 0.001) than in the remaining 31 instances in which normal pretransfusion values of CIC were measured. We conclude that CIC were an important factor in rapidly removing transfused platelets from the circulation, thereby, adversely affecting the benefit of platelet transfusions.
Differences in synthesis of membrane proteins by leukemic cells from spleen and peripheral blood indicate distinct subsets of malignant cells in a patient with prolymphocytic leukemia
Authors: Spiro Robert C. , Ansell Jack , Katayama Isao , Muralidharan Kamala , Sullivan John L. , Humphreys Robert E.
Published: February 2007

Morphological and biochemical differences were demonstrated between prolymphocytic leukemia cells obtained from the spleen and peripheral blood of one patient. Peripheral blood prolymphocytes had consistently smaller nuclear‐cytoplasmic ratios than did splenic prolymphocytes. Percoll gradient‐purified prolymphocytes from the spleen synthesized abundant amounts of some membrane proteins which were hardly expressed by peripheral blood prolymphocytes. Peripheral blood prolymphocytes did not change their expression of membrane proteins during three days in culture. These findings are consistent with the view that prolymphocytic leukemia cells from the spleen exist, on the average, at an earlier stage of differentiation than do circulating leukemic cells, and that peripheral blood leukemic cells are frozen at a specific phase of differentiation.
Problems in measurement of erythrocyte calcium
Authors: O'Rear Ed. A. , Udden Mark M. , McIntire Larry V. , Lynch Edward C.
Published: February 2007

As calcium has increasingly been the object of study in erythrocyte physiology, we reviewed the current methodologies for determination of calcium by atomic absorption spectrometry. The published normal values for erythrocyte calcium vary from 5 to 500 μmol/liter of packed cells. A method based on Harrison and Long's determination of calcium is presented and shows normal red cell calcium concentration to be 0.0149 ± 0.0023 μmol/ml of packed red cells. The influence of temperature and type of crucible used in ashing red cells is assessed. The method of additions is employed to corroborate our results.
Primary splenic hairy cell leukemia — remission for 21 years following splenectomy
Authors: Myers Thomas J. , Ikeda Yasuo , Schwartz Stanley , Pharmakidis Angelo B. , Baldini Mario G.
Published: February 2007

A patient with primary splenic hairy cell leukemia is reported. This patient presented with massive splenomegaly and pancytopenia due to hypersplenism. Exploratory laparotomy failed to demonstrate any disease outside the spleen and splenic hilar lymph nodes. Splenectomy was the only form of treatment. During the following 21 years, no recurrent hairy cell leukemia has been found. This case allows speculation that hairy cell leukemia may originate in the spleen and that prolonged survivial or cure of the disease after Splenectomy alone may be due to removal of stem cells in the spleen.
Diagnosis and treatment of anticoagulant‐related adrenal hemorrhage
Authors: Anderson Kenneth C. , Kuhajda Francis P. , Bell William R.
Published: February 2007

Anticoagulant‐related adrenal hemorrhage, as a cause of adrenal insufficiency, is rarely suspected and diagnosed during life. We report three patients in whom prompt diagnosis allowed successful treatment with replacement doses of corticosteroids. In all three patients, a hypotensive episode preceded the signs and symptoms of acute adrenal insufficiency. The symptoms and signs of adrenal hemorrhage were nonspecific and variable, making clinical diagnosis difficult. In none of our patients was the prothrombin time, partial thromboplastin time, or Lee White clotting time prolonged beyond the therapeutic range, and in none were excessive doses of anticoagulant therapy employed. Characteristic serum chemistry abnormalities, serum ACTH stimulation tests, and daily urinary collections for free cortisol, 17‐OH ketosteroids, and 17‐ketosteroids confirmed the clinical suspicion of primary adrenal insufficiency in all cases. Diagnosis was aided in one patient by abdominal computerized axial tomographic scan. In all patients, response to replacement doses of corticosteroids was prompt and gratifying, underlining the importance of suspecting this complication of anticoagulant therapy and instituting prompt lifesaving therapy.

Adrenal hemorrhage is a complication of anticoagulant therapy that has been diagnosed during life in only 11 reported cases. We describe three patients in whom early diagnosis allowed prompt medical therapy and favorable outcome. The clinical presentation, diagnostic laboratory features, and therapeutic modalities are reviewed. Prolongation of prothrombin time, high levels of stress, and hypotension predispose to this complication in a patient maintained on anticoagulant therapy.
Two cases of red cell aldolase deficiency associated with hereditary hemolytic anemia in a japanese family
Authors: Miwa Shiro , Fujii Hisaichi , Tani Kenzaburo , Takahashi Keisuke , Takegawa Susumu , Fujinami Noboru , Sakurai Minoru , Kubo Masako , Tanimoto Yasuo , Kato Tomoaki , Matsumoto Noboru
Published: February 2007

Two cases of red cell aldolase deficiency associated with congenital nonspherocytic hemolytic anemia are reported. The proband is a fourteen‐month‐old Japanese boy. Consanguineous marriage was not proven but probable in this family, as the parents were born in the same small island. The proband had moderate to mild anemia aggravated by upper respiratory infections, 1 cm hepatomegaly and 2.5 cm splenomegaly, but was unremarkable in other respects and has thus far not shown mental or growth retardation. He did not have dysmorphic features. The red cell aldolase activity was 6% of the normal mean. The enzyme was unstable with respect to heat, and Km for fructose 1,6‐diphosphate (F‐1,6‐DP) was high. The parents and other heterozygotes showed intermediate activity between that of the proband and that of normal subjects. Red cell F‐1,6‐DP concentration in this case was remarkably increased. Red cell glucose consumption, and lactate formation, as well as hexose monophosphate shunt activity, were decreased as compared with a comparable reticulocyte‐rich hereditary spherocytosis patient. Hexose monophosphate shunt activity was assumed to be decreased due to inhibition of glucose 6‐phosphate dehydrogenase by a high concentration of F‐1,6‐DP in his red cells.

As a result of family study, another homozygous aldolase deficiency case associated with hemolytic anemia was found. He is 13 years old and a nephew of the proband's paternal grandmother. His hemolytic anemia also is moderate to mild and aggravated by upper respiratory infections. He does not seem to have mental or growth retardation, nor does he possess dysmorphic features.
Studies of nicotinamide adenine dinucleotide methemoglobin reductase activity in a jewish population
Authors: Moore Michael R. , Conrad Marcel E. , Bradley Edwin L. , Prchal Josef T.
Published: February 2007

In the original study of the deleterious effects of oxidant drugs on persons heterozygous for nicotinamide adenine dinucleotide methemoglobin reductase (NADH MetHb R) deficiency, several Jewish physicians used as controls had decreased enzyme activity. We collected blood samples from 555 Jewish persons to 1) estimate the heterozygote frequency for NADH MetHb R deficiency; and 2) determine if external variables such as age, gender, and ingested medications may alter the NADH MetHb R activity and thereby alter estimates of heterozygosity. Two persons possibly heterozygous for the deficiency were identified (carrier incidence = 1/277; calculated homozygote incidence = 1/309,000 Jewish births). A difference in enzyme activity was found between males and females (P < 0.03). We have examined the influence of the mean red cell age on the activity of erythrocyte NADH MetHb R and found it not to be materially influenced by mean red cell age; thus the possible intersex differences of mean red cell age cannot explain this observation. No significant difference (P > 0.1) was seen between persons ingesting the following drugs and those not: Dilantin; antidepressants/sedatives; cardiac, antineoplastic, or antigout/antiinflammatory drugs; thyroid supplements, estrogen‐containing preparations, and antihistamines. NADH MetHb R activity did not correlate with age or spontaneous abortion. We conclude that heterozygosity for NADH MetHb R deficiency is not prevalent among Ashkenazic Jewish persons, and that external variables should be considered to ensure accurate interpretation of NADH MetHb R activity for correct estimation of heterozygosity.
Existence of tartrate‐resistant acid phosphatase activity in differentiated lymphoid leukemic cells
Authors: Usui Tadao , Konishi Hiroshi , Sawada Hiroyoshi , Uchino Haruto
Published: February 2007

Acid phosphatase (AcP) in neoplastic cells from various lymphoid leukemias was examined. In the cytochemical studies, tartrate‐resistant AcP (T‐rAcP) activity was observed in the neoplastic cells from well‐differentiated lymphoid leukemias such as adult T‐cell leukemia (ATL), B‐cell chronic lymphocytic leukemia (B‐CLL), T‐cell chronic lymphocytic leukemia (T‐CLL), and hairy‐cell leukemia (HCL). T‐rAcP activity was also detected in a small number of leukemic cells obtained from T‐cell acute lymphoblastic leukemia (T‐ALL), while it was not detected in the neoplastic cells from null‐ALL, macroglobulinemia, and multiple myeloma (MM). In the electrophoretical studies, fraction 1 (F‐1), F‐3, F‐3b, and F‐4 were completely tartrate‐sensitive, while F‐2 was partially resistant and F‐5 was completely resistant. T‐rAcP activity (F‐5) was observed in ATL cells, B‐CLL cells, and HCL cells, while it was not detected in ALL cells, macroglobulinemia cells, and MM cells. The present study indicates that T‐rAcP activity is observed not only in HCL cells but also in the well‐differentiated lymphoid cells such as ATL cells, B‐CLL and T‐CLL cells except the most highly differentiated forms of B‐cells of MM and macroglobulinemia.
The effect of a low‐phosphate diet on hematocrit and oxygen transport in uremic rats
Authors: Connelly Thomas J. , Caro Jaime , Erslev Allan J. , Silver Ruth
Published: February 2007

Dietary restriction of phosphate was found to prevent the development of anemia in partially nephrectomized rats. In an attempt to examine the reason for this beneficial effect, hematologic and nephrologic studies were carried out on normal and on partially nephrectomized rats fed either a normal or a low‐phosphate diet. It was first found that a low‐phosphate diet ameliorates the degree of renal impairment found four weeks after partial nephrectomy. Nevertheless, it did not eliminate it, and the degree of uremia actually observed should have been associated with a significant reduction in hemoglobin and red cell mass. However, it did reduce serum phosphate and red cell 2,3‐diphosphoglycerate (2,3 DPG) levels and increase hemoglobin affinity for oxygen to a degree that should impair oxygen transport to the tissues. That the low phosphate actually caused tissue hypoxia with increased stimulation of the bone marrow was furthermore suggested by the observation that normal rats fed a low‐phosphorous diet developed a significant increase above normal in hemoglobin concentration and red cell mass. It was concluded that the effect of a low‐phosphate diet on the anemia of uremia is caused by a combination of reduced renal failure and increased tissue hypoxia.
Morphologic observations of contact‐induced lysis of EBV‐infected B lymphocytes by autologous hand mirror T cells
Authors: Thomas William J. , Duval‐Arnould Bertrand , Creegan William J. , Schumacher Harold R. , Forman David S. , Strong Douglas M.
Published: February 2007

To study the possible immunologic role of hand mirror lymphocytes (HML) in the control of Epstein‐Barr virus (EBV)‐infected B lymphocytes in patients with infectious mononucleosis (IM), we studied the in vitro interactions of these cells by time‐lapse video light microscopy. Peripheral blood T lymphocytes isolated from three patients in the early recovery phase of IM were mixed with their autologous EBV‐infected B cells. Motile lymphocytes with their characteristic hand mirror shape were observed to attach by their uropods to the B cells. The HML remained attached for variable periods ranging from 45‐75 min. Following detachment, B cells that were in contact with HML underwent lysis. Mixtures of T cells from healthy donors and EBV‐infected cell lines exhibited only rare uropod formation with no attachment or lysis of B cells. The present experiment indicates that contact‐induced lysis of EBV‐infected B lymphocytes is operative in IM and that this process is mediated by the HML.
The uptake of etiocholanolone by human bone marrow immature granulocytes
Authors: Minguell José J. , Lee S. , Helmer R. E. , Gardner F. H.
Published: February 2007

A cell fraction enriched in immature granulocytes was obtained after fractionation of human bone marrow by a three‐step procedure. This fraction sediments at an average density of 1.073 and has a high content of myelocytes and metamyelocytes. It showed the highest specific uptake of etiocholanolone when compared to other bone marrow cell fractions. Autoradiographic studies demonstrated a higher number of silver grains over myelocytes and metamyelocytes than in other cell types. These results suggest the presence in human bone marrow of a target cell for etiocholanolone, located in the late mitotic or postmitotic pool or granulocytes.
Kinetics and sites of destruction of 111indium‐oxine‐labeled platelets in idiopathic thrombocytopenic purpura: A quantitative study
Authors: Heyns A. Dup. , Lötter M. G. , Badenhorst P. N. , De Kock Felicity , Pieters H. , Herbst C. , van Reenen O. R. , Kotze H. , Minnaar P. C.
Published: February 2007

Kinetics andw quantification of the sites of destruction of 111‐Indium‐oxine‐labeled autologous platelets were investigated in eight patients with idiopathic thrombocytopenic purpura. The mean platelet count was 17 ± 9 × 109/liter; platelets were separated by differential centrifugation and labeled with 5.6 ± 2.5 MBq 111In. Whole body and organ 111In‐platelet distribution was quantitated with a scintillation camera and a computer‐assisted imaging system acquisition matrix. Areas of interest were selected with the computer and organ 111In‐radioactivity expressed as a percentage of whole body activity. Mean platelet survival was 49.5 ± 29.6 hr and the survival curves were exponential. Equilibrium percentage organ 111In‐radioactivity was (normal values in parentheses): spleen 33.7 ± 8.8(31.1 ± 10.2); liver 16.1 ± 9.5(13.1 ± 1.3); thorax 22.8 ± 3.7(28.2 ± 5.6). Percentage organ 111In‐activity at the time when labeled platelets had disappeared from the circulation was: spleen 44.5 ± 16.4 (40 ± 16); liver 16.0 ± 11.5 (32.4 ± 7.2); thorax 19.7 ± 6.0 (17.7 ± 10.3). Thorax activity corresponds to bone marrow radioactivity. Three patterns of platelet sequestration were evident. Three patients had mainly splenic sequestration, two mainly hepatic sequestration, and three diffuse reticuloendothelial system sequestration with a major component of platelets destroyed in the bone marrow. Splenectomy was performed in two patients. The pattern of 111In‐platelet sequestration was not predictive of response of glucocorticoid therapy or indicative of the necessity for splenectomy. Quantitative 111In‐labeled autologous platelet kinetic studies provide a new tool for the investigation of platelet disorders.