bluuebunny/crossref_metadata_2025 huggingface dataset:
Lymphocyte phenotype and function in chronically transfused children with sickle cell disease
Authors: Wang Winfred , Herrod Henry , Presbury Gerald , Wilimas Judith
Published: February 2007

Immunologic studies were performed on mononuclear cells from ten chronically transfused children with sickle cell disease, and the results were compared with those from five other groups: 21 sickle cell patients who were not receiving regular transfusions, 6 chronically transfused children with other forms of refractory anemia, 10 previously splenectomized children, 5 boys with hemophilia A, and 27 normal adult controls. The helper:suppressor T cell ratios (T4:T8) in all groups except hemophiliacs were normal, were unrelated to the number of units transfused, and were not suggestive of findings reported for patients with the acquired immune deficiency syndrome (AIDS) or groups at risk for the syndrome. Percentages of T3, T4, and T8 cells were low in sickle cell and splenectomized patients, but not in chronically transfused patients with other anemias. Serum IgG was frequently elevated, and IgG synthesis in vitro was increased relative to IgM synthesis in sickle cell patients. Coculture experiments indicated that such findings may stem from a selective increase in IgG synthesis by B cells. Thus, transfused sickle cell patients have a particular pattern of immunologic abnormalities that is distinct from that seen in AIDS.
Altered spectrin association and membrane fragility without abnormal spectrin heat sensitivity in a case of congenital hemolytic anemia
Authors: Ravindranath Yaddanapudi , Johnson Robert M.
Published: February 2007

In hereditary pyropoikilocytosis (HPP) and one type of hereditary elliptocytosis (HE), spectrin self‐association is abnormal [5,7]. Spectrin extracted from normal erythro‐cyte membranes at 0°C is nearly all tetrameric, while in HPP and HE (type 1) a substantial amount of the extracted spectrin is dimeric. Abnormal reassociation of spectrin dimers to tetramers can also be demonstrated. We here report the case of a family in which the child has moderately severe hemolysis, with extreme microcytosis and poikilocytosis. The spectrin extracted at 0°C was predominately dimer. Parents had levels of dimer intermediate between patient and control values. The temperature dependence was normal for (a) erythrocyte fragmentation; (b) spectrin extractability; and (c) circular dichroism of purified spectrin. Neither the patient nor either parent had elliptocytic red cells as judged from smears and scanning electron microscopy. The presence of substantial amounts of dimeric spectrin in the parents is consistent with a model in which each parent is heterozygous for a different nonassociating mutant spectrin, while the child has inherited a nonassociating molecule from each parent. In each individual, the degree of mechanical stability of the erythrocyte membrane, determined by ektacytometry, was proportional to the amount of tetramer found in the membrane. The description of this case is consistent with either HPP or a form of homozygous HE which is asymptomatic in the carrier state.
Granulopoiesis in patients with congenital neutropenia
Authors: Kawaguchi Yoshinori , Kobayashi Masao , Tanabe Akio , Hara Michimaru , Nishi Yoshikazu , Usui Tomofusa , Nagai Shinya , Nishibayashi Yoh‐Hei , Nagao Kenji , Yokoro Kenjiro
Published: February 2007

Granulopoiesis was investigated in five patients with congenital neutropenia (CN) (one Kostmann type, four benign forms). In semisolid agar culture, the marrow cells of all five patients produced normal numbers of CFU‐c (colony‐forming unit‐culture). The size and classification of colonies were normal. In suspension culture in vitro with exogenous colony‐stimulating factor (CSF) generated from omental‐conditioned medium (OMCM), the myeloid precursors of all patients could proliferate and differentiate into normal polymorphonuclear neutrophils (PMNs). But in the absence of exogenous CSF, myeloid precursors of the patient with Kostmann‐type CN did not proliferate or differentiate into PMNs at all. In the four patients with benign neutropenia, however, PMNs were found even without exogenous CSF similar to normal individuals. These results suggest that patients with CN may have normal granulopoietic stem cells with normal proliferative and differentiating capacity in response to exogenous CSF.

When a small amount of normal human serum was added to normal marrow cultures stimulated by exogenous CSF, the colony growth increased in a superadditive manner. The enhancing activity of serum from neutropenic patients differed from that of normal serum. Especially, the addition of serum from the patient with Kostmann type CN to normal marrow cultures did not show this enhancement effect. The sera of patients with benign neutropenia had less enhancement effect than did normal control serum. These findings might be interpreted as showing an imbalance between CSF enhancer and inhibitors in the patients' serum.
T‐cell subpopulations in patients with monoclonal gammopathies: Essential monoclonal gammopathy, multiple myeloma, and Waldenstrom macroglobulinemia
Authors: Miguel J. F. San , Caballero M. D. , Gonzalez M.
Published: February 2007

T‐cell subsets defined by monoclonal antibodies (OKT3, OKT4, and OKT8) were analyzed in 117 patients with monoclonal gammopathies—69 multiple myeloma (MM) (30 untreated and 39 treated), 14 Waldenström's macroglobulinaemia (WM), and 34 essential monoclonal gammopathy (EMG) patients. The percentage and absolute numbers of total T‐lymphocytes (E+, OKT3+ cells) were within the normal range in all groups except for the treated MM patients, in which a decrease in the absolute number could be observed. The percentages of OKT4+ cells were significantly lower in MM (35 ± 1.7) than in EMG patients (43 ± 2) and controls (50 ± 2). In contrast, OKT8 cells correspondingly increased in MM (38 ± 1.6) compared with EMG patients (29 ± 1) and controls (27 ± 1). The OKT4/0KT8 ratio was lower in MM than that in EMG patients and controls (p < 0.01) and was shown to be one of the four most significant variables in a linear discriminant analysis used to distinguish between MM and EMG groups. The MM patients in clinical stage III as well as Bence‐Jones myeloma patients showed a more pronounced OKT4/0KT8 imbalance. The treatment did not influence the percent distribution of T‐cell Subpopulations.

The patients with WM exhibit an alteration in the distribution of the T‐cell subsets similar to the MM patients with a T4/T8 ratio of 1.1±0.1. This imbalance was more pronounced in WM patients with monoclonal B‐lymphocytes in peripheral blood (leukaemic phase of WM).

The functional significance of the altered T‐cell subsets in MM and WM patients remains to be established, though it is probable that such an imbalance plays an important role in regulating these B‐cell proliferations.
Combined factor V/VIII deficiency: A case report including levels of factor V and factor VIII coagulant and antigen as well as protein C inhibitor
Authors: Brown Janet M. , Selik Nancy R. , Voelpel Mary Jo , Mammen Eberhard F.
Published: February 2007

Comprehensive coagulation studies were performed on members of a family with combined factor V/VIII deficiency. The purpose of these studies was to investigate the hypothesis that combined factor V/VIII deficiency is due to a lack of the inhibitor to activated protein C. The analyses performed included routine APTT and PT, factor V and VIII coagulant activity and antigen levels, van Willebrand factor levels, protein C antigen assay, and both protein C inhibitor activity and antigen levels. Three of the 19 family members studied were found to have a deficiency of both factors V and VIII. These three individuals showed prolonged APTTs and PTs and decreased levels of factor V and factor VIII coagulant activity and antigen. Factor VIII related antigen and ristocetin cofactor (von Willebrand factor) levels were normal. Protein C and both protein C inhibitor activity and antigen levels were also found to be normal. These findings confirm the results of other recent investigators and indicate that the autosomal, inherited combined factor V/VIII deficiency is not due to a protein C inhibitor deficiency. The real defect in this combined deficiency remains to be determined.
Impaired IgM antibody responses to an influenza virus vaccine in adults with sickle cell anemia
Authors: Ballester Oscar F. , Abdallah Jørge M. , Prasad Ananda S.
Published: February 2007

Type‐specific IgM and IgG antibody responses to a polyvalent influenza vaccine were evaluated in 16 adults with sickle cell anemia, with the use of an enzyme‐linked immunosorbent assay. When compared to healthy controls, 8 out of the 16 patients had decreased or undetectable postvaccination anti‐influenza IgM antibody levels. These patients were found to have significantly lower serum IgM levels and nondetectable splenic tissue (by 99Tc scans), as compared to those with normal IgM responses. Impaired IgM antibody primary immune responses may playa role in the pathogenesis of infectious complications seen in adult patients with sickle cell anemia.
Lymphocyte subpopulation abnormalities in sickle cell anemia: A distinctive pattern from that of AIDS
Authors: Ballester Oscar F. , Abdallah Jorge M. , Prasad Ananda S.
Published: February 2007

In this study, we evaluated lymphocyte subpopulations in 23 adults with SCA. When compared to controls, SCA patients had higher lymphocyte counts with normal numbers of T101+ cells (T‐lymphocytes) and T4 + cells. T8+ cells were significantly increased in SCA patients in comparison to controls (1684 ± 243 vs 980 ± 367, p < .001). This increment was largely dependent on a T101 ‐, T8+ cell population. The SCA patients as a group had significantly decreased T4/T8 ratio (p < .0001). The SCA patients with history of blood transfusions had higher T4+ cells numbers and higher T4/T8 ratio, but no other significant differences from nontransfused patients were noted. Our results are different from those reported for nontransfused children with SCA who showed normal T4/T8 ratio.

Thus, a distinct pattern of abnormalities is seen in the lymphocyte subpopulations of adult SCA patients, unrelated to their exposure to blood transfusions.
Acute nonlymphocytic leukemia with eosinophilic differentiation
Authors: Gabbas Attilio G. , Li Chin‐Yang
Published: February 2007

Four cases of de novo acute nonlymphocytic leukemia (ANLL) with early eosinophilic differentiation are described. The clinical course did not differ from that of the usual forms of ANLL. Morphologic and cytochemical features that can support this diagnosis are discussed. Particularly, the cyanide‐resistant peroxidase stain appeared to be a specific marker of eosinophilic differentiation. Acute eosinophilic leukemia is a distinct entity, and this unusual subtype of ANLL can be set apart from other forms of ANLL characterized by hypereosinophilia.
Decreased adenosine deaminase (ADA) and 5′nucleotidase (5NT) activity in peripheral blood T cells in hodgkin disease
Authors: Murray James L. , Perez‐Soler Roman , Bywaters Daniel , Hersh Evan M.
Published: February 2007

The purine metabolic enzymes adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and 5′nucleotidase (5NT) play an important role in normal lymphocyte differentiation. Abnormal levels of one or all of these enzymes have been associated with immunodeficiency diseases and lymphoproliferative disorders. ADA, PNP, and 5NT activity was measured in peripheral blood T cells from 24 patients with Hodgkin disease (HD) (12 in complete remission and 12 with active disease) to determine whether an association existed between enzyme abnormalities and the decreased cellular immune function previously described in this disorder. HD patients had a significantly decreased absolute lymphocyte count (1,618 ± 1107/mm3; mean ± SD) compared to controls (2,320 ± 980; p <.001). ADA, PNP, and NT activity was assessed in lymphocyte extracts by measuring the conversion of radiolabeled substrates to products over time. ADA activity expressed as mean ± SEM nanomoles/106 lymphocytes/hr was significantly decreased in T cells from HD patients (84.6 ± 7.5) compared to controls (128 ± 12.3; p < 0.025). Likewise, 5NT was significantly decreased in HD patients (12.7 ± 1.3) compared to controls (24.0 ± 3.6; p <.005). There was not a significant difference in PNP activity between both groups. Low 5NT activity was present irrespective of whether patients had active disease (12.1 ± 1.5) or were in unmaintained complete remission (14.5 ± 2.4). These findings suggest that biochemical abnormalities may be responsible for or related to the persistent abnormalities in T‐cell function noted throughout the clinical course of HD.
Influence of the poly (ADP‐ribose) polymerase inhibitor 3‐aminobenzamide on macrophage and granulocyte differentiation of HL‐60 cells
Authors: Damji Nazim , Khoo Kong Eng , Booker Lynda , Browman George P.
Published: February 2007

We investigated the influence of the poly (ADP‐ribose) polymerase inhibitor 3‐aminobenzamide (ABA) on induction of phenotypic markers of granulocyte differentiation by retinoic acid and markers of macrophage differentiation by TPA in HL‐60 cells. The differentiation of HL‐60 cells towards the granulocyte lineage was assessed by hexose monophosphate shunt activity, proportion of cells capable of reducing NBT dye, and the appearance of recognizable neutrophils and bands. The effect of ABA and retinoic acid on NBT dye reduction and appearance of mature neutrophils and bands was synergistic, whereas the effects of these agents on hexose monophosphate shunt activity were additive. The differentiation inducing capacity of ABA in the presence of retinoic acid was dose‐related. The influence of ABA on TPA‐induced markers of macrophage differentiation was assessed by determining the proportion of adherent cells produced after treatment and by measuring acid phosphatase activity in the adherent cell fraction. In the presence of ABA, the number of cells adhering to plastic declined after day 2 of exposure to TPA, and acid phosphatase activity in adherent cells was inhibited fourfold (p = 0.01). The influence of ABA on the phenotypic markers of granulocyte and macrophage differentiation was detectable at concentrations that were not cytotoxic. The influence of ABA on HL‐60 differentiation is similar to that previously reported for human bone marrow CFU‐GM. Our data suggest that poly (ADP‐ribose) polymerase plays a role in differentiation of HL‐60 cells and that HL‐60 might provide a useful model for evaluating control mechanisms involved in the differentiation of CFU‐GM.
Slow infusion of vincristine in the treatment of idiopathic thrombocytopenic purpura
Authors: Manoharan Arumugam
Published: February 2007

Ten patients with idiopathic thrombocytopenic purpura (ITP) were treated wtih vincristine (VCR), given as a slow infusion over a 4‐hr period, at weekly intervals for 4 weeks. The VCR therapy achieved a sustained recovery of platelet count in five patients with ITP of 2 weeks to 5 months duration; a transient recovery was observed in four patients with ITP of 6 months to 5 years duration. Therapy had no effect at all in one patient who had ITP of 10 years duration. Vincristine therapy appears to be therapeutically more beneficial when given as a slow infusion and can achieve sustained recovery of platelet count in patients with ITP of less than six months duration.
A naturally occurring, warm‐reactive macroglobulin specific for papain‐treated human platelets: Preliminary characterization
Authors: Gill Joan C. , Carlson Patricia , Kunicki Thomas J. , Aster Richard H.
Published: February 2007

Sera from 28 of the 113 normal children and adults (25%) studied were found to contain an immunoglobulin capable of causing complement‐dependent lysis of normal platelets treated with small quantities of papain. This factor reacts equally well at 4°C and at 37°C with a determinant induced on platelets from normal subjects by treatment with papain or bromelain, but not by trypsin, chymotrypsin, or neuraminidase. It does not bind to red cells treated with any of these enzymes. The site(s) for which the factor was specific could not be induced on platelets from six patients with type I Glanzmann's thrombasthenia (lacking glycoproteins IIb and IIIa), in contrast to platelets from each of 20 normal donors. Isolation and characterization of the factor has been difficult because of its intolerance to chemical and physical manipulation. In 11 of the 20 individuals studied, however, it was found to have the properties of an IgM immunoglobulin. The factor appears to be different from any previously described, naturally occurring human immunoglobulin. It has not yet been shown to be associated with any disease state, but in the presence of complement, it is capable of causing profound damage to platelets previously subjected to minimal proteolysis, and the possibility that it can provoke platelet destruction in some conditions deserves further study.
Hb bicětre or α2β263(E7)His → pro in a white male: Clinical observations over a period of 25 years
Authors: Miller D. R. , Wilson J. B. , Kutlar A. , Hulsman T. H. J.
Published: February 2007

A description is given of clinical observations over a period of 25 years for a 26‐year‐old white male with a heterozygosity for the β‐chain variant, Hb Bicětre, in which β63His is replaced by Pro. The variant was identified through analysis of tryptic peptides from a digest of the βX‐chain isolated by PCMB precipitation and purified by chromatography. Similarity between the clinical histories of the first (French) heterozygote and our patient was most striking, including improvement after splenectomy.
The effect of erythrocyte membrane on the birefringence formation of sickle cell hemoglobin
Authors: Mizukami H. , Bartnicki D. E. , Burke S. , Brewer G. J. , Mizukami I. F.
Published: February 2007

The birefringence formation of sickle cell hemoglobin (HbS) in a thin liquid layer was observed while its environment was deoxygenated at different rates, and the effect of membrane was examined.

Under slow rate of deoxygenation at 37°C, at pH 7.4, the birefringence of purified HbS appeared at a concentration higher than 24% and its relative magnitude increased as the concentration was increased. Similarly, the partial presure of oxygen, at which the birefringence formation was evident, increased from 0 to 27 torr as the concentration of HbS was increased from 24 to 28%, but it remained the same above this protein concentration.

In all the samples tested relative birefringence was largest at the slow rate of deoxygenation (30 torrO2/min) and the magnitude decreased as the rate of deoxygenation was increased. The samples showed different sensitivity to the rate of deoxygenation. For example, while the total untreated hemolysate made by freeze‐thawing of packed sickle cells was most resistant to the increased rates of deoxygenation, purified HbS was not. Washed open ghosts partially restored the birefringence formation pattern of purified HbS.

The results indicate that the inner surface of the membranes of erythrocytes could behave as a template for large HbS polymer formation at relatively higher rates of deoxygenation.
Nuclear bridging of erythroblasts in acquired dyserythropoiesis: An early and transient preleukemic marker
Authors: Bethlenfalvay Nicholas C. , Phaure T. A. J. , Phyliky Robert L. , Bowman Robert P.
Published: February 2007

The clinical, hematologic, and histological characteristics of two patients who progressed from refractory anemia to acute leukemia are described. When first studied, nuclear bridging of erythroblasts, similar to that seen in congenital dyserythropoietic anemia type I and megakaryocytic dysplasia, were the only abnormalities. Within 6 years, both patients died, the first of acute nonlymphocytic leukemia, the second of erythroleukemia. Nuclear bridging of erythroblasts in the marrrow of these patients was an early and transient phenomenon and was not observed during the terminal phase of leukemia.
Deoxythymidine and deoxycytidine incorporation into DNA in B leukemic cells
Authors: Vertongen F. , Lebacq A.‐M. , Fondu P. , Mandelbaum I. M.
Published: February 2007

Measurements of radioactive DNA after incubation of normal and B leukemic peripheral mononuclear cells, from c‐ALL and CLL with labeled deoxythymidine (dTh) and deoxycytidine (dCt) showed that for dTh, incorporation into DNA was similar for normal and c‐ALL cells but lower in B‐CLL cells and that for dCt, incorporation was highest in c‐ALL and lowest in CLL cells. These results contrast with those of dTh and dCt kinase activities; the former has been previously found elevated in c‐ALL cells, and the latter is found, in the present study, similar in the three groups tested.
Vitamin E consumption by human blood platelets activated by latex particles
Authors: Principe Domenico Del , Menichelli Adriana , Lubrano Riccardo , Bandino Donatella , Giulio Stefano Di , Di Corpo Maria Luisa , Giardini Omero
Published: February 2007

Human blood platelet activation elicited by latex particles is associated to a 30% decrease in the cellular content of vitamin E. The vitamin E consumption is inhibited by the addition of catalase (500 U/ml) and azide (1 mM), but it is not affected by potassium cyanide (1 mM). It may be proposed that the challenge of platelets with particulate stimuli causes generation of oxygen reduction products, which leads to vitamin E depletion.
Studies on the effects of primary therapy for DIC following circulatory arrest
Authors: Takemoto Yoshinobu , Tanaka Shigeru , Tanabe Jun , Nakamura Yoshihiro , Kohama Akitsugu , Shibata Susumu
Published: February 2007

Comparative studies in early treatment for DIC caused by circulatory arrest were carried out on three groups of dogs during 300 min from the recovery of circulatory arrest: a heparin group, a FOY (gabexate mesilate) group, and a FUT‐175 (nafamostat mesilate) group. The parameters employed were platelet count, prothrombin time (PT), activated partial prothrombin time (APTT), fibrinogen, antithrombin‐III (AT‐III), and fibrin or fibrinogen degradation products (FDP).

In the heparin group, there was less of a drop in the platelet count and the level of AT‐III than in the control group, but the FDP levels were the same as in the control group. The PT and APTT remained within normal limits in the FOY group and no decrease was observed in either platelet count or AT‐III levels. In addition, FDP levels were kept within normal limits. In the FUT‐175 group, prolongation of APTT, no decline in the platelet count and AT‐III levels, and normal levels of FDP were observed.

The results of these experiments indicate the importance of early treatment for DIC. Judging from the parameters, better results were obtained in the FOY and FUT‐175 group than in the heparin group.
Teardrop‐shaped red cells in autoimmune hemolytic anemia
Authors: Farolino Deborah L. , Rustagi Pradip K. , Currie Mark S. , Doeblin Thomas D. , Logue Gerald L.
Published: February 2007

The presence of teardrop‐shaped red cells in peripheral blood has traditionally been felt to reflect altered marrow architecture, namely myelofibrosis. We evaluated two patients with splenomegaly, moderately severe hemolytic anemia due to warm‐reactive IgG anti‐red cell autoantibody, and bone marrow erythroid hyperplasia without myelofibrosis. A striking predominance of teardrop‐shaped red cells was noted upon examination of their blood films. Removal of a spleen containing extramedullary hematopoiesis in one and resolution of splenomegaly in the other were accompanied by disappearance of these cells. Our observations support a role for the spleen and for extramedullary hematopoiesis in the pathogenesis of this distinctive red cell morphologic abnormality.
Immunoregulatory abnormalities in myelodysplastic disorders
Authors: Baumann Michael A. , Milson Thomas J. , Patrick Catherine W. , Libnoch Joseph A. , Keller Robert H.
Published: February 2007

Immunoregulation was assessed in a group of patients with myelodysplasia (MDS) by flow cytometric analysis of peripheral blood lymphocyte subsets and in vitro studies of mitogen‐stimulated T‐lymphocyte blastogenesis. Mitogenesis was significantly depressed in MDS patients compared to controls (p < .001) and a similar defect was found in a small group of patients with untreated acute nonlymphocytic leukemia (ANLL) (p < .005). The impaired mitogenic response ability of T‐cells in these patients did not appear to be the result of alteration in lymphocyte subpopulation ratios. The observed defect might result from defective cooperation between Tlymphocytes and abnormal myeloid elements. Alternatively, the lymphocytes themselves could be derived from the abnormal clone and thus be functionally abnormal.
Protein C levels in nephrotic syndrome: Use of a new enzyme‐linked immunoadsorbent assay for protein C antigen
Authors: Soff Gerald A. , Sica Domenic A. , Marlar Richard A. , Evans Herbert J. , Qureshi G. Dastgir
Published: February 2007

Activated protein C is a potent, physiologic anticoagulant that inactivates the activated forms of factors V and VIII as well as facilitates in vivo fibrinolysis. We developed a competitive protein‐binding enzyme‐linked immunoadsorbent assay (ELISA) for protein C that was utilized to investigate if the hypercoagulability of the nephrotic syndrome is related to a deficiency of circulating plasma protein C. Protein C was measured in plasma of 11 patients with nephrotic syndrome (24 hr protein 8.4 ± 1.6 g, SEM; serum creatinine 4.2 ± .74 mg/dl, SEM). Ten azotemic nonnephrotic patients were employed as controls (serum creatinine 6.0 ± 1.25 mg/dl, SEM). No significant reduction of protein C values was observed (mean 108%, range 65–200%) in nephrotic patients when compared with the controls (mean 127%, range 100–200%) even though protein C antigen was present in all nephrotic urine samples tested. Also, no correlation existed between blood levels of urea nitrogen, creatinine, albumin, total protein, or 24‐hr urine protein excretion and the observed protein C values. These results suggest that in patients with the nephrotic syndrome, a hyper‐coagulable state may not be related to a deficiency of protein C and that the level of this zymogen in nephrotic syndrome reflects a dynamic balance between urinary losses and systemic production.
A patient with the inability to maintain in vivo levels of bound cobalamin (Cbl) and manifestations of tissue deficiency of Cbl
Authors: Begley James A. , Burkart Peter T. , Hall Charles A.
Published: February 2007

A 39‐year‐old woman presented with mild anemia, glossitis, an increased MCV, a low serum cobalamin (Cbl) (vitamin B12), mild tissue deficiency of Cbl, but with neither malabsorption of Cbl, impaired intake, nor deficiency of or inactivity of transcobalamin II (TC II). Because of a persistently low holo‐TC II (TC II carrying Cbl as the circulating complex of TC II‐Cbl), much of the evaluation was focused on the patient's TC II. Her TC II promoted the uptake of Cbl, reacted with anti‐TC II, and bound Cbl in vitro. A test dose of 200 μg of cyanocobalamin (CN‐Cbl) i.m. increased her holo TC II to levels higher than those in healthy persons, but with a much more abrupt fall to a subnormal level. Two milligrams of CN‐Cbl i.m. followed by 100 μg i.m. monthly failed to maintain normal amounts of circulating TC II‐Cbl or to overcome the tissue deficiency of Cbl. One milligram i.m. weekly or daily p.o. corrected both. The low holo TC II was considered to be responsible for the clinical expression and may have been primary to the reduced amounts of total and holo R binder of Cbl in the circulation. This study of a newly recognized defect points out the need for circulating holo TC II, a rational use of pharmacologic amounts of Cbl, and a possible interrelationship between TC II and the R binder of Cbl.
The effect of acute and chronic leukapheresis on the natural killer (NK) cell function of normal human volunteers
Authors: Stevenson Henry C. , Beman Joann S. , Crisp Beth , Coggin David , Kanapa D. J. , Miller Paul , Herberman Ronald B. , Maluish Annette E.
Published: February 2007

Twenty‐two normal volunteers had approximately eight, 2‐hr‐long leukapheresis procedures over a 2‐year period and their natural killer (NK) cell function was prospectively measured. The NK activity of the preprocedure peripheral blood (pre‐PB) was found to correlate well with the NK activity of the inital leukocytes removed by leukapheresis (I‐Leuk). When the I‐Leuk specimens were compared with the leukapheresis specimens removed at the termination of leukapheresis (T‐Leuk), T‐Leuk showed a consistent 10% increase in NK activity. When the pre‐PB and the I‐Leuk values were analyzed for each donor over the 2 years of the study, 18 donors revealed no significant change from their baseline NK activity, two donors showed a minimal increase in NK cell activity, and two donors displayed a minimal decrease in NK cell activity. We conclude that although leukapheresis appears acutely to boost NK cell activity, this increase is transient and small in magnitude. Most importantly, repeated leukapheresis does not appear adversely to effect this important effector function in normal donors.
Changes in platelet β‐thromboglobulin, fibrinogen, albumin, 5‐hydroxytryptamine, ATP, and ADP during and after surgery with extracorporeal circulation in man
Authors: Mezzano Diego , Aranda Eduardo , Urzúa Jorge , Lema Guillermo , Habash Jeanette , Irarrázabal Manuel J. , Pereira Jaime
Published: February 2007

Surgery with extracorporeal circulation (ECC) is associated with transient alterations of platelet function whose pathogenic mechanisms are not completely understood. To define further the platelet abnormalities, we determined the changes in platelet content of several granule‐associated substances during and after ECC in patients subjected to aortocoronary bypass surgery.

Platelet β‐thromboglobulin (β‐TG) decreased to 79.8% of the preoperative level at the end of ECC (p < 0.01) and, as expected, did not recover 1 hr after the end of surgery. Platelet fibrinogen and albumin decreased to 67.9% (p < 0.01) and to 29.8% (p < 0.01) of baseline, respectively. However, 1 hr after surgery, platelet fibrinogen rose to 92% and albumin to 55.5% of baseline, denoting that during the recovery from ECC, platelets incorporate some plasma proteins.

During ECC, platelet 5‐hydroxytryptamine (5‐HT) and total ATP and ADP decreased to 50.8% (p < 0.01), 63.2% (p < 0.01), and 69.9% (p < 0.01) of their respective preoperative values, indicating dense body release. One hour after surgery, ATP recovered to 83.8%, suggesting that previous depletion compromised also the metabolic pool of adenine nucleotides.

In summary, 1) Our results confirm and extend previous observations demonstrating α‐granule release during ECC. 2) Platelets undergoing ECC can incorporate plasma proteins as evidenced by the rapid increase of platelet fibrinogen and albumin after bypass. Although the mechanisms of this increase and site of storage of the exogenous substances are unknown, this observation justifies further studies to determine if internalization of plasma proteins, especially fibrinogen, may take place in physiological conditions. 3) Dense body depletion with transient storage pool deficiency appears to be a component of the reduced platelet function during ECC. 4) Consumption of metabolic ATP with alteration of platelet energy metabolism may further impair platelet function, contributing to the bleeding episodes observed during surgery with ECC.
Burkitt's cells can be triggered by teleocidin to secrete interferon‐γ
Authors: Benjamin David , Hartmann Dan P. , Bazar Leonard S. , Jacobson Robert J.
Published: February 2007

The secretion of interferon (IFN)‐γ by T lymphocytes is mediated by the synthesis of interleukin 2 (IL‐2) and the availability of IL‐2 receptors. Since some Burkitt's lymphoma lines express Tac antigen and can be triggered to secrete IL‐2 following activation with the new tumor promoter teleocidin, we addressed the question of whether the induction of IL‐2 by B lymphocytes is accompanied by the induction of IFN‐γ. IFN‐γ has not been detected in any of the 25 cell lines studied, and following stimulation with teleocidin, we triggered the synthesis of IFN‐γ in JLP(C), a pre‐Burkitt's cell line.

The mechanism of IFN‐γ secretion by B lymphocytes is not clear. Our findings demonstrate that the synthesis of IL‐2 by B cells is not accompanied by IFN‐γ and suggest that the synthesis of IFN‐γ is not mediated by IL‐2 or IL‐1 or B‐cell growth factor. Neutralization studies have shown that IFN‐γ secretion is not accompanied by the induction of IFN‐α or IFN‐β.

Our data imply that B cells can be triggered to secrete IFN‐γ under certain circumstances. Whether similar function occurs in vivo is not known.
Terminal deoxynucleotidyl transferase activity in acute leukemia: A study of 100 cases comparing an immunoperoxidase (PAP) vs immunofluorescent method
Authors: Krause John R. , Brody Judith P. , Kaplan Sandra S. , Penchansky Lila
Published: February 2007

A comparison between the immunofluorescent (IF) method for terminal deoxynucleotidyl transferase (TdT) activity and the immunoperoxidase (IP) method by peroxidase‐anti‐peroxidase (PAP) technique was done for 100 cases of acute leukemia. For the acute lymphoblastic leukemias (ALL) there was agreement in 93% of the cases. However, the IP method detected 51/55 (93%) TdT+ cases versus 47/55 (85%) by the IF method. For the acute nonlymphocytic leukemias (ANLL), there was an agreement in 89% of the cases. The IP method detected 8/36 (22%) TdT‐positive cases while IF detected 4/36 (11%) positive cases. If a figure of 10% TdT+ cells is considered significant in the marrow of the ANLLs, then the IP method would detect eight additional cases for a total of 16/36 (44%) TdT+ cases. This latter figure questions the ability of the IP TdT assay as a single test adequately to determine the lineage of a cell line. It may be rather that TdT is a marker that is expressed in a stem cell.
The β0‐thalassemia in an American black family is due to a single nucleotide substitution in the acceptor splice junction of the second intervening sequence
Authors: Padanilam B. J. , Huisman T. H. J.
Published: February 2007

An AG dinucleotide is an invariant feature of all acceptor splice sites, and deletion or substitution of (one of) these nucleotides will result in abnormal processing of the β‐globin mRNA. Restriction endonuclease mapping of DNA from an American black patient with Hb S‐β0‐thalassemia failed to detect any deletion in the β0‐globin gene region, but cloning and sequencing of the β0‐globin gene showed a point mutation (A → C) in the highly conserved dinucleotide AG of the acceptor splice junction of the IVS‐2. Blot hybridization analysis of RNA prepared from the erythroid cells of the patient showed only RNA of normal size. The patient and her daughter, who has the same condition, have high levels of Hb F (27%–35%); the mechanism responsible for the greatly increased γ chain production remains unclear.
Acute lymphocytic leukemia in children presenting with bone marrow necrosis
Authors: Macfarlane S. D. , Tauro G. P.
Published: February 2007

Bone marrow necrosis has been regarded as an indicator of very poor prognosis in malignant disease. The cause and incidence are unknown, and reports of treatment response are few. We describe four children with marrow necrosis at presentation with acute lymphocytic leukemia (ALL), all of whom entered remission with standard treatment showing complete marrow healing. The bleak outlook for patients with marrow necrosis based on early experience in adults with disseminated malignancy does not appear to apply to children with ALL. The incidence of marrow necrosis at diagnosis of childhood ALL is 1%.
Zinc inhibits FPA release and increases fibrin turbidity
Authors: Marx Gerard , Hopmeier Pierre
Published: February 2007

Physiologic concentrations of Zn(II) (4–40 μM) can increase the rate of thrombin‐induced fibrin clot formation (decreased clotting time, CT) and increase the turbidity of the fibrin gel. Both the initial and ultimate turbidity (Abs600 mm) of fibrin gels are increased in the presence of Zn(II). Two techniques were used to elaborate the mechanisms of Zn+2 procoagulant effect. Analytical ultracentrifugation indicates that Zn(II) does not induce the formation of fibrinogen multimers. Radioimmunoassay for FPA indicates that thrombin activation of fibrinogen is decreased by Zn(II), with 50% inhibition of FPA release observed at 35 μM Zn(II). These experiments indicate that the critical feature of Zn(II) procoagulant effect is not due to the induction of fibrinogen proteolysis by thrombin, which is actually decreased. Rather, it appears that Zn(II) accelerates the polymerization step of fibrin assembly and concomitantly modifies fibrin gel structure.
Complete recovery of pure red cell aplasia by intramuscular gammaglobulin therapy in a child with hypoparathyroidism
Authors: Etzioni A. , Atias D. , Pollack S. , Levy J. , Vardi P. , Hazani H. , Benderly A.
Published: February 2007

A 10‐year‐old girl with multiple endocrinopathies, hypoparathyroidism, and pernicious anemia subsequently developed pure red cell aplasia, which responded only temporarily to cytotoxic therapy. Because of transfusion dependency, resistance to steroids, plasmapheresis, and Cytoxan, she was given injections of intramuscular gammaglobulin. The injections caused a prompt and sustained rise in the child's erythropoiesis.
Lymphoblastic transformation of myelodysplastic syndrome
Authors: Ascensao Joao L. , Kay Neil E. , Wright John J. , Arthur Diane , Finkel Barbara , Rydell Robert , Kaplan Manuel E.
Published: February 2007

Acute myeloblastic leukemia occurs as a complication of myelodysplastic syndromes, but the appearance of an acute lymphoblastic leukemia (ALL) has only been reported once. We describe a case in which lymphoblastic transformation occurred in the setting of a dysmyelopoietic syndrome. This leukemia was characterized by lymphoid morphology, terminal deoxynucleotidyl transferase (TdT) positivity, cytogenetic abnormalities, and immunoglobulin gene rearrangements. The patient responded to conventional therapy for this leukemia (vincristine and prednisone). Our case supports the hypothesis of a common lymphohemopoietic progenitor and suggests that in vitro tests may help identify a subset of these patients and be important in selecting appropriate therapy.
Suppression of chemotactic activity of human polymorphonuclears by monoclonal IgMs with and without biological activity
Authors: Pruzanski W. , Nitzan D. W. , Saito S.
Published: February 2007

The influence of monoclonal IgMs on migration of human polymorphonuclears was studied at various temperatures by the use of 19 sera with monoclonal IgMs from patients with macroglobulinemia of Waldenstrom (MW) without obvious biological activity, 29 sera with monoclonal IgM cold agglutinins (18 with anti‐I and 11 with anti‐i IgMs) and 3 sera with monoclonal IgM rheumatoid factor (RF). Under‐agarose migration method and modified Boyden chamber method with double filters and 51Cr‐PMNs were used. In under‐agarose method, chemotactic differentials for controls, MW, anti‐I, and anti‐i groups were, respectively, 57 ± 8 mm, 39 ± 9 mm, 44 ± 14 mm, and 32 ± 16 mm at 37°C and 47 ± 18 mm, 22 ± 11 mm, 17 ± 9 mm, and 15 ± 12 mm at 24°C. All three sera with IgM RF inhibited chemotaxis. The differences between all groups and controls were significant at p < 0.01. Random migration was inhibited at 24°C (p < 0.01) but not at 37°C. Inhibitory concentrations of IgM in the sera tested were equal or less than 0.5 mg/ml. Thirteen sera were tested by the modified Boyden chamber method. At 37°C 8 of 13 sera and at 24°C 11 of 13 sera inhibited significantly chemotaxis at a concentration of IgM of 1 mg/ml. The lowest inhibitory concentration of IgM was 25 μg/ml.

Eleven chromatographically pure IgMs were tested in the under‐agarose assay. At concentrations of 0.4‐3.7 mg/ml, eight IgMs inhibited chemotactic differential at 37°C and nine inhibited it at 24°C. At concentrations of 0.6‐2.0 mg/ml, all seven pure IgMs tested by the Boyden chamber method significantly inhibited chemotaxis at 24°C and 37°C. Some IgMs inhibited chemotaxis at concentrations as low as 25 μ/ml. Ten IgM CA were eluted from the red blood cells. Eluates inhibited strongly chemotaxis at 24°C and 37°C. Heat inactivation did not alter inhibitory activity of IgM, however pepsin digestion or reduction and alkylation of purified IgMs did abolish their inhibitory activity. Inhibition of chemotaxis was not related to the light chain type, the titre, or the thermoamplitude of cold agglutination. However, monoclonal IgMs with anti‐i cold agglutinin activity were stronger inhibitors than anti‐I. Since 75% of IgMs tested inhibited chemotaxis at 37°C, it is possible that monoclonal IgMs, especially those with anti‐i cold agglutinin activity, inhibit PMN migration in vivo.
Disseminated intravascular coagulation in lupus erythematosus responding to prednisone therapy
Authors: Riddell Stanley R. , Shojania A. Majid
Published: February 2007

A 48‐year‐old man with systemic lupus erythematosus (SLE) developed disseminated intravascular coagulation (DIC) with clinical bleeding. Since no other cause for DIC could be demonstrated, he was treated with prednisone, which rapidly corrected his DIC.

This case demonstrates the need for a complete hematological investigation of patients with SLE who present with hemostatic abnormalities.
Comparative activity of erythrocyte adenosine deaminase and orotidine decarboxylase in Diamond‐Blackfan anemia
Authors: Glader Bertil E. , Backer Karen
Published: February 2007

It previously has been reported that red blood cells (RBC) of patients with Diamond‐Blackfan syndrome (DBS) have increased activity of orotidine decarboxylase (ODC) and adenosine deaminase (ADA). The studies reported here compared the activity of these two enzymes in DBS erythrocytes, cord blood, and reticulocytes. The activity of ODC, although increased in some DBS erythrocytes, was not significantly different from that seen in cord RBC or reticulocytes. In contrast, RBC‐ADA activity was increased in 23 of 26 DBS patients; and this enzyme elevation was distinct from that seen in cord blood and reticulocytes. Moreover, ADA activity was normal in 26 of 27 patients with transient erythroblastopenia of childhood (TEC). Taken together, these data indicate RBC‐ADA activity is more sensitive than ODC as a marker of DBS. In addition, RBC‐ADA activity continues to be useful for distinguishing DBS and TEC in most patients with RBC hypoplasia.
A serum ferritin assay for prevalence studies of iron deficiency
Authors: Flowers Carol A. , Kuizon Miriam , Beard John L. , Skikne Barry S. , Covell M. , Cook James D.
Published: February 2007

A specialized serum ferritin assay has been developed for the detection of iron deficiency in epidemiologic studies. An enzyme immunoassay (EIA) was employed to climinate the need for radioisotopes. The problem of low sensitivity inherent with the EIA for serum ferritin was eliminated by the use of monoclonal immunologic reagents. The working range of the assay is 1–100 μg/L with a sensitivity of 0.5 μg/ L. Excellent agreement in serum ferritin levels was observed between the present method and the two‐site immunoradiometric assay (IRMA), while the variability at low ferritin concentrations was significantly less with the EIA. Because only 10 μl of serum is required for each assay, duplicate measurements can be performed on a single capillary tube of blood. When an automatic microtiter plate reader for optical density measurements is used, 80‐100 duplicate determinations can be completed by one technologist in a single working day.
Autoreactive erythroid progenitor‐T suppressor cells in the pure red cell aplasia associated with thymoma and panhypogammaglobulinemia
Authors: Mangan Kenneth F. , Volkin Robert , Winkelstein Alan
Published: February 2007

In vitro erythroid culture studies and lymphocyte markers were performed in a patient with a spindle cell thymoma who developed red cell aplasia, panhypogammaglobulinemia, and multiple opportunistic infections. At the time of presentation, erythroid progenitor cells (CFUe, BFUe) were markedly reduced when cultured from marrow mononuclear cells. Removal of T cells from bone marrow mononuclear cells by Erosetting or complement‐mediated lysis with OKT3 pan T cell monoclonal antibody increased growth of erythroid progenitor cells in vitro. Readdition of bone marrow or pleural fluid T cells derived from the thymoma suppressed autologous, but not allogenic, erythroid progenitor cell (CFUe, BFUe) proliferation in vitro. The erythroid progenitor suppressor T cells were predominantly OKTH +, OKT3 +, OKT8 + and Ia + consistent with activated suppressor T cells. Treatment of the patient with cyclophsphamide and corticosteroids reduced marrow lymphocytes fourfold, and a prompt reticulocytosis ensued. After recovery, erythroid progenitor cells were easily detectable. These studies provide new evidence for T cell‐mediated suppression of erythropoiesis in a unique subset of patients with red cell aplasia associated with thymoma and hypogammaglobulinemia.
Treatment of myelodysplastic syndrome with 1.25‐dihydroxy‐vitamin D3
Authors: Richard C. , Mazo E. , Cuadrado M. A. , Iriondo A. , Bello C. , Gandarillas M. A. , Zubizarreta A.
Published: February 2007

1.25‐dihydroxy‐vitamin D3 (1.25 (OH)2D3) was tested in seven patients with myelodysplastic syndrome. The study was undertaken because 1.25 (OH)2D3 promotes differentiating myeloid cells in vitro and because of a prior report of potential benefit in a clinical study. The drug was given orally at a dose of 2.5 μg/day for a minimum of 8 weeks (range 8–28). After therapy, there were no significant changes in any of the parameters observed in peripheral blood or bone marrow. We did not observe any feature of granulocytic‐monocytic differentiation. Treatment was well tolerated. One patient died because of bone marrow failure. Survivors have persisting myelodysplastic syndrome and continue to be transfusion dependent. 1.25 (OH)2D3 has no beneficial effect in patients with myelodysplastic syndrome with this dose regimen.
Monoclonal antibodies to CALLA do not alter polymorphonuclear functions
Authors: Kaplan S. S. , Pesando J. M. , Basford R. E. , Zdziarski U. E.
Published: February 2007

The common acute lymphoblastic leukemia antigen (CALLA) is present on the malignant cells of most patients with acute lymphoblastic leukemia (ALL). Monoclonal antibodies (MoAbs) to CALLA have been useful for differentiating lymphoblastic from nonlymphoblastic leukemias as well as for serotherapy in ALL. Since this MoAb also reacts with polymorphonuclear (PMN) leukocytes, it was of interest to determine whether or not MoAbs to CALLA affected PMN function. We therefore evaluated four MoAbs to CALLA for their effect on PMN function. Two of the MoAbs were IgG and two were IgM. Chemotaxis was studied using Micro‐Boyden chambers. Intraleukocyte bacterial killing was studied using Staph: PMN of 2: 1 and 10: 1, and metabolic activation was evaluated by hexose monophosphate shunt activity. The results showed that these MoAbs to CALLA did not impair the parameters of PMN function studied. These findings have relevance to the usefulness of MoAbs to CALLA for serotherapy and also as a probe for understanding the surface molecule properties that have a bearing on PMN host defense functions.