bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of platelet factor 4 (PF4) on assays of plasma heparin
Authors: Levine Shirley P. , Sorenson Richard R. , Harris Margaret A. , Knieriem Linda K.
Published: March 2008

Summary.Platelet factor 4 (PF4) is a potent antiheparinin vitro. In view of the large amount of PF4 secreted from platelet α‐granules during routine blood collection and processing techniques, the potential significance of this release was investigated using three measurements of heparin activity: the activated partial thromboplastin time (aPTT), the thrombin time, and factor Xa inactivation using the chromogenic substrate S2222 for assay of factor Xa. The results demonstrate that purified PF4 neutralizes heparin activity when added in increasing amounts to normal platelet‐poor plasma containing a fixed concentration of commercial porcine gut mucosal heparin. This effect was seen when assaying heparin activity by all three methods. In addition, when heparin was added in increasing concentrations to pooled plasma samples that were collected from normal volunteers, there was neutralization of heparin activity in blood samples collected by routine citrate anticoagulation (CIT60) in comparison to blood samples collected simultaneously with platelet secretion inhibiting agents added to the anticoagulant (CIT+). This effect was seen when assaying heparin by the aPTT and thrombin time. These data confirm that both purified and secreted PF4 have significant antiheparin activity when heparin is addedin vitroto normal plasma.

Neutralization of circulating heparin by PF4 secreted during blood collection from anticoagulated patients could result in underestimation of the actualin vivoheparin concentration. In order to evaluate the significance of this effect, purified PF4 was added to plasma collected from heparinized patients and again PF4 neutralized heparin activity. This was seen, however, only when heparin activity was measured by the thrombin time or Xa inactivation assays. There was minimal shortening of the aPTT when PF4 was added in final concentrations up to 1000 ng/ml. When blood samples were simultaneously collected from anticoagulated
Hybridoma antibodies to human von Willebrand factor
Authors: Meyer Dominique , Zimmerman T. S. , Obert Bernadette , Edgington T. S.
Published: March 2008

Summary. Twenty‐seven stable subclones of seven independent cellular hybrids producing murine monoclonal antibodies to human von Willebrand factor (vWF) have been established. The specificity of the hybridoma antibodies for vWF has been substantiated by a variety of methods including binding to highly purified vWF, absence of binding to plasma or cryoprecipitate from severe von Willebrand's disease, binding to different size multimers in normal plasma, and binding to low molecular weight multimers in type IIA von Willebrand's disease plasma. Monoclonality of the hybridoma derived antibodies has been sought by repeated cloning at limiting dilutions. All seven specificities of hybridoma antibodies cross‐react to a variable degree with porcine vWF and all but one with bovine vWF, indicating that they bind to structural loci that are relatively though not identically conserved between species. Hybridoma antibodies bind native as well as denatured vWF, suggesting that the epitopes may be determined to a considerable extent by primary structure rather than entirely by tertiary or quaternary conformation. The hybridoma antibodies define precise epitopes in the vWF molecule, extend the horizon of direct analysis of vWF to the intramolecular level, and may be useful in assigning functional loci to parts of the molecule.
Oral administration of cyclosporin A for recipients of allogeneic marrow transplants: implications of clinical gut dysfunction
Authors: Atkinson K. , Biggs J. C. , Britton K. , Short R. , Mrongovius R. , And A. Concannon , Dodds A.
Published: March 2008

Summary. Cyclosporin A (CyA) was used to minimize graft‐versus‐host disease (GVHD) in 28 recipients of allogeneic marrow transplants. When given orally, the absorption of CyA was markedly dependent on normal gut function. Patients without gut dysfunction showed normal serum concentration‐time curves while those with diarrhoea from any cause (chemo‐radiation enteritis, acute GVHD of the gut, infectious enteritis) showed minimal absorption of the drug. These data indicate the desirability of the intravenous administration of CyA during periods of gut dysfunction in marrow transplant recipients.
Relationship between the per cent of marrow cells in S phase and the outcome of remission‐induction therapy for acute nonlymphocytic leukaemia
Authors: Preisler H. D. , Azarnia N. , Raza A. , Grunwald H. , Vogler R. , Browman G. , Goldberg J. , Chervenick P. , Miller K. , Brennan J. , Winton L. , And R. Joyce , Gottlieb(Intergroup Leukemia Study) A.
Published: March 2008

Summary. The relationship between the pretherapy cell cycle characteristics of leukaemic marrow cells and the outcome of remission‐induction therapy for acute nonlymphocytic leukaemia was studied in newly diagnosed and relapsed patients who were then treated with either combination chemotherapy consisting of cytosine arabinoside/anthracycline antibiotic 6 thioguanine or with single agent high‐dose cytosine arabinoside therapy. The outcome of high‐dose cytosine arabinoside therapy was highly dependent upon the per cent of pretherapy cells in S phase with no remissions occurring in patients in whom the 3H‐TdR labelling index was <6%. In contrast, the outcome of cytosine arabinoside/anthracycline antibiotic therapy was independent of the pretherapy cell cycle characteristics of the leukaemic cells.
Hereditary factor XIII deficiency: report of four families and definition of the carrier state
Authors: Berliner Sh. , Lusky A. , Zivelin A. , Modan M. , Seligsohn U.
Published: March 2008

Summary. Definition of the carrier state of hereditary factor XIII deficiency was attempted by a study of four unrelated families with patients affected by the disease. Ten homozygotes, 14 obligatory carriers and 20 suspected carriers were available for determination of plasma factor XIII subunits A and B levels (by electroimmunoassay) and for assay of factor XIII enzymatic activity (by 14C‐putrescine incorporation into casein). Evaluation of the ability of seven variables related to these tests to discriminate between healthy controls and obligatory carriers disclosed that two variables related to the enzymatic assay classified correctly 92.9% of the carriers and 73.3% of the controls, whereas the variables related to subunit A had a lower discriminative ability. Entering both enzymatic and immunologic variables into the discriminant function did not improve the result of the former. A classification function was established from which the posterior probability of suspected carriers can be determined by measuring the incorporation of 14C‐putrescine into casein at 90 min after adding the labelled amine.
Simultaneous detection of membrane markers with monoclonal antibodies and peroxidatic activities in leukaemia: ultrastructural analysis using a new method of fixation preserving the platelet peroxidase
Authors: Breton‐Gorius J. , Vanhaeke D. , Pryzwansky K. B. , Guichard J. , Tabilio A. , Vainchenker W. , Carmel R.
Published: March 2008

Summary. Simultaneous detection of specific surface markers by immunogold and intracellular peroxidase activity was determined ultrastructurally in normal and leukaemic progenitors of platelets, erythrocytes and granulocytes. A new method of fixation was employed to preserve platelet peroxidase activity. Monoclonal antibodies to platelet glycoproteins labelled exclusively platelet peroxidase (PPO) positive cells, i.e. platelets, megakaryocytes and promegakaryoblasts (PMKB). In acute megakaryoblastic leukaemia, most PMKB possessed both markers while a few PMKB identified by PPO did not bind monoclonal antibodies. This result suggests that PPO appears earlier in maturation than platelet glycoproteins. Although all glycoproteins (GP) displayed fewer sites in PMKB than platelets, GP lb was often observed in more mature megakaryocytes. Surface (glycophorin A) and intracytoplasmic markers including ferritin, intra‐mitrochondrial iron and diffuse peroxidase activity due to haemoglobin of erythroid progenitors, appeared simultaneously. The number of glycophorin A sites increased with maturation. In leukaemia involving PMKB and proerythroblasts, the surface markers were coincident with the localization of peroxidase activity; glycophorin A was always absent from blasts which exhibited PPO activity localized in endoplasmic reticulum. Platelet glycoproteins were never expressed in any other cell lineage. The myeloid surface antigen was present on normal late neutrophilic promyelocytes after the cessation of myeloperoxidase synthesis. In some cases of Ml and M2 AML (FAB classification), labelling was identical to normal cells while in others the antigen appeared earlier than normal. Our findings show that the surface phenotype of blasts from non‐lymphoid leukaemia and the intracellular peroxidase activity of a given cell type can be simultaneously demonstrated and analysed by electron microscopy.
Immunohistological study of transferrin receptor expression in non‐Hodgkin's lymphoma
Authors: Pileri Stefano , Gobbi Marco , Teresa Maria , Ivano R , Martinelli Giuseppe
Published: March 2008

Summary. Lyophilized frozen sections of 24 lymph nodes from patients with different types of non‐Hodgkin's malignant lymphoma, diagnosed according to the Kiel Classification, were tested by an immunoperoxidase‐ABC‐method, in order to assess the expression of the transferrin receptors, recognized by the OKT9 monoclonal antibody. Our results support the hypothesis that 0KT9 reactivity of lymphomatous tissues might reflect the histological grading, offering a new important prognostic parameter, and stress the concept that immunological studies in frozen sections allow a more precise definition of the lymphoma phenotype.
The effect of low dose Ara‐C in acute non‐lymphoblastic leukaemias and atypical leukaemia
Authors: Ishikura Hiroto , Sawada Hiroyoshi , Okazaki Toshiro , Mochizuki Toshihiro , Izumi Yoichiro , Yamagishi Morihisa , Uchino Haruto
Published: March 2008

Nine patients with nonlymphocytic leukaemia and one patient with refractory anaemia with excess of blasts (RAEB) were treated subcutaneously with Ara‐C at a low dose of 10 mg/m2/12 h; complete remission was obtained in seven patients. In all cases severe pancytopenia was observed during treatment. We measured the concentration of Ara‐C in serum, and found that the maximum concentration reached a peak (52‐132 ng/ml; mean 84‐2 ng/ml) at 15 min following injection. These concentrations can be considered sufficient to inhibit DNA synthesis. Primary short‐term culture of human leukaemic cells with low dose Ara‐C was also performed, and differentiation of leukaemic cells was observed for several types of leukaemic cells. The in vitro findings corresponded to the observed clinical effects. From the above results, the action of low dose Ara‐C may have resulted from a combination of two different mechanisms, its cytostatic effect and its differentiation‐inducing effect.
Prediction of response of acute nonlymphocytic leukaemia to therapy with ‘high dose’ cytosine arabinoside
Authors: Preisler H. D. , Epstein J. , Barcos M. , Priore R. , Raza A. , Browman G. P. , Vogler R. , Winton E. , Grunwald H. , Rai K. , Brennan J. , Bennett J. , Goldberg J. , Gottlieb A. , Chervenick P. , Joyce R. , Miller K. , Larson R. , D'Arrigo P. , Doeblin T. , Stein M. , Bloom M. , Steele R. , Lee H.
Published: March 2008

The Leukemia Intergroup Study has treated 110 patients with acute nonlymphocytic leukaemia with ‘high dose’ cytosine arabinoside remission induction therapy and studied the factors which were related to the outcome of therapy. With respect to death during remission induction therapy, only patient age was of prognostic significance. Treatment failure due to resistant leukaemia was associated with a high pretherapy leukaemic cell mass, the presence of few cells in S phase, and insensitivity of DNA synthesis to cytosine arabinoside. If, after 6 d of therapy, more than 40% of the marrow cells were leukaemic, the patient almost invariably failed to enter complete remission because of persistent leukaemia. Simultaneous consideration of the pretherapy labelling index and the per cent abnormal cells in the day 6 marrow permitted a distinction to be made between almost all patients who would enter remission or fail therapy because of persistent leukaemia.
Sequential karyotypic evolutions and bone marrow aplasia preceding acute myelomonocytic transformation from myelodysplastic syndrome
Authors: Tomonaga M. , Tomonaga Y. , Kusano M. , Ichimaru M.
Published: March 2008

Serial haematopathological and cytogenetic studies disclosed three distinct clinical phases in a case of refractory anaemia (RA), a subtype of myelodysplastic syndrome (MDS; FAB group, 1982): first, chronic MDS phase (1 year 10 months) with karyotypic abnormality (45, XY, –7) (Clone I); second, hypo‐aplastic phase concurrent with first clonal evolution (45, XY, –7, 12p –) (Clone II); third, acute myelomonocytic leukaemia phase (6 months) with second clonal evolution (45, XY, ‐ 7, t (lq‐; Bq +), Bq ‐, 12p ‐) (Clone III). In the second phase the bone marrow became almost aplastic as Clone II expanded progressively, indicating simultaneous occurrence in Clone II stem cells of growth advantage for self‐renewal function over Clone I and normal stem cells, and arrest of differentiation. These observations support the hypothesis that leukaemic change in MDS, at least in RA, occurs by stepwise clonal evolution(s), not by progressive arrest of differentiation in original MDS clone.
The severity of immune neutropenia correlates with the maturational specificity of antineutrophil antibodies
Authors: Harmon David C. , Weitzman Sigmund A. , Stossel Thomas P.
Published: March 2008

Sera from 15 patients with antineutrophil antibodies against peripheral blood neutrophils were incubated with normal bone marrow, and the reaction with marrow precursors was determined. Serum from controls did not deposit IgG on either peripheral blood or marrow myeloid cells. Serum from patients with moderate immune neutropenia with myeloid hyperplasia deposited IgG on neutrophils, bands and metamyelocytes, but not on earlier precursors. Serum from one patient with severe neutropenia and myeloid hypoplasia deposited IgG on myelocytes and promyelocytes as well as more mature cells. Serum from a patient with antineutrophil antibodies which impaired chemotaxis but who did not have neutropenia deposited IgG only on neutrophils and occasional bands. Immune neutropenia may become increasingly severe as antibodies become directed against earlier precursors.
Low dose cytosine arabinoside (Ara C) in myelodysplastic syndromes
Authors: Tricot G. , Bock R. De , Dekker A. W. , Boogaerts M. A. , Peetermans M. , Punt K. , Verwilghen R. L.
Published: March 2008

Myelodysplastic syndromes (MDS) are acquired bone‐marrow disorders, characterized by a decreased ability of the haemopoietic cell to differentiate, resulting in peripheral cytopenias. The majority of patients will die either from acute myeloid leukaemia or from infection and/or haemorrhage. Thirty‐eight courses of low dose Ara C were administered in 26 MDS patients. Nineteen courses (50%) were associated with good (12) or partial (7) response. Three complete remissions were observed. The median duration of response overall was 19‐5 weeks, 26 weeks for the good and 10 weeks for the partial responders. A high incidence of treatment failure was seen in patients treated after transition to AML. Major complications were observed during 14 courses and mortality was directly related to therapy in five patients. Platelet transfusions were required during 26 courses. Aggravation of peripheral‐blood cytopenia during the first weeks and hypocellularity of bone‐marrow aspirates at the end of therapy suggest that low‐dose Ara C exerts its main activity by suppression of leukaemic growth, rather than by induction of differentiation in malignant cells. Our results in MDS patients demonstrate that low‐dose Ara C can be of value in severe cytopenia and can decrease the proportion of leukaemic cells in the bone marrow, but the danger of treatment should not be underestimated.
Separation of human bone marrow by counterflow centrifugation monitored by DNA‐flowcytometry
Authors: Witte T. DE , Plas A. , Koekman E. , Blankenborg G. , Salden M. , Wessels J. , Haanen C.
Published: March 2008

Human bone marrow was fractionated by counterflow centrifugation into 16 fractions with increasing cell size. Three distinct subpopulations could be recognized: small lymphocytic cells, medium‐sized nucleated erythroid cells and large myeloid elements. DNA‐flowcytometry and 3H‐thymidine uptake showed that within the erythroid and myeloid cell populations counterflow centrifugation separates each population according to the cell cycle phase. Hypotonic treatment of bone marrow for removal of the erythroid nucleated cells resulted in a complete abrogation of the proliferating erythroid cell population. Counterflow centrifugation also separates the small non‐proliferating myeloid and erythroid committed stem cells from the larger proliferating stem cells. It appeared feasible to separate the small lymphocytic cells from the majority of BFU‐E and CFU‐GM, due to the larger size of the proliferating normoblasts and the committed progenitor cells. Elimination of the mature lymphocytes from the haematopoietic stem cells by counterflow centrifugation may offer an alternative approach to the prevention of graft versus host disease (GvHD).
Inverse correlation between age related abnormalities of T‐cell immunity and circulating thymosin α1 levels in haemophilia A
Authors: Kessler Craig M. , Schulof Richard S. , Alabaster Oliver , Goldstein Allan L. , Naylor Paul H. , Phillips T. M. , Luban Naomi L. C. , Kelleher Jack F. , Reaman Gregory H.
Published: March 2008

T‐cell immunity and serum levels of thymosin α1, β2‐microglobulin, circulating immune complexes, serum immunoglobulin levels, antibodies to hepatitis surface or core antigen, and to cytomegalovirus, and Epstein‐Barr virus were investigated in 51 patients with haemophilia A ranging in age from 2 to 52 years. All patients had received commercial U.S. lyophilized concentrates of antihaemophilic factor (AHF). The mean helper/cytotoxic‐suppressor (0KT4/0KT8) ratio of 11 pre‐adolescents (1‐6 ±0‐4 SE) was not significantly different from that of age matched normal controls. In contrast, the mean 0KT4/0KT8 ratios of 13 adolescent (1‐2 ±0‐2 SE) and 23 adult (0‐8 ±0‐1 SE) haemophiliacs were significantly reduced. Abnormalities of lymphocyte mitogenic responses were found only in adult haemophiliacs. Nine individuals treated with commercial U.S. prothrombin complex concentrates for antibodies directed against AHF or for haemophilia B had normal mean OKT4/OKT8 values. The mean serum thymosin α1 levels for each age category was similar to that of age matched controls; however, regression analysis revealed a significant relationship between elevated thymosin α1 levels and decreased OKT4/OKT8 ratios in adult haemophiliacs (P= 0‐012). Although the mean serum level of β2‐microglobulin was significantly increased in the adult haemophiliac group, there was no correlation between OKT4/OKT8 ratios and any of the other serologic parameters studied.
Expression of SSEA‐I antigen (3‐fucosyl‐N‐acetyl‐lactosamine) on normal and leukaemic human haemopoietic cells: modulation by neuraminidase treatment
Authors: Tabilio A. , Canizo M. C. Del , Henri A. , Guichard J. , Mannoni P. , Civin. C.I. , Testa U. , Rochant H. , Vainchenker W. , Breton‐Gorius J.
Published: March 2008

Several mouse monoclonal antibodies (MoAbs) considered specific for the myeloid lineage recognize the same carbohydrate structure (3‐fucosyl‐N‐acetyl‐lac‐tosamine) which is similar to the murine antigen SSEA‐I. We have investigated the expression of this antigen with six different well‐characterized murine IgM MoAbs on normal, leukaemic, and cultured cells by immunofluorescence and immuno‐electron microscope cytochemistry. The cells were studied before and after neuraminidase treatment since epitopes recognized by these MoAbs may be masked by sialic acid. Among the recognizable normal marrow or blood cells, all these MoAbs specifically labelled the granulocytic lineage from the promyelocyte to the polymorph. After neuraminidase treatment, monocytes became labelled. All the other lineages remained unstained. Several cell lines were studied. Six of eight lymphoblastoid cell lines were stained by these MoABs; reactivity was increased by neuraminidase. One Burkitt cell line and two T cell lines were also found to be positive. These antibodies were tested on leukaemic cells. In acute non‐lymphocytic leukaemia they usually labelled promyelocytes, more mature granulocytic and monocytic precursors but did not label myeloblasts; after neuraminidase treatment, these myeloblasts became stained. No labelling was observed on leukaemic proerythroblasts and promegakaryoblast before and after neuraminidase treatment except in one case of promegakaryoblastic leukaemia in which the SSEA‐I antigen and platelet peroxidase were expressed in the same cell. In addition, six cases of common acute lymphoblastic leukaemia were studied; the blasts became positive after desialylation. Two examples of T cell acute leukaemia were essentially negative. We conclude, therefore, that the reactivity of haemopoietic cells with these MoAbs alone does not represent a criterion sufficient to sustain their myeloid origin since the SSEA‐I antigen may be expressed at the surface of all cell lineages in the early phases of haemopoietic differentiation.
Multimarker phenotypic characterization of adult and childhood acute lymphoblastic leukaemia: an Italian multicentre study
Authors: Foá R. , Baldini L. , Cattoretti G. , Foa P. , Gobbi M. , Lauria F. , Madon E. , Masera G. , Miniero R. , Paolucci P. , Perona G. , Pizzolo G. , Saitta M. , Gavosto F.
Published: March 2008

Summary. A multicentre phenotypic study was carried out in Italy combining conventional immunological techniques with monoclonal antibody (MoAb) analysis in 190 cases of adult and childhood acute lymphoblastic leukaemia (ALL), in an attempt to define better the lineage relationship of the neoplastic cells. Of the 140 children evaluated, 79·3% expressed the common ALL (cALL) antigen (all analyses performed by MoAb), 11·4% were T‐ALL and 9·3% were non‐T, non‐B, non‐common (‘null’) ALL. The proportion of adult cALL cases was slightly lower (64% of the 50 cases studied) than that of childhood ALL, whilst the incidence of T‐ALL was significantly higher in adults than in children (26%v. 11·4%, P <0·05). Because of the high proportion of cALL cases, the incidence of ‘null’ ALL in adult patients was similar (10%) to that of children, and lower than previously reported. The recognition of early pre‐T‐ALL cases (T1+, RFT2+, T10+, T6‐, T11‐, E‐) contributed to the overall low proportion of ‘null’ ALL; prior to the use of MoAb, such cases would probably have been classified as undifferentiated acute leukaemia or ‘null’ ALL. The search for B‐cell‐related markers showed that the incidence of pre‐B‐ALL cases (cytoplasmic immunoglobulin positive cases) was similar in adults and in children (25·6% and 32%, respectively). Furthermore, the great majority of cases studied expressed the BA‐1 antigen (92·8% of adults and 79% of children), whilst the BA‐2 antigen was found in 53% of cases (tested only in children), confirming a hierarchy in the expression of B‐cell related markers in CALL: BA‐1, BA‐2, Cylg. Several of the ‘null’ cases also expressed the BA‐1 antigen on a variable proportion of cells, pointing to a possible B‐cell origin of the blasts.

This multicentre study shows that both in adult and in childhood ALL thecombined use of conventional immunological markers and of a panel of MoAb allows identification of the cell lineage of the great majority of cases, thus reducing the number of ‘null’ ALL. Furthermore, these findings suggest that practically all cases of ALL belong either to the T or to the B cell compartment and that the neoplastic cells appear blocked at different levels along the lymphoid differentiation pathway.
Regulation of erythropoiesis during rapid growth
Authors: Sanengen Truls , Halvorsen Sverre
Published: March 2008

Summary. High levels of plasma erythropoiesis stimulating factor(s) (ESF) have been found in neonatal WLO‐mice during the period of rapid growth. If the high ESF activity is due to the concomitant physiological anaemia of infancy alone, it should be possible to block erythropoiesis by hypertransfusion. Mice were hypertransfused starting on day 14, and killed on day 20. Although hypertransfusion reduced the ESF levels by approximately 55% (P<0·001), ESF levels were still detectable in the cell culture assay used (P<0·001). Moreover, hypertransfused mice showed active erythropoiesis in the bone marrow, and none had a reticulocyte count below 2%. No correlation was found between PCV and ESF in the hypertransfused animals (r= 0·07, P>0·5), nor was there any difference in weight gain between control and hypertransfused mice (P>0·5). These results show that hypertransfusion did not totally supress erythropoiesis in neonatal WLO‐mice, which is different from hypertransfused adult mice. The data indicate that the high plasma ESF found in neonatal WLO‐mice during the growth period are not due to the anaemia alone. These findings support studies indicating that regulation of erythropoiesis in the neonate differs from the adult. Factors related to growth per se could be responsible for this difference.
Human mononuclear phagocyte differentiation: a study of the U‐937 cell line by ultrastructural cytochemistry and surface antigen analysis
Authors: Gourdin M. F. , Vasconcelos A. W. , Tabilio A. , Divine M. , Farcet J. P. , Reyes F.
Published: March 2008

Summary. U‐937 represents a well‐established permanent human haematopoietic cell line, which exhibits characteristics of the monocyte/macrophage series. U‐937 cells were investigated by peroxidase ultrastructural cytochemistry in order to determine the normal developmental stage to which they correspond. This study was performed in non‐ and TPA‐stimulated cells, in conjunction with surface analysis by monoclonal antibodies. It is concluded: (1) peroxidase‐positive U‐937 cells are monoblasts and promonocytes involved in myeloperoxidase synthesis; (2) TPA‐stimulation caricatures transformation of these cells into monocytes but not into resident macrophages, as far as peroxidase cytochemistry is concerned; (3) the reactivity of myeloperoxidase present in the endoplasmic reticulum of synthesizing cells is inhibited by glutaraldehyde fixation.
In vitro erythropoietin assay based on erythroid colony formation in fetal mouse liver cell culture
Authors: Sakata Susumu , Enoki Yasunori , Tomita Susumu , Kohzuki Hisaharu
Published: March 2008

Summary. Critical studies were made on erythroid colony formation from cultured fetal mouse liver cells in an attempt to develop a simple and sensitive erythropoietin (Epo) assay procedure. The maximum colony formation was observed 24 h after plating of the cells when an evident dose–response relation was found for Epo added. The colony forming ability decreased steadily as the gestational age of the fetus advanced and was gradually lost by postnatal days 10–11. By morphological and cytochemical criteria almost all the colonies were found to be erythroid. 59Fe‐labelling experiments revealed a fairly good correlation between the colony number and 59Fe incorporation into both cells and haem. Dose–response curves for plasma were parallel to the Epo standard curve. Based on these findings we developed a procedure which could measure as little as 0·4 mU of Epo without requiring 59Fe. Using this method, plasma Epo titres were determined in 16 normal and 69 anaemic subjects.
Counterflow centrifugation of red cell populations: a cell age related separation technique
Authors: Van Der Vegt S. G. L. , Ruben A. M. Th. , Werre J. M. , Palsma D. M. H. , Verhoef C. W. , de Gier J. , Staal G. E. J.
Published: March 2008

Summary. Red cell populations were fractionated on the basis of differences in density by a centrifugation method described by Murphy and on the basis of differences in mean cell volume (MCV) by counterflow centrifugation. By 59Fe‐incorporation and determination of the HbA1c content, both methods were studied for their ability to separate red cell populations into fractions of different mean cell age. It can be concluded that separation on the basis of differences in cell volume results in a linear separation according to age whereas separation on the basis of density only results in an accumulation of very young cells in the top fractions. A gradual decrease in cell volume with age, combined with a constant haemoglobin concentration in the cells, indicates release of haemoglobin from the red cells during their lifespan.
Use of a monoclonal antibody against human heart ferritin for evaluating acidic ferritin concentration in human serum
Authors: Cazzola Mario , Arosio Paolo , Bellotti Vittorio , Bergamaschi Gaetano , Dezza Laura , Iacobello Carmelo , Ruggeri Giuseppina
Published: March 2008

Summary. Immunoassays for acidic ferritins rich in H subunits have shown that these isoferritins are predominant in some cells such as monocytes and red blood cells but have provided conflicting results about their presence in human serum. We have used an immunoradiometric assay based on a monoclonal antibody against human heart ferritin (monoclonal 2A4) for evaluating acidic ferritin concentration in human serum. This assay proved to be highly specific for acidic isoferritins having more than 60% H subunits. Heart‐type ferritin was detected in only one fifth of normal sera and sera from patients with iron overload; values were very low compared with those for basic ferritin. Acidic ferritin was found in relatively high concentrations in most patients with iron deficiency anaemia. In other disease states characterized by increased serum concentrations of basic ferritin, acidic ferritin was always less than 21% of the total ferritin. Dialysis in low‐ionic‐strength buffer showed that both normal and pathological sera had binding factors for human heart ferritin. We conclude that: (i) human serum contains low concentrations of acidic isoferritins which, at variance with basic ferritin, do not appear to be directly related to the amount of storage iron; (ii) the findings of the present study reinforce the opinion that basic and acidic ferritins have different functional behaviours.
Blood coagulation and fibrinolysis in heat stroke
Authors: Mustafa Khatim Y. , Omer O. , Khogali M. , Jamjoom A. , Gumaa K. A. , El‐Nasr N. Abu , Gader M. A.
Published: March 2008

Summary. Blood coagulation and fibrinolysis were assessed in 55 cases of heat stroke who presented with or without bleeding tendencies during the Makkah pilgrimage of 1983. 17 patients were identified to have evidence of disseminated intravascular coagulation (DIC). Bleeders with DIC had a higher incidence of shock and a higher mortality when compared to non‐bleeders. Thrombocyto‐penia and liver cell damage were not limited to cases with DIC. Coagulation factors and serum enzyme studies suggested non‐specific tissue damage as the trigger mechanism for DIC possibly proceeding through the extrinsic system of blood clotting.

We conclude that the breakdown of haemostasis in heat stroke is multifactorial: thrombocytopenia, liver cell damage and DIC.
Acute biphenotypic leukaemia (myeloid and null‐ALL type) supervening in a myelodysplastic syndrome
Authors: Eridani S. , Chan L. C. , Halil O. , Pearson T. C.
Published: March 2008

Summary. An acute leukaemia was seen in a 72–year‐old patient with a myelodysplastic syndrome (MDS) of 14 months duration, who had been treated only with steroids. The morphological appearance of the blast cells suggested a poorly differentiated cell type. Surface marker analysis, including double staining studies, showed the coexistence of a population of null acute lymphoblastic leukaemia (ALL) blast cells together with acute myeloid leukaemia type cells. No chromosomal alterations were detected.

The change from a chronic MDS to an acute leukaemia of mixed (myeloid and null ALL) type suggests either transformation of a pre‐existing abnormal clone or de novo appearance of two separate leukaemic clones.
Post‐transfusion purpura responding to high dose intravenous IgG: further observations on pathogenesis
Authors: Berney S. I. , Metcalfe P. , Wathen N. C. , Waters A. H.
Published: March 2008

Summary. A typical case of post‐transfusion purpura (PTP) is described in a 61‐year‐old woman. Treatment with steroids corrected the purpuric manifestations, but the patient remained thrombocytopenic in spite of steroid therapy and plasma exchange. However, an intravenous infusion of high dose IgG led to a rapid and sustained increase in the platelet count.

The mechanism of thrombocytopenia in PTP is still a matter for speculation, as the platelet specific antibody usually cannot be shown to react with the patient's own platelets in vitro. Therefore, the finding of a coexistent platelet specific IgM autoantibody in this patient, first detected during the early recovery phase, is of particular interest and suggests a possible autoimmune basis for the profound thrombocytopenia.
Electrophoretic analysis of red blood cells in polycythaemia
Authors: Brown K. A. , Guthrie D. L. , Pearson T. C. , Roche C. , Crawford N.
Published: March 2008

Summary. In contrast with previously published observations, this study has found no increase in the net negative surface charge of the red blood cells (RBC) from patients with various forms of polycythaemia when compared with normal subjects.

Two sub‐populations of RBCs with different electrophoretic mobilities (EPM), termed fast and slow, were present in samples from both patients and normal individuals. Frequently, the slow cell population in RBCs from patients with polycythaemia, particularly the apparent polycythaemia sub‐group, was of a lower EPM and contained more cells than the corresponding population in normal subjects.

The membrane‐bound sialic acid content of RBCs from patients with primary proliferative polycythaemia, associated with iron deficient RBC changes, was increased if the results were expressed per unit volume of RBCs and decreased if the sialic acid concentration was presented per number of RBCs.

These results imply that differences in surface charge of RBCs are of no value in discriminating between primary and secondary polycythaemia.
The kinetics of 111Indium distribution following injection of 111Indium labelled autologous granulocytes in man
Authors: Saverymuttu S. H. , Peters A. M. , Keshavarzian A. , Reavy H. J. , Lavender J. P.
Published: March 2008

Summary. The kinetics of human autologous granulocytes, separated and labelled with 111In without isolation from plasma, have been studied in subjects with and without sepsis with the aim of identifying the fate and sites of destruction of granulocytes in man.

In subjects without inflammatory disease, 111In granulocyte recovery in faeces, urine and saliva over 4 d was less than 1% of the dose, so that the activity visualized by the gamma camera represented almost 100% of the dose. On images taken at 24 and 48 h, this activity was distributed between spleen, bone marrow and liver, with foci of additional abnormal activity in subjects with inflammatory disease.

Splenic activity fell between 40 min and 24 h, consistent with the presence of a splenic granulocyte pool, but remained constant after 24 h. Since granulocyte clearance from the blood was predominantly completed by 24 h, the residual splenic activity at that time reflected splenic granulocyte destruction. In patients with sepsis, the fall in splenic activity was greater than in those without, implying diversion of granulocytes from splenic destruction to tissue utilization when inflammation is present. Bone marrow activity increased between 40 min and 24 h and then remained stable. Granulocytes that were extensively manipulated in saline prior to labelling failed to localize in marrow, suggesting that visualization of the latter reflected destruction of intact, normal granulocytes. Although the changes in splenic and marrow activities terminated at 24 h, at which time granulocyte clearance from blood was at least 80% completed, plasma 111In remained essentially unchanged between 40 min and 48 h at less than 5% of the dose, discounting it as the source of splenic and marrow activities.
The relevance of α‐naphthyl acetate esterases to various monocyte functions
Authors: Oertel J. , Hagner G. , Kastner M. , Huhn D.
Published: March 2008

Summary. Human monocytes contain a series of α‐naphthyl acetate (αNA) esterases which are not present in other blood cells and which can be specifically inhibited by bis(4‐nitrophenyl)‐phosphate (BNPP). This inhibitor is non‐toxic at the concentration used and thus enabled studies on the possible significance of this enzyme towards various monocyte functions. BNPP has no noticeable influence on adhesion and spreading of monocytes on glass surfaces, nor does it inhibit the phagocytosis of IgG‐coated latex particles. BNPP does, however, diminish the spontaneous cytotoxicity of freshly isolated monocytes towards the erythroleukaemic cell line K562. In the single cell assay in agarose, BNPP treatment of monocytes leads to a decrease in the number of lytic and non‐lytic effector‐target cell conjugates. In contrast, the antibody‐dependent cell‐mediated cytotoxicity (ADCC) of monocytes as well as the natural cytotoxicity of lymphocytes towards K562 cells are not influenced by BNPP. The present investigations show that monocyte specific αNA esterases are involved in the spontaneous cytotoxicity of monocytes toward tumour cells.
Natural killer cell activity in childhood acute lymphoblastic leukaemia in remission
Authors: Dickinson Anne M. , Proctor S. J. , Jacobs Elizabeth , Reid M. M. , Walker W. , Craft A. W. , Kernahan Jennifer
Published: March 2008

Summary. Fifteen children with acute lymphoblastic leukaemia (ALL) in remission receiving maintenance chemotherapy and 12 ALL patients off treatment and in remission were tested for natural killer (NK) cell activity in vitro. Compared with a control population the children with ALL receiving maintenance chemotherapy had low levels of NK cell activity. This effect was not due to a specific reduction in NK cell numbers since proportions of mononuclear cells detected by the monoclonal antibodies HNK‐1 (Leu‐7) and Leu‐11a were normal. Furthermore NK cell activity in patients could only be partially increased by pre‐incubation of effector cells with interferon (αIFN). These studies confirm the lack of NK cell activity in children with ALL and show that this phenomenon is directly related to functional NK cell impairment. Our study has further shown that this effect is transient since ALL patients off treatment and in remission showed normal levels and augmentation of NK cell activity.
The association of chronic lymphocytic leukaemia and multiple myeloma: a study of eleven patients
Authors: Brouet J. C. , Fermand J. P. , Laurent G. , Grange M. J. , Chevalier A. , Jacquillat C. , Seligmann M.
Published: March 2008

Summary. We present haematological and immunological data obtained in 11 patients with the rare association of chronic lymphocytic leukaemia (CLL) and multiple myeloma (MM). CLL occurred before MM in six cases and both diseases were diagnosed simultaneously in five. With regard to CLL, no peculiar clinical feature was found; in six patients, MM was characterized by extra‐osseous lesions. A significant remission (3–7 years) was obtained in three patients. In six patients, CLL and plasma cells synthesized Ig molecules with different light chains, probably indicating the coexistence of two distinct clonal proliferations. In two cases where monoclonal Ig synthesized by both types of cells had the same type of light chains, studies with anti‐idiotypic antibodies showed that the two proliferations were also distinct. The pathophysiology of this rare association is discussed in the light of these findings and of data from the literature.
Effects of hyperthermia on the myelograms of adult and fetal guinea‐pigs
Authors: Edwards Marshall J. , Penny Richard H. C.
Published: March 2008

Summary. The myelograms of adult guinea‐pigs at various stages after brief exposure to hyperthermia were compared with those of control animals. The proportions of neutrophil and eosinophil metamyelocytes and of intermediate normoblasts were significantly reduced for at least 24 h after exposure and the proportions of neutrophil polymorphs and lobulated eosinophils were significantly increased over this period. The mitotic index was reduced for up to 5 h. Morphological changes in cells included hypersegmentation of nuclei, pyknosis, necrobiotic changes, development of pseudopodia, and abnormalities of mitosis. The myelograms of newborn guinea‐pigs which had been heated on days 20–23 of pregnancy showed no significant changes.
Book Reviews
Authors: Luzzatto L. , Gaffney Patrick J. , Gordon M. Y. , Weatherall D. J. , Machin S. J. , Bloom A. L. , Rampling Mike , Poller Leon , Epenetos A. A.
Published: March 2008

Advances in Human Genetics

Clinical Aspects of Fibrinolysis and Thrombolysis. Edited by T. Jespersen, C. Kluft and C. Korsgaard.

Normal and Neoplastic Hematopoiesis. UCLA Symposium on Molecular and Cellular Biology, New Series Volume 9. Edited by D. W. Golde and S. A. Mares.

Malaria and the Red Cell. Edited by J. W. Eaton and G. J. Brewer.

Thrombosis and Cardiovascular Diseases. Advances in Experimental Medicine and Biology, Volume 164. Edited by A. Strano.

Factor VIII Inhibitors. By L. W. Hoyer.

Red Cell Deformability and Filtration. Edited by J. Dormandy.

Thromboplastin Calibration and Oral Anticoagulant Control. Edited by A. M. H. P. van den Besselaar, H. R. Gralnick and S. M. Lewis.

The Reticuloendothelial System. A Comprehensive Treatise to Cancer. Ed. Herberman & Friedman.
Thrombocytopenia in SLE and related autoimmune disorders: association with anticardiolipin antibody
Authors: Harris E. N. , Asherson R. A. , Gharavi A. E. , Morgan S. H. , Derue G. , Hughes G. R. V.
Published: March 2008

Summary

Anticardiolipin antibody levels were determined in 116 patients with systemic lupus erythematosus and related autoimmune disorders. Forty‐three of these patients had a history of thrombocytopenia—36 of whom had SLE, three primary Sjögren's syndrome, two rheumatoid arthritis and two mixed connective tissue disease. IgG anticardiolipin antibody levels were raised in 31 (72%) of the 43 patients and IgM anticardiolipin antibody levels were raised in 19 (44%). There was a strong statistical correlation between thrombocytopenia and raised anticardiolipin antibody levels of both the IgG (P < 0.001) and IgM (P < 0.01) immunoglobulin classes. Of the 20 patients with the highest IgG anticardiolipin antibody levels 16 had a history of thrombocytopenia. We suggest that anticardiolipin antibodies may play a direct role in mediating platelet destruction in autoimmune disorders.
Heterogeneity of hereditary and acquired factor X deficiencies by combined immunochemical and functional analyses
Authors: Fair Daryl S. , Edgington Thomas S.
Published: March 2008

Summary

Limited information is available regarding molecular abnormalities associated with derived factor X deficiencies. In order to assess the types of molecular aberrations that may occur in this group of haemostatic diseases we have analysed plasmas of 33 individuals from 28 kindred exhibiting factor X deficiency. These included those of hereditary type, transiently acquired deficiency as well as factor X deficiency associated with amyloidosis. Plasmas were analysed by one‐stage assays for factor X activation by the extrinsic coagulation pathway, intrinsic coagulation pathway and Russell's viper venom. Selected plasmas were analysed in two‐stage assays. Normal factor X concentration by specific radioimmunoassay was 6.38 ± 1.29 μg/ml. Factor X associated with factor X deficiency were grouped by their specific activities measured for the three activation pathways and antigen concentration. The results suggest that a broad spectrum of molecular aberrations exist in the factor X deficiency states. The most common group of factor X deficiency was associated with abnormal activation of factor X by all three pathways. Variants of factor X associated with primary amyloidosis and transient acquired deficiency appeared to be abnormal molecules and not just reduced factor X concentration. Hereditary abnormal factor X molecules include the spectrum of potential defective molecules. The relationship of factor X structure to function and the heterogeneity of these defective molecules is discussed.
The sequence of the Aγ globin gene in a Gγβ+ type of hereditary persistence of fetal haemoglobin
Authors: Jones R. W. , Goodbourn S. E. Y. , Old J. M. , Weatherall D. J.
Published: March 2008

Summary

The sequence of the Aγ globin gene from the affected chromosome of an individual heterozygous for the Gγβ+ type of hereditary persistence of fetal haemoglobin (Gγβ+ HPFH) is reported. With two exceptions, it is identical to one of the two allelic Aγ globin genes already sequenced and there is therefore no change which could explain the absence of its expression in the face of the persistent Gγ globin expression which is a feature of this condition.
The haematology of steady state homozygous sickle cell disease: frequency distributions, variation with age and sex, longitudinal observations
Authors: Hayes R. J. , Beckford M. , Grandison Y. , Mason K. , Serjeant B. E. , Serjeant G. R.
Published: March 2008

Summary

The steady state haematological characteristics observed in 1071 patients with homozygous sickle cell (SS) disease aged 5–66 years are presented.

Cross sectional studies indicated that HbA2 levels were consistently higher in males but no age related change was apparent. Fetal haemoglobin levels were consistently higher in females and fell significantly in males between the 5–9 and 10–14 year age groups. Total haemoglobin was significantly higher in females before age 15 and higher in males after 20 years, a dramatic age related rise occurring in males between the 10–14 and 25–29 year age groups, and a fall in patients aged 40 years and over. The mean cell volume was consistently greater in females after 15 years and a marked age related rise occurred in both sexes between the 5–9 and 25–29 year age groups. Counts of irreversibly sickled cells were consistently higher in males. Reticulocytes fell significantly with age, while platelets and total bilirubin fell significantly after the age of 15 years.

Longitudinal studies confirmed the increase in total haemoglobin levels in males over the ages 10–14 years, and a significant fall in males after the age of 30 years. Such studies also confirmed the fall in HbF in males aged 5–14 years, the increase in MCV in both sexes aged 5–29 years, and the fall in platelet counts in both sexes over the age of 20 years.

These observations provide ‘normal’values for patients seen elsewhere and also contribute to an understanding of factors determining the haemoglobin indices in SS disease.