bluuebunny/crossref_metadata_2025 huggingface dataset:
Ultrastructure and cell cycle distribution of erythropoietic cells in heterozygotes and homozygotes for haemoglobin E
Authors: Wickramasinghe S. N. , Hughes Madeleine , Wasi P. , Fucharoen S. , Litwinczuk R. A. C.
Published: March 2008

Summary. Marrow aspirates from heterozygotes and homozygotes for haemoglobin E (HbE) have been studied by electron microscopy and by the technique of combined Feulgen microspectrophotometry and 3H‐thymidine autoradiography. The erythropoietic cells of heterozygotes did not contain any precipitated globin chains and the proliferating erythroblasts of such individuals showed no abnormality in their distribution in the different stages of interphase. By contrast, 0–1.5% of late erythroblast profiles and 3.1–12.8% of marrow reticulocyte profiles of homozygotes contained intracellular inclusions resembling precipitated α‐chains. Although precipitated globin chains were not seen in the early polychromatic erythroblasts of homozygotes, the number of these cells in the G2 phase relative to that in the S phase was increased. These data indicate that there is (1) probably little or no imbalance of globin chain synthesis in heterozygotes, (2) a substantial degree of imbalance in homozygotes, and (3) a disturbance of erythroblast proliferation in homozygotes which cannot be attributed to the deleterious effects of detectable intracellular α‐chain precipitates. The electron microscope and cell cycle distribution data in the homozygotes for HbE were similar to those in two heterozygotes for β thalassaemia.
Normal glutathione content and some related enzyme activities in the fetal erythrocytes
Authors: Lestas A. N. , Rodeck C. H.
Published: March 2008

Summary. Pure fetal blood was obtained by direct‐vision fetoscopy from 66 fetuses at 17–24 weeks gestation. The concentration of GSH and the activities of the enzymes γ‐glutamylcysteine synthetase (GCS), glutathione synthetase (GS), glutathione reductase (GR) and glutathione peroxidase (GPx) were analysed by established techniques to find the normal ranges for this gestational age. The ranges were relatively narrow and could serve as reference values for the prenatal diagnosis of defects in the GSH metabolism of erythrocytes. The results were compared with those obtained from 38 normal adults and with published values on neonatal blood. In the case of GR a comparison was also made with maternal blood.

In comparison with adults, fetal erythrocytes showed higher GSH concentration and GCS activity and lower GS and GPx activities. This pattern resembled that found in neonatal erythrocytes except for the GCS activity, which was higher in the fetal cells. Furthermore the differences between fetal and adult erythrocytes were more pronounced than those between neonatal and adult cells. The GR activity of fetal erythrocytes was also higher than that of either normal adult or maternal blood. This difference, however, was reduced to an insignificant level when the enzyme was activated in vitro by flavin adenine dinucleotide (FAD) because of a relatively low per cent activation of the GR in the fetal erythrocytes.
8;21 translocation in acute granulocytic leukaemia: cytological, cytochemical and clinical features
Authors: Swirsky D. M. , Li Y. S. , Matthews J. G. , Flemans R. J. , Rees J. K. H. , Hayhoe F. G. J.
Published: March 2008

Summary. Thirty patients with the 8;21 translocation and three with closely related variants have been studied. Ages ranged from 3 to 64 years (mean 28±3). Thirty‐one were entered into the MRC's 8th Acute Myeloid Leukaemia Trial. Twenty‐nine (88%) achieved complete remission. Marrow smears from most patients showed granulocytic maturation (M2, FAB classification) with characteristic abnormalities, but at least six showed predominantly myeloblastic (M1) morphology. The blast cells were markedly heterogeneous with regard to size and nuclear cytoplasmic ratio. Typical staining patterns were observed in the blast cells using Sudan black B and diaminobenzidine peroxidase stains, and to a lesser extent with periodic acid‐Schiff and chloroacetate esterase. Butyrate esterase was negative in all cases. Auer rods were present in the granulocyte precursors in 31 cases and in eosinophil precursors in two cases. In most cases the existence of the translocation was predicted from the cytological and cytochemical findings. Seven patients developed solid leukaemic deposits, principally in the mastoid cavities, orbital cavities or thoracic spine (extradural).
Effect of pure erythropoietin on DNA‐synthesis by human marrow day 15 erythroid burst forming units in short‐term liquid culture
Authors: Dessypris Emmanuel N. , Krantz Sanford B.
Published: March 2008

Summary. The effect of pure erythropoietin (EP) on human marrow day 15 burst‐forming units‐erythroid (BFU‐E) was studied using a short‐term liquid culture system containing 30% human serum. Non‐adherent marrow cells were cultured in liquid medium for 0–48 h and then the number of BFU‐E was assayed by the use of the plasma clot method. The addition of 1 U/ml of EP into the liquid culture medium resulted in maintenance of the number of BFU‐E assayed after 24–48 h of incubation. The number of BFU‐E recovered after 24–48 h culture was directly proportional to the concentration of EP present in the liquid medium. In addition, the proliferative status of BFU‐E before and after exposure to EP was studied by 3H‐thymidine and hydroxyurea suicide. It was found that EP doubles the percentage of BFU‐E in DNA synthesis after 24–48 h of incubation in the liquid medium. This effect of EP on DNA synthesis by bone marrow day 15 BFU‐E is detectable as early as 6 h after the onset of incubation and at EP concentrations as low as 0.2 U/ml of medium, a concentration present in the serum of moderately anaemic patients. The human marrow day 15 BFU‐E is an EP‐responsive cell and pure EP can induce it into DNA synthesis.
Thalassaemia intermedia: a new molecular basis
Authors: Thein S. L. , Al‐Hakim I. , Hoffbrand A. V.
Published: March 2008

Summary. A 5‐year‐old child heterozygous for β thalassaemia has the clinical picture of thalassaemia intermedia. Restriction endonuclease mapping shows that the child is homozygous for a triplicated α gene complex. The greater degree of globin chain imbalance resulting from two additional α chain genes is the likely mechanism for this unusually severe clinical phenotype.
Rapid platelet morphological changes visualized by scanning‐electron microscopy: kinetics derived from a quenched‐flow approach
Authors: Gear Adrian R. L.
Published: March 2008

Summary. Platelet activation is accompanied by distinct morphological changes from disc‐shaped cells to more rounded particles with multiple blebs or pseudopodia. A quenched‐flow approach has been used to follow the kinetics and nature of these morphological events. Scanning‐electron micrographs revealed very rapid alterations in platelet shape. At 0.5 s after activation with ADP or thrombin, the number of resting disc‐shaped particles was nearly halved and short blebbed forms were maximal at 0.5 s. By 1.7 s about 60% of particles were in the fully activated or multiple‐blebbed form. The calcium ionophore A‐23187 caused slightly slower effects. Adrenalin was much less potent, with about 14% of platelets becoming fully activated by 10 s. Control experiments showed only small changes in particle morphology when unactivated platelets were pumped through the reaction tubing and then quenched in glutaraldehyde. The resistive volume of platelets increased by 0.42 fl at 0.5 s after ADP stimulation and was essentially independent of variations in particle shape. These results show that the quenched‐flow approach can provide new information about platelet function and that morphological changes begin earlier than previously thought.
Bone marrow transplantation from identical twins in the treatment of aplastic anaemia: implication for the pathogenesis of the disease
Authors: Champlin Richard E. , Feig Stephen A. , Sparkes Robert S. , Gale Robert Peter
Published: March 2008

Summary. Treatment of aplastic anaemia by bone marrow transplantation from a syngeneic (identical twin) donor has provided insights into the pathophysiology of the disease.

We report from patients with severe anaemia who were treated by syngeneic bone marrow transplantation. None of the patients had sustained recovery of peripheral blood counts. All four received second transplants from the same twin donor after immunosuppressive conditioning treatment. Each had prompt recovery of haematopoiesis. A review of the literature indicates that failure of syngeneic bone marrow transplantation in patients with aplastic anaemia is not uncommon. These data indicate that aplastic anaemia may be caused by a mechanism other than an absence or intrinsic abnormality of haematopoietic stem cells in many patients.
Isolation of normal human megakaryocytes
Authors: Sitar Giammaria
Published: March 2008

Summary. This paper reports a simple procedure for obtaining human megakaryocytes with a high purification and high recovery yield. Bone marrow cells, obtained from surgically removed ribs, were separated by a two‐step procedure. Initially, a single cell suspension was enriched in megakaryocytes by equilibrium density centrifugation, the low density cell fraction was subsequently layered over a shallow continuous albumin gradient in a glass sedimentation chamber. Megakaryocytes averaged 0.03 ± 0.02% of all nucleated cells in the starting marrow cell suspension, after this procedure an average 80 ± 15% of the initial megakaryocyte population was recovered with a purity of 94 ± 4%. Previous methods, based upon the use of a two‐step procedure, are reviewed. The theory of velocity sedimentation is discussed with regard to the differences in the methodology used, which account for the different results I obtained.
Natural killer‐interferon system in patients with preleukaemic states
Authors: Takagi Shojiro , Kitagawa Seiichi , Takeda Akira , Minato Nagahiro , Takaku Fuminaro , Miura Yasusada
Published: March 2008

The natural killer (NK)‐interferon (IFN) system was investigated in patients with preleukaemic states. Endogenous NK cell activity was markedly reduced in all 12 patients studied. No significant correlation was observed between the activity of NK cells and the percentage of blast cells in the peripheral blood or bone marrow. The frequency of large granular lymphocytes (LGL) in lymphocytes of the peripheral blood was not reduced in most patients. The percentage of target binding cells in patients was essentially the same as that in normal controls, and the activity of NK cells from normal donors was not affected by the coexistence of mononuclear cells obtained from the peripheral blood of patients.

Furthermore, the α‐IFN production in response to HeLa cells persistently infected with measles virus was reduced in all patients studied, except for one case with acquired idiopathic sideroblastic anaemia. The augmented activity of NK cells induced by α‐IFN was variable, but remained at lower levels than the endogenous activity of NK cells in normal controls.

These findings suggest that not only the intrinsic defect and reduced number of NK cells but also the dysfunction of the NK‐IFN system may be responsible for the reduced activity of NK cells in patients with preleukaemic states.
Analysis of lymphocyte subsets in patients with aplastic anaemia
Authors: Zoumbos Nicholas C. , Ferris Warren O. , Hsu Su‐Ming , Goodman Steven , Griffith Patricia , Sharrow Susan O. , Humphries R. Keith , Nienhuis Arthur W. , Young Neal
Published: March 2008

Lymphocyte subsets have been measured in the blood of 28 patients with aplastic anaemia. The mean helper/inducer:suppressor/cytotoxic T lymphocyte ratio (1.24 ± 0.74) was significantly decreased in the total population in comparison to mean ratios in a normal population (1.78±0‐57) and in patients with other haematological diseases (l‐82±0‐92). A reversed ratio (< 1) was present in a large proportion (53%) of aplastic patients, due both to an absolute deficiency of helper/inducer lymphocytes and an increase in suppressor/cytotoxic lymphocytes. Increased HLA‐DR expression, evidence of lymphocyte activation, was present in seven of 12 patients evaluated and was confined to the suppressor/cytotoxic lymphocyte population. In the bone marrow, the percentage of lymphocytes was increased two‐fold compared to normal bone marrow but the ratio of T cell subsets was not abnormal. Two of four patients evaluated after haematopoietic recovery following ATG treatment showed a return to normal of the T4/T8 ratio; in two others a low T4/T8 ratio persisted despite recovery. These results indicate that lymphocyte subset imbalance and lymphocyte activation are present in many patients with aplastic anaemia, either as a contributing factor or as a result of bone marrow failure.
Arachidonic acid and leukotriene B4 induce aggregation of human peripheral blood mononuclear leucocytes in vitro
Authors: Villa Silvia , Colotta Francesco , Gaetano Giovanni DE , Semeraro Nicola
Published: March 2008

Peripheral human blood mononuclear leucocytes (MNL) aggregated in response to arachidonic acid (AA) in vitro. This phenomenon was similar to that already described for polymorphonuclear cells (PMN). The effect of AA was concentration‐dependent and was shared only by the structurally related di‐homo‐y‐linolenic acid among the other fatty acids tested. A number of agents able to induce platelet aggregation such as ADP, collagen, serotonin and a stable prostaglandin analogue all failed to stimulate MNL or PMN aggregation. Inhibitors of cyclo‐oxy‐genase activity such as acetylsalicylic acid and indomethacin not only did not prevent AA‐induced aggregation, but even potentiated it. In contrast, both nordihydroguaiaretic acid and BW 755C, two inhibitors of cyclo‐oxygenase and lipoxygenase, strongly prevented MNL aggregation. Thus AA seems to aggregate MNL through the mediation of lipoxygenase products. This is supported by the observation that leukotriene B4 (LTB4) also induced MNL aggregation. When highly purified lymphocyte and monocyte preparations were assessed separately, the latter responded to AA similarly to mixed MNL whereas lymphocyte aggregation was inconsistent, small and reversible even at high concentrations of AA. Although the pathophysiological significance of the MNL aggregation described here is still obscure, assembly of these cells‐particularly monocytes‐at the site of injury might be a crucial event.
GM 58/8: a monoclonal antibody that identifies a new lineage‐specific determinant expressed by myeloid progenitors (CFU‐GM) and their progeny
Authors: Gupta Amar Das , Brice Martha , Yokochi Takashi , Papayannopoulou Thalia , Stamatoyannopoulos George
Published: March 2008

A cytotoxic (IgM) monoclonal antibody (McAb) raised against human erythroleukaemia cell line HEL appears specific for the myelomonocytic lineage. Indirect immunofluorescence and fluorescence‐activated cell sorter analyses of peripheral blood and bone marrow cells showed that this McAb (designated GM 58/8) reacts exclusively with more than 90% of the granulocytes, monocytes and myeloid precursors. The positive (fluorescent) fraction sorted from McAb‐treated bone marrow buffy coat cells showed a marked enrichment of myeloid precursors, while nonmyeloid cells were recovered in the negative fraction. GM 58/8 specifically inhibited the growth of 90‐100% of the committed myelomonocytic progenitors (CFU‐GM) in five complement‐mediated cytotoxicity experiments. GM 58/8 had virtually no effect on the erythroid progenitor (BFU‐E and CFU‐E) growth. Cell sorting experiments using this McAb resulted in recovery of 84% of CFU‐GM among the fluorescent fractions. Comparisons with other anti‐myelomonocytic McAbs reported so far suggest that GM 58/8 has more restricted reactivity and shows more efficient CFU‐GM cytotoxicity. Patterns of reactivity with normal cells and established lines indicate that the antigenic determinant identified by GM 58/8 is different from that recognized by previously described anti‐myelomonocytic McAbs. McAb GM 58/8 could be used to isolate or remove pure populations of progenitors and/or more differentiated cells of the myelomonocytic lineage for in vitro studies. This study also provides evidence that the HEL cell line, derived originally from an erythroleukaemia patient, expresses an antigenic determinant shared by the normal cells of myelomonocytic lineage.
Monoclonal antibody specific for granulocytic‐lineage cells and reactive with human pluripotent and committed haematopoietic progenitor cells
Authors: Janowska‐Wieczorek Anna , Mannoni Patrice , Turner A. Robert , McGann Locksley E. , Shaw Andrew R. E. , Turc Jean‐Michel
Published: March 2008

A murine monoclonal antibody (82H5, IgM class) has been developed that detects an antigenic determinant expressed by ± 90% of normal granulocytes and 60‐80% of light‐density normal bone‐marrow cells, including human pluri‐potential progenitors (colony‐forming‐unit‐granulocyte, erythroid, macrophage, megakaryocyte; CFU‐GEMM) and committed progenitors: granulocyte‐macrophage (CFU‐GM), erythroid (BFU‐E), and megakaryocytic (CFU‐MK). This antibody did not react with erythrocytes, monocytes, platelets, lymphocytes from normal peripheral blood, lymphoblasts from patients with acute lymphoblastic leukaemia, or with lymphoid cell lines. The 82H5‐defined antigenic determinant was expressed on ±90% of leukaemic cells of promyelocytic, myelomonocytic and monocytic morphology, and cell lines KG.1, ML.1, HL.60, K562 and U.937. Cortical thymocytes were unreactive with 82H5. Treatment of human bone‐marrow cells with granulocytic‐specific monoclonal antibody 82H5 plus complement significantly inhibited colony formation (48‐74%; P±0.05) of CFU‐GEMM, CFU‐GM, BFU‐E, CFU‐MK, whereas treatment with control monoclonal anti‐la antibody plus complement caused 79‐89% inhibition. This antibody reacted strongly with 3‐fuc‐NAc lactosamine when tested with a panel of synthetic carbohydrate structures. We conclude that 82H5 may be a useful probe for phenotypic analysis of leukaemic cells and investigation of haematopoiesis.
Neutrophils with ring‐shaped nuclei in myeloproliferative disease
Authors: Langenhuijsen M. M. A. C.
Published: March 2008

Neutrophils with ring‐shaped nuclei were found in 12 out of 28 patients with myeloproliferative disease and only rarely in patients with toxaemic granulocytosis and in healthy controls. Nuclear ring formation and hyposegmentation appeared to be significantly related.
Normal content of brain spectrin‐like protein in sph/sph mice
Authors: Casoria Linda A. , Zagon Ian S. , Bernstein Seldon E. , Shohet Stephen B. , McLaughlin Patricia J. , Goodman Steven R.
Published: March 2008

In the erythrocytes of WBB6F1‐sph/sph mice spectrin constitutes only ‐ 1% of the total sph/sph membrane protein compared to ‐ 23% in WBB6F1‐+/+ controls. No increase in proteolytic degradation of spectrin in sph/sph erythrocyte membranes could be detected with antibodies directed againt mouse erythrocyte spectrin or mouse brain spectrin‐like protein. As attachment of normal spectrin to the erythrocyte membrane of these animals appeared to be normal, and as spectrin not detected when whole sph/sph erythrocytes are solubilized in SDS for SDS PAGE, the deficient erythrocyte spectrin was probably due to diminished production. Brain spectrin‐like protein, a nonerythroid spectrin analogue, is antigenically, morphologically and functionally related to erythrocyte spectrin, but appears by peptide mapping analysis to be a distinct gene product. It was found by protein‐ and antibody‐staining of brain membranes to be present in normal concentrations in sph/sph animals. Indirect immunofluorescence of mouse brain tissue with anti‐brain spectrin‐like protein IgG or anti‐erythrocyte spectrin IgG indicated that the distribution of brain spectrin‐like protein was normal in sph/sph brain. Therefore the mutation causing diminished production of sph/sph erythrocyte spectrin does not affect the expression of this nonerythroid spectrin analogue.
Depletion of excessive liver iron stores with desferrioxamine
Authors: Cohen Alan , Martin Marie , Schwartz Elias
Published: March 2008

TO determine the therapeutic effect of long‐term, intensive iron chelation therapy, we studied liver iron content and histology in four children with thalassaemia major during 52‐83 months of intensive therapy with desferrioxamine. The initial biopsies obtained prior to or within 21 months after beginning chelation therapy had Grade IV iron staining, with heavy iron deposition present in parenchymal and reticuloendothelial cells. Subsequent biopsies, obtained when serum ferritin levels had fallen to 71‐246 μg/1, contained Grade 0 or Grade I stainable iron. Little or no iron was present in parenchymal or reticuloendothelial cells. The liver iron concentration, measured by magnetic susceptibility, returned to normal or nearly normal levels. Hepatic fibrosis did not progress during treatment with desferrioxamine. These findings demonstrate that intensive and sustained chelation therapy with desferrioxamine will remove excessive liver iron and preserve hepatocellular structure.
Autologous bone marrow transplantation contributes to haemopoietic recovery in children with solid tumours treated with high dose melphalan
Authors: Kingston Judith E. , Malpas J. S. , Stiller C. A. , Pritchard J. , McElwain T. J.
Published: March 2008

During a 5 year period, 64 children with advanced stage malignant solid tumours have been treated with high dose melphalan (140‐220 mg/m2) combined with non‐cryopreserved autologous marrow transplantation. All children developed a profound neutropenia and thrombocytopenia following high dose melphalan. Both the duration of neutropenia (median l1d) and the duration of thrombocytopenia (median 18 d) were related to the nucleated cell count of the reinfused marrow but neither appeared to be affected by the dose of melphalan. This suggests that the initial recovery of the peripheral blood count results from the engrafted autologous marrow. We conclude that autologous marrow transplantation accelerates haemopoietic recovery in children with solid tumours treated with high dose melphalan.
Actin pools and actin microfilament organization in cultured human endothelial cells after exposure to thrombin
Authors: Galdal Kjell Sverre , Evensen Stein A. , Høglund Anna‐Stina , Nilsen Erling
Published: March 2008

Human umbilical vein endothelial cells (HUVEC) in primary confluent cultures lost their normal polygonal shape and assumed a ‘contracted’ appearance as judged by phase contrast microscopy when exposed to highly purified bovine thrombin (2 N.I.H. u/ml). Total actin in thrombin‐exposed cells did not differ from that of control cells, as measured by the deoxyribonuclease I inhibition assay. However, the monomeric actin pool (unpolymerized actin) in thrombin‐treated HUVEC was c. 15% smaller (P<0.01) than in control HUVEC (in which it represented approximately 50% of total actin). Transmission electron microscopy showed that thrombin‐stimulated HUVEC contained more and thicker bundles of filamentous actin than control cells. Polymerization of actin and reorganization of actin microfilaments may contribute to the shape changes of HUVEC induced by thrombin.
Heat production rate in polymorphonuclear granulocytes from patients with acute myelogenous leukaemia and healthy individuals
Authors: Faldt R. , Ankerst J , Monti M.
Published: March 2008

Heat production rate in polymorphonuclear granulocytes (PMN) from 18 AML patients and 21 healthy individuals was measured using microcalorimetry. An increased PMN heat production rate was found in most AML patients at diagnosis and during the first 6 months of remission. In 7/7 patients showing values above the normal mean level the heat production rate decreased during sequential analysis. The increased PMN heat production rate is serum dependent and leukaemic sera induced increased calorimetric values also in PMN from healthy donors. This effect was not due to heat labile complement factors.
Measurement of platelet life‐span in normal subjects and patients with myeloproliferative disease with indium oxine labelled platelets
Authors: Bautista A. P. , Buckler P. W. , Towler H. M. A. , Dawson A. A. , Bennett B.
Published: March 2008

The use of “‘Iridium oxine as a platelet label for the performance of platelet life‐span studies has been examined. Platelet life‐span in normal subjects varied between 8‐10 and 10‐36 d. Patients with primary thrombocythaemia had clearly reduced platelet life‐span whether or not they presented with vascular occlusion and this abnormality persisted after reduction of the platelet count to normal by busulphan therapy. Patients with similarly elevate platelet counts due to chronic granulocytic leukaemia or after splenectomy had platelet life‐span values in the normal range. Plasma β‐TG levels could not be used to predict platelet life‐span in these groups of patients. Measurement of platelet life‐span using 111 Indium labelled platelets is a useful technique in the examination of platelet function in occlusive vascular disease.
Low plasma cell 3(H) thymidine incorporation in monoclonal gammopathy of undetermined significance (MGUS), smouldering myeloma and remission phase myeloma: a reliable indicator of patients not requiring therapy
Authors: Boccadoro Mario , Gavarotti Paolo , Fossati Giorgio , Pileri Alessandro , Marmont Filippo , Neretto Giorgio , Gallamini Andrea , Volta Carla , Tribalto Maurizio , Testa Maria G. , Amadori Sergio , Mandelli Franco , Durie Brian G. M.
Published: March 2008

The kinetics of bone marrow plasma cells were evaluated by means of in vitro 3(H) thymidine incorporation in 143 patients with monoclonal gammopathies. Fifty‐three patients had symptomatic multiple myeloma (MM) at diagnosis, nine were in stable remission, six in unstable remission, and 16 in the relapse phase. Thirty‐seven patients were classified has having monoclonal gammopathy of undetermined significance (MGUS) and 22 as smouldering myeloma (SM). A thymidine labelling index (LI%) of > 3 at initial diagnosis predicted a very short survival. High LI% values (median 2.8 ± 1.1) were also seen at relapse. However, the major new finding was that the LI% could be used to discriminate precisely between the SM‐MGUS group and the MM patients including stage I disease (P < 0‐0001). Only one patient developed MM during follow up, that being 8 months after the initial diagnosis of SM. During the unmaintained stable remission (plateau) phase a low proliferative activity was also observed (LI%= 0.6 ± 0.2).

Thus the LI% was extremely useful in identification of both poor risk groups with a LI% > 3 and stable patients requiring no immediate therapy with a LI% < 1. The ability to discriminate between MGUS and SM and stage I MM should prove particularly useful clinically.
Successful chemotherapy with deoxycoformycin in adult T‐cell lymphoma‐leukaemia
Authors: Daenen S. , Rojer R. A. , Smit J. W. , Halie M. R. , Nieweg H. O.
Published: March 2008

A patient from the Caribbean area with active T‐cell lymphoma‐leukaemia was primarily treated with deoxycoformycin (DCF), 5 mg/m2 i.v. on 3 consecutive days, followed by 5 mg/m2 i.v. weekly. A complete remission was attained and maintained during several weeks with DCF. A single consolidation course with other cytostatics was then given. The patient continues in complete remission without further therapy, 24 months after diagnosis, 17 months after the last cytostatic drugs. T‐cell lymphoma‐leukaemia has a bad prognosis with conventional anti‐lymphoma therapy but was exquisitely sensitive to DCF in this patient.
Bone marrow biopsy findings in angioimmunoblastic lymphadenopathy
Authors: Ghani Ahmed M. , Krause John R.
Published: March 2008

Summary. Bone marrow biopsies from 13 cases of lymph‐node‐biopsy‐proven angioimmunoblastic lymphadenopathy (AILD) were studied. Eight cases (62%) showed a characteristic ‘granulomatoid’ lymphoreticular infiltrate composed of a mixture of lymphocytes, epithelioid histiocytes, immunoblasts, plasma cells and eosinophils in varying proportions. Proliferation of blood vessels, although much less prominent than that described in lymph nodes, and a marked degree of reticulin fibrosis were also noted. The pattern of involvement was mainly focal and rarely diffuse. The foci were either single or multiple, and were distributed throughout the marrow but were only rarely paratrabecular. The associated haematopoietic marrow findings were nonspecific regardless of degree of bone marrow involvement. The histopathology of involved bone marrows in angioimmunoblastic lymphadenopathy is such that in some cases it should strongly suggest the diagnosis even in the absence of a prior definitive lymph node biopsy. Differentiating features from other disorders that might be confused with AILD are discussed. Survival rate was adversely affected by bone marrow involvement.
Lineage specific classification of leukaemia: results of the analysis of sixty cases of childhood leukaemia
Authors: Ha Kyungsae , Hozumi Nobumichi , Hrincu Angela , Gelfand Erwin W.
Published: March 2008

Summary. In addition to conventional morphological, histochemical and immunological marker studies, cells from 60 children with leukaemia were further analysed using the Southern blot hybridization technique to look at differences in the organization of immunoglobulin (Ig) genes. Of the 60 patients studied by conventional means, 47 were diagnosed as acute lymphocytic leukaemia (ALL) and 13 as non‐lymphocytic leukaemia. Seven patients were initially classified as T ALL and 40 as non‐T, non‐B ALL. Further subclassification of the 40 patients with non‐T, non‐B ALL indicated three pre‐B ALL and 29 patients diagnosed as common ALL, expressing Ia and CALLA antigens. All 29 patients with common ALL demonstrated Cμ gene rearrangements with or without light chain (κ and λ) genes rearrangement. Based on the developmental hierarchy of Ig gene rearrangements, it was possible to further subclassify the patients with common ALL into different stages of B cell development. Eight (of the 40) patients with non‐T, non‐B ALL were identified as CALLA‐ but further analysis indicated T‐lineage origin in two patients and three patients were reclassified as acute undifferentiated leukaemia (AUL). Cμ gene rearrangements were detected in two patients with T ALL, two patients with AUL and one patient with acute myelogenous leukaemia (AML). In contrast to the patients with common ALL, Ig gene rearrangement observed in these non‐B‐lineage cells was restricted to a single Cμ gene while retaining germ‐line configuration of the other allele of the Cμ gene and both light chain genes.
A novel form of factor VII in plasma from men at risk for cardiovascular disease
Authors: Dalaker Knut , Hjermann Ingvar , Prydz Hans
Published: March 2008

Summary. In the Northwick Park Heart Study high activity of the coagulation factor VII was associated with cardiovascular deaths. We confirm here that men at high risk for such disease have increased levels of factor VII activity and demonstrate for the first time that this is due to a phospholipid–factor VII complex in their plasma. This complex increases the specific activity of its factor VII significantly. It is not inhibited by a neutralizing antiserum to thromboplastin and thus does not represent a factor VII‐thromboplastin complex. Gel filtration data suggest an apparent Mr of 66–72 000 for the complex. Factor VII in this phospholipid complex is more sensitive than native factor VII to the serine protease inhibitor diisopropyl‐fluorophosphate, consistent with a more accessible active site conformation (and a higher specific activity) for factor VII in the complex. The level of phospholipid–factor VII complex showed a significant positive correlation with plasma triglycerides and correlation of borderline significance with cholesterol.
Membrane characteristics and osmotic fragility of red cells, fractionated with anglehead centrifugation and counterflow centrifugation
Authors: Van Der Vegt S. G. L. , Ruben A. M. Th. , Werre J. M. , de Gier J. , Staal G. E. J.
Published: March 2008

Summary. Red cell populations were separated on the basis of differences in density using anglehead centrifugation and on the basis of differences in mean cell volume using counterflow centrifugation. In the different fractions, mean surface area was calculated, phospholipid and cholesterol content determined as well as the osmotic behaviour in hypotonic salt solutions. Older red cells appeared to be more resistant to hypotonic salt solutions, due to favourable surface area to volume ratio.
The immunodepletion of factor VII from human plasma using a monoclonal antibody
Authors: Cerskus A. L. , Ofosu F. A. , Birchall K. J. , Clarke B. J. , Modi G. J. , Johnston M. , Blajchman M. A.
Published: March 2008

Summary. A murine hybridoma cell line which secretes monoclonal antibody to factor VII has been prepared to facilitate the immunodepletion of this clotting factor from plasma. Specific monoclonal antibody was purified from mouse ascites tumours by protein A‐Sepharose chromatography and shown to be of the IgG1 immunoglobulin subclass. On immunoblotting, this antibody reacted with a single protein band identical to purified factor VII. The purified monoclonal antibody was coupled to Sepharose 4B and was used to immunodeplete factor VII from pooled normal human plasma. The prothrombin time of plasma immunodepleted in this way was 35 s compared to 12 s for the starting plasma. Specific factor assays of the immunodepleted plasma showed factor VII activity to be less than 1% while the levels of the other clotting factors were unchanged. The immunodepleted plasma was equivalent to severe congenital factor VII deficient plasma as a substrate for factor VII assays. Bound factor VII could be eluted from the immunoaffinity column with citrate buffer, pH 6·0, with good recovery.
Changes in plasma fibronectin during donor apheresis and therapeutic plasma exchange
Authors: Norfolk D. R. , Bowen M. , Cooper E. H. , Robinson E. A. E.
Published: March 2008

Summary. Changes in plasma levels of the opsonic glycoprotein fibronectin were studied during machine apheresis in 32 healthy plasma donors and 18 patients undergoing a total of 86 therapeutic plasma exchanges. Donation of 500 ml of plasma at 3‐monthly intervals produced a 10% immediate decrease in plasma fibronectin with no evidence of a long‐term affect on levels of this protein. One plasma volume exchange with fibronectin‐poor plasma protein fraction produced subnormal plasma fibronectin levels in 98% of post‐exchange samples. Efficiency of exchange was significantly lower in patients with paraproteinaemia, probably reflecting increased plasma volume in such patients. Daily plasma exchange produced progressive depletion of plasma fibronectin (index of recovery 0·68) whereas levels were maintained close to the initial value with longer exchange intervals, except in the more severely ill patients. Reduction of plasma fibronectin by plasma exchange may increase susceptibility to infection and reduce the efficacy of this procedure when the object of exchange is to reverse reticuloendothelial blockade.
Treatment of primary proliferative polycythaemia by venesection and low dose busulphan: retrospective study from one centre
Authors: Messinezy M. , Pearson T. C. , Prochazka A. , Wetherley‐Mein G.
Published: March 2008

Summary. Sixty‐five patients with primary proliferative polycythaemia (polycythaemia rubra vera) were followed during the period 1962–83 and analysed retrospectively. Primary control of PCV was by venesection only with low dose busulphan solely as required to keep the platelet count below 400 × 109/l. Median survival was 11·1 years from diagnosis which is equal to or marginally better than with other reported regimens. Vascular causes of death were only a little higher than expected in a comparable normal population. Only deaths from acute leukaemia and myelofibrosis were significantly increased above the normal population incidence. There was no evidence to suggest that these transformations were busulphan induced. Analysis of the incidence of occlusive vascular lesions lends support to an earlier recommendation that the PCV level be maintained below 0·45. No support was found for the possible disadvantages of a predominantly venesection regimen, such as iron deficiency and reactive thrombocytosis. The case is put for this use of low dose busulphan. The data presented would warrant the future inclusion of this therapeutic regime as one limb of a controlled trial.
Episodes of increased fibronectin level observed in a patient suffering from recurrent thrombosis related to congenital hypodysfibrinogenaemia (fibrinogen Malmoe)
Authors: Soria J. , Soria C. , Hedner U. , Nilsson I. M. , Bergqvist D. , Samama M.
Published: March 2008

Summary. A study has been conducted in a Swedish patient with severe thrombotic disease and repeated miscarriages related to a hypodysfibrinogenaemia with defective thrombin binding to the abnormal fibrin. The hypodysfibrinogenaemia was found in several members of the family. The patient also had an increased concentration of fibronectin in her plasma at two different occasions. This would appear to be unrelated to the abnormal fibrinogen since a normal concentration of fibronectin has been found in her relatives presenting the same fibrinogen anomaly, and in the patient at two other times. In conclusion, the thrombotic disorder in this patient presenting a congenital hypodysfibrinogenaemia may be explained by the defective thrombin binding to fibrin.
Circulating autologous stem cells collected in very early remission from acute non‐lymphoblastic leukaemia produce prompt but incomplete haemopoietic reconstitution after high dose melphalan or supralethal chemoradiotherapy
Authors: Juttner C. A. , To L. B. , Haylock D. N. , Branford A. , Kimber R. J.
Published: March 2008

Summary. Haemopoietic reconstitution (HR) using autologous peripheral blood stem cells (PBSC) was attempted after intensive chemotherapy or chemoradiotherapy in two patients with relapsed acute non‐lymphoblastic leukaemia (ANLL). The PBSC were collected by leukapheresis very early in first remission and cryopreserved in liquid nitrogen. Both patients demonstrated early evidence of trilineage engraftment. The first patient received melphalan 200 mg/m2 followed by rescue with 1·3 × 108 mononuclear cells/kg body weight containing 29 × 104 granulocytemacrophage progenitor cells (CFU‐GM)/kg, and HR was evident by Day 14. The second patient was treated with supralethal chemoradiotherapy followed by rescue with 3·0 × 108 mononuclear cells/kg containing 23 × 104 CFU‐GM/kg. He demonstrated early engraftment with near normal peripheral blood counts by Day 16. There was a subsequent fall in both bone marrow cellularity and peripheral blood counts to a level of low but persistent activity. There was a further phase of haematological recovery from 8 weeks following transplantation with an increase in peripheral blood counts and bone marrow cellularity until final relapse at 13 weeks. This study demonstrates that circulating stem cells have haemopoietic reconstitutive capacity, previously only shown with buffy coat cells from chronic granulocytic leukaemia. The minimum number of PBSC required for satisfactory engraftment remains unknown, although it seems probable that the ratio of pluripotent stem cells to committed progenitor cells is lower in very early remission peripheral blood than in either allogeneic normal bone marrow or autologous bone marrow collected laterin stable remission. The question of leukaemic contamination of the PBSC remains to be answered.
Expression of human c‐fes one‐gene occurs at detectable levels in myeloid but not in lymphoid cell populations
Authors: Ferrari Sergio , Torelli Umberto , Selleri Licia , Donelli Amedea , Venturelli Donatella , Moretti Luigi , Torelli Giuseppe
Published: March 2008

Summary. Total cellular RNA from a variety of myeloid and lymphoid cell populations, normal and leukaemic, was analysed for the expression of a human cellular one‐gene, c‐fes, by northern blot hybridization assays. The probe used was a molecularly cloned human DNA sequence homologous to the 5′ terminal sequence of v‐fes. All the myeloid cellular populations expressed the c‐fes gene. In some cell populations at an advanced stage of differentiation (circulating leucocytes from Chronic myeloid leukaemia (CML), HL60 cells induced to differentiate by retinoic acid) the level of expression was even higher than in early stages of the myeloid lineage (blast cells from AML, uninduced HL60 cells). No transcript of the c‐fes gene was detected in the different lymphoid populations studied. The occurrence of an RNA complementary to the c‐fes sequence appears sufficiently characteristic of a myeloid population to distinguish it from a lymphoid population, normal and leukaemic.
Detection of the blastic crisis cell clone in chronic myeloid leukaemia
Authors: Löwenberg Bob , Hagemeijer Anne , Abels Johannes
Published: March 2008

Summary. Three patients with Ph1 positive chronic myeloid leukaemia (CML) were followed until the occurrence of blastic transformation by means of cytogenetic analysis of whole marrow or blood cells, of blast cell enriched fractions from marrow or blood and of colonies grown from these cell fractions in two culture systems. Karyotypic evolution was used to indicate the development of new clones from pre‐existing cell lines either as transient phenomena during the chronic phase or as the ultimate indicators of a blastic crisis (BC). The clones which were to lead to a BC were detected some months before the clinical diagnosis of the BC. At a time before overt BC had become clinically manifest, the BC clone outgrew the other ones in culture and thus also in vitro showed specific proliferative advantages over the cell lines characteristic of the chronic phase of CML.
Stability of azure B–eosin Y staining solutions
Authors: Bins M. , Huiges W. , Halie M. R.
Published: March 2008

Summary. The stability of azure B–eosin Y staining solutions of varying composition and of a routine May Grunwald Giemsa (MGG) stain were studied by analysis of the density histogram of white blood cells obtained by an image analysis computer. The stability appeared to be variable and depended on the concentration of the dyes, the molarity of the buffer solutions and the presence of dimethylsulfoxide (DMSO) as a stabilizer. Although most staining solutions including the routine MGG stain showed marked loss of staining capacity soon after preparation, it was possible to obtain an azure B–eosin Y mixture with very satisfactory staining properties which did not decrease during 8 h after its preparation.
Serial incidence of bone marrow GM‐CFC prior to the development of acute non‐lymphoblastic leukaemia in a child treated for non‐Hodgkin's lymphoma
Authors: Haworth Catherine , Stevens R. F. , Testa Nydia E. G.
Published: March 2008

Summary. A child treated for NHL developed acute non‐lymphoblastic leukaemia 27 months after stopping treatment. Serial in vitro bone marrow studies showed a normal incidence of GM‐CFC following treatment. However, GM‐CFC incidence dropped at least 15 months prior to the development of leukaemia. This was associated with an asymptomatic neutropenia but no disturbance of bone marrow morphology. It is concluded that sub‐clinical disturbances of bone marrow function may play an important part in leukaemogenesis.
Anticardiolipin antibodies in autoimmune thrombocytopenic purpura
Authors: Harris E. N. , Gharavi A. E. , Hegde U. , Derue G. , Morgan S. H. , Englert H. , Chan J. K. H. , Asherson R. A. , Hughes G. R. V.
Published: March 2008

Summary

Using a recently devised solid phase radioimmunoassay to detect anticardiolipin antibodies, we report the presence of these antibodies in 30 of 96 patients with chronic autoimmune thrombocytopenic purpura (AITP). IgG anticardiolipin antibody levels were elevated in 14 patients and IgM anticardiolipin antibody levels were elevated in 27 patients.

We suggest that these antibodies may mediate peripheral platelet destruction by binding to phospholipids in the platelet membrane. It is also conceivable that the presence of anticardiolipin antibodies may select a subpopulation of patients with chronic AITP who may go on to develop other autoimmune disorders, such as systemic lupus erythematosus.
Immunoassays of factor IX antigen using monoclonal antibodies
Authors: Yoshioka A. , Giddings J. C. , Thomas J. E. , Fujimura Y. , Bloom A. L.
Published: March 2008

Summary

Monoclonal antibodies to factor IX were produced by immunization of balb/c mice with purified factor IX and fusion of spleen cells with SP‐1 murine myeloma cells. Antibody producing hybrids were detected by an enzyme linked immunoassay (ELISA) and by coagulation inhibitor (Bethesda type) methods. Monoclonal antibodies with titres of > 1 x 105 tested by the ELISA and 5000–8000 inhibitor units were obtained. A new ELISA method was developed using one of these monoclonal antibodies to quantitate factor IX antigen (IXAg) in plasma samples from patients with hereditary factor IX deficiency. In addition a two‐site solid phase immunoradiometric assay (IRMA) for factor IX was established. The results of these new methods were compared with those obtained on the same plasma samples using conventional factor IX coagulation assays and the Laurell rocket method. The lower limit for the detection of IXAg by the ELISA was approximately 0.01 unit per ml whilst that for the IRMA was about 0.001 unit per ml (normal plasma = 1 unit per ml). The lower detection limit for IXAg using the Laurell rocket method was about 0.06 units per ml. The improved sensitivity of the new immunoassays enabled quantitation of low levels of IXAg in patients with moderately severe factor IX deficiency and confirmed the presence of excess IXAg compared to IX activity (IXC) in a relatively high proportion of cases (28 out of 51 tested). Results of testing plasma from obligate carriers confirm the suggestion that measurements of IXAg and IXC may improve the classification of carrier status in these kindred.
Characterization of blast cells in chronic granulocytic leukaemia in transformation, acute myelofibrosis and undifferentiated leukaemia
Authors: Polli N. , O'Brien M. , Tavares de Castro J. , Matutes E. , San Miguel J. F. , Catovsky D.
Published: March 2008

Summary

A systematic analysis of the blast cell population was carried out on samples from 50 patients suffering from blast transformation of chronic granulocytic leukaemia (CGL) (31) and of myelofibrosis (4), acute myelofibrosis (AM) (11) and undifferentiated acute leukaemia (4). Transmission electron microscopy (TEM), used in 41 samples, included: morphology and techniques for myeloperoxidase (MPO), platelet‐peroxidase (PPO) and acid phosphatase (AP). The majority of cases were also studied by light microscopy cytochemistry and with a battery of cell markers which are reported in the accompanying paper (San Miguel et al, 1985). The characterization of the type(s) of proliferating blasts was made from the integration of ultrastructural and immunological data. TEM morphology allowed the precise recognition of specific granules in basophil and mast‐cell precursors and of ferritin particles in blasts of erythroid lineage; these rare cell types were not adequately characterized by other methods. The PPO reaction made possible the identification of pure megakaryoblastic proliferations in 38% of cases, including eight of the 11 with AM; megakaryoblasts were also present in nine of 12 cases with mixed blast cell types. The MPO and AP reactions were useful for the characterization of myeloblasts and monoblasts, respectively. Lymphoblasts could be distinguished from other cell types by TEM morphology and negative MPO and PPO reactions. TEM techniques were valuable for diagnosing correctly the type of blast cell in this study in which only four cases (8%) remained unclassifiable.