bluuebunny/crossref_metadata_2025 huggingface dataset:
Abnormal low and high density lipoproteins in homozygous beta‐thalassaemia
Authors: Goldfarb Ada W. , Rachmilewitz Eliezer A. , Eisenberg Shlomo
Published: March 2008

The levels, structure and composition of plasma lipoproteins were determined in 67 patients with homozygous beta‐thalassaemia and compared to healthy or heterozygous members of the same families and to patients with either sickle cell or iron deficiency anaemia. Plasma total and LDL and HDL cholesterol levels were low in patients with homozygous beta‐thalassaemia and with sickle cell anaemia. Plasma triglycerides did not differ between subjects. The low plasma and lipoprotein cholesterol was independent of age, transfusion requirements and splenectomy.

Abnormal structure and composition of lipoproteins was found in homozygous beta‐thalassaemia. The LDL was of higher density and was triglyceride‐rich and cholesterol ester‐poor. HDL separated to three populations. HDL2was prominent (in spite of low plasma HDL cholesterol). HDL2–3was of normal density and an intermediate HDL population, not found in normal subjects, was identified and designated HDL2–3. All three HDL populations were enriched with triglycerides and poor in cholesterol ester content. The modified LDL and HDL particles may then be possibly cleared rapidly from the plasma by activated monocytes and macro‐phages.
Clonality of cell populations in refractory anaemia using combined approach of gene loss and X‐linked restriction fragment length polymorphism‐methylation analyses
Authors: Abrahamson Gail , Boultwood Jacqueline , Madden Jacqueline , Kelly Susan , Oscier D. G. , Rack Katrina , Buckle Veronica J. , Wainscoat J. S.
Published: March 2008

We have used X‐linked restriction fragment length polymorphism (RFLP)‐methylation and gene deletion analyses to investigate the nature of the progenitor cell of origin in the myelodysplastic syndromes (MDS). Gene deletion studies were performed on the granulocyte and T‐lymphocyte fractions of six women with refractory anaemia (RA) and either a partial deletion of the long arm of chromosome 5 (5q‐) or monosomy 7. All six showed gene loss in the granulocyte but not the T‐lymphocyte fractions, indicating monoclonality of the granulocytes but not the T‐lymphocytes. In order to further investigate this finding, we subsequently performed X‐RFLP‐methylation studies using the probe M27β, and also a probe for the phosphoglycerate kinase (PGK) gene. These studies have confirmed the monoclonality of the granulocytes and the polyclonality of the T‐lymphocytes in these cases. Our findings suggest that in this group of patients with MDS the T‐lymphocytes were not involved in the disorder, and furthermore, in the one case where B‐lymphocytes were also available, that the progenitor cell of origin was restricted to the myeloid lineage.

This work was supported by the Leukaemia Research Fund of the United Kingdom. We thank Dr I. Craig and Professor A. Jeffreys for the probes M27β and pγ3, respectively. The probe for the FMS gene was kindly donated by Dr N. Spurr. The probes for the HPRT and PGK gene regions were used with the permission of Dr B. Vogelstein. We thank Dr D. Y. Mason for use of the monoclonal antibodies CD22 and CD68.
Second transplants for leukaemic relapse after bone marrow transplantation: high early mortality but favourable effect of chronic GVHD on continued remission. A REPORT BY THE EBMT LEUKAEMIA WORKING PARTY
Authors: Barrett A. J. , Locatelli F. , Treleaven J. G. , Gratwohl A. , Szydlo R. , Zwaan F. E.
Published: March 2008

Ninety second bone marrow transplants (BMT) for relapsed leukaemia were carried out in 30 European BMT centres. At second BMT, after further treatment in 64 cases, 43 patients were in complete remission or in chronic phase of CML, and 47 were in continuing relapse, accelerated phase or blast crisis of CML. Seventy patients died, 37 from early transplant‐related toxicity and relapse or failure to eradicate leukaemia which occurred in 23. There were 20 survivors. The actuarial disease‐free survival was 11% with a relapse probability of 69% at 3 years. Associated with reduced graftversus‐host disease (GVHD) prophylaxis during second BMT, the incidence and severity of acute and chronic GVHD, was increased when compared with the first BMT (P= 0.02, and 0.002 respectively for acute and chronic GVHD).

In multivariate analysis survival was shown to be favoured by a prolonged interval between first and second BMT (relative risk 1.3/year,P= 0.02), and no or mild chronic GVHD following first BMT (relative risk 2.3,P= 0.02). Continuing remission was favoured by chronic GVHD occurring after second BMT (relative risk 8.1,P= 0.004).

These results confirm the high treatment‐related mortality following second BMT, but identify superior survival in selected patients. Improved results might be achieved by further reduction in preparative regimen intensity, and increasing graft‐versus‐leukaemia reactivity.
In vitro Ig‐synthesis and proliferative activity in multiple myeloma are stimulated by different growth factors
Authors: Sonneveld Pieter , Schoester Martijn , Leeuw Kees de
Published: March 2008

Plasma cells isolated from bone marrow (BM) aspirates of 15 patients with active multiple myeloma (MM) were cultured and analysed for in vitro proliferative response and Ig‐synthesis upon stimulation with interleukin‐3 (IL‐3), interleukin‐4 (IL‐4) and interleukin‐6 (IL‐6). The proliferative response, determined as Ki‐67 positivity in concentrated plasma cells, was increased by IL‐6 (Stimulation Index, SI=1.77±0.21 (M±SEM)) but not by IL‐3 or IL‐4. This proliferation could be blocked by anti‐IL‐6. In vitro Igsynthesis was stimulated by IL‐4 (SI = 1.62±0.12 (M±SEM) P<0.05) but not by IL‐6 or IL‐3. This effect was not antagonized by anti‐IL‐6. An inverse correlation was found in this group of patients between the IL‐6 induced stimulation of plasma cell proliferative activity and the IL‐4 induced increase of Ig‐synthesis (P=0.027).

These data indicate in MM that Ig‐synthesis and the in vitro proliferative activity may be stimulated by different haematopoietic growth factors, which may reflect the involvement of different responding cells.
An in‐vitro direct chemiluminescence assay for assessment of platelet‐bound antibody in thrombocytopenic patients
Authors: Kazemi A. , Singh A. K. , Slater N. G. P.
Published: March 2008

A direct chemiluminescence test (CLT) employing autologous monocytes and granulocytes was used to assay platelet‐bound antibody in comparison to the direct platelet suspension immunofluorescence test (PSIFT) in 50 patients. Twenty‐six of the 30 immune thrombocytopenic purpura (ITP) patients, including all 21 chronic ITP patients with platelet counts below 100 × 109/1, showed positive results with at least one type of phagocytic cell; so did seven of the eight patients with other presumed autoimmune thrombocytopenias, and three out of 12 patients with thrombocytopenia associated with non‐immune disorders. This test appears potentially valuable in the clinical diagnosis of immune thrombocytopenias.
Signal transduction pathway activating interferon‐alpha‐stimulated gene expression
Authors: Veals Susan A. , Kessler Daniel S. , Josiah Serene , Leonard DebraG. B. , Levy David E.
Published: March 2008

Interferon‐alpha (IFNα) causes profound physiological changes following binding to susceptible target cells. These changes, which include induction of an antiviral state, inhibition of cellular proliferation, and modulation of differentiation, require the transcriptional activation of a set of genes. We have characterized the macromolecular components required for this stimulation of gene expression in order to define the biochemical mechanism of IFNα signal transduction. IFNα stimulated genes (ISGs) are immediate response genes which utilize a pre‐existing set of proteins to mediate their induction. A 15 bp IFNα‐inducible enhancer element present in the promoters of IFNα‐stimulated genes, termed the IFNα stimulated response element (ISRE), is the genetic target for activation of ISGs. This DNA sequence is both necessary and sufficient for transcriptional activation and is the target for action of a positive transcription factor termed ISGF3. The active, DNA‐binding form of ISGF3 is only found in cells which have been exposed to IFNα; however, it is activated from a silent form present in all responsive cells. ISGF3 is a multimeric complex assembled from cytoplasmic precursors which are translocated to the nucleus in response to IFNα. Assembly and translocation of ISGF3 is the earliest defined event in the IFNα response pathway.
Hairy cell leukaemia: review of treatment
Authors: Kurzrock Razelle , Talpaz Moshe , Gutterman Jordan U.
Published: March 2008

The cardinal features of hairy cell leukaemia are: (i) cytopenias, (ii) splenomegaly, and (iii) mononuclear cells of B‐cell origin with cytoplasmic projections and tartrate‐resistant acid phosphatase‐positivity. The most common complication is infection. In the past, the mainstay of therapy has been splenectomy, and this procedure is still often suggested as a first‐line approach. However, research during the last decade has resulted in three new, highly effective therapies for hairy cell leukaemia: interferon‐alpha (IFN‐alpha), 2′‐deoxycoformycin (DCF) and 2‐chlorodeoxyadenosine (2CDA). IFN‐alpha is currently approved for this indication. About 90% of patients have a durable haematologic recovery, and complete remission rates range from < 5% to > 40% in different series. It should be noted that patients with partial remissions generally have normal or near‐normal blood counts, and can live indefinitely without disease‐related problems, despite a few remaining hairy cells in the bone marrow. In this paper we will discuss the various therapeutic modalities available for patients with hairy cell leukaemia.
Treatment of multiple myeloma with interferon alpha: the Scandinavian experience
Authors: Mellstedt H. , ÖSterborg A. , Björkholm M. , Björeman M. , Brenning G. , Gahrton G. , Grimfors G. , Gyllenhammar H. , Hast R. , Juliusson G. , Jaurnmark M. , Killander A. , Kimby E. , Lerner R. , Merk K. , Ohrling M. , Simonsson B. , Smedmyr B. , Stalfelt A. M. , Strander H. , Svedmyr E. , Udean A. M. , Wadman D. , ÖSby E.
Published: March 2008

IFNα is a biologic therapeutic agent with documented antitumoural effect in multiple myeloma. 15% of previously untreated myeloma patients achieve a clinical response to IFNα alone with a possible dose‐response relationship. A particularly good effect is noted in IgA myelomas treated with natural IFNα.

A randomized study was started in April 1986 comparing melphalan/prednisone (MP) therapy with MP plus natural IFNα (MP/IFN) in untreated patients with multiple myeloma stages II and III. 220 patients had entered the study by autumn 1989. An interim report is given here. The response frequency was 48% in the MP group and 66% in the MP/IFN group (P<0·02). Stage II patients responded better to MP/IFN (76%) than to MP alone (48%) (P<0·01). No significant difference was noted for stage III patients. 91% of all IgA myelomas responded to MP/IFN and 52% to MP (P<0·01). The difference in response frequency of IgG and BJ myelomas between the two treatment groups was not statistically significant. The observation period is still too short to draw firm conclusions on survival. However, a statistically significant longer response duration time and survival from response (P<0·01) was noted for stage II patients.
Interferon alfa‐2b therapy in untreated early stage, B‐chronic lymphocytic leukaemia patients: one‐year follow‐up
Authors: Boussiotis V. A. , Pangalis G. A.
Published: March 2008

Recent reports have shown that interferon alpha is effective in B‐chronic lymphocytic leukaemia (B‐CLL). In a previous study from our unit we obtained a 50% response rate with interferon alfa‐2b in B‐CLL in early stages. Subsequently, we performed the present study randomizing 34 Binet stage AI, B‐CLL patients who were not previously treated (eight controls and 26 who were to receive interferon alfa‐2b (IFNα‐2b) (Intron‐A) according to a randomized schedule as follows: 1·5 MU/d in eight patients, 1·5 MU three times a week in 10 patients and 3·0 MU three times a week in eight patients). At 3 months complete response (CR) was obtained in one, partial haematologic response (PHR) in nine and minor haematologic response (PHR) in seven patients with an overall response rate 17/26 or 65%. No bone marrow changes regarding the pattern or degree of infiltration was noticed in any of the responding patients except for the complete responder. Treatment was continued in 15 patients (reduced to two‐thirds of the initial dose in the PHR and to one third of the initial dose in the MHR) while in 11 patients it was discontinued because of no response, negative response or toxicity. At 6 months sustained response was observed in 10 patients, while in the remaining five an increase of blood lymphocytes was seen at numbers similar or higher to those noted prior to interferon alfa‐2b therapy. Two of the 10 patients in whom blood lymphocytes were still reduced to less than 50% of the pretreatment values, developed gradual lymph node enlargement and therapy had to be discontinued. During the following 6 months the interferon alfa‐2b dose was further reduced in all remaining responders to 1·5 MU once or twice a week and their counts remained at the same level as at 6 months of therapy. At 12 months, 18 of our patients were still in stage A while seven had progressed (five to stage B and two to stage C). The overall response after 12 months of therapy was sustained in 8/17 responders. We conclude from our study that interferon alfa‐2b is effective in untreated B‐CLL patients in early stages and therefore should be investigated in combination with conventional chemotherapy. Clinical trials utilizing interferon alfa‐2b and chlorambucil in combination in untreated B‐CLL patients are currently in progress in our unit.
Interferon alpha in the therapy of CML
Authors: Talpaz M. , Kantarjian H. , Kurzrock R. , Gutterman J. U.
Published: March 2008

This paper summarizes the experience with interferon alpha (IFNα) as single‐agent therapy and in combination gathered at M. D. Anderson Cancer Center. It also covers some of the laboratory research associated with these clinical studies.
Interferon alpha in the treatment of AIDS‐related Kaposi's sarcoma
Authors: Mitsuyasu Ronald T.
Published: March 2008

Interferon alpha was one of the first drugs tested for the treatment of patients with AIDS‐related Kaposi's sarcoma based on its known antiviral properties and its abilities to modulate immune function and inhibit neoplastic cell proliferation. In vitro studies demonstrated defective production of interferon by blood cells of HIV‐infected individuals and suppression of HIV replication by interferons alpha and beta. Interferons have also been shown to inhibit angiogenesis induced by tumour cells or by allogeneic lymphocytes in mice. The efficacy of recombinant interferon alpha for the treatment of AIDS‐related Kaposi's sarcoma has been well documented with antitumour responses seen in approximately 30% of all patients treated in single agent efficacy trials with doses of at least 20 MU/m2. In several uncontrolled studies, response of Kaposi's sarcoma to treatment with interferon alpha was associated with longer survival and few opportunistic infections. Tumour response appears to be correlated with an absence of opportunistic infection and with CD4 cell numbers. Several studies using high interferon alpha doses have demonstrated decreases in serum HIV P24 core antigens which appear to be confined to patients whose tumours also regressed. The use of interferon alpha in HIV‐infected patients with or without Kaposi's sarcoma have demonstrated in vivo anti‐HIV activity. Studies have recently evaluated the tolerance and therapeutic potential of interferon alpha in combination with the reverse transcriptase inhibitor, zidovudine (azidothymidine AZT). Synergistic suppression of HIV replication in vitro has been demonstrated with the combination of interferon alpha and zidovudine. The description of HIV isolates with reduced sensitivity to zidovudine following prolonged treatment, and the finding that interferon alpha, but not zidovudine, prevents HIV expression in chronically infected cell lines, suggests that this combination might be useful in long‐term treatment of patients with HIV infection.
Interferon alpha in combination with other biologics: the scientific rationale
Authors: Herberman Ronald B. , Ernstoff Marc S. , Kirkwood John M.
Published: March 2008

Interferon alpha (IFNα) has widely pleiotropic effects, on both the immune system and tumour cells, and any of these effects might provide the basis for additive or synergistic effects when administered in combination with other biologics. Some of the main sites of action to consider include the ability of IFNα to: inhibit the growth of tumour cells; induce differentiation, and/or MHC expression on tumour cells; and stimulate the activation and/or differentiation of natural killer (NK) cells, cytotoxic T lymphocytes, and cytotoxic macrophages. Other biologics with similar sites of action might be expected to provide additional effects when combined with IFNα, and factors with other, complementary anti‐tumour effects might synergize. Potentially important therapeutic effects have been observed when IFNα has been combined with IFNγ, interleukin 2, tumour necrosis factor, interleukin 1, monoclonal antibodies, and retinoids. The development of therapeutic strategies on the basis of hypotheses as to how IFNα might effectively interact with another biologic, coupled with careful assessment of effects on the tumour cells and/or serial monitoring of immunologic parameters, might be expected to lead most expeditiously to successful therapeutic combinations of IFNα with other biologics.
Combination of cytokines: current status and future prospects
Authors: Kanz L. , Brugger W. , Bross K. , Mertelsmann R.
Published: March 2008

Clinical trials with individual cytokines and extensive in vitro studies have provided the basis for the in vivo use of these molecules in combination. Animal models, with haemopoietic growth factors as well as preliminary studies in humans—as shown by our studies with the sequential use of IL‐3 and GM‐CSF in patients receiving intensive chemotherapy—indicate that the selection of the appropriate cytokines could optimize haematological responses according to particular clinical requirements. That immunotherapy with IL‐2 can induce regression of disseminated human malignancies serves as an encouraging starting point for combinations with other cytokines with the goal of improving the therapeutic efficacy and reducing toxicity. Future prospects of combination therapy will be discussed.
Immunomagnetic bead isolation of megakaryocytes from guinea‐pig bone marrow: effect of recombinant interleukin‐6 on size, ploidy and cytoplasmic fragmentation
Authors: Leven Robert M. , Rodriguez Adrian
Published: March 2008

Guinea‐pig bone marrow megakaryocytes were isolated using an antibody to platelet glycoprotein Ib and a second antibody conjugated to magnetic beads. The procedure yielded an average of 644 800 megakaryocytes from two guinea‐pigs with an average viability of 83%. All of the platelet glycoprotein Ib positive cells also expressed the platelet glycoprotein IIb–IIIa complex. The size and ploidy of megakaryocytes isolated by this technique were analysed in the presence of 10 ng/ml of interleukin‐6 (IL‐6). Without IL‐6 megakaryocyte size increased significantly after 24 h. but an even larger increase in size occurred in the presence of IL‐6. The modal ploidy class was 16N with an average of 19% 2N. 2·6% 4N, 16·4% 8N. 50·8% 16N and 11·1% 32N cells as determined by flow cytometry. Measurements made by microspectrophotometry were in close agreement. After 24 h incubation there was a significant rise in the percentage of 2N and 32N cells. The ploidy distribution after 24 h with IL‐6 was the same as the control. Megakaryocytes cultured in the absence of serum on collagen gels did not form pseudopods and fragment, as occurs with serum (Leven et al. 1987). Addition of IL‐6 to the serum‐free cultures caused megakaryocytes to form extensive proplatelet extensions. We conclude that large numbers of pure guinea‐pig bone marrow megakaryocytes can be isolated by immunomagnetic bead selection, including low ploidy immature megakaryocytes. Spontaneous maturation occurred as evidenced by the increase in megakaryocyte size and ploidy. IL‐6 altered megakaryocyte size and morphology but not ploidy, indicating that these different characteristics of megakaryocytes may be regulated separately.
Difference in activity properties and subcellular distribution of neutrophil alkaline phosphatase between normal individuals and patients with trisomy 21
Authors: Grozdea J. , Vergnes H. , Brisson‐Lougarre A. , Bierme R. , Bourrouillou G. , Duchayne E. , Martin J. , Colombies P.
Published: March 2008

Biochemical, cytochemical characteristics and electron microscopy subcellular distribution of neutrophil alkaline phosphatase (NAP) were analysed in blood and/or smear samples from 39 trisomy 21 patients (Down's syndrome) aged 11·5‐18 years (mean 15·5 years) and 55 normal subjects aged 12‐20·5 years (mean 17 years). All patients were karyotyped.

NAP cytochemical procedures were carried out on all subjects: biochemical NAP determinations were made in 10 patients and 20 controls: ultrastructural electron microscopy of AP was performed in three patients and four normal subjects.

Neutrophil alkaline phosphatase from patients with trisomy 21 displayed the following changes: (1) a significant increase of enzyme activity, (2) a high thermal lability of enzyme. Electron microscope morphology exhibited large deposits of NAP reaction product associated with the plasma membrane and intracellular main organelles, like phosphasomes. The NAP biochemical and cytochemical characteristics suggest that trisomy 21 neutrophils contain a nonspecific AP isoenzyme, closely related to the early placental form.
Variable X‐chromosome DNA methylation patterns detected with probe M27β in a series of lymphoid and myeloid malignancies
Authors: Hodges E. , Howell W. M. , Boyd Y. , Smith J. L.
Published: March 2008

In this study the X chromosome probe M27β was used to investigate DNA methylation at the DXS255 locus and hence X inactivation status and determination of tumour clonality in blood, bone marrow and biopsy tissue involved with morphologically and phenotypically defined lymphoid and myeloid disease from 14 female patients along with uninvolved bone marrow from two control individuals. Thirteen out of 16 individuals (81%) were restriction fragment length polymorphism (RFLP) heterozygous for DXSZ 55. DNA methylation status could not be assessed in the three DXS255 homozygous individuals. In eight DXS255 heterozygous individuals clonality was clearly demonstrated using M27β and in six of these cases independent analysis using T cell receptor (TcR) and immunoglobulin (Ig) gene probes confirmed the presence of clonal tumour cell populations. In the two controls, polyclonality was inferred from M27β probe analysis. In the remaining three cases (all acute lymphoblastic leukaemia (ALL)) both DXSZ55 X chromosome sequences appeared to be methylated. Clonality in these cases was demonstrated by TcR or Ig monoclonal gene rearrangements. These data demonstrate the value of the M27β probe for determining tumour clonality in a number of cases with lymphoid and myeloid disease but indicate that there may not always be a complete correlation between DNA methylation, X inactivation status and tumour clonality in certain lymphoid neoplasms, restricting the use of this probe in clonality studies. Correlations between DNA methylation, X inactivation status and stage of normal and neoplastic T and B cell development require further investigation.
The fibrinolytic response to venous occlusion and the natural anticoagulants in patients with antiphospholipid antibodies both with and without systemic lupus erythematosus
Authors: Keeling D. M. , Campbell S. J. , Mackie I. J. , Machin S. J. , Isenberg D. A.
Published: March 2008

Patients with systemic lupus erythematosus (SLE) have an increased risk of thrombosis and this is increased in the presence of antiphospholipid antibodies (APA). These APA are also associated with thrombosis in patients who do not have SLE. We compared haemostatic parameters in SLE patients with and without APA, and also compared patients who had APA but not SLE with healthy normal controls. No relationships between the natural anticoagulants. anti‐thrombin III, heparin cofactor II, protein C and protein s. and the presence of APA were found. In the patients with SLE both tissue plasminogen activator antigen and plasminogen activator inhibitor (PAI) were increased, but these changes were not due to APA which had no effect on fibrinolysis in these patients. In the patients with APA who did not have SLE the fibrinolytic response to venous occlusion was reduced due to raised levels of PAI; similar changes have, however, been reported in some patients with idiopathic thrombosis.
The incidence of dysfunctional antithrombin variants: four cases in 210 patients with thromboembolic disease
Authors: Harper P. L. , Luddington R. J. , Daly M. , Bruce D. , Williamson D. , Edgar P. F. , Perry D. J. , Carrell R. W.
Published: March 2008

210 patients, with a history of venous thrombosis, have undergone prothrombotic investigations. In nine cases a consistent deficiency of antithrombin was identified. In five there was a reduction in the plasma antigenic concentration of antithrombin and in a further four cases deficiency was due to the presence of a dysfunctional antithrombin variant. The variants have all been characterized by DNA analysis and in three the mutations have been confirmed by peptide sequencing (antithrombin Basel (41 Pro to LEU), Hamilton (382 Ala to Thr). Cambridge I (384 Ala to Pro) and Cambridge II (384 Ala to Ser). The incidence of antithrombin deficiency in patients with a history of venous thrombosis has previously been quoted at between 2% and 3%: there is no published data available on the incidence of antithrombin variants. In our series 5% of patients who presented before the age of 40 years had antithrombin deficiency, and 2% of the total number of patients investigated had a dysfunctional variant. Our figures indicate that a significant number of cases of antithrombin deficiency are due to dysfunctional variants and that the true incidence of antithrombin deficiency in patients with a history of venous thrombosis is in the order of 5%.
Erythrocyte size, number and haemoglobin content in vertebrates
Authors: Hawkey C. M. , Bennett P. M. , Gascoyne S. C. , Hart M. G. , Kirkwood J. K.
Published: March 2008

Kelationships between erythrocyte number, size and haemoglobin content were examined in 441 species (101 families) of mammals, birds and reptiles. Whereas the total number of red cells (RBC), the mean cell volume (MCV) and mean cell haemoglobin (MCH) showed much variation, the haemoglobin level (Hb), packed cell volume (PCV) and mean cell haemoglobin concentration (MCHC) were relatively constant in all adult mammals and birds but lower in reptiles. There was a strong positive correlation between MCV and MCH and a strong negative correlation between MCV and RBC across all species. Mammals had more, smaller erythrocytes per unit volume of blood than birds, which, in their turn, had more, smaller erythrocytes than reptiles. The findings confirm that the oxygen‐carrying capacity of the blood is highly conserved in birds and mammals but is lower in exothermic groups such as reptiles. Although the significance of the observed species differences in RBC and MCV has yet to be explained, the findings have considerable evolutionary, physiological and diagnostic interest.
Responsiveness of bone marrow erythroid progenitors (CFU‐E and BFU‐E) to recombinant human erythropoietin (rh‐Ep) in vitro in multiple myeloma
Authors: Aoki Isao , Nishijima Kosuke , Homori Masashi , Nakahara Kazuhiko , Higashi Katsumi , Ishikawa Kyozo
Published: March 2008

The responsiveness of bone marrow erythroid progenitors (CFU‐E and BFU‐E) to recombinant human erythropoietin (rh‐Ep) was investigated in vitro in 21 patients with multiple myeloma to assess the clinical usefulness of rh‐Ep in this disease. CFU‐E and BFU‐E assays were performed by methylcellulose culture methods. The myeloma patients were divided into two groups according to the percentage of plasma cells in the bone marrow (over 50% and under 50%). Among the patients with few plasma cells, some revealed normal CFU‐E and BFU‐E growth at 2 units of rh‐Ep, and no further increase was observed even with an increasing dose of rh‐Ep. Among the other patients, more than half demonstrated a good response to rh‐Ep. Among the patients with a high percentage of plasma cells, some revealed no response to rh‐Ep, but there were patients with a high percentage of plasma cells in the bone marrow who had a good response to rh‐Ep. High doses of rh‐Ep may be clinically effective in some patients with multiple myeloma independently of the level of plasma cells in the bone marrow.
Purification of human monocyte‐specific esterase (MSE): molecular and kinetic characteristics
Authors: Scott Colin Stephen , Patel Divyen , Keen Jeffrey Norman
Published: March 2008

Human monocyte‐specific esterase (MSE) derived from leukaemic AMoL‐M5 blast cells was purified to homogeneity by the sequential application of anion‐exchange, hydrophobic interaction, affinity and gel filtration chromatographic procedures. The resulting enzymatically active MSE primarily existed as an apparent trimer which, under both reducing and non‐reducing conditions, dissociated to an inactive 63·4 kD glycoprotein monomer. Electrophoretic studies further confirmed that purified MSE comprised a narrow series of pI (5·5–6·1) forms and one main charge species. Neuraminidase failed to modify observed pI values for individual MSE isoenzymes, and endoglycosidase H treatment revealed that the deglycosylated form of MSE had an apparent molecular weight of 60·1 kD. In support of the known cytochemical characteristics of human MSE, substrate kinetic studies demonstrated that purified enzyme hydrolysed esters of higher acyl chain length (butyrate > propionate > acetate) but did not show peptidase activity. Amino acid sequencing of the MSE N‐terminus further revealed that there was almost complete identity with human alveolar macrophage esterase and close similarities with rat and rabbit liver carboxylesterases. These kinetic and molecular studies are particularly important in elucidating the biological and functional role(s) of one of the few haemopoietic cell enzymes that can be considered truly lineage‐specific.
Myelodysplastic syndromes in childhood: a population based study of nine cases
Authors: Hasle Henrik , Jacobsen Bendt Brock , Pedersen Niels Tinggaard
Published: March 2008

Nine cases of de novo myelodysplastic syndromes (MDS) in childhood from a population based study are presented. The annual incidence of MDS was 3·4/1000 000 in children less than 15 years old, corresponding to 8·7% of all haematological malignancies in childhood. Two patients had Down's syndrome. None of the remaining patients had constitutional anomalies.

All patients were classified according to the FAB classification. Five patients presented with refractory anaemia (RA), only one of these did not progress, one showed clonal evolution, and the remaining three patients all progressed to refractory anaemia with excess of blasts (RAEB). Three patients presented with RAEB. Two progressed to overt leukaemia. The last patient was classified as chronic myelomonocytic leukaemia (CMML). Clonal cytogenetic abnormalities were detected in five patients, in three of them as monosomy 7.

Five patients have died; two of progressive disease, two of infections, and one of haemorrhage, two of the latter three patients died during therapy induced cytopenia.

Of the four patients still alive, one patient showed a complete remission after cyclosporine and later immunoglobulin therapy, one patient is a long‐term survivor after allogeneic bone marrow transplantation, and one patient apparently obtained a spontaneous remission several months after chemotherapy.
Extended cytogenetic follow‐up of patients with myelodysplastic syndrome (MDS)
Authors: White A. D. , Culligan D. J. , Hoy T. G. , Jacobs A.
Published: March 2008

The prognostic significance of clonal karyotype status in myelodysplastic syndrome (MDS) is assessed after an extended follow‐up period of 5 years. There are three prognostic sub‐groups according to the presence of a normal karyotype, single abnormalities or multiple abnormalities at the time of referral. However, there is no correlation between the size of the abnormal clone and prognosis. Karyotype status has independent prognostic significance in ‘high risk’MDS so that patients with a refractory anaemia with excess of blasts (RAEB)/RAEB in transformation (RAEB‐t) and a normal karyotype survive significantly longer than those with an abnormal karyotype (P<0·001) and do not differ significantly from patients with refractory anaemia (RA). Significant differences in survival according to karyotype status are also seen in patients with chronic myelomonocytic leukaemia (P<0·001) but not in those with primary acquired sideroblastic anaemia and RA. Among patients studied sequentially, those who retained a normal karyotype survived significantly longer than those who developed an abnormality on follow‐up (P<0·001). The risk of leukaemic transformation was also increased in patients who presented with or subsequently developed a clonal karyotype abnormality compared with those who remained normal (P<0·05).
A predictive model for the clinical response to low dose ara‐C: a study of 102 patients with myelodysplastic syndromes or acute leukaemia
Authors: Hellström‐Lindberg Eva , Robèrt Karl‐Henrik , Gahrton Gösta , Lindberg Greger , Forsblom Ann‐Mari , Kock Yvonne , Öst Åke
Published: March 2008

The response to treatment with low‐dose ara‐C was studied in 102 consecutive patients; 79 with myelodysplastic syndromes (MDS) and 23 with acute myelogenous leukaemia (AML) following MDS. The aim was to find variables that could predict the response to treatment. All patients had clinical symptoms related to cytopenia. Peripheral blood values, bone marrow morphology histology and chromosomes were analysed before the start of treatment. The median survival of the patients was 9 months and a poor survival was predicted by advanced age, low platelet counts, the presence of pseudo‐Pelger morphology and <2 chromosomal aberrations.

Thirty patients (29%) responded with either a complete remission or a significant increase in haemoglobin level. For the remaining 71%, the treatment was ineffective and in some cases hazardous. The factors associated with a poor response to treatment could be divided into two groups: one included low platelet counts and the presence of chromosomal aberrations, both signs of progressive MDS with a short survival, and the other comprised morphological findings, indicating ineffective haemopoiesis. Patients with platelet counts > 150 × 109/l had a response rate of 55% compared to 23·5% in patients with subnormal platelet counts. Logistic regression identified low bone marrow cellularity, absence of ring sideroblasts and <2 chromosomal aberrations as predictors of a favourable response in patients with platelet counts <150 × 109/l. These factors and the platelet count were combined in a predictive model which can divide patients into three groups with different probabilities of response: a favourable group, 38·6% of the patients, with a response rate of >50%, an intermediate group, 32·7% of the patients, with a response rate of 24%, and an unfavourable group, 28·7% of the patients, with only 3% responses.

While low‐dose ara‐C is an effective treatment for some patients, it is ineffective and hazardous for others. We present a model that can facilitate therapeutic decision making in two‐thirds of patients with MDS and MDS‐AML by identifying patients who should not be treated with low‐dose ara‐C as well as patients with a relatively high probability of response.
Mixed phenotype of blasts in acute megakaryocytic leukaemia and transient abnormal myelopoiesis in Down's syndrome
Authors: Yumura‐Yagi Keiko , Hara Junichi , Kurahashi Hiroki , Nishiura Tetsuo , Kaneyama Yoshio , Osugi Yuko , Sakata Naoki , Inoue Masami , Tawa Akio , Okada Shintaro , Kawa‐Ha Keisei
Published: March 2008

Blasts from eight cases with acute megakaryoblastic leukaemia (AMKL) and seven with transient abnormal myelopoiesis in Down's syndrome (TAM) were investigated to clarify their phenotypic characteristics. CD41 and CD7 were the most frequently expressed in both disorders. CD41 was positive in six TAM and five AMKL cases, and CD7 was positive in five TAM and five AMKL cases, respectively. CD33 was detected in four TAM and five AMKL cases. Other myeloid‐lineage associated antigens such as CD13 and CD11b could not be found in TAM but were expressed in five AMKL cases. Interestingly, CD56, a neural adhesion molecule, was expressed in three of four TAM and one of five AMKL cases. Cytoplasmic CD3 antigen was also noted in three of five examined cases.

A short‐term culture study was conducted on blasts from two TAM cases and five AMKL cases. In two cases in which CD41 was not expressed before culture, the expression of CD41 was enhanced after culture with or without 12–0‐tetradecanoyl‐phorbol‐13‐acetate (TPA). The expression of CD7 remarkably was depressed, while that of CD13 was enhanced after culture with TPA.

These findings suggest that blasts of TAM and AMKL originate from very immature cells and represent a mixed phenotype. In the present study, distinction of pheno‐typical differences between blast in TAM and AMKL was not possible.
Red blood cells from patients with sickle cell disease exhibit an increased adherence to cultured endothelium pretreated with tumour necrosis factor (TNF)
Authors: Vordermeier S. , Singh S. , Biggerstaff J. , Harrison P. , Grech H. , Pearson T. C. , Dumonde D. C. , Brown K. A.
Published: March 2008

Red blood cells (RBCs) from 24 patients with sickle cell disease were more adherent to cultured endothelium pretreated with the inflammatory cytokine, tumour necrosis factor (TNF) than RBCs from 22 healthy subjects. The enhanced sticking was apparent in RBC preparations from patients who were in crisis (mean 190% increase from controls) and out of crisis (mean 220% increase) and was not related to the number of circulating RBCs, reticulocytes, platelets, leucocytes or haemoglobin levels. When irreversibly sickled RBCs, enriched by centrifugation on density gradients, were added to TNF‐treated endothelium they were found to be significantly more adherent (mean 411% increase; P<0·001) than the unfractionated RBCs from the same patients. There was no difference between the adherent properties of sickle RBCs and normal RBCs for untreated endothelium.

Contributing factors to the enhanced adhesion to TNF‐treated endothelium may be the low surface change of sickle RBCs, and increased levels of fibrinogen and von Willebrand's factor (vWF) in the patients’plasma. By acting on vascular endothelium to increase its adhesiveness for sickled RBCs, it is concluded that inflammatory cytokines such as TNF may have a prominent role in mediating the events that lead to microvascular occlusions in sickle cell disease.
Sequential administration of recombinant interferon alpha and deoxycoformycin in the treatment of hairy cell leukaemia
Authors: Habermann Thomas M. , Andersen Janet W. , Cassileth Peter A. , Bennett John M. , Oken Martin M.
Published: March 2008

Summary Both recombinant interferon alpha and deoxycoformycin (dCF) are effective in the treatment of hairy cell leukaemia. In an attempt to reduce the complications from dCF therapy, a pilot study of the Eastern Cooperative Oncology Group (ECOG) first treated patients with interferon to improve peripheral blood cell counts before dCF treatment began. Thirty‐four patients were treated for 3 months with recombinant interferon alpha‐2a (rIFN alpha‐2a), 3 × 106 IU subcutaneously three times a week for 3 months, and then by dCF, 4 mg/m2 intravenously every 2 weeks for a maximum of 12 months. The overall response rate was 94% (32/34); 76% of patients (26/34) had complete response (CR) (90% confidence interval, 62–88%) and 18% (6/34) partial response. One patient was found to have a Mycobacterium avium infection while receiving rIFN alpha‐2a. Without specific antimycobacterial therapy and with continued administration of rIFN alpha‐2a and dCF, the infection resolved and he achieved CR. Three patients had culture‐negative febrile episodes during the dCF phase of treatment. Non‐disseminated herpes zoster developed in four patients, but three of the episodes occurred only after treatment was discontinued. Sequential administration of rIFN alpha‐2a and dCF resulted in fewer infections (P=0.027) than in ECOG's previous study of dCF used alone. Two patients died, one of combined hairy cell leukaemia and non‐Hodgkin's lymphoma of intermediate histologic type 17 months after entry into the study and the other of cardiac arrest 20 months after entry. Thirty‐two patients were alive with a median follow‐up of 21 months (range 13–31 months). This combination produces durable CRs with a low incidence of infection.
α‐ and β‐haemoglobin chain induced changes in normal erythrocyte deformability: comparison to β thalassaemia intermedia and Hb H disease
Authors: Scott Mark D. , Rouyer‐Fessard Philippe , Ba Mariegme Soda , Lubin Bertram H. , Beuzard Yves
Published: March 2008

Summary The α and β‐thalassaemias are characterized by decreased erythrocyte deformability. To determine what effects excess α and β‐haemoglobin (globin) chains have on cellular and membrane deformability, purified haem‐con‐taining α and β‐chains were entrapped within normal erythrocytes. Entrapment of purified α‐chains in normal erythrocytes resulted in a significant decrease in cellular and membrane deformability similar to that observed in β‐thalassaemia intermedia. The decreased deformability was correlated with α‐chain membrane deposition, an alteration in membrane proteins and a decrease in membrane reactive thiol groups. These changes in membrane and cellular deformability were time dependent and closely correlated with membrane α‐chain deposition. The membrane changes and the loss of membrane deformability appeared to account for the loss of cellular deformability in the α‐chain loaded cells. While both β‐chain loaded and Hb H erythrocytes demonstrated a significant loss of cellular deformability, this loss was less pronounced than in the α‐chain loaded and β‐thalassaemic cells and may arise from either the increased intracellular viscosity of the β‐chain loaded cells or to the smaller amount of membrane bound globin. In summary, these studies demonstrate that alteration of cellular and membrane deformability occurs very rapidly and as a direct consequence of the autoxidation and membrane binding of the unpaired globin chains.
Human serum stimulates the production of G‐CSF, IL‐1, IL‐6 and IL‐8 by human peripheral blood leucocytes
Authors: Quesniaux Valerie F. J. , Wehrli Susi , Ziegler Irma , Legendre Beatrice , Wishart William , Fagg Barbara , Schreier Max H. , Nissen Catherine
Published: March 2008

Summary Human serum induces human peripheral blood leucocytes (PBL) to release an activity stimulating neutrophil colony formation (G‐CSA) from human bone marrow cells. By titrating individual growth factors and using specific neutralizing antibodies we showed that: human serum contains very low levels of G‐CSF which are by themselves insufficient to stimulate myeloid colony formation in primary human bone marrow cultures and cannot account for the serum releaser activity; that although no detectable levels of IL‐1, IL‐2, IL‐3, IL‐4, IL‐6 or IL‐8 are found in the serum, anti IL‐1 antibodies partially block the release of G‐CSA when added early during PBL incubation; that PBL incubated in the absence of serum for 2 d produce small amounts of IL‐1, IL‐6, IL‐8 and G‐CSF and this is increased 6–16 fold in the presence of human serum; and that the neutrophil colony‐stimulating activity released by PBL incubated with human serum is G‐CSF.
Variable expression of features of normal and neoplastic stem cells in patients with thrombocytosis
Authors: Turhan Ali G. , Cashman. Johanne D. , Eaves Connie J. , Humphries R. Keith , Eaves Allen C.
Published: March 2008

Summary Essential thrombocytosis (ET) is currently diagnosed by histopathologic assessment of the marrow after exclusion of a secondary cause or another myeloproliferative disorder. To evaluate the potential of more direct diagnostic methods, we compared the frequency and association of several abnormal features characteristic of neoplastic precursors in 32 patients presenting with platelet counts >500.109/1. Assays for erythropoietin (Ep)‐independent erythroid progenitors were performed on all patients, determination of the cycling status of circulating progenitors on 27. and assessment of granulocyte clonality on 15. In most, but not all, patients deregulated progenitor turnover. Ep‐independent progenitors and clonal granulocytes were concordant findings. The presence of polyclonal granulocytes and lack of evidence of abnormalities in Ep‐dependence or progenitor cycling were also concordant findings in most, but not all patients. Thus, normal (i.e. polyclonal) granulocytes may be produced in occasional patients in spite of the presence of a neoplastic clone. Interestingly, one third of patients thought to have ET on the basis of blood and marrow histopathology showed no abnormalities previously associated with neoplastic progenitors. These findings suggest variability in dominance of the neoplastic clone in some ET patients and the potential utility of a multifaceted laboratory approach to investigate the underlying pathology in patients with thrombocytosis.
Defects in type IIA von Willebrand disease: a cysteine 509 to arginine substitution in the mature von Willebrand factor disrupts a disulphide loop involved in the interaction with platelet glycoprotein Ib‐IX
Authors: Lavergne J.‐M. , DePaillette L. , Bahnak B. R. , Ribba A.‐S. , Fressinaud E. , And D. Meyer , Pietu G.
Published: March 2008

Summary Type IIA von Willebrand disease (vWD) is characterized by the loss of high and intermediate weight multimers of von Willebrand factor (vWF) from plasma. The 3’end of exon 28 in the vWF gene from four type IIA vWD patients was amplified by the polymerase chain reaction, cloned and sequenced. Sequencing identified two potential missense mutations resulting in the amino acid substitutions Arg 834Gln and Glu 875Lys in the mature vWF subunit within an area of vWF where mutations in type IIA vWD have been reported. Neither of these amino acid substitutions was found in over 100 normal alleles tested by allele specific oligonucleotide hybridization. A polymorphism (Val 802Leu) was identified in another patient. Other areas of exon 28 were analysed by denaturing gradient gel electro‐phoresis (DGGE) and DNA from one patient demonstrated an irregular DGGE pattern on the 5’end of the exon. Sequencing demonstrated an amino acid substitution of an arginine for cysteine at position 509 adjacent to an area of vWF where defects associated with type IIB vWD have been found. This substitution was not found in 100 normal chromosomes tested by restriction enzyme digestion. The Cys 509Arg substitution eliminates an intramolecular disulphide bridge formed by Cys 509 and Cys 695 which is important to maintain the configuration of vWF functional domains that interact with platelet glycoprotein Ib‐IX.