bluuebunny/crossref_metadata_2025 huggingface dataset:
Resistance of leukemic blasts to lymphokine activated killer (LAK)‐mediated cytotoxicity is not related to their adhesion properties
Authors: Palucka A. Karolina , Porwit Anna , Reizenstein Peter
Published: July 2010

Abstract: The expression of adhesion molecules on blasts from 14 patients with acute myeloid leukemia (AML) was investigated by immunofluorescence and flow cytofluorometry. All tested blast populations expressed CD18/CDlla complex {leukocyte function antigen‐1 (LFA‐1)} and CD29 (very‐late antigen (VLA)) and the majority were positive for CD54 {intercellular adhesion molecule‐1 (ICAM‐1), 78.6%} and CD56 {neural cell adhesion molecule (NCAM), 64.3%}. The expression of two other alpha chains of CD18/CDllb and CDllc varied considerably (64.3% and 42.8% of positive cases, respectively). Only one case (AML‐M4) showed a weak expression of the activated platelet antigen CD41b. None of the tested blasts expressed the vitronectin receptor (CD61/CD51). No significant correlation between the expression of adhesion molecules and the FAB type of leukemia could be found. All tested blast populations were completely resistant to NK‐mediated cytotoxicity and relatively resistant to LAK‐mediated cytotoxicity in the standard 51Cr release assay. While no statistically significant correlation of the results in cytotoxicity assays with the expression of adhesion molecules or the expression of HLA‐DR antigen could be observed, 2 out of 3 completely resistant cases lacked ICAM‐1. These results show that even leukemic blasts which express all of the tested adhesion molecules can still be resistant to LAK‐mediated cytotoxicity.
Constitutive production of granulocyte colony‐stimulating factor and interleukin‐6 by a human lung cancer cell line, KSNY: Gene amplification and increased mRNA stability
Authors: Matsuguchi Tetsuya , Okamura Seiichi , Kawasaki Chiyuki , Shimoda Kazuya , Omori Fusayuki , Hayashi Shin , Kimura Nobuhiro , Niho Yoshiyuki
Published: July 2010

Abstract: A human lung squamous cell carcinoma cell line designated KSNY was established from a patient suffering from marked neutrophilia and polyclonal hyper‐γ‐globulinemia. In our previous report, we demonstrated colony‐stimulating activities in the culture supernatant of this cell line. To determine the exact molecules for the activities, we investigated the gene expression of various cytokines in KSNY cells and showed the mRNA expression of both granulocyte colony‐stimulating factor (G‐CSF) and interleukin‐6 (IL‐6). We also detected substantial amounts of G‐CSF and IL‐6 in the culture supernatant with sensitive enzyme‐linked immunosorbent assays (ELISA). The amplification of the gene locus for G‐CSF, but not for IL‐6 was shown by Southern blot analysis. Furthermore, we also showed that the mRNAs for G‐CSF and IL‐6 were relatively stable in KSNY cells. These findings are thought to be related to the constitutive production of the cytokines in KSNY cells.
Influence of blood donation on iron stores assessed by serum ferritin and haemoglobin in a population survey of 1433 Danish males
Authors: Milman Nils , Kirchhoff Marianne
Published: July 2010

Abstract: The general impact of blood donation on iron status has been studied in Danish males. Iron stores were assessed by serum (S‐) ferritin and haemoglobin (Hb) in a population survey comprising 1433 males in age cohorts of 30, 40, 50, and 60 years; 389 (27%) were blood donors and 1044 (73%) non‐donors. Hb levels were identical in donors and non‐donors, mean 155 ± 11 (SD) g/1 (9.6 ± 0.7 mmol/1); values< 129 g/1 (8.0 mmol/1) were observed in 1.3% of donors vs 1.9% of non‐donors. Correlations between S‐Ferritin and Hb were slight and without practical clinical relevance: rs = 0.13, p<0.01 in donors vs rs = 0.16, p<0.0001 in non‐donors. Donors had lower S‐Ferritin, median 95 μg/1, than non‐donors, median 136 μg/1 (p<0.0001). S‐Ferritin values < 15 μg/1 (i.e. depleted iron stores) were seen in 3.3% of donors vs 0.4% of non‐donors, and S‐Ferritin values of 15–30 μg/1 (i.e. small iron stores) in 9.8% of donors vs 1.4% of non‐donors. Iron‐deficiency anaemia (i.e. S‐Ferritin <15 μg/1 and Hb<129 g/1) was seen in 0.26% of donors vs 0.10% of non‐donors; employing the 5th percentile for Hb (137 g/1 (8.5 mmol/1)) as discriminatory value increased the percentage of iron‐deficiency anaemia to 0.51% in donors vs 0.10% in non‐donors. Blood donation had a marked influence on iron status in the adult male population. The frequency of phlebotomy should be adjusted according to S‐Ferritin as well as Hb levels. If Hb is used as single criterion for donation, only donors with pre‐donation values > 135–137 g/1 should be allowed phlebotomy. Optimal donation standards should include monitoring of iron status through measurement of S‐Ferritin and Hb, combined with individualised postdonation iron supplementation.
Expression of cell surface transferrin receptor and intracellular ferritin after in vitro stimulation of peripheral blood T lymphocytes
Authors: Pattanapanyasat Kovit , Hoy Terry G.
Published: July 2010

Abstract: Measurements of peripheral blood lymphocyte surface transferrin receptor using flow cytometry show that phytohaemagglutinin (PHA) stimulation causes a marked increase in both the number of cells bearing receptors and the absolute number of receptors. This increase is accompanied by the interleukin‐2 receptor and there is a progressive increase of cells in the S phase of their cycle. Furthermore, ferritin synthesis in proliferating cells, as determined by immunoprecipitation techniques, increases significantly compared to non‐stimulated cells. Intracellular concentrations of both spleen‐type and heart‐type ferritin are also increased with a preferentially high proportion of heart‐type ferritin. These results suggest that expression of transferrin receptor and synthesis of ferritin is regulated by a complex mechanism. Cellular proliferative activity increases the expression of transferrin receptor and could modulate the biosynthesis of intracellular ferritin that is normally controlled by iron. These two processes do not appear to be linked.
Absence of correlation between cytotoxicity and drug transport by P‐glycoprotein in clinical leukemic cells
Authors: Kato Shoko , Ideguchi Hiroshi , Muta Koichiro , Nishimura Junji , Nawata Hajime
Published: July 2010

Abstract: Development of resistance to cytotoxic agents is a common problem in the treatment of acute leukemia. In cell lines having multidrug resistance (MDR) phenotype, a decrease in the intracellular accumulation of drugs has been closely related to the overexpression of P‐glycoprotein/mdrl genes. We analyzed the relationship between the cytotoxicity of adriamycin (ADR) in vitro, intracellular accumulation of ADR, and the expression of P‐glycoprotein on fresh leukemic cells from 19 patients at their initial presentation and from 9 relapsed patients. Pretreatment patients showed significantly higher ratio of complete remission than relapsed patients, and mean value of IC50 for adriamycin in initial presentation was higher than at relapse. But we found no significant relationship between in vitro cytotoxicity and drug transport. In addition, only 2 of the 5 relapsed patients examined by monoclonal antibody C219 expressed the P‐glycoprotein. These results suggest that the acquisition of clinical drug resistance may involve various mechanisms other than the reduction of drug accumulation with P‐glycoprotein expression.
The effect of interleukin‐1 on iron metabolism in rats
Authors: Uchida Tatsumi , Yamagiwa Akio , Nakamura Kenichi
Published: July 2010

Abstract: The effect of interleukin‐1 on iron metabolism in rats was evaluated. Plasma iron decreased from 184 ± 16 μg/dl (mean ± SE) to 24 ± 12 at 6 hours after interleukin‐1 intramuscular administration in non‐fasting rats and 109 ± 6 μg/dl to 12 ± 1 μg/dl in fasting rats, which was significantly lower than in control rats. Ferrokinetic studies showed a more rapid disappearance rate and lower iron turnover in interleukin‐1‐injected rats. The release of iron from the mononuclear phagocyte system to plasma was studied at 3 h after interleukin‐1 administration. Although the percent of radioactivity in plasma of the total injected dose was 3.2 ± 0.6% in interleukin‐1, which was significantly lower than in the control rats (5.4 ± 0.6%) at 9 h after intravenous injection of 59Fe chondroitin ferrous sulfate, there was no difference between the amount of 59Fe released from the mononuclear phagocyte system over the first 9 h in interleukin‐1 and control rats. These data appear to imply that iron release is unimpaired but that, for some reason, there is an enhanced rate of clearance of the 59Fe once it has been released from the mononuclear phagocyte system into the plasma.
Dipyridamolein vitrosuppresses the generation of T‐cell cytotoxic functions: Synergistic activity with cyclosporine
Authors: Massaia Massimo , Bianchi Alberto , Attisano Carmela , Dianzani Umberto , Boccadoro Mario , Pileri Alessandro
Published: July 2010

Abstract:The pharmacological activity of dipyridamole has been related to its ability to increase intracellular cAMP. Elevated cAMP concentrations can tone down T‐cell effector functions. The aim of this study was to evaluate the dipyridamole effectin vitroon the generation of alloreactive cytotoxic and lymphokine‐activated killer cells in normal T‐cell subpopulations. Dipyridamole suppressed T‐cell cytotoxic functions in a dose‐dependent way. The kinetics of suppression showed that dipyridamole prevented the first step of cytotoxicity, i.e. activation of the lytic program following allogeneic or interleukin‐2 stimulation. The ability of dipyridamole to interact with the immunosuppressive activity of cyclosporine was also investigated. By itself, cyclosporine suppressed the generation of alloreactive cytotoxicity, but not the generation of lymphokine‐activated killer cells. A synergistic immunosuppressive activity between dipyridamole and cyclosporine was observed on the generation of alloreactive cytotoxic T lymphocytes.
Treatment of advanced myelodysplastic syndromes with recombinant interferon‐alpha2b
Authors: Aul Carlo , Gattermann Norbert , Schneider Wolfgang
Published: July 2010

Abstract: We examined the efficacy and toxicity of recombinant interferon‐alpha2b (rIFN‐α2b) in 10 previously untreated patients with advanced myelodysplastic syndromes. Morphological subtypes were refractory anaemia with excess of blasts (RAEB) in 4, RAEB in transformation (RAEB/T) in 3 and chronic myelomonocytic leukaemia (CMML) in 3 cases. IFN was administered subcutaneously at increasing doses of 1 to 3 times 106 IU per day. The median duration of therapy was 6 months (range, 3 to 14). 2 patients, both with a diagnosis of CMML, achieved a complete and partial remission, respectively. In the complete responder, remission could be maintained for 9.5 months by daily administration of 1 times 106 IU IFN. The other patients were classified as failures, although in 4 cases a decrease of bone marrow blasts was noted and none of the patients progressed to overt leukaemia while being treated with IFN. During the study, all patients with RAEB and RAEB/T showed a moderate to severe reduction in peripheral leukocyte and platelet counts, requiring premature termination of IFN therapy in 5 cases. Despite adequate supportive measures, 2 patients died of pneumococcal pneumonia and gastrointestinal bleeding, respectively. In 1 patient, IFN therapy had to be stopped because of neurologic toxicity (polyneuropathy). From these data we conclude that rIFN‐α2b at the doses and schedule tried is not a useful treatment for advanced myelodysplastic syndromes. Patients with CMML, however, may be an exception and should further be considered as candidates for therapeutic trials with rIFN‐α2b.
Influence of interferon on antibody production and viability of malignant cells from patients with multiple myeloma
Authors: Grandér Dan , Einhorn Stefan , von Stedingk Lars‐Victor , von Stedingk Marit , Wasserman Jerzy
Published: July 2010

Abstract: The influence of interferon (IFN) on antibody production and viability of malignant cells from patients with multiple myeloma was evaluated. Following incubation of bone marrow cells with IFN‐α (5000 units/ml) for 7 days) a decreased production of monoclonal immunoglobin (mlg) was detected in all experiments except one. IFN induced a > 50% decrease in myeloma cell viability in 11 patients and a ≥ 25 % decrease in 4 patients. whereas in myeloma cells from 8 patients IFN had no or only minor effects. The observed effect was not due to an inhibition of proliferation since <5% of the myeloma cells were labeled with [3H]‐thymidine during 7 d of culture. There was no statistically significant correlation between decreases in myeloma cell viability and effects on mlg production, exemplified by the fact that mlg production was decreased also in patients showing no sensitivity to IFN's cytotoxic action. Depletion of autologus T‐cells, NK‐cells and/or monocytes did not abrogate the effects observed. We conclude that IFN can reduce the viability of myeloma cells and the production of Ig from these cells and that the latter can be exerted without an antitumor effect.
Regulation of autologous T‐lymphocyte clones on in vitro growth of BFU‐e: Enhancing effects of both CD4‐ and CD8‐positive clones
Authors: Nielsen B. , Jessen P. , Hokland P.
Published: July 2010

Abstract: In order to characterize the effect of T lymphocytes on the in vitro growth of erythroid progenitor cells (EPC), we have optimized an assay for evaluating the influence of autologous T‐ lymphocyte clones on BFU‐e, grown from PBMNCs depleted of T lymphocytes. Testing a large number of T‐lymphocyte clones from 2 individuals we found that, irrespective of immunological phenotype, all clones tested were found to enhance the growth of BFU‐e. Thus, no evidence for a functional dichotomy between CD4+ and CD8+ T lymphocytes was found in this system. The enhancing effect worked across a histocompatibility barrier and was, at least in part, mediated by soluble factors. We therefore conclude that the role of the majority of cloned normal T lymphocytes in the regulation of the in vitro growth of EPC is a supportive one.
Mitoxantrone, etoposide, cytarabine and prednisone as salvage therapy for refractory non‐Hodgkin lymphoma (NHL) and alternated with CHOP in previously untreated patients with NHL
Authors: Merk Karl , Ideström Kajsa , Johansson Bo , Kimby Eva , Lindemalm Christina , Ösby Eva , Björkholm Magnus
Published: July 2010

Abstract: The treatment of relapsing or refractory high‐grade malignant non‐Hodgkin lymphoma (NHL) following CHOP chemotherapy remains a challenge for the clinician. In this study, 29 patients with relapsing or refractory high‐ or refractory low‐grade malignant NHL received a combination of mitoxantrone 12 mg/m2 i.v. on days 1–2, cytarabine 100 mg/m2 i.v., b.d. d 1–2, etoposide 100 mg/m2 i.v. d 1–3 and prednisone 100 mg/m2 orally d 1–3 (ENAP). An overall response rate of 55% encouraged us to use ENAP alternated with conventional CHOP chemotherapy in 45 previously untreated NHL patients (35 with high‐grade and 10 with “aggressive” low‐grade malignant NHL). All patients responded with a complete remission rate (CR) of (27%) and a partial remission rate (PR) of 73% after only one course of ENAP. After a median number of 3.5 ENAP/CHOP courses, the CR and PR rate was 69 and 22%, respectively. Myelo‐suppression was pronounced and fever of unidentified origin and documented infections followed 59% of all cases given ENAP courses. In the last 19 previously untreated patients mitoxantrone was given at a dose of 10 mg/m2 on d 1 and cytarabine 100 mg i.v., b.d. during d 1–2. Non‐hematologic toxicity was mild. We conclude that this novel chemotherapy program is effective both as first‐line and salvage treatment in patients with high‐grade malignant NHL. Furthermore, ENAP appears clinically to be partly non‐cross resistant with CHOP chemotherapy. The dose‐limiting toxicity is myelosuppression. The combination should be explored as primary therapy in combination with other chemotherapy or radiotherapy programs.
Clinical features and response to treatment of infradiaphragmatic Hodgkin's disease
Authors: Villamor Neus , Reverter Joan C. , Martí Josep M. , Montserrat Emili , Rozman Ciril
Published: July 2010

Abstract: Among 247 patients with Hodgkin's disease, initial disease presentation was restricted to infradiaphragmatic sites in 17 (6.9%). Advanced age, B symptoms, increased ESR, low lymphocyte and platelet counts, as well as advanced pathological stage and lymphocyte depletion histology were common presenting features of these patients. 7 patients with infradiaphragmatic disease had isolated involvement of inguinofemoral nodes (“peripheral” group) and 10 had only intrabdominal disease (“central” group). Clinical characteristics of patients with “central” forms were different from those with supradiaphragmatic disease, but no differences were observed between “peripheral” infradiaphragmatic and supradiaphragmatic groups. Complete remission was achieved in the 82.2% of patients with infradiaphragmatic disease. Overall survival was 68% at 5 years, and disease‐free survival was 74%. No statistically significant differences were observed in complete remission rates, survival, and disease‐free survival when supradiaphragmatic, “central” infradiaphragmatic and “peripheral” infradiaphragmatic forms were compared.
Imipenem‐cilastatin for empirical therapy in neutropenic patients with fever: An open study in patients with hematologic malignancies
Authors: Huijgens P. C. , Ossenkoppele G. J. , Weijers T. F. , van Loenen A. C. , Simoons‐Smit A. M. , Wijermans P. W. , van Pampus E. C. M. , Langenhuijsen M. M. A. C.
Published: July 2010

Abstract: In adult neutropenic patients with hematological malignancies, we explored imipenem‐cilastatin as empirical antibiotic therapy in a dose of 500 mg four times daily. Changing to second‐line treatment was only resorted to if clinical deterioration, new infections or recurrence of fever occurred. A clinically or microbiologically documented infection was apparent in 115 of 150 episodes studied (76.7%). Imipenem‐cilastatin was successful in 70.7% of all episodes and in 67.8% of all proven infections. Modification was necessary and successful in 22.0% of all episodes. 11 patients died while still febrile, 6 of them by infection (4%) and 5 because of progressive disease (3.3%). Imipenem‐cilastatin is safe initial therapy in neutropenic febrile patients.
Immune activation and the anaemia associated with chronic inflammatory disorders
Authors: Fuchs Dietmar , Hausen Arno , Reibnegger Gilbert , Werner Ernst R. , Werner‐Felmayer Gabriele , Dierich Manfred P. , Wachter Helmut
Published: July 2010

Chronic inflammatory disorders are associated with an increased risk of patients developing anaemia. There is some evidence that cytokines released during cell‐mediated immune responses are capable of inhibiting bone marrow haematopoiesis. In vitro, interferon gamma and tumournecrosis factor alpha inhibit growth of erythroid precursor cells. The mode of action of these cytokines is probably associated with their antiproliferative capacity. Decrease of serum iron and increase of storage iron in patients appears to be a consequence of the defense strategy of macrophages during long‐lasting inflammatory disorders. Decreased serum iron correlates to decreased haemoglobin concentrations. In view of this, the development of anaemia seems likely to result from the altered iron metabolism induced by stimulated macrophages. Low haemoglobin levels and associated hypoxia up‐regulate the release of erythropoietin, which can explain why increased circulating erythropoietin is usually found in patients with anaemia.
Dual rearrangement of immunoglobulin and T‐cell receptor gene in a case of T‐cell hairy‐cell leukemia
Authors: Palumbo Antonio P. , Corradini Paolo , Battaglio Silvano , Omedè Paola , Coda Renato , Boccadoro Mario , Pileri Alessandro
Published: July 2010

We report a case of T‐cell hairy‐cell leukemia with a dual rearrangement of Ig‐ and T‐cell receptor genes. The cytochemical, transmission electron microscopy, and surface antigens data (CD3 +, CD8 +, CD11 +, HLA‐DR +, CD19 ‐, CD20 ‐) were consistent with a T‐cell hairy‐cell leukemia. Molecular analysis according to Southern revealed a dual rearrangement of immunoglobulin heavy‐chain (JH) and T‐cell receptor beta (TcRβ) chain genes. Our findings suggest that the coexistence of JH and TcR gene rearrangements, frequently detected in acute leukemia, may also be observed in hematologic malignancies derived from more differentiated cells.
Chronic granulocytic leukemia: Reassessment of morphologic and cytogenetic characteristics in Ph1‐positive and Ph1‐negative cases
Authors: Crisan Domnita , Mattson Joan C. , Al‐Saadi A.
Published: July 2010

33 cases of chronic granulocytic leukemia (CGL) were reassessed to determine if, by strict morphologic criteria, Philadelphia chromosome (Ph1)‐negative CGL exists as a diagnostic entity and if Ph1‐positive CGL could be distinguished from Ph1‐negative CGL. Cases were reassessed using published criteria and, of 11 Ph1‐negative cases, only 4 could be reclassified as myelodysplastic syndromes or undifferentiated chronic myeloproliferative disorder. Of the morphologic parameters evaluated, peripheral blood basophilia and bicytopenia proved to be good discriminators between Ph1‐positive and Ph1‐negative cases. As a group, Ph1‐negative cases were more heterogeneous and tended to have lower hemoglobin, WBC, platelet count and absolute eosinophilia. Chromosomal abnormalities other than Ph1 were seen only in the Ph1‐positive cases. Based on these findings, we conclude that Ph1‐negative CGL constitutes a heterogeneous group, a subgroup of which is morphologically identical with the Ph1‐positive CGL. The parameters that best discriminate between Ph1‐positive and Ph1‐negative cases are peripheral blood absolute basophilia and bicytopenia.
A liquid culture method for the in vitro growth of hemopoietic progenitor cells from normal human adult peripheral blood allowing for analysis by multiparameter flow‐cytometry
Authors: Serke Stefan , Säuberlich Sabine , Huhn Dieter
Published: July 2010

A liquid culture method has been developed allowing for the in vitro growth of peripheral blood‐derived hemopoietic progenitor cells of myeloid, erythroid, monocytic and megakaryocytic lineages. Adherent cell‐ and CD21‐positive cell‐depleted PBMC from normal subjects have been cultured in the presence of rhEPO, rhGM‐CSF or rhII‐3. Culturing cells in liquid cultures and in plasma clots, a similar dose‐response was observed for granulocytic cells/liquid culture and granulocytic colonies/plasma clot with rhGM‐CSF, and also for erythroid cells/liquid culture and erythroid colonies/plasma clot with rhEPO. Comparing serum‐ liquid cultures to serum+ liquid cultures, the ratio of CD13+ cells to CD15+ cells was higher in serum‐ cultures, indicating a maturation arrest of myeloid cells with serum deprivation. Using dual‐colour flow‐cytometry, cell‐cycle analysis of CD13+ cells, comparing the effects of rhGM‐CSF to those of rhII‐3, have been performed. The liquid culture method promises to be a useful tool for the study of in vitro differentiation and proliferation of hemopoietic progenitor cells.
Retinoic acid inhibits sodium butyrate‐induced monocytic differentiation of HL60 cells while synergistically inducing granulocytoid differentiation
Authors: He Ru‐Yi , Breitman Theodore R.
Published: July 2010

The human myeloid leukemia cell line HL60 is an in vitro model to study myeloid differentiation. HL60 cells differentiate along different cell type lineages in response to a variety of compounds. The direction of differentiation is usually inducer‐specific. However, the response of HL60 cells to sodium n‐butyrate (NaB) is pleiotropic. NaB induces HL60 along the monocytic, neutrophilic, eosinophilic, and basophilic pathways. In this study we saw that physiologic concentrations of all‐trans‐retinoic acid (RA) switched the direction of NaB‐induced differentiation from monocytic to granulocytic. We showed previously (Breitman & He, Cancer Res 1990: 50: 6268–6273) that combinations of RA and NaB synergistically induce HL60 to cells that reduce nitroblue tetrazolium. The present study shows that this synergy was even greater if the parameter measured was mature granulocytes. Our results raise the possibility that the endogenous RA in the serum used to grow cells in culture may affect the direction of differentiation of HL60 cells induced by NaB. Furthermore, our results may provide additional rationale for the use of combinations of RA and NaB in the treatment of some leukemias.
Influences of antiplatelet autoantibodies on platelet function in immune thrombocytopenic purpura
Authors: Yanabu Mutsumasa , Suzuki Masahiko , Soga Tetsuji , Sone Naoaki , Nagata Hirokazu , Nomura Shosaku , Kokawa Terutoshi , Yasunaga Kojiro
Published: July 2010

We investigated the characteristics of the antiplatelet autoantibodies in 60 patients with ITP. Using flow cytometry, the binding of monoclonal antibodies to the platelet glycoprotein (GP) IIb/IIIa complex and to GPIb was examined in these patients. The extent of binding was decreased in 15 patients (anti‐GPIIb/IIIa in 12 patients and both anti‐GPIIb/IIIa and anti‐GPIb in 3 patients). Western blotting revealed that 10 of these 15 patients had either anti‐GPIIb or anti‐GPIIIa and 2 had anti‐GPIb autoantibodies. ADP‐induced aggregation of normal platelets was inhibited by autoantibodies in 12 of 60 patients, and 11 of these had anti‐GPIIb/IIIa antibodies. Ristocetin‐induced aggregation was inhibited in 4 of these patients, and 2 with prominent inhibition had anti‐GPIb antibodies. There was a significant relationship between platelet‐associated IgG value and ATP secretion. These results suggest that some antiplatelet autoantibodies can affect platelet function and thus have an influence on the pathophysiology of ITP.
Regulation of mRNA levels of alkaline phosphatase gene in neutrophilic granulocytes by granulocyte colony‐stimulating factor and retinoic acid
Authors: Sato Noriharu , Mizukami Hiroaki , Tani Kenzaburo , Asano Shigetaka
Published: July 2010

We examined steady‐state levels of mRNA for alkaline phosphatase in neutrophils (NAP) treated with granulocyte colony‐stimulating factor (G‐CSF). The amount of mRNA for NAP was shown to increase after 6 hours of culture with G‐CSF when no increase in NAP activity was yet observed, and the transcript was the greatest after 20–24 h of culture with G‐CSF. Treatment of neutrophils with both G‐CSF and retinoic acid augmented the amount of mRNA for NAP over the amount obtained by G‐CSF alone, which was most marked at 24 h. These results show that both G‐CSF‐mediated NAP induction and its potentiation by retinoic acid are due to the increased levels of mRNA for NAP.
In vitro functions of neutrophils induced by treatment with rhG‐CSF in severe congenital neutropenia
Authors: Roesler J. , Emmendörffer A. , Eisner J. , Zeidler C. , Lohmann‐Matthes M. , Welte K.
Published: July 2010

Neutrophils (and monocytes) from 5 patients suffering from severe congenital neutropenia (SCN) were investigated in vitro after induction of this cell type by treatment with recombinant human granulocyte colony‐stimulating factor (rh G‐CSF) in vivo. Some abnormal morphological features were seen, such as anomalies of nuclei of phagocytes. No limitations were found 1) in the ability of neutrophils and monocytes to produce reactive oxygen intermediates (ROI) following stimulation with phorbolmyristate‐acetate (PMA), 2) in the ability of neutrophils to phagocytose particles of zymosan A and to produce ROI simultaneously and 3) in the ability to kill bacteria of the species staphylococcus aureus. However the specific migration of neutrophils in a gradient of FMLP under soft agar was decreased to approximately 50% in all patients tested as compared to healthy controls. In addition, spontaneous motility was decreased in one patient. Nevertheless, the good clinical improvements of patients suffering from SCN after treatment with rh G‐CSF appeared to be due to induction of neutrophils displaying overall good functional activities with respect to natural defense.
Erythropoietin deficiency in the anaemia of chronic disorders
Authors: Boyd H. Kathryn , Lappin T. R. J.
Published: July 2010

Abstract: Defective iron metabolism, mild haemolysis and impaired erythropoiesis contribute to the anaemia of chronic disorders (ACD), but evidence for a deficiency of circulating erythropoietin (Epo) is equivocal. We have examined serum Epo in moderately anaemic patients with Hb < 10 g/dl — 41 patients with ACD (23 associated with rheumatoid disease and 18 with malignancy), 17 with uncomplicated iron‐deficiency anaemia and 33 with chronic renal failure (CRF). In ACD the serum Epo (mean (confidence limits)) results of 41 (31, 54) mU/ml for the rheumatoid group and 63 (49, 80) mU/ml for the malignancy group, were significantly lower than the Epo of 104 (78, 136) mU/ml for the iron‐deficiency group. The CRF group with more severe anaemia had serum Epo of 27 (19, 35)mU/ml. Thus, recombinant human erythropoietin (rHu Epo) should be considered for the treatment of ACD associated with rheumatoid disease and malignancy.
Gliotoxin treatment selectively spares M‐CSF‐ plus IL‐3‐responsive multipotent haemopoietic progenitor cells in bone marrow
Authors: Kobayashi Masanobu , Müllbacher Arno , Waring Paul , Hapel Andrew J.
Published: July 2010

Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. Gliotoxin has previously been used to treat murine allogeneic bone marrow prior to transplantation into irradiated recipients, and in this situation the drug prevents development of graft‐versus‐host disease, and permits the establishment of allogeneic bone marrow chimeras. We have examined the nature of the cells that survive gliotoxin treatment and report here that gliotoxin selectively spares a unique class of haemopoietic stem cell that forms large (HPP) colonies in the presence of mixtures of M‐CSF and IL‐3. We confirm that the cells which survive gliotoxin treatment are capable of reconstituting the haemopoietic system in allogeneic lethally irradiated mice.
Severe aplastic anemia (SAA): Response to cyclosporin A (CyA) in vivo and in vitro
Authors: Tong J. , Bacigalupo A. , Piaggio G. , Figari O. , van Lint M. T. , Occhini D. , Lamparelli T. , Gualandi F. , Marmont A. M.
Published: July 2010

Abstract: The aim of the present study was to test the effect of cyclosporin A (CyA) in vitro on CFU‐GM growth from patients with severe aplastic anemia (SAA). For this purpose, bone marrow (BM) cells from 9 SAA patients and 5 healthy individuals were incubated with or without CyA and then cultured for CFU‐GM growth in the presence of exogenous recombinant human GM‐CSF (30 ng/ml). SAA patients were tested before or after treatment with CyA, or after treatment with antilymphocyte globulin (ALG). In 3 patients responding to CyA, the addition of CyA in vitro enhanced colony growth from 13 ± 10 to 40 ± 20/105BM cells (p = 0.01) — the median increment of colony formation was 2.4‐fold. In 5 ALG responders, CyA produced no increment of CFU‐GM growth (from 14 ± 26 to 15 ± 16/105BM cells, p = 0.1). CyA did not enhance significantly CFU‐GM growth in normal controls (from 57 ± 45 to 58 ± 81/105BM cells, p = 0.9). In conclusion, it would appear that some patients with SAA can respond to CyA in vivo and in vitro, and ALG responders are not necessarily among these. This is in keeping with different mechanisms of action of CyA and ALG and possibly with the existence of distinct pathogenetic pathways in SAA.
Postoperative erythroderma with change of HLA phenotypes from heterozygotes to homozygotes
Authors: Ito K. , Fujita M. , Norioka M. , Yoshida H. , Arii S. , Tanaka J. , Tobe T. , Kakuyama M. , Maruya E. , Saji H. , Hosoi T.
Published: July 2010

Abstract: Fatal “postoperative erythroderma” (POE) developed in 2 patients treated with liver lobectomy and the transfusion of fresh blood. Their clinical features and skin‐histological findings were indicative of acute graft‐versus‐host disease (GVHD). The HLA phenotypes of circulating lymphocytes of the 2 patients were heterozygous but became homozygous late in the clinical course and were identical with those of the blood donors. One of the patient's haplotypes was identical with the donor's homozygous haplotype. These findings suggest the mechanism of development of POE in apparently immunocompetent patients. The donor's T lymphocytes are histocompatible with the patient's tissues, are not rejected, and become engrafted. The patient's tissues are not histocompatible with the donor's, so that GVHD develops.
Human granulocyte colony‐stimulating factor receptors in acute myelogenous leukemia
Authors: Kondo Seiji , Okamura Seiichi , Asano Yoshinobu , Harada Mine , Niho Yoshiyuki
Published: July 2010

Abstract: Human granulocyte colony‐stimulating factor (G‐CSF) receptors on human acute leukemia cells were investigated using human G‐CSF iodolabeled by the lactoperoxidase method. Among various human leukemic cell lines, only cells of myelogenous lineage including HL‐60, THP‐1 and U937 had one type of high‐affinity receptor for G‐CSF, as shown by Scatchard analysis. Fresh leukemia cells from 19 patients with acute myelogenous leukemia (AML) were then studied. Specific receptors for G‐CSF were demonstrated on blast cells in all 19 cases, the mean number of G‐CSF receptors per AML cell ranging from 95 to 1436. G‐CSF receptors on AML cells appeared to be a single affinity type, although some variations were observed. The mean number of G‐CSF receptors on leukemic cells from patients with either FAB M3 or FAB M2 was greater than that of cells from patients with Ml (p < 0.01, p < 0.10, respectively). Moreover, the mean number of receptors for G‐CSF on CD13‐and CD34‐positive AML cells was higher than that on CD13‐negative and CD34‐positive AML cells (p < 0.01), and the mean number of G‐CSF receptors on CD7‐positive AML cells was lower than that for CD7‐negative AML cells (p < 0.10). Since the FAB classification and surface phenotypes reflect maturation stages, our findings indicate that the distribution of G‐CSF receptors, even on AML cells, may be related to the maturation process.
Recombinant interferon‐ γ inhibits the growth of IL‐6‐dependent human multiple myeloma cell lines in vitro
Authors: Jernberg‐Wiklund H. , Pettersson M. , Nilsson K.
Published: July 2010

Abstract: Recombinant human IFN‐γ (100–1000 U/ml) inhibited the IL‐6‐induced growth of 2 human IL‐6‐dependent multiple myeloma (MM) cell lines U‐1958 and U‐266–1970 in vitro. In contrast, the U‐1996 line, independent of IL‐6 for maintenance at a slow growth rate but responding to IL‐6 by increased proliferation, and the IL‐6‐independent U‐266–1984 were refractory to the anti‐proliferative effect of IFN‐γ. The effect of IFN‐γ in the sensitive MM cell lines was cytostatic in U‐266–1970, and cytostatic and cytotoxic in U‐1958. Northern blot analysis revealed that the growth inhibition of the IL‐6‐dependent MM cell line U‐1958 was not due to down‐regulation of IL‐6 receptor mRNA expression and that the differential sensitivity to IFN‐γ was not due to differences in IFN‐γ receptor expression. The growth inhibition was not a consequence of an IFN‐γ‐induced terminal differentiation as flow cytometric analyses demonstrated an arrest in all phases of the cell cycle. IFN‐α inhibited the growth in 3 of the 4 cell lines tested. The results thus suggest that the particular MM phenotype, which includes IL‐6 dependency for survival and growth, may also be characterized by IFN‐γ sensitivity. Furthermore, the study demonstrates that MM cell lines are not simultaneously sensitive to IFN‐γ and α, indicating that the mechanisms of action of the two types of IFN are distinct.
Remission rate and survival in acute myeloid leukemia: Impact of selection and chemotherapy
Authors: Wahlin Anders , Hörnsten Per , Jonsson Håkan
Published: July 2010

Abstract:113 patients with acute myelogenous leukemia (AML), representing 82% of the total cohort of AML patients within the geographical area of northern Sweden, were recorded. The total complete remission (CR) rate was 47.8 %, and median survival was 4 months. The probability of long‐term survival for all patients without exclusions was only 5%. Thus, the results from this study differ strongly from data on patient outcome in most therapy studies in AML, where the influence of patient selection on the results is larger. The median age in our patients was 63 years, which is also higher than in most other studies. Elderly patients had a low CR rate (24% in patients >70 yr), but remission duration was similar in the different age groups. Patients treated according to “high‐dose” protocols had a CR rate of 64%, while only 14% of less aggressively treated patients achieved remission. A better response rate after more aggressive chemotherapy was evident also in elderly patients. CR rate was 81% in patients below 60 yr of age who had no antecedent blood disorder and who had had symptoms for less than 3 months. Other variables with prognostic implications were: cytogenetic subgroup, antecedent hematological disease, and level of serum ferritin. High serum ferritin was associated with short CR duration. Ferritin is produced by the leukemic cells and could be regarded as a marker for leukemic activity.
Serum cobalamins in the elderly: A longitudinal study of a representative population sample from age 70 to 81
Authors: Nilsson‐Ehle H. , Jagenburg R. , Landahl S. , Lindstedt S. , Svanborg A. , Westin J.
Published: July 2010

Abstract: In a representative population sample (n = 973) born 1901–1902 and examined at the ages of 70, 75, 79, and 81, the change in serum cobalamins with increasing age was studied by trend analysis using values obtained in single individuals at all four examinations. In subsamples without definable disorders, the mean annual decline was: among men 3.4 pmol/l (p < 0.05), among women 3.2 pmol/l (n.s.). The decline was possibly more pronounced among individuals with low and intermediate concentrations. The health‐related lower reference limits (the 2.5 percentile values of subsamples without definable disorders) did not differ significantly between sexes and age groups, but low concentrations or ongoing cobalamin medication became more common with advancing age. The results indicate a slight fall in serum cobalamins between age 70 and 81 but do not call for age‐related lower reference limits.
Cytogenetics of secondary myelodysplasia (sMDS) and acute nonlymphocvtic leukemia (sANLL)
Authors: Johansson B. , Mertens F. , Heim S. , Kristoffersson U. , Mitelrnan F.
Published: July 2010

Abstract: 76 cases of secondary myelodysplasia (sMDS) and acute nonlymphocytic leukemia (sANLL) were cytogenetically analyzed. Among the 36 sMDS patients, 13 (36%) had only normal karyotypes whereas 23 (64%) displayed clonal chromosomal abnormalities. The most common aberrations were −7, 5q‐, −5, and +8. In 10 patients (43% of the cytogenetically aberrant cases), clones with only one anomaly, mostly 5q −or − 7, were found. Of the 40 sANLL patients, normal karyotypes were detected in 10 (25%). Among the 30 (75%) abnormal cases, the most frequent aberrations were −7, −5, +8, 7q‐, − 17, and +21. 12 patients (40%) had clones with single abnormalities, most often − 7. In 4 sANLL patients cytogenetically unrelated clones were detected. A survey of all previously published secondary hematologic neoplasias reveals that the most frequent abnormalities in sMDS are −7 (41%), 5q‐ (28%), and − 5 (11x), followed by der(21q), + 8, 7q‐, der(12p), t(1;7), − 12, − 17, der(17p), der(3p), der(6p), and − 18. Clones with single aberrations have been found in 45 % of the cases and cytogenetically unrelated clones have been described in 6%. The most common abnormalities in sANLL are −7 (38%), 5q‐ (17%), −5 (15%), +8 (13%), and − 17 (llx), followed by der(3q), der(11q), der(12p), −21, 7q‐, − 18, der(3p), der(17p), +21, der(21q), der(6p), and − 16. 38% of the sANLL patients have had clones with only one aberration and 3% have had unrelated clones. The frequencies of these nonrandom abnormalities in sMDS and sANLL are thus remarkably similar ‐ the only exception appears to be 5q ‐, which is more common in sMDS. Also the mean number of abnormalities per case is similar − 5.3 in sMDS and 5.6 in sANLL. When the incidences of characteristic cytogenetic abnormalities were correlated with the type of previous therapy, − 7 was found to be more frequent in sMDS and sANLL patients who had been exposed to chemotherapy whereas 5q ‐was associated with previous exposure to ionizing radiation in sMDS patients.
Distinct morphophenotypic features of chronic B‐cell leukaemias identified with CDlc and CD23 antibodies
Authors: Orazi Attilio , Cattoretti Giorgio , Polli Nicoletta , Delia Domenico , Rilke Franco
Published: July 2010

Abstract: Morphological criteria usually applied to diagnose various subtypes of B‐cell chronic lymphoid leukaemia are largely subjective. Immu‐nophenotyping of 61 relevant cases using a selected panel of monoclonal antibodies (mAb), showed that CDlc and CD23 mAb were able to separate B‐cell chronic lymphocytic leukaemia (B‐CLL) from other chronic B‐cell lymphoproliferative diseases. Lymphocytes of B‐CLL were CDlc‐, CD23+, whereas those of other types of chronic B‐cell leukaemia were CDlc+/‐, CD23‐, and CD38 +/‐. Non‐B‐CLL cases had a significantly higher amount of large peroxidase‐negative (unstained) cells analyzed with an automated blood cell counter (Technicon H6000). This type of volumetric assessment allowed a separation between typical and “atypical” B‐CLL, which otherwise were both CDlc‐, and CD23+. These combinations of phenotypic markers corresponded to well‐defined haematopathologic entities, conventionally diagnosed on peripheral blood (PB) and bone marrow smears, and on histologic sections of lymph nodes and spleen.
Non‐lymphoid blast crisis of CML with rearrangement of immunoglobulin and T‐cell receptor delta genes
Authors: Bashey Asad , Vulliamy Tom , Cai Bi Su , Schwarer Anthony P. , Goldman John M.
Published: July 2010

Abstract: We report a patient with chronic myeloid leukaemia (Philadelphia‐positive with M‐BCR rearrangement) in transformation whose blast cells had myelomonocytic morphology, absent terminal deoxynucleotidyl transferase expression and non‐lymphoid cell surface markers (CD10‐, CD19‐, CD33+, CD14+, CD11+). Leukaemia cell DNA showed rearrangement of both immunoglobulin heavy chain and T‐cell receptor δ genes. Such rearrangements may be a feature of a small proportion of patients with non‐lymphoid transformation of CML as they are in a minority of cases of de novo acute non‐lymphoblastic leukaemia.
Occurrence and characteristics of hereditary spherocytosis in Algeria
Authors: Zerhouni F. , Guetarni D. , Henni T. , Colonna P.
Published: July 2010

Abstract: In a survey of more than 12000 persons referred to a hematological outpatient clinic in Algiers, we estimated that the incidence of hereditary spherocytosis (HS) is 1/1000. Another 9 cases were found in nine of the corresponding families. Anemia was present in a total of 44 subjects (81 %). The transmission was dominant in five of eight informative families (63%). No firm conclusion could be reached concerning the amount of spectrin and ankyrin in nine families; however two‐dimensional peptide maps ruled out any alpha11 domain abnormality in these families. We estimate that HS has roughly the same incidence and features among Algerians as in Europeans or people of European descent.
Inhibition of hexose monophosphate shunt in young erythrocytes by pyrimidine nucleotides in hereditary pyrimidine 5‘ nucleotidase deficiency
Authors: David Onorata , Ramenghi Ugo , Camaschella Clara , Vota Maria Grazia , Comino Luisella , Pescarmona Gian Piero , Nicola Paolo
Published: July 2010

Abstract: Recent reports have suggested that haemolytic anaemia in pyrimidine 5′ nucleotidase (P5′N) deficiency might be due to impaired erythrocyte hexose monophosphate shunt (HMS). To investigate the relationship between pyrimidine accumulation, HMS impairment and shortened red‐cell survival, we tested glucose 6‐phosphate deydrogenase (G‐6PD), HMS, P5′N activities and the UV spectrum in whole red cells and in red cells of different age from 2 P5′N‐deficient patients with different degrees of haemolytic anaemia. In whole red cells we found a reduction of both GdPD and stimulated HMS activity in the presence of a variable amount of pyrimidine nucleotides (37.79 and 17.88 pmol/gHb respectively). A drastic inhibition of stimulated HMS activity was already present in the lightest red‐cell fractions from patient 1, who presented a more severe haemolytic anaemia. The variable degree of pyrimidines found among red cell fractions, with a minor accumulation in the older red cells, supports the hypothesis that pyrimidine accumulation and HMS impairment occur in the younger erythrocytes of P5′N‐deficient patients.
Influence of cytostatic treatment on the coagulation system and fibrinolysis in patients with non‐Hodgkin's lymphomas and acute leukemias
Authors: Zurborn K.‐H. , Gram J. , Glander K. , Delbrück K. , Pelzer H. , Loffler H. , Bruhn H.D.
Published: July 2010

Abstract: Cytostatic therapy is known to aggravate tumor‐induced coagulopathy. Therefore, we have studied the effect of different chemotherapeutic regimens on the activation of coagulation and fibrinolysis in patients with non‐Hodgkin's lymphomas or acute leukemias. In non‐Hodgkin's lymphoma patients treated with an aggressive protocol (COL‐BLAM) and in leukemia patients (TAD‐9) fibrinopeptide A, prothrombin fragment (F1 + 2) and thrombin antithrombin I11 complexes (TAT) increased (Tables 4 and 6), while D‐dimer did not deviate significantly. The ratio D‐dimer/TAT consequently showed a significant decrease, indicating increased formation of thrombin after release of procoagulant factors, which is not paralleled by an activation of fibrinolysis. Both these groups were also characterized by an increase in uric acid and in C‐reactive protein and plasminogen‐activator inhibitor, two acute‐phase reactants. In contrast, patients with non‐Hodgkin's lymphomas treated with a less aggressive protocol (COP) showed no significant changes in hemostatic variables, uric acid, or acute‐phase reactants. The release of procoagulant factors relates to the cytostatic sensitivity of the tumor and to a high tumor‐cell destruction. Our results further emphasize the need for large‐scale studies on antithrombotic prophylaxis in patients undergoing cytostatic treatment.
Vitamin B‐12 abnormalities in HIV‐infected patients
Authors: Remacha Angel F. , Rierasp Anna , Cadafalch Josep , Gimferrer Enric
Published: July 2010

Abstract: A prospective study of 60 consecutively admitted patients with HIV infection was performed to document the prevalence, etiology and manifestations of low serum vitamin B‐12 in such patients. Low serum B‐12 levels were found in 10 patients (16.7%). In 6, vitamin B‐12 absorption was impaired and hog intrinsic factor addition did not improve it. Patients with low vitamin B‐12 levels showed lower hemoglobin, leukocytes, lymphocytes, CD4 lymphocytes and CD4/CD8 lymphocyte ratio than HIV patients with physiological serum vitamin B‐12 levels. However, bone marrow megaloblastosis was found in only 3 low vitamin B‐12 patients and the deoxyuridine suppression test was pathological in only 1 case. In 7 patients, parented treatment was begun with variable response despite serum vitamin B‐12 correction. In conclusion, low serum vitamin B‐12 is often found in HIV‐infected patients and it could be related to malabsorption, but clear megaloblastic abnormalities and treatment response could not be demonstrated. A decreased concentration of the serum binders due to disturbances in the leukocytes and related immunocompetent cell may play an additional role.