bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic and genotypic switch in Philadelphia‐positive, BCR‐positive blast crisis of chronic myeloid leukemia
Authors: Callea V. , Morabito F. , di Celle P. Francia , Ronco F. , Carbone A. , Nobile F. , Foa R.
Published: July 2010

Abstract:  We report a case of Ph1‐positive, bcr‐positive chronic myeloid leukemia blast crisis (CML‐BC) which at presentation showed a mixed myeloid/B‐lymphoid immunophenotype along with TdT positivity and, at the molecular level, an oligoclonal rearrangement of the immunoglobulin heavy chain (IgH) gene region. After obtaining a successful remission, at the time of relapse the patient underwent a phenotypic and genotypic switch from mixed to myeloid phenotype, characterized by the loss of the lymphoid markers and TdT expression and by a germline configuration of the IgH gene region. The same bcr rearrangement was, however, found in both phases of the disease, supporting the suggestion of a true phenotypic and genotypic conversion. This report confirms that the neoplastic event in CML may take place at an early multipotent stem‐cell level, prior to a well‐defined phenotypic and genotypic lineage expression. Moreover, it is suggested that different factors (chemotherapy? growth factors?) may have either eradicated the bcr+ /IgH+ clone and promoted the growth of bcr+ /IgH− leukemic cells or, alternatively, supported the lymphoid differentiation program and induced a myeloid lineage shift.
Occurrence of monoclonal components in general practice: Clinical implications
Authors: Aguzzi F. , Bergami M. R. , Gasparro C. , Bellotti V. , Merlini G.
Published: July 2010

Abstract:  We report on the occurrence of monoclonal components observed in a provincial hospital in northern Italy from 1987 to 1990. The monoclonal components were detected by visual inspection of high‐resolution acetate serum electrophoreses and typed by immunofixation. The percentage of monoclonal components increases steeply with age, and reaches a plateau of 7–8% in individuals over 55 years old. Besides the high percentage of monoclonal component, the other relevant finding of this study is that ˜80% of monoclonal components are of low concentration (<5 g/l). Most of these subjects with small monoclonal component passed undetected in the previous studies on the prognostic significance of monoclonal gammapathy. These findings indicate the need for a revision of the current concepts on the biological and clinical significance of MC discovered by chance.
Transient calcium elevation in polymorphonuclear leukocytes triggered by thrombin‐activated platelets
Authors: Takahashi A. , Yamamoto K. , Okuma M. , Sasada M.
Published: July 2010

Abstract:  The effect of thrombin‐activated platelets and their release products on the intracellular free calcium concentration ([Ca2+]i) of human polymorphonuclear leukocytes (PMNs) was studied by loading PMNs with a fluorescent indicator of calcium, fura‐2. [Ca2+]i of PMNs was transiently elevated by thrombin‐activated platelets. The supernatant of thrombin‐activated platelets also elicited a transient elevation of [Ca2+]i in PMNs. Pretreatment of the supernatant with hexokinase caused a decrease in the transient [Ca2+]i elevation of PMNs, while hexokinase abrogated the [Ca2+]i elevation of PMNs elicited by 80 μmol/1 adenosine triphosphate (ATP). Pretreatment of the supernatant with trypsin also decreased the magnitude of the elevation, while trypsin had no effect on the response to ATP. These findings suggest that thrombin‐activated platelets induce a transient [Ca2+]i elevation in PMNs by releasing ATP and some trypsin‐sensitive factor(s).
In vivo kinetics of 111Indium‐labelled autologous granulocytes following i.v. administration of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF)
Authors: Hovgaard D. , Schifter S. , Rabøl A. , Mortensen B. T. , Nissen N. I.
Published: July 2010

Abstract:  Administration of both glycosylated and non‐glycosylated recombinant human granulocyte‐macrophage colony‐stimulating factor (rhGM‐CSF) induces an immediate transient granulocytopenia of 1–3 hours' duration. In order to explore this phenomenon, granulocytes were labelled with 111Indium and the effect on the kinetics of granulocytes after administration of rhGM‐CSF was studied in 10 previously untreated patients with malignant lymphoma. For both types and doses of rhGM‐CSF, a significant and dramatic accumulation of the 111Indium‐labelled granulocytes was observed in the lung within a few minutes after i.v. injection of rhGM‐CSF. The accumulation of radioactivity coincided with the pronounced and transient granulocytopenia in peripheral blood. The 111Indium‐labelled granulocytes later reappeared in the peripheral blood, indicating reversible pulmonary vascular margination of the granulocytes. Half‐life of labelled granulocytes after reappearance was comparable to half‐life values under normal conditions. The transient accumulation of granulocytes in the pulmonary vessels seems not to be of clinical importance in the management of patients, but it may to some degree explain previously described side‐effects, such as transient hypoxemia (“first‐dose” reaction) following administration of rhGM‐CSF.
Myelofibrosis in primary myelodysplasic syndromes
Authors: Maschek H. , Georgii A. , Kaloutsi V. , Werner M. , Bandecar K. , Kressel M.‐G. , Choritz H. , Freund M. , Hufnagl D.
Published: July 2010

Abstract:  In a retrospective study of 352 patients with primary myelodysplastic syndromes, 61 (17.3%) revealed myelofibrosis in bone marrow biopsies. The fibrosis was observed to occur mostly focally (41/61 cases), and collagen deposits were found very rarely (4/61). The histopathology of bone marrow biopsies revealed hyperplasia and disturbed differentiation in megakaryopoiesis; the frequency and grade of dysplasia in megakaryopoiesis increased with advancing myelofibrosis. Reticulin fibrosis occurred in all subtypes of MDS; however, there was a higher incidence in chronic myelo‐monocytic leukaemia (CMMoL). The frequency of cytogenetic aberrations was significantly higher in the MDS cases with myelofibrosis, compared to the cases without fibrosis. Clinical data showed significantly lower values of haemoglobin and lower platelet counts in MDS with myelofibrosis. Life expectancy was reduced to 9.6 months, compared with 17.4 months in MDS without fibrosis. In refractory anaemia, the survival times were 10.0 months in MDS with myelofibrosis, compared to 28.9 months in MDS without myelofibrosis. 36.6% of the patients with MDS and myelofibrosis developed a transformation into ANLL during the course of the disease. Myelofibrosis therefore seems to herald a poor prognosis.
High remission rate in acute myeloblastic leukemia in children treated with high‐dose methylprednisolone
Authors: Hicsönmez Gönül , Karadeniz Nilufer , Zamani V. Prozorova , Tuncer A. Murat , Gümrük Fatma , Erturk Gülyüz , Gurgey Aytemiz , Ozsoylu Sinasi
Published: July 2010

Abstract:  Since the differentiating effect of high‐dose methylprednisolone (HDMP) on myeloid leukemic cells has been shown in one of our patients with acute myeloblastic leukemia (AML‐M4), 27 previously untreated children with AML were given HDMP (20–30 mg/kg per day) combined with cytosine arabinoside (Ara‐C; 3 mg/kg) for the first 2 weeks of induction therapy. Marked clinical improvement was observed in all patients with the exception of one who died within 24 hours of the treatment. Enlarged liver and spleen (>5 cm) became nonpalpable in 3 (37%) out of 8 and 5 (100%) out of 5 patients, respectively, and bone marrow blasts decreased below 5% in 7 patients (27%) within 2 wk of HDMP and Ara‐C treatment. Adriamycin (1 mg/kg) was added 2 wk after initiation of induction therapy. Twenty‐two (84.6%) of the 26 patients achieved complete remission, 3 (11.5%) had partial remission and no response was obtained in one. Treatment was well tolerated. The addition of HDMP as a differentiating and/or cytolytic agent to conventional anti‐leukemic chemotherapy increased the complete remission rate and prolonged the duration of remission of our AML patients.
Cobalamin binding proteins in patients with HIV infection
Authors: Hansen Mads , Gimsing Peter , Ingeberg Steen , Jans Henning , Nexø Ebba
Published: July 2010

Abstract:  P‐Cobalamins have been reported to be decreased in patients with HIV infection. Because of this, we found it of interest to examine both cobalamin‐saturated binding proteins (holo‐transcobalamin, holo‐TC and holo‐haptocorrin, holo‐HC) and cobalamin unsaturated binding proteins (apo‐transcobalamin, apo‐TC and apo‐haptocorrin, apo‐HC). The results are given as range and (median). Eighteen male HIV‐infected patients with plasma cobalamins below 200 pmol/1 were studied. We found low concentrations of holo‐TC (37–88 (47.5) pmol/1) and holo‐HC (64–184 (135.5) pmol/1). The concentration of apo‐TC and apo‐HC was increased (480–1730 (1025) pmol/1; 70–800 (235) pmol/1). It is concluded that, in HIV‐infected patients, low plasma cobalamin does not reflect a low concentration of transcobalamin or haptocorrin. In 20 HIV‐infected patients and 31 patients with malignant haematological diseases, the TC isopeptide patterns were determined. In the HIV group, an increased frequency of TC isopeptide X was found and the overall distribution of TC isopeptides was significantly different from the reference population (p < 0.05). There was no difference between the group of patients with malignant haematological diseases and the reference group.
The receptors for regulatory molecules of hematopoiesis
Authors: Olsson I. , Gullberg U. , Lantz M. , Richter J.
Published: July 2010

Abstract: Proliferation and differentiation of hematopoietic cells are controlled by pleiotropic regulatory molecules. While the sequences of these factors are not related, their membrane receptors are restricted to two gene families with homologous domains. The members of the hematopoietic (or cytokine) receptor family (for erythropoietin, interleukins‐2, ‐3, ‐4, ‐6 and ‐7, granulocyte‐macrophage and granulocyte colony‐stimulating factor) are composed of multiple subunits necessary for high‐affinity binding and cell signalling. Signal transducing mechanisms are largely unknown. The occurrence of variant signal transducers in different tissues could explain the pleiotropy of these regulatory molecules. Members of the receptor tyrosine kinase family bind dimeric forms of macrophage colony‐stimulating factor, stem cell factor and platelet‐derived growth factor leading to kinase activation and phosphorylation of many substrates involved in production of second messengers. Soluble forms (binding proteins) exist for members of both families. These may be proteolytic cleavage products of transmembrane receptors or naturally secreted products. Such binding proteins can potentially function as inhibitors in feedback regulation and in protection and transport of cytokines and would provide a rational therapy when cytokines are produced in excess. Knowledge of signal transduction mechanisms and of the three‐dimensional structure of ligands and receptors can lead to the design of drugs with cell‐specific effects.
CD 1‐reactive leukemic cells in bone marrow: Presence of Langerhans cell marker on leukemic monocytic cells
Authors: Misery Laurent , Campos Lydia , Dezutter‐Dambuyant Colette , Guyotat Denis , Treille Danielle , Schmitt Daniel , Thivolet Jean
Published: July 2010

Abstract: Langerhans cells originate in bone marrow and probably belong to the monocyte‐macrophage lineage. CD1 is a specific marker of Langerhans cells. By immunofluorescence and immunoelectron microscopy, CD1a antigen and myeloid markers (CD11, CD13, CD14, CD15, CD33, HLA‐DR) were studied in 53 cases of acute myeloid leukemias (AML) and 3 acute lymphoblastic leukemias (ALL). The 11 ANLL without monocytic component were CD1a negative. 2/5 of acute myelomonocytic leukemias (AML4) and 9/37 of acute monocytic leukemias (AML5) were positive. All 3 ALL were negative. No correlation was found between CD1a and myeloid markers. CD1a+ AML did not differ from CD1a‐ AML with regard to cytogenetics or response to therapy. The CD1a positive cells may originate from an abnormal proliferation of CD1a positive cells which are present in bone marrow and which may differentiate into Langerhans cell precursors.
Remission may continue after termination of rIFNα ‐2b treatment for essential thrombocythemia
Authors: Kasparu H. , Bernhart M. , Krieger O. , Lutz D.
Published: July 2010

Abstract: Essential thrombocythemia, a myeloproliferative disorder of clonal origin, is often associated with various clinical manifestations resulting from thromboembolic or hemorrhagic complications. The long‐established successful method of treatment with cytotoxic agents or radioactive phosphorus has recently been superseded by interferon alpha. We treated 14 symptomatic patients with 5 times 106 IU recombinant interferonα‐2b s.c. daily. 12/14 pts responded within 14–75 days. When platelet counts decreased to below 450 g/l the frequency of administration was reduced stepwise. 7 patients remained in CR during this reduction phase and treatment was stopped in 5 pts after 12–32 months. Until now, 3 of them are still in continuous good PR without any drug therapy and free of symptoms for 3+, 19+ and 36+ months. Continuous response during maintenance was associated with age, initial platelet count and time required to reduce platelet counts to <450 g/l.
Combination chemotherapy MOCCA in resistant and relapsing multiple myeloma
Authors: Palva I.P. ,   Finnish Leukaemia Group
Published: July 2010

Abstract: 80 patients with resistant or relapsing multiple myeloma received a combination of vincristine, cyclophosphamide, lomustine, melphalan and methylprednisolone (MOCCA) as a second‐line chemotherapy. 27 of them were resistant to primary chemotherapy with alkylating agents, and 53 had relapsed after initially responding to these drugs. An objective response was achieved in 39 patients (49%): in 14 patients who were primarily resistant (52%) and in 25 patients who had a relapse (47%). Of 41 patients relapsing during maintenance chemotherapy 14 (34%) responded, while 11 of 12 patients (92%) treated for a relapse off‐therapy responded. The median duration of response was 22 months. Severe complications, in most cases infections, occurred in 30% of patients, and were fatal in 9% of the cases. According to our experience the five‐drug combination MOCCA is an effective second‐line chemotherapy for myeloma patients primarily resistant to or relapsing after therapy with single alkylating agents.
Bilineage response in refractory aplastic anemia patients following long‐term administration of recombinant human granulocyte colony‐stimulating factor
Authors: Sonoda Yoshiaki , Yashige Hiromi , Fujii Hiroshi , Tsuda Shouichiro , Maekawa Taira , Misawa Shinichi , Abe Tatsuo
Published: July 2010

Abstract: 5 patients with refractory aplastic anemia (AA) received long‐term administration (2–11 + months) of recombinant human G‐CSF (rhG‐CSF) in doses from 250–500 μg/body/day by intravenous infusion or 75–300 μg/body/d by subcutaneous injection. All 5 evaluable patients showed a substantial increase in absolute neutrophil count (ANC) with a recovery of myeloid components in the bone marrow after 1 to 2 months of treatment. Interestingly, 2 out of the 5 patients showed a dramatic improvement in severe anemia after 2 to 4 months of treatment accompanying a recovery of erythroid components in the bone marrow. In addition, there was no serious infection before or during therapy. Long‐term administration of rhG‐CSF was well tolerated because of its minimal toxicity. Clonal assay revealed a recovery of myeloid progenitors in all patients and a recovery of erythroid progenitors in 3 out of the 5 patients. These results suggest that long‐term administration of rhG‐CSF at least mobilizes residual myeloid as well as erythroid progenitor cells and induces a bilineage response in severe refractory AA.
Comparison of poly‐ and monoclonal antibodies for determination of B‐cell clonal excess in an routine clinical laboratory
Authors: Agrawal Y. P. , Hämäläinen E. , Mahlamäki E. K. , Aho H. , Nousiainen T. , Lahtinen R. , Penttilä I. M.
Published: July 2010

Abstract: Flow cytometry (FCM) has gained wide use in the determination of clonality in B‐cell lymphoproliferative diseases and many methodological variations exist. We have compared the suitability of a) dual fluorochrome (FITC/PE)‐labelled monoclonal antibodies, b) single fluorochrome (FITC)‐labelled monoclonal antibodies and c) F(ab')2 fragments of FITC‐labelled polyclonal antibodies for flow cytometric determination of clonality using commercially available software and a short sample preparation protocol. The FCM method was validated by analysis of immunoglobulin heavy chain and light chain gene rearrangements. We recommend the use of FITC‐labelled monoclonals to obtain three parameters, the k/λ ratio, D and D/S(n) values (Kolmogorov‐Smirnov statistics) instead of the commonly used k/λ ratio and D values only. This allows the use of a rapid sample preparation protocol to blood and bone marrow aspirates without sacrificing sensitivity or specificity obtained by the usual FCM method.
Mechanisms accounting for the impaired natural‐killer cell activity in refractory anaemia with excess of blasts
Authors: Eliopoulos G. D. , Coulocheri S. A. , Eliopoulos A. G. , Katrinakis G. , Karkavitsas N. , Kyriakou D. , Vaiopoulos G. , Foudoulakis A.
Published: July 2010

Abstract: Natural killer (NK) cells were analyzed in 38 untreated patients with refractory anaemia with excess of blasts (RAEB), using cytotoxicity assays and immunofluorescence with monoclonal antibodies. We found that patients with RAEB have normal numbers of peripheral blood and bone marrow NK cells. NK cells from RAEB patients express very low natural‐killer cell activity (NKa) which may be increased significantly with recombinant a‐interferon and recombinant interleukin‐2, although it remains below the lower limit of the control range. The cells exhibit normal tumour cell binding capacity, but fail to release sufficient amounts of natural‐killer cytotoxic factors (NKCFs) upon their interaction with NK‐sensitive K562 cell targets or their stimulation with phytohaemagglutinin. Our results suggest that defective NKa in RAEB patients may be due, at least in part, to impaired release of functionally active NKCFs. This disturbance is probably the result of some intrinsic defect of RAEB NK cells in NKCF production, storage, and/or release. The possibility of an impairment in the activation signal provided by the stimulatory K562 cells cannot be excluded, although it seems unlikely. We postulate that this abnormality might represent a manifestation of dysplastic haemopoiesis. Further studies are certainly needed to investigate whether other defective mechanisms are also implicated in the determination of the low NKa in patients with RAEB.
Association between the activation of macrophages, changes of iron metabolism and the degree of anaemia in patients with malignant disorders
Authors: Denz H. , Huber P. , Landmann R. , Orth B. , Wachter H. , Fuchs D.
Published: July 2010

Abstract: In this study, we investigated a possible association between the degree of macrophage activation ‐ as measured by serum neopterin concentrations ‐ and disturbances of iron metabolism, determined by the concentrations of ferritin and serum iron, in patients with malignant disorders. Additionally we evaluated correlations between these factors and the degree and type of anaemia. Seventy‐three patients, who suffered from non‐Hodgkin's lymphoma (NHL) (n = 43), Hodgkin's disease (n = 11), myeloma or monoclonal gammopathy of unknown significance (n = 9), myelodysplastic syndrome (n = l), and solid tumours (n = 9), were examined. Mean neopterin levels were raised in all groups, patients with NHL showing the highest concentrations. Ferritin but not neopterin concentrations were higher in males than in females. A significant correlation was found between neopterin and ferritin concentrations (p<0.01). Considering only female patients the strength of the correlation was the same (p<0.02). In addition, we found inverse correlations of neopterin with haemoglobin and iron concentrations (all p< 0.01). Similar relationships existed in patients during follow‐up. Our results support the hypothesis of an association between the degree of activation of macrophages and the development of anaemia by a shift or iron towards the storage sites.
Donor alloreactivity may predict acute graft‐versus‐host disease in HLA‐matched bone marrow transplantation for leukemia in early remission
Authors: Johnsen H. E. , Beatty P. G. , Michelson E. , Hansen J. A. , Thomas E. D.
Published: July 2010

Abstract: The pretransplant alloantigen‐dependent responding and stimulating capacity of donors and of recipients was studied retrospectively, in a study of prediction of acute graft‐versus‐host disease. Donor responding capacity (DRC) and host stimulating capacity (HSC) were defined by mixed lymphocyte culture (MLC) and normalized by help of the pool response. High and low DRC and strong and weak HSC was defined by distribution plots. Kaplan‐Meier estimates of the risk of developing Grade I1 or higher aGvHD showed that first remission patients (N= 125) had a significantly different risk if transplanted with marrow from a donor with high (N = 54) or low (N = 71) DRC (χ2 = 9.49; d.f. = 1; p<0.002). This was not the case for patients transplanted in later remissions. Host SC status had no significant influence on the aGvHD status (χ2 = 1.75 and 2.40; d.f. = 1; p = 0.19 and 0.12, defined by normal controls A and B respectively). In conclusion, the results indicate that pretransplant donor alloreactivity may predict aGvHD, confirming the results from a Scandinavian study. The results need to be confirmed in prospective studies of alloreactivity as risk factor for aGvHD.
Correlation of MDR1 /P‐170 expression with daunorubicin uptake and sensitivity of leukemic progenitors in acute myeloid leukemia
Authors: Campos Lydia , Guyotat Denis , Jaffar Christine , Solary Eric , Archimbaud Eric , Treille Danielle
Published: July 2010

4Service d'Hématologie, Hôpital Edouard Herriot, Lyon, France

Abstract: Prior studies have shown that multidrug resistance gene products may be detected in acute myeloid leukemia (AML) cells, and are associated with poor response to therapy. We studied whether P‐170 expression was associated with in vitro daunorubicin (DNR) accumulation and sensitivity of leukemic clonogenic cells (CFU‐L) to DNR in 16 newly diagnosed AML samples. P‐170 expression was assessed by indirect immunofluorescence using the monoclonal antibody MRK16. DNR cellular content was measured by flow cytometry after short incubation with increasing concentrations of DNR, and was not correlated with P‐170 expression, although there was a trend for higher values in P‐170‐negative samples. The sensitivity of CFU‐L was studied in a semisolid culture assay by calculating the dose of DNR inhibiting the growth of 90% of CFU‐L (D90). The D90 was significantly higher in P‐170‐positive than in P‐170‐negative samples (mean = 1.68 ± 0.42 pg/ml versus 0.97 ± 0.35 μg/ml respectively, p< 0.005). Eight of 9 cases achieving complete remission (CR) after intensive chemotherapy were P‐170‐negative, whereas 7 of 7 nonresponders were P‐170‐positive (p< 10−5). D90 was significantly lower for patients achieving CR than for those who did not achieve CR (1.12 ± 0.55 μg/ml versus 1.59 ± 0.37 μg/ml, p = 0.04). It is concluded that P‐170 expression is correlated with in virto resistance of clonogenic cells to DNR and may be one mechanism of resistance to chemotherapy.
Identification of a normal human bone marrow cell population co‐exmessin‐g megakaryocytic and erythroid markers in culture
Authors: Bellucci Sylvia , Han Zhong C. , Pidard Dominique , Caen Jacques Philippe
Published: July 2010

Abstract: A population of haematopoietic cells co‐expressing glycoprotein IIIa (GPIIIa), which has been shown to be present in the megakaryocyte‐platelet lineage, and glycophorin A, which has been shown to be specific for the erythroid lineage, has been identified in normal bone marrow cultures using double immunofluorescence staining. The cells showing this phenotype have the size of lymphocytes and appear at an early time (day 1 to d 6) of culture. We have also detected a type of mixed megakaryocyte (MK) cluster containing this phenotype on d 3 or 4 in most normal marrow samples. Such a cell phenotype was not detected after 6 d of culture. It is thus the first time that this phenotype, although already described for several human neoplastic cell lines, has been observed in several normal human bone marrow cultures. Cells expressing this phenotype may represent haematopoietic cells, common to megakaryocytic and erythroid lineages, at a very early stage of differentiation.
Infections and serum IgG levels in patients with chronic lymphocytic leukemia
Authors: Itala M. , Helenius H. , Nikoskelainen J. , Remes K.
Published: July 2010

Abstract: To review our policy of prophylactic treatment with intravenous immunoglobulin (i.v.IG) in chronic lymphocytic leukemia (CLL), we analyzed the infection history, serum IgG levels (S‐IgG) and disease stage of 146 patients who were treated and followed at our institution in 1980–1989. S‐IgG was available for 98 patients: 55% were hypogammaglobulinemic and 56% had had at least one severe infection. There were significant associations between S‐IgG and the occurrence of infections (p<0.01) and disease stage (p < 0.02). There was also a significant association between disease stage and occurrence of infections (p< 0.001). Severe infections tended to accumulate in patients with subnormal S‐IgG and advanced disease stage. Totally, 292 infections were recorded, and the incidence of moderate to severe infections was 0.47 per patient year. Infection mortality was high: 42 patients died of a severe infection (46% of all causes of death). Patients with a low S‐IgG and advanced disease stage are the most susceptible to death from infection and would be most likely to benefit most from i.v.IG prophylaxis; however, the cost of this therapy is so high that strict individual consideration still remains crucial for treatment decisions.
Synovial‐type (group II) phospholipase A2 in serum of febrile patients with haematological malignancy
Authors: Rintala Esa M. , Nevalainen Timo J.
Published: July 2010

Abstract: Elevated concentrations of synovial‐type (group II) phospholipase A2 (PLA2‐II) in serum are associated with septic bacterial infections. We measured the concentrations of PLA2‐II in serum in 24 fever episodes involving patients suffering from haematological malignancies and having fever after cytotoxic treatment. We applied a novel time‐resolved fluoroimmunoassay using a polyclonal antibody raised against recombinant human synovial‐type PLA2. The concentrations of PLA2‐II in serum were 194.7 ± 204.4 μg/l (mean ± SD, median 141.9, range 4.6–931.5 μg/l). The concentrations of PLA2‐II correlated well to the concentrations of C‐reactive protein (CRP) in serum (r = 0.688, p<0.001). The PLA2‐II concentrations increased faster than the corresponding CRP values and began to decrease 12 hours after the beginning of antimicrobial treatment. Inverse correlations were found between the concentrations of PLA2‐II and blood neutrophil and platelet counts. No correlation was found between the concentrations of PLA2‐II and the duration of the time interval from the onset of preceding cytotoxic and corticosteroid treatment to the first blood sample. The concentration of pancreatic PLA2 was within the reference interval in all samples. The present results indicate that PLA2‐II resembles an acute‐phase protein and is not of blood cell or pancreatic origin.
Long‐term treatment with zidovudine in patients with human immunodeficiency virus (HIV)‐associated thrombocytopenia: Modes of response and correlation with markers of HIV replication
Authors: Cinque Paola , Landonio Guiseppe , Lazzarin Adriano , Nosari Anna Maria , Ruggieri Alessandro , Coen Massimo , Meraviglia Paolo , Gringeri Alessandro , Gallo Luciana , Quirino Tiziana , Santagostino Elena , Cargnel Maria Antonietta
Published: July 2010

Abstract: The effects of zidovudine on the platelet count were studied in 152 patients with HIV‐related thrombocytopenia of severe grade (platelet count <50 × 109/1) and moderate grade (platelet count <100 and >50 × 109/1). In both groups of patients there was a significant increase in the mean platelet count from the baseline value, after 2 weeks (from 21 × 109/1 to 48 × 109/1 and from 75 × 109/1 to 97 × 109/1) and 3 months of therapy (to 59 × 109/1 and to 144 × 109/1). Sixty‐five and 39 patients were followed up for 12 and 18 months, respectively, and the mean platelet values after 12 and 18 months of therapy were still significantly increased, compared to the respective mean baseline values, in both groups of patients. Clinical progression of the disease was observed in 23 treated patients, none of them showing concomitant reductions of the platelet number. An increase in the mean CD4+ cell count after 3 months of therapy was followed by a progressive decline in the 65 patients with a 12‐month follow‐up, while no significant changes of the p24 antigenemia rates were observed after 1 year of therapy in 53 patients evaluated. The long‐term effects of zidovudine on the platelet count, but not on other parameters of clinical outcome, might be explained by the involvement of specific mechanisms in the pathogenesis of this kind of thrombocytopenia and of its response to zidovudine.
On the interaction between cytosine arabinoside and etoposide in vivo and in vitro
Authors: Liliemark Jan , Knochenhauer Eva , Gruber Astrid , Pettersson Birgitta , Björkholm Magnus , Peterson Curt
Published: July 2010

Abstract: Cytosine arabinoside (ara‐C) and etoposide are often used in combination in the treatment of acute myelocytic leukemia (AML). The intracellular phosphorylation of ara‐C to its 5′‐triphosphate (ara‐CTP) is a prerequisite for its cytotoxic effects. It has been shown in vitro that etoposide can impair the formation of ara‐CTP in leukemia cells. The present study was undertaken in order to elucidate whether this interaction may be of clinical importance. Leukemia cells were isolated from 3 patients with acute myelocytic leukemia and incubated in medium (RPMI‐1640) with or without 10% fetal calf serum or in human plasma. When the cells were incubated in RPMI‐1640 with ara‐C (10 μmol/l) and etoposide during 2 h, the formation of ara‐CTP was decreased to 71 ± 18 (mean ± S.D.) and 30 ± 15% of control at 1 and 10 μg/ml etoposide, respectively. When the cells were incubated in human plasma, the formation of ara‐CTP was not influenced by the presence of etoposide (101 ± 6 and 103 ± 20% at 1 and 10 μg/ml etoposide). When incubated in RPMI supplemented with 10% fetal calf serum, the corresponding figures were 81 ± 8 and 70 ± 20%. Six patients with AML were therefore treated with ara‐C 0.5 or 1.0 g/m2 as a 2‐h infusion every 12 h and, during 1 h before the second ara‐C infusion, 100 or 200 mg/m2 etoposide was administered. The median change in the AUC of cellular ara‐CTP between the first and second ara‐C dose was 0% (‐37 to +21%). The corresponding median change in rate of accumulation of ara‐CTP in leukemia cells was 12% (‐26 to +110%). The concentration of etoposide in plasma during the ara‐C infusion was 18.7 ± 5.1 μg/ml while the non‐protein bound etoposide was 0.73 ± 0.34 μg/ml. Thus, despite exposure to higher etoposide concentrations in vivo than in vitro, no impairment of ara‐CTP formation was seen in the patients. This corresponds to the results obtained when leukemic cells were incubated in plasma. It is concluded that the inhibition of ara‐CTP formation by etoposide seen in vitro is offset by the high protein binding of etoposide in plasma (96%) and that etoposide does not impair the formation of ara‐CTP in leukemia cells in vivo during treatment with standard‐dose etoposide.
Red cell destruction by human monocytes — changes in intracellular ferritin concentration and phenotype
Authors: Raha‐Chowdhury R. , Williams B. J. , Worwood M.
Published: July 2010

Abstract: Mononuclear cells from 5 normal men and 5 patients homozygous for hereditary haemochromatosis (HFE) have been incubated for 18 h with or without the addition of sheep red blood cells coated with antibody (SRBC). In the absence of SRBC mean H type ferritin concentrations were greater than L type (normals: mean L type 11.6 ng/106 cells, H type 15.5; patients, L type 23.5 ng/106 cells, H type 41.6). In the presence of SRBC, monocyte L type ferritin concentrations increased considerably (76 ng/106 cells in normals and 141 ng/106 cells in patients) but H type ferritin concentrations were the same or decreased compared with incubation in medium only. Incubation with additional iron (ferric ammonium citrate, 2.5 μg Fe/ml) increased both H and L type ferritin concentrations. Erythrophagocytosis thus appears to cause differential regulation of H and L ferritin subunit synthesis or breakdown. Normal subjects and patients do not differ in this response to erythrophagocytosis.
Comparative pharmacokinetics of single‐dose administration of mammalian and bacterially‐derived recombinant human granulocyte‐macrophage colony‐stimulating factor
Authors: Hovgaard Doris , Mortensen Børge Thing , Schifter Søren , Nissen Nis I.
Published: July 2010

Abstract: Pharmacokinetics of recombinant human non‐glycosylated bacterially‐synthesized (E. coli) granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) were studied following single intravenous (i.v.) and subcutaneous (s.c.) bolus injection, and compared to equivalent doses of glycosylated mammalian‐derived CHO‐GM‐CSF. Each route of administration gave a different GM‐CSF concentration‐time profile. The highest peak serum concentrations (Cmax) were observed following i.v. bolus injection. After i.v. administration, a two‐phase decline in concentration was noted for both types of GM‐CSF with a significantly shorter ***t1/2 α of 7.8 minutes for the E. coli GM‐CSF versus 20.0 min for the CHO‐GM‐CSF, while no significant difference was observed for the terminal phase. Following s.c. administration of equivalent doses, a higher peak serum concentration was observed in the E. coli‐treated patients and, again, a faster elimination where pretreatment serum levels were reached after 16–20 h, versus more than 48 h after administration of CHO‐GM‐CSF. Although the non‐glycosylated E. coli GM‐CSF thus seems to undergo a faster elimination that the glycosylated CHO‐GM‐CSF no significant difference could be demonstrated in the in vivo effect of corresponding doses of the two compounds with respect to stimulation of granulopoiesis — with reservation for small patient numbers and a large individual variations in response.
Lomustine, etoposide, methotrexate and prednisone (LEMP) therapy for relapsed and refractory non‐Hodgkin's lymphoma
Authors: Dorigo A. , Mansberg R. , Kwan Y. L.
Published: July 2010

Abstract: Twenty‐two patients with non‐Hodgkin's lymphoma (NHL) were treated with a regimen aimed at administering 6–9 cycles at 6‐weekly intervals of oral lomustine (CCNU) 50 mg/m2 on day 2, i.v. etoposide 50 mg/m2 d 1 and 8, methotrexate 30 mg/m2 d 1 and 8, prednisone 60 mg/m2 d 1–10 (LEMP). The patients had a median age of 65 years (range 34–81) at diagnosis and comprised 14 males and 8 females. Eighteen patients had had prior chemotherapy (5 patients received more than two combinations). Seven had also received prior radiotherapy. Five patients achieved a complete remission (CR) with a median duration of 18 months (range 4–37). Twelve patients achieved a partial remission with a median survival of 8 months (range 2–45). Responses were seen in 6/6 nodular small cleaved cell, 1/1 diffuse small cleaved cell, 0/2 nodular mixed small and large cell, 3/4 diffuse mixed small and large cell, 7/9 diffuse large cell. One patient with diffuse large cell histology was treated initially with LEMP, achieving and maintaining CR for 18 months. Five patients did not respond to LEMP. This regimen was well tolerated requiring only 2 injections every 6 wk, there was minimal toxicity, no alopecia or cardiotoxicity, and little myelosuppression. There were no treatment‐related deaths. This regimen has a useful role in inducing partial or complete remission in patients who have relapsed or progressed following previous intensive chemotherapy. It may also be used as first‐line therapy in patients who may not tolerate more intensive regimens. These results are encouraging and warrant further investigation.
Potential of bone marrow biopsy in chronic myeloproliferative disorders (MPD)
Authors: Bartl Reiner , Frisch Bertha , Wilmanns Wolfgang
Published: July 2010

Abstract: The chronic myeloproliferative disorders (MPD) comprise polycythemia vera (PV), idiopathic thrombocythemia (IT), chronic myeloid leukemia (CML) and myelofibrosis/osteomyelosclerosis (MF/OMS). Bone marrow biopsies of 3500 patients with known or suspected MPD were studied, and the clinical and morphologic variables registered were utilized for multivariate data analysis by selected BMD computer programs. The histologic criteria and the histologic subdivisions, as well as the evolution and prognosis of disease are outlined for each of the clinical entities. The results show that a bone marrow biopsy provides independent diagnostic and prognostic data in this group of hematologic malignancies and therefore constitutes an additional parameter in the diagnostic investigation of patients with suspected or established MPD.
Misincorporation of uracil into the DNA of folate‐ and B12‐deficient HL60 cells
Authors: Wickramasinghe S. N. , Fida S.
Published: July 2010

Abstract: HL60 cells were cultured for 10 days under various experimental conditions. They were then incubated with 1 μmol/l [5‐3H] uridine for 2 hours and their DNA extracted. The DNA was hydrolysed to deoxyribonucleosides with phosphodiesterase and alkaline phosphatase and the hydrolysate subjected to Aminex A6 chromatography. The elution profiles showed that, when compared with control cells, DNA from cells grown in medium deficient in folate, B12 or both folate and B12 contained increased amounts of deoxyuridine (dU) and increased radioactivity in the dU peak. The data demonstrate that misincorporation of uracil into DNA occurs in a myeloid cell line cultured in growth medium deficient in folate, B12 or both folate and B12.
Interleukin‐4 induces proliferation of adult T‐cell leukemia cells
Authors: Mori Naoki , Yamashita Uki , Tanaka Yoshiya , Nakata Koichi , Oda Susumu , Morimoto Isao , Eto Sumiya
Published: July 2010

Abstract: To evaluate the effect of IL‐4 on the growth of leukemic cells from adult T‐cell leukemia (ATL) patients (ATL cells) and determine whether the IL‐4 autocrine mechanism is involved in the growth of ATL cells, we studied the proliferative response of ATL cells, from 11 patients, cultured in the presence or absence of IL‐4 in vitro. Leukemic cells from 10 of the 11 patients examined proliferated in response to both IL‐2 and IL‐4 in a dose‐dependent manner. The proliferative response to IL‐4 was higher than that obtained with IL‐2 in 8 patients. The expression of the IL‐2 receptor (IL‐2R) αα‐chain in leukemic cells from some patients was also enhanced by IL‐4. The IL‐4 receptor was demonstrated by flow cytometry on the surface of ATL cells. Neither IL‐4‐induced proliferation of ATL cells nor IL‐4‐induced IL‐2R expression on ATL cells was inhibited by anti‐Tac or anti‐IL‐2 antibody and, therefore, these effects of IL‐4 are considered independent of endogenous IL‐2 activity. However, IL‐2 and IL‐4 were undetectable in the culture supernatants of ATL cells from any patient by enzyme‐linked immunosorbent assay. Interferon‐γ (IFN‐γ) partially inhibited IL‐2‐ or IL‐4‐induced proliferation of ATL cells. These results suggest that leukemic cells from ATL patients proliferate by an IL‐2 or IL‐4 paracrine mechanism in lymphoid tissue in vivo and that IFN‐γ inhibits IL‐2‐ or IL‐4‐induced proliferation of ATL cells.
Immunological detection of myeloperoxidase in poorly differentiated acute leukemia
Authors: Lepelley P. , Preudhomme C. , Sartiaux C. , Ghevaert C. , Lai J. L. , Iaru T. , Fenaux P. , Cosson A.
Published: July 2010

Abstract: We performed immunocytochemical detection of myeloperoxidase (MPO), using monoclonal antibody MPO‐7, in 15 consecutive cases of adult acute leukemia (AL) unclassified by conventional cytological and cytochemical criteria and 7 AML‐M, with less than 10% of cytochemically MPO‐positive blasts. In AL with negative MPO cytochemistry the anti‐MPO reaction was positive in 5 of the 15 patients with 3, 3, 7, 11 and 45% positive blasts respectively. In AML‐M1, immunocytochemistry was positive in a larger percentage of blasts than cytochemistry in 2 cases. Immunological detection of myeloid surface markers was positive in all 15 cases of unclassified AL (including the 10 AL with negative anti‐MPO reaction). Eleven of the 22 patients from this study had mixed lymphoid‐myeloid phenotype. Discrepancy between immunological MPO detection and light cytochemistry was more frequent in patients with mixed immunophenotype than in patients without lymphoid markers. No relationship between MPO‐antigen positivity and clinical or biological features was seen. These findings confirm immunological detection of MPO as useful for the diagnosis of poorly differentiated AL. The high incidence of inactive MPO detectable only by immunocytochemistry in mixed lineage AL needs to be confirmed.
A retrospective analysis of bleeding complications in 438 patients with acute leukaemia during the years 1972–1991
Authors: Törnebohm E. , Lockner D. , Paul C.
Published: July 2010

Abstract: The incidence and mortality of bleeding complications have been investigated in 438 patients with acute leukaemia consolidated either by chemotherapy (n = 241) or by bone marrow transplantation (n = 197). Bleeding signs on admission were found in 38% of the chemotherapy‐treated group. Haemorrhagic deaths during the 1st month were seen in 10%. The majority of the major bleedings were localized intracranial, but gastrointestinal haemorrhages were also common. The platelet count was significantly lower (40 × 109/1 versus 69 × 109/1, p < 0.001) and the leukocyte count significantly higher (31.2 × 109/1 versus 11.6 × 109/1, p<0.001)in the group with bleeding complications than in those without. The haemorrhagic mortality in patients consolidated with chemotherapy compared with transplant patients was similar, 23% and 19%. The majority of the lethal haemorrhages in the latter group were observed in patients undergoing allogenic bone marrow transplantation after engraftment. Septicaemia, graft‐versus‐host and venous occlusive disease were contributing factors.
Improved survival and marrow engraftment of mice transplanted with bone marrow of GM‐CSF‐treated donors
Authors: Ballin Ami , Sagi Orit , Schiby Ginette , Meytes Dina
Published: July 2010

Abstract: Recombinant granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) administered to bone marrow (BM) transplant recipients is associated with earlier recovery. We have investigated the possibility of stimulating normal donor mice in vivo with GM‐CSF. Donor balb/c mice were injected i.p. with GM‐CSF (5000 u) or saline. Seventy‐two hours later 5 × 105 BM cells from either GM‐CSF‐treated or control donors were infused into lethally irradiated (850 R) recipients. In the recipients of BM from GM‐CSF‐treated donors, significantly higher CFU‐S and significantly higher survival rate (57% [n = 65]; vs. 30% [n = 63]; p<0.05) were noted. Donor mice of the GM‐CSF group did not differ in bone‐marrow cellularity and composition from their controls. However, recipients of BM from GM‐CSF‐treated mice had higher blood counts of haemoglobin, leukocytes and platelets compared to controls. These data demonstrate that pretreatment of BM donors with GM‐CSF may be of benefit in improving survival and marrow engraftment in mice.
Failure of stem cell factor to ameliorate AZT‐induced anemia in immunodeficient mice
Authors: Hamburger Anne W. , Zsebo Krisztina , Chow Fung‐Phing Rose
Published: July 2010

Abstract: Recent attempts to reduce 3′azido‐3′deoxythymidine (AZT)‐induced anemia in AIDS patients have focused both on AZT dose reduction and on the use of recombinant cytokines. The newly cloned cytokine stem cell factor (SCF) is a potent regulator of hematopoietic progenitor cell proliferation. Therefore, we attempted to ameliorate AZT‐induced anemia using stem cell factor (SCF) in the LP‐BM5 murine leukemia virus‐induced model of AIDS (MAIDS). SCF was administered with oral AZT for up to 1 month, and effects on erythropoiesis examined. SCF alone increased both splenic BFU‐E and CFU‐E. AZT alone also increased the number of splenic BFU‐E and CFU‐E. SCF, administered to AZT‐treated MAIDS mice, did not further enhance these increases. SCF increased bone marrow cellularity in AZT‐treated MAIDS mice. However, the total number of bone marrow BFU‐E was unaffected. In contrast, AZT, SCF, and the combination significantly decreased bone marrow CFU‐E. SCF alone increased the absolute numbers of peripheral blood reticulocytes in MAIDS mice, but did not increase reticulocyte numbers in AZT‐treated mice. SCF did not significantly increase hematocrits in either control or AZT‐treated mice. Further studies are needed to maximize the differentiating capacity of the enlarged erythroid progenitor cell pool induced by SCF.
Detection of platelet antigen for antiplatelet antibodies in idiopathic thrombocytopenic purpura by flow cytometry, antigen‐capture ELISA, and immunoblotting: a comparative study
Authors: Kokawa Terutoshi , Nomura Shosaku , Yanabu Mutsumasa , Yasunaga Kojiro
Published: July 2010

Abstract:  We compared three methods of detecting platelet antigens for antiplatelet antibodies in patients with idiopathic thrombocytopenic purpura (ITP), i.e., a microtiter well antigen‐capture enzyme‐linked immunosorbent assay (AC‐ELISA), a platelet suspension immunofluorescence test using flow cytometry (PSIFT‐FCM), and Western blotting. Using PSIFT‐FCM, the reactivity of NNKY1–32, an anti‐glycoprotein (GP) IIb/IIIa antibody, and of NNKY5‐5 (anti‐GPIb) to platelets from 60 ITP patients were examined. By PSIFT‐FCM, both the peak channel and the relative fluorescence value were below the mean‐2SD for healthy control platelets in 15 patients when NNKY1–32 was used and in 2 patients when NNKY5‐5 was used. Western blotting gave an apparent molecular weight for GPIb of 160000, while GPIIb was 135000 and GPIIIa was 88000. By the AC‐ELISA, 12 patients were positive for NNKY1–32 and 4 for NNKY5‐5. Although NNKY1‐32 binding was detected by PSIFT‐FCM in 15 of the ITP patients using platelets, only 3 were positive using plasma. By AC‐ELISA and Western blotting of plasma, 12 and 10 of the patients were positive for NNKY1–32 and NNKY5‐5, respectively. Our results suggest that none of the three methods is good enough to stand alone and that they should be used together in the analysis of platelet antigens for antiplatelet antibodies in ITP.
9‐nitro‐camptothecin delays growth of U‐937 leukemia tumors in nude mice and is cytotoxic or cytostatic for human myelomonocytic leukemia linesin vitro
Authors: Pantazis P. , Mendoza J. T. , Early J. A. , Kozielski A. J. , Natelson E. A. , Giovanella B. C.
Published: July 2010

Abstract: The camptothecin derivatives 9‐nitro‐camptothecin (9NC) and 9‐amino‐camptothecin (9AC) inhibit similarly growth of HL‐60, KG‐1, and U‐937cells in vitro, whereas growth of THP‐1 cells is inhibited by 9AC, but not by 9NC. Growth inhibition is accompanied by enlargement of cells which contain one (HL‐60, THP‐1) or more (KG‐1, U‐937) nuclei. Flow cytometry studies showed that 9NC‐treated HL‐60 and U‐937 cells accumulate in the S and G2phases of the cell cycle; then they die by apoptosis, with the HL‐60 cells being more sensitive than U‐937 cells to 9NC. In contrast, 9NC‐treated KG‐1 and THP‐1 cells accumulate in S and G2phases, but resist death by apoptosis. Of the cell lines tested, only U‐937 cells xenografted in nude mice generated subcutaneous myeloid tumors, which exhibited a delayed growth in the presence of 9NC. Further, 9NC‐treated advanced U‐937 tumors regressed temporarily, indicating that U‐937 cells consist of 9NC‐sensitive and 9NC‐resistant populations.