bluuebunny/crossref_metadata_2025 huggingface dataset:
The red cell mass, plasma erythropoietin and spleen size in apparent polycythaemia
Authors: Carneskog Jan , Safai‐Kutti Soodabeh , Suurküla Madis , Wadenvik Hans , Bake Björn , Lindstedt Göran , Kutti Jack
Published: July 2010

Abstract: It has been shown previously that measurement of the spleen size and plasma erythropoietin (EPO) concentration are valuable adjuncts in the diagnostic work‐up of patients with polycythaemia vera. The aim of the present work was to evaluate their value in the assessment of apparent polycythaemia (AP). Therefore, over a 24‐month period we routinely performed bone marrow biopsies, measurement of red cell mass (RCM) and plasma volume (PV), spleen size determination by gamma camera scintigraphy and determination of the plasma EPO concentration in consecutive patients referred to us because of elevated values for packed cell volume (>0.48 in females and >0.51 in males). After having excluded patients with clonal and secondary polycythaemias we were left with 38 patients (27 males and 11 females) with AP. In all of them the measured RCM was within normal range, i.e. <36 ml/kg for males and <32 ml/kg for females. The subjects were characterized by moderate increase in RCM and a concomitant moderate decrease in PV. Thus, as an average the measured RCM exceeded the predicted values by 14% in males and by 12% in females; conversely, as compared to the predicted values the average measured value for PV was reduced by 17% in males and by 8% in females. The average RCM for males was 29 ± 3 ml/kg; the corresponding figure for females was 23 ± 4 ml/kg. It was shown that 86% of the subjects had plasma EPO concentrations within the control range; the remaining had values slightly above or below the control range. The mean posterior spleen scan area was 57 ± 16 cm2 and mean left lateral area 57 ± 17 cm2; the reference value for spleen scan area (for both projections) is 57 ± 12 cm2. Of the patients 35/38 (92%) had a spleen scan area within the mean + 2SD for controls and 38 subjects (100%) had values within the mean + 3SD. It is concluded that measurement of plasma EPO and a careful assessment of the spleen size should always be considered in the evaluation of patients with elevated values for venous packed cell volume.
2‐Chlorodeoxyadenosine (cladribine) in the treatment of hairy cell leukemia and hairy cell leukemia variant: 7‐year experience in Poland
Authors: Robak T. , Błasińska‐Morawiec M. , Błasiński J. , Hellmann A. , Hałaburda K. , Konopka L. , Kotlarek‐Haus S. , Potoczek S. , Hansz J. , Dmoszyn̊ska A. , Urasiński I. , Zdziarska B. , Dwilewicz‐Trojaczek J. , Hołowiecki J. , Skotnicki A. B.
Published: July 2010

Abstract: Between January 1991 and December 1997, 103 patients, 97 with typical hairy cell leukemia (HCL) and 6 with HCL‐variant (HCL‐V) were treated with 2‐chlorodeoxyadenosine (2‐CdA) given as 2‐h infusion for 5 consecutive d at a daily dose 0.12 mg/kg. To our knowledge this is the largest cohort of HCL patients treated with this type of regimen. Median follow‐up amounted to 36 months. Fifty‐six of 97 patients with typical HCL were newly diagnosed and 41 were relapsed after previous treatment. Splenectomy as a first‐line therapy was performed in 23 patients and 18 remaining patients received prednisone, chlorambucil or interferon‐α (IFN‐α) alone or in combinations. Seventy‐five (77.3%) patients entered CR and 18 (18.6%) achieved PR, giving an overall response rate of 95.9%. The mean time of first CR duration amounting to 32 months (range 3–72) did not correlate to the number of 2‐CdA cycles. 2‐CdA was equally effective in treatment of newly diagnosed patients and patients who relapsed after previous therapeutic procedures. Relapse of the disease occurred in 20 of 75 patients who achieved CR after 2‐CdA and was usually manifested by very discrete changes in peripheral blood counts (neutropenia and/or relative lymphocytosis). The mean progression‐free survival (PFS) time in this group was 37.4 (range 10–66) months. Ten of 20 relapsed patients were retreated with 2‐CdA given an identical course to the first one. Seven patients entered second CR lasting 19+ (range 8–47) months and 3 experienced PR. This confirms the previous observations that 2‐CdA gives no resistance to leukemic clone. Ten remaining patients have not required retreatment so far and remain in a good clinical and hematological state. The results of HCL‐V treatment with 2‐CdA were poor. Only 2 patients achieved PR and 4 patients did not respond to this drug. Seven patients (5 with typical HCL and 2 with HCL‐V) died, 3 of causes unrelated to the disease. Second neoplasms were noted in 5 patients. 2‐CdA‐related side effects resulted mainly from myelosuppression and infectious complications. In conclusion we confirm the effectiveness of 2‐CdA in inducing CR in patients with typical HCL, but this drug is unable to completely eradicate the leukemic clone which results in the relapse of the disease. The real incidence of the relapse rate may be underestimated unless bone marrow biopsy is performed. The results of our study indicate that a 2‐h infusion of 2‐CdA in HCL patients is at least as effective as a 24‐h infusion but more convenient to the patients, and may be given on an outpatient basis.
Antibodies reactive with neutrophils following allogeneic haematopoietic stem cell transplantation
Authors: Tazzari P. L. , Rondelli D. , Re F. , Bandini G. , Ricci F. , Tassi C. , Bonini A. , Stanzani M. , Laudadio M. A. , Babini L. , Conte R. , Tura S.
Published: July 2010

Abstract: Allogeneic haematopoietic stem cell transplants might induce immunological alterations leading to autoimmune‐like syndromes. In particular neutrophil‐associated antigens could represent the target for autoantibodies against neutrophils in patients receiving an allogeneic peripheral stem cell or bone marrow transplantation, giving rise to granulocytopenia. With this aim we studied prospectively 43 allotransplanted patients for the presence of antibodies reacting with neutrophils (ARN), looking for a correlation with a post‐engraftment neutropenia. Our data showed that the direct test for ARN was positive in 30 patients. Interestingly, 7/7 patients who received a T‐cell‐depleted marrow transplant developed ARN. Antibodies with a specific neutrophil‐antigen reactivity were detected in 4 patients, 1 with an anti‐CD16/FcγRIIIb receptor reactivity and 3 with anti‐NA1 reacting patterns, respectively. From a clinical point of view, it was not possible to demonstrate a close and significant relationship between neutropenia and ARN, although patients showing ARN had slightly lower absolute levels of peripheral neutrophils until 6 months after BMT. In conclusion, ARN may be detected in the majority of patients following allogeneic stem cell transplantation; in addition, since ex vivo or in vivo T‐cell‐depletion leads to a higher percentage of patients positive for ARN, it could be hypothesized that “autoimmune‐like” disorders in transplanted patients might be related to a T‐cell derangement due to different numbers and subsets of T lymphocytes.
A staging system for multiple myeloma based on the morphology of myeloma cells
Authors: Murakami Hirokazu , Kawada Tomoyuki , Saitoh Takayuki , Uchiumi Hideki , Moridaira Kazuaki , Matsushima Takafumi , Tsukamoto Norifumi , Tamura Jun'ichi , Morita Kimio , Sawamura Morio , Karasawa Masamitsu , Miyawaki Shuichi , Shinonome Shogo , Shimano Shunichi , Sato Sadao , Ogawara Hatsue , Tsuchiya Jun
Published: July 2010

Abstract: The morphology of myeloma cells is reported to be one of the prognostic factors in multiple myeloma (MM) patients. We analyzed the prognostic factors, including morphological classification, in 292 patients with MM in order to select poor‐risk patients who should be considered candidates for early intensive chemotherapy, including stem cell transplantation. Multivariate analysis was applied to 90 patients diagnosed between 1989 and 1996, because serum beta‐2‐microglobulin (β2M) has been measured regularly since 1989, and showed that serum albumin, serum β2M, and the morphology of myeloma cells predicted survival. According to these factors, patients were divided into 3 risk groups; a high‐risk group (14%), a intermediate‐risk group (46%) and a low‐risk group (40%). There were significant differences between survival times in these 3 groups (median survival: high‐risk, 16; intermediate‐risk, 22; and low‐risk, 44 months).
Effect of recombinant human erythropoietin in preterm infants
Authors: Krallis Nikolaos , Cholevas Vasilis , Mavridis Anestis , Georgiou Ioannis , Bourantas Konstantinos , Andronikou Styliani
Published: July 2010

Twenty‐five premature infants (mean gestational age±SD, 31.4±1.9 weeks) were administered subcutaneously recombinant human erythropoietin (rHuEpo) at a dose of 300 u/kg of body weight three times a week beginning on the third day of life and continuing for 6 weeks. The controls (n=23) were premature infants with a mean gestational age of 32.2±2.3 weeks who did not receive rHuEpo. Haematological indices, haemoglobin and serum phosphate (Pi), and red blood cell (RBC) phosphate metabolites (ATP, 2,3‐DPG, RBCPi) were tested monthly until the 6th month and thereafter at the 9th and 12th months of life. The level of serum soluble transferrin receptors (sTfR) correlated significantly with rHuEpo (p<0.05). The ratio of sTfR to log (ferritin) was significantly higher (p<0.001) in the infants treated with rHuEpo than the controls. Intracellular organic and inorganic Pi changes were not affected by the Epo administration. The RBC 2,3‐DPG seemed adequate in infants receiving rHuEpo.
Wilms' tumour gene (wt1) expression at diagnosis has no prognostic relevance in childhood acute lymphoblastic leukaemia treated by an intensive chemotherapy protocol
Authors: Gaiger A. , Linnerth B. , Mann G. , Schmid D. , Heinze G. , Tisljar K. , Haas O. A. , Gadner H. , Lion T.
Published: July 2010

Abstract: Expression of the Wilms' tumour gene (wt1) has been demonstrated in a large proportion of human acute leukaemias and is thought to play a role in leukaemogenesis. Recent observations in adult patients with acute leukaemia suggest that wt1 gene expression is a poor prognostic factor. In childhood acute leukaemia, the clinical role of wt1 gene expression has not been established. We have therefore investigated bone marrow samples from 50 children with acute lymphocytic leukaemia at the time of diagnosis for the presence of wt1 transcripts to determine whether wt1 gene expression is associated with specific characteristics of leukaemic cells and whether it is predictive of response to treatment. All patients were treated according to the ALL‐BFM 90 protocol. The median observation time was 30 months. Wt1 transcripts were detected by RT‐PCR in 60% of the diagnostic samples. Wt1 PCR positive patients showed a higher median leukocyte and peripheral blast cell count than wt1 negative patients. High and intermediate risk patients more frequently displayed wt1 transcripts than low risk patients. No correlation between wt1 gene expression and FAB type, immunophenotype, co‐expression of myeloid antigens or karyotype has been observed. Furthermore, there was no correlation between wt1 gene expression at diagnosis and achievement of complete remission (CR) and no difference in disease‐free survival (DFS) or overall survival (OS) between wt1 positive and negative patients (p >0.1). These data indicate that (1) wt1 gene expression at diagnosis is detected more frequently in patients with high leukocyte and peripheral blast cell counts, but is not associated with specific characteristics of leukaemic cells, (2) wt1 gene expression is not an independent prognostic factor for CR, DFS or OS in childhood ALL treated by an intensive therapy protocol.
Induction of cytosolic phospholipase A2 and prostaglandin H2 synthase‐2 by ipopolysaccharide in human polymorphonuclear leukocytes
Authors: Zaitsu Masafumi , Hamasaki Yuhei , Matsuo Muneaki , Miyazaki Michiko , Hayasaki Rika , Muro Eriko , Yamamoto Shuichi , Kobayashi Ikuko , Ichimaru Tomohiro , Miyazaki Sumio
Published: July 2010

Abstract: Polymorphonuclear leukocytes (PMNs) produce arachidonic acid (AA) metabolites including thromboxane A2 (TXA2). These cells are the first line of defense against bacterial invasion, which often causes endotoxin shock. TXA2 which plays an important role in the pathogenesis of endotoxin shock is synthesized by three consecutive enzyme activation, cytosolic phospholipase A2 (cPLA2), prostaglandin H2 synthase (PHS type 1 and type 2) and TXA2 synthase. Among them, cPLA2‐ and PHS‐2 activity is known to be transcriptionally and/or post‐transcriptionally up‐regulated by various bioactive substances including lipopolysaccharide (LPS), a bacterial endotoxin, in many cell types. We investigated the action of LPS on TXA2 synthesis in human PMNs. A23187‐stimulated production of thromboxane B2 (TXB2, a stable metabolite of TXA2), assayed by specific radioimmunoassay (RIA), was significantly increased from 566.7±44.1 pg/106 cells to 966.7±44.1 pg/106 cells (p <0.05) after 6 h‐exposure to LPS at the concentration of 100 ng/ml. Messenger RNA for PHS‐2, PHS‐1, TXA2 synthase and cPLA2, which was assessed by reverse transcription‐polymerase chain reaction (RT‐PCR), was expressed in PMNs without LPS stimulation. Although PHS‐2 was putatively an inducible enzyme, abundance of mRNA for PHS‐2 in PMNs without LPS stimulation was detectable. Messenger RNA abundance for PHS‐2 and cPLA2, but not for PHS‐1 and TXA2 synthase, was enhanced by LPS‐treatment, indicating that the increased production of TXB2 was attributable to the up‐regulation of cPLA2 and PHS‐2. We conclude that (1) PHS‐2 plays a more important role than PHS‐1 in the production of TXA2 in human PMNs and (2) TXA2 synthesis in human PMNs is transcriptionally up‐regulated by new induction of cPLA2 as well as PHS‐2, when the cells encounter endotoxin producing bacteria.
Low‐dose oral iron absorption test in anaemic patients with and without iron deficiency determined by bone marrow iron content
Authors: Jensen Nanna M. , Brandsborg Margrethe , Boesen Anne M. , Yde Hans , Dahlerup Jens F.
Published: July 2010

Abstract: The low‐dose oral iron absorption test (OIAT) was performed in 85 consecutive anaemic patients referred for bone marrow examination in order to investigate the ability of the test to predict bone marrow iron stores and to differentiate between different categories of anaemia. Eight patients were excluded for technical reasons. Test results from 77 patients are presented as Cmax (μmol/l): the maximum increase in S‐iron measured during a 3 h period after administration of 10 mg oral iron sulfate. Iron deficiency was defined as the absence of stainable iron in bone marrow aspirates. Cmax was higher in 46 iron deficient patients [3 (median); 0 and 13 (1st and 3rd quartiles); 0–40 (range)] than in 31 non‐iron‐deficient patients (0; 0 and 2; 0–4) (p <0.01). 27 patients had primary bone marrow disease, 25 patients had absent bone marrow iron stores accompanied by inflammation, 17 patients had anaemia of chronic disease (ACD) and 8 patients had uncomplicated iron deficiency anaemia (IDA). Patients with IDA had higher Cmax (15; 13 and 28; 6–40) than patients with ACD (1; 0 and 2; 0–3), and than the 69 non‐IDA patients (1; 0 and 3; 0–19) (p<0.001). Cmax values above 5 μmol/l always indicated absent bone marrow iron stores.
Induction of differentiation in U‐937 and NB4 cells is associated with inhibition of tissue factor production
Authors: Tenno Taavo , Öberg Fredrik , Nilsson Kenneth , Siegbahn Agneta
Published: July 2010

Tissue factor (TF) production is under strict control in mature monocytic cells. However, constitutive expression of TF can be found in myelomonocytic cells and in haematopoietic cells arrested at an early stage of differentiation. In this paper we show that TF expression is down‐regulated during the monocyte/granulocyte differentiation process, using the human monoblastic U‐937 and the acute promyelocytic leukaemia NB4 cell lines as models. Expression of TF mRNA, protein and procoagulant activity (PCA) was constitutively high in untreated cells. Exposure of U‐937 cells to 1 α,25‐dihydroxycholecalciferol (VitD3) and all‐trans retinoic acid (ATRA) resulted in down‐regulation of TF expression and PCA. In NB4 cells induction by ATRA, but not VitD3, resulted in the down‐regulation of TF expression and PCA. Consistent with this, induction of terminal differentiation, as confirmed by the expression of differentiation associated antigens and cell cycle arrest, was inversely correlated to TF expression in U‐937 and NB4 cells. Moreover, terminally differentiated U‐937 cells retained the capacity to respond to inflammatory mediators, i.e. lipopolysaccharide and interferon‐γ, by a rapid increase in TF expression. In conclusion, we show that not only ATRA but also VitD3 is a potent suppressor of monocytic TF expression and thus might have potential clinical use for the treatment of coagulopathies.
High incidence of maternal HLA A, B and C antibodies associated with a mild course of haemolytic disease of the newborn
Authors: Neppert J. , Witzleben‐Schürholz E. v. , Zupanska B. , Bartz L. , Greve O. , Eichler H. , Kerowgan M. , Wichmann M. G.
Published: July 2010

Abstract: Mild courses of haemolytic disease of the foetus or newborn (HDN) due to Rh (D) blood group antibodies are associated with and may therefore be ameliorated by maternal antibodies reacting with human leucocyte antigens (HLA) of the child, an observation drawn from our own earlier data (Neppert J, Kissel K. Lancet 1992;339:1481). This study (i) corroborates this association; (ii) reveals shortcomings in the published data; and (iii) examines the characteristics of HDN cases when these shortcomings have been rectified. Samples from 51 women with antibodies against their child's blood group antigens of the Rh system were analysed for HLA A, B, C and DR antibodies during parturition. The mothers were divided into two groups, either severe or mild, dependent upon the clinical course of the HDN, and the incidence of HLA antibody production was determined for each group. HLA A, B, C and/or DR antibodies were detected in 85.2% of those women whose children had a mild course of HDN prenatally or perinatally (n=27). This is statistically greater than the incidence of 50.0% (Fisher's exact test: p=0.014) found in the group of women whose children had a severe HDN either pre‐ or perinatally (n=24) and is greater than the 35% (n=20; p=0.0001) found in women without Rh or other irregular antibodies. HLA DR antibodies were detected in three cases. The findings support our hypothesis that maternal anti HLA A, B and C antibodies may protect against a potential severe HDN. We therefore assume that those women will benefit who have already had a child with a severe HDN and in whom HLA antibodies were not previously detected, if HLA antibody production is provoked by subcutaneously inoculating with the father's leucocytes before or at the beginning of the new pregnancy.
Long‐term results of 60 patients with pathologic stage I & IIA Hodgkin's disease treated with exclusive mantle radiation therapy
Authors: Enrici Riccardo Maurizi , Osti Mattia Falchetto , Zurlo Alfredo , Anselmo Anna Paola , Iacari Valter , Mandelli Franco
Published: July 2010

Abstract:Between January 1972 and December 1982 60 patients with pathological stage IA and IIA Hodgkin's disease (HD) were submitted to Mantle irradiation only. Twenty‐five were in stage I (32.1%) and 35 in stage II (67.9%). All patients were submitted to staging laparotomy. Cases with large mediastinal mass were excluded from this series. Delivered doses were 44 Gy in involved areas, 40 Gy on the mediastinum and 36 Gy on uninvolved sites. Twenty‐four patients in stage I (96%) and 33 in stage II (94.2%) obtained complete remission. Actuarial 10‐ and 20‐yr overall (OS) rates were 86% and 79.1%, respectively. Event‐free (EFS) and relapse‐free (RFS) survival rates at 10 and 20 yr were 67.5% and 62.1%, respectively. The occurrence of disease relapse resulted in the only statistical significant prognostic factor for OS in both univariate and multivariate analysis. Distant and extranodal recurrences were significantly (P<0.01) related to a reduced OS. On multivariate analysis stage was the only determinant factor for increased RFS. Extended field RT proved to be an effective curative modality for stage I HD patients, whereas 15 out of 33 patients in stage II relapsed requiring salvage therapy. Long‐term analysis of survival and treatment‐related morbidity rates will improve our knowledge and assist the physicians to choose the therapeutic option to offer to HD patients.
Arterial elastorrhexis: manifestation of a generalized elastic tissue disorder in β‐thalassaemia major
Authors: Tsomi K. , Karagiorga‐Lagana M. , Fragodimitri C. , Karabatsos F. , Katsiki V.
Published: July 2010

Abstract: Pseudoxanthoma elasticum‐like lesions of the eye and skin are frequently observed in β‐thalassaemia, and there has been speculation about associated vascular lesions. This led to our study of histopathological material from thalassaemic patients.

Histological re‐examination was made of a series of 45 spleens and 45 surgical liver biopsies from 45 patients with β‐thalassaemia major, aged 6–25 yr and treated over the 20‐yr period 1975–95. Correlations between clinical, laboratory and histological findings were demonstrated by statistical analysis.

Arteriopathy characterized by fragmentation and multiple defects of the internal elastic lamina (‘arterial elastorrhexis’), with deposits of iron and calcium salts, was found in the hilar arteries of the spleen with a frequency of 96%. Similar lesions were observed in the parenchymal arteries and the stromal elastic tissue (‘stromal elastorrhexis’) of the spleen, liver and lymph nodes. Arterial and elastic tissue alterations appear in the first decade of life and become generalized over the course of the disease, independent of the time of onset of transfusion and iron chelation therapy.

Arterial elastorrhexis is the earliest and most frequent manifestation of a systemic elastic tissue disorder in β‐thalassaemia major. It appears to be an acquired pseudoxanthoma elasticum‐like syndrome, related primarily to tissue hypoxia and disturbance of elastin metabolism.
Pathophysiological significance of simultaneous measurement of reticulated platelets, large platelets and serum thrombopoietin in non‐neoplastic thrombocytopenic disorders
Authors: Koh Ki‐Ryang , Yamane Takahisa , Ohta Kensuke , Hino Masayuki , Takubo Takayuki , Tatsumi Noriyuki
Published: July 2010

Abstract: An automated reticulocyte counter using flowcytometric analysis, the R‐3000 (Sysmex Inc. Kobe, Japan), has recently been modified to determine reticulated platelets (RPs) and large platelets (LPs). We measured frequencies of RPs, LPs in total platelet count and serum thrombopoietin concentration comprehensively in non‐neoplastic thrombocytopenic patients with immune thrombocytopenic purpura (ITP, n = 23), aplastic anemia (AA, n = 21), liver cirrhosis (LC, n = 17), and hematologically normal subjects (control, n = 151). ITP was characterized as high frequencies of both RP and LP, AA as high RP frequency and elevated thrombopoietin concentration, and LC as no difference compared with control. Interestingly, the frequency of RP appeared to depend on total platelet count rather than the cause of thrombocytopenia, while the frequency of LP appeared to depend much less on total platelet count. Furthermore, significant positive correlations were observed between frequencies of RP and LP in control, ITP and LC subjects, in whom bone marrow stem cells are intrinsically normal. However, there was no such correlation in AA patients with stem cell deficiency, suggesting that this correlation might be a useful new parameter for detecting qualitatively abnormal platelets. Measurement of RP and LP is thus useful for elucidating the pathophysiology of thrombocytopenic disorders.
Erythropoietin induces the expansion of c‐kit+ progenitors for myeloid and erythroid cells, but not for lymphoid cells, in the bone marrow and liver
Authors: Koya Toshiyuki , Narita Junichi , Honda Shigeru , Watanabe Hisami , Arakawa Masaaki , Abo Toru
Published: July 2010

Abstract: In humans, the numbers of erythrocytes and granulocytes, but not that of lymphocytes, tend to increase in parallel. To determine the mechanism, we investigated how the administration of erythropoietin induces the expansion of erythroid cells and other lineage cells in the bone marrow, liver, and other organs of mice. When mice were injected twice (days 1 and 2) with erythropoietin at a dose of 20 or 200 IU/day/mouse, a prominent expansion of TER119+ (erythroid cells) and Gr‐1high cells (granulocytes) occurred in the liver, spleen, and bone marrow day 3 after the initial injection. On the other hand, lymphoid cells, including NK cells, extrathymic T cells, and conventional T cells, did not expand. In parallel with the expansion of erythroid cells and granulocytes, the levels of c‐kit+Lin− cells increased in the liver and bone marrow. Despite the increase in the proportion of c‐kit +Lin− cells, the generation of lymphocytes (e.g., T cells) decreased when such bone marrow cells were injected to scid mice. These results suggest that erythropoietin has the ability to induce the expansion of not only erythroid cells but also granulocytes in the liver, spleen, and bone marrow. Furthermore, c‐kit+ progenitors which may commit themselves to erythroid and myeloid cells, but not to lymphoid cells, were also activated in the liver and bone marrow of mice treated with erythropoietin.
Ectopic haematopoiesis in the mouse: roles of stroma and cytokines in granulocytopoiesis in in vivo diffusion chamber cultures
Authors: Wang Xiuli , Strøm‐Gundersen Inger , Fjerdingstad Hege , Benestad Haakon B.
Published: July 2010

Abstract: We have examined a possible role of two different types of irradiated stromal cells, i.e. murine bone marrow (BM) stromal cells and stromal cell line MS‐5R, when cocultured with murine blood‐borne progenitors or sorted Lin −Sca‐1+ bone marrow cells in vivo in peritoneal diffusion chambers (DC). Retrieval and quantification of the cultured cells were performed after 4, 7, and 14 d. Granulocyte and/or macrophage colony‐forming cells (G/M‐CFC) were enumerated in subcultures from the DC. G/M‐CFC production was not enhanced in the stroma‐contact cultures, in comparison with the standard stroma‐non‐contact cultures, but early granulocytopoiesis was stimulated. Perturbation of the humoral environment of DC was investigated in a number of ways, for example with continuous infusion of rhG‐CSF from a subcutaneous implanted minipump to DC host mice, with DC host mice carrying a transplantable leukaemia, secreting interleukin 3 (IL‐3), and with injections of various cytokines. None of these interventions sustained the expansion of the G/M‐CFC population. In conclusion, for ectopic haematopoiesis to take place, several requirements must be met. Relevant stromal cells apparently affect haematopoiesis both via direct cell‐cell interactions and via humoral mediators (viz. cytokines) which they secrete.
Combination of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and erythropoietin (EPO) for the treatment of advanced non‐responsive chronic lymphocytic leukemia
Authors: Russo Filippo , Guadagni Stefano , Mattera Gioacchino , Esposito Giuseppe , Abate Giuseppe
Published: July 2010

Abstract: We report the use of a colony‐stimulating granulocyte‐macrophage factor (GM/CSF) and erythropoietin (EPO) combination as salvage treatment in four heavily‐pretreated patients with refractory/recurrent B‐CLL. Induction therapy was subcutaneous GM‐CSF 2.5 μg/kg, and EPO, 150 units/kg both daily for the first 14 d. Maintenance therapy was GM‐CSF on days 1, 3 and 5 and Epo on days 2, 4 and 6 at the same doses with weekly recycling. All the patients responded favourably. A significant reduction of lymphocytosis, lymphoadenomegaly, and organomegaly was obtained within one month of therapy. The number of infections and transfusional requirement decreased dramatically. The hemoglobin increased to over 11 g/dl in 3 out of 4 patients. With a median follow‐up of 11 months (range 5–13) we observed 4 partial responses (NCI/IWCLL) and only one progression after a 10‐month partial response.

This combination regimen seems very active, safe and easy to administer. It may represent a promising therapeutical option in heavily pretreated patients. Further clinical and biological studies on a larger cohort of patients are needed to confirm these preliminary data, to clarify the hypothetical interactions between these cytokines and B‐CLL cell proliferation pathways, and to establish if this therapy may have an impact on survival.
Incidence of nucleoli in erythroblasts in patients suffering from refractory anemia of myelodysplastic syndrome
Authors: Smetana Karel , Jirásková Nona , Cermák Jaroslav
Published: July 2010

Abstract: Nucleoli of erythroblasts have been studied in patients suffering from refractory anemia (RA) of myelodysplastic syndrome (MDS) and in control patients without a disturbed erythropoiesis in order to provide information on the incidence of nucleoli and micronucleoli in these cells. Nucleoli in erythroblasts were visualized by a simple cytochemical procedure for the demonstration of RNA which facilitated the visualization not only large nucleoli but also micronucleoli in advanced stages of the erythroblastic maturation. In control patients nucleoli were detected in all stages of erythroblastic development. In patients suffering from RA of MDS, a relatively large population of polychromatic and orthochromatic erythroblasts was characterized by a loss of nucleoli accompanied by the decreased incidence of micronucleoli characteristic of these cells. In contrast to control patients, in patients suffering from RA of MDS the number of nucleoli expressed by the values of the nucleolar coefficient of erythroblasts was smaller, particularly in both the early and terminal stages of erythroblastic development. Thus in patients with RA of MDS both the abnormal loss of nucleoli and decreased number of nucleoli in erythroblasts apparently represent and reflect a further abnormality of disturbed erythropoiesis.
Response and adverse drug reactions to combination chemotherapy in elderly patients with aggressive non‐Hodgkin's ymphoma: comparison of CHOP, COP‐BLAM, COP‐BLAM III, and THP‐COPBLM
Authors: Niitsu Nozomi , Umeda Masanori
Published: July 2010

Abstract: We retrospectively compared therapeutic results and adverse events in 198 elderly patients (≥70 yr old) with aggressive non‐Hodgkin's lymphoma diagnosed between 1981 and 1995 who underwent CHOP, COP‐BLAM, COP‐BLAM III, or THP‐COPBLM chemotherapy. Complete remission (CR) was achieved in 138 patients (69.7%). The CR rate was 47.0% for CHOP, 76.3% for COP‐BLAM, 67.9% for COP‐BLAM III, and 74.4% for THP‐COPBLM therapy (p = 0.013). The 5‐yr survival rate was 37.0% for CHOP, 49.0% for COP‐BLAM, and 53.5% for COP‐BLAM III. The event‐free survival rate showed no significant differences between the four treatments. Adverse events of Grade 3 or worse were commonly anemia or granulocytopenia in patients receiving THP‐COPBLM therapy. Cardiac sympathetic dysfunction and cardiac mitochondrial damage were less common with pirarubicin than with doxorubicin.

For elderly patients, it is better to select therapy with as few adverse events as possible based on the complications and medical history of the individual patients.
The tumor necrosis factor family and correlation with disease activity and response to treatment in hairy cell leukemia
Authors: Barak Vivian , Nisman Benjamin , Polliack Aaron
Published: July 2010

Abstract:  Hairy cell leukemia (HCL) is a well recognized indolent B‐cell lymphoproliferative disorder. HCL cell proliferation is regulated by growth factors and cytokines, of which tumor necrosis factor‐α (TNF‐α) may be one of the most important. The mechanism of TNF‐α‐induced HCL cell growth is mediated via 2 receptors, which are present in both cellular and soluble forms. In this study we determined the serum levels of TNF‐α and their soluble receptors — sTNF‐R60 and sTNF‐R80 — in 23 HCL patients and correlated them with clinical parameters before and after therapy. Patients were classified according to their clinical status as either “active” at diagnosis or during relapse and “non‐active” (responding to therapy with partial and complete remission). Most patients were treated with 2‐CDA, following which serum levels of TNF‐α, sTNF‐R60 and sTNF‐R80 were significantly decreased, particularly in CR. Significant differences in paired observation values were noted for TNF‐α and sTNF‐R80, indicating a good correlation with the clinical status of disease, but this was not the case for sTNF‐R60 (coefficients of correlation between levels of TNF‐α and sTNF‐R80 and of TNF‐α and sTNF‐R60 were r = 0.85 and r = 0.64, respectively). These results suggest that decreases in both TNF‐α and sTNF‐R80 are indicative of response to treatment, while increased levels accompany active disease. Accordingly, we conclude that serum levels of the TNF family, as for the sIL‐2R and IL‐1 family, may also be used as sensitive markers for monitoring HCL status. Levels may be indicative of the clinical efficacy of therapy and can be used as an indicator of the presence of residual disease.
IDEC‐C2B8 anti‐CD20 (Rituximab) immunotherapy in patients with low‐grade non‐Hodgkin's lymphoma and lymphoproliferative disorders: evaluation of response on 48 patients
Authors: Nguyen D.T. , Amess J. A. , Doughty H. , Hendry L. , Diamond L.W.
Published: July 2010

Abstract:  This study focused on the efficacy of IDEC‐C2B8 (chimeric anti‐CD20) immunotherapy relative to specific subtypes of low‐grade lymphoproliferative disorders/non‐Hodgkin's lymphomas (LPD/NHL). Forty‐eight patients with resistant or relapsed disease completed the IDEC‐C2B8 infusion schedule of 375 mg/m2/wk × 4 wk. The LPD/NHL subtypes included: (a) follicular centre cell lymphoma (FCC) in 22 patients; (b) mantle cell lymphoma (MCL) in 10; (c) 1 diffuse large cell lymphoma (DLCL); and (d) the category of small lymphocytic lymphoma/chronic lymphocytic leukaemia (SLL/CLL) and related disorders in 15 patients. No patient obtained a complete remission. Ten patients (21%) achieved partial remission: 6 FCC, 2 MCL, 1 DLCL and 1 patient from the SLL/CLL group. Twenty‐eight patients had stable disease and 10 progressed during immunotherapy. In patients with CLL and MCL in leukaemic phase, there was no correlation between the marked decrease in circulating neoplastic cells following antibody infusions and amelioration of the tumour burden. The results suggest that the subtype of LPD/NHL and the intensity of CD20 on the tumour cells influence the effectiveness of IDEC‐C2B8. The antibody was most efficacious against FCC lymphoma. The efficacy (at the dose schedule of 375 mg/m2/wk × 4) against MCL and SLL/CLL appeared to be limited, however.
A single‐center population‐based consecutive series of 1500 cytogenetically investigated adult hematological malignancies: karyotypic features in relation to morphology, age and gender
Authors: Mauritzson N. , Johansson B. , Albin M. , Billström R. , Ahlgren T. , Mikoczy Z. , Nilsson P. G. , Hagmar L. , Mitelman F.
Published: July 2010

Abstract:  During the 18‐yr period 1976–93, a population‐based series of 1586 adults with suspected or confirmed hematological malignancies were successfully cytogenetically investigated at a single center. Eighty‐six cases were excluded due to unretrievable medical records or if analyzed only in remission or at relapse. The remaining 1500 medical records were reviewed regarding morphology and clinical parameters in order to investigate possible associations between karyotypic pattern (normal, 1, 2 or complex anomalies; specific abnormalities) and gender, age and morphological subgroups. The impact of time‐period, i.e. 1976–87 vs. 1988–93, and referring center on cytogenetic findings was also studied. A total of 372 acute myeloid leukemias (AML), 389 myelodysplastic syndromes (MDS), 64 acute lymphoblastic leukemias (ALL) and 262 chronic myeloid leukemias (CML) were identified, altogether 1087 cases. Patients with other (n = 261) or no hematological malignancies (n = 152) were excluded from the present analysis. Cytogenetic abnormalities were detected in 52% AML, 51% MDS, 68% ALL and 97% CML, frequencies that did not differ significantly between the 2 time periods or referring centers. No significant age‐ or gender‐related differences in karyotypic patterns were discerned in AML, MDS, ALL or CML, whereas the karyotypic patterns varied among the FAB groups in both AML (p = 0.001) and MDS (p< 0.001). The specific abnormalities t(8;21), t(15;17) and inv(16) were more common (p<0.001) in younger AML patients and 5q‐ was more frequent in females with MDS (p< 0.001). These findings indicate, in contrast to previous series, that neoplasia‐associated karyotypic aberrations are not more common among older patients or in males.
Flt‐3 ligand does not induce the growth of peripheral blood granulocyte‐macrophage colony‐forming cells in myeloproliferative disorders
Authors: Siitonen T. , Savolainen E.‐R. , Koistinen P.
Published: July 2010

Abstract:  Flt‐3 ligand (FL) is a growth factor (GF) which might have clinical use as a mobilizer of stem and progenitor cells into peripheral blood (PB) in autologous transplantations of various malignant haematological diseases, unless FL stimulates the growth of malignant cells in these diseases. The present study evaluated the effects of FL on the proliferation of granulocyte‐macrophage (GM) progenitor cells collected from PB of 24 patients with chronic myeloproliferative disorders (MPDs) by using a methylcellulose assay in serum‐free culture conditions. It was shown that FL as a single factor had no stimulatory effect on GM colony formation either in the whole MPD group or in the MPD subgroups, which comprised 9 patients with essential thrombocythaemia, 7 with polycythaemia vera and 8 with chronic myelogenous leukaemia. No increase in GM colony formation was observed, either, when FL was used in combination with other GFs, such as mast cell growth factor (MGF), granulocyte‐colony stimulating factor (G‐CSF), GM‐CSF or interleukin‐3 (IL‐3). GM‐CSF and IL‐3 were the only single GFs which significantly increased GM colony formation in the whole MPD group. As a conclusion, FL does not seem to induce GM colony formation of MPDs alone or in combination with G‐CSF in in vitro colony assays.
Response level and survival after conventional chemotherapy for multiple myeloma: a Finnish Leukaemia Group study
Authors: Oivanen Tiina M. , Kellokumpu‐Lehtinen Pirkko , Koivisto Anna‐Maija , Koivunen Elli , Palva Ilmari ,   Finnish Leukaemia Group
Published: July 2010

Abstract:  The relation between overall survival time (OS) and response level (CR, 75% R, PR, MR, SD) was analysed in 432 multiple myeloma patients from 4 prospective Finnish Leukaemia group trials, treated with conventional chemotherapy. The primary regimen was either melphalan and prednisolone or combination chemotherapy with melphalan as a main component. Both the influence of the pre‐treatment factors in patients with different levels of responses and the aspects related to chemotherapy were analysed. The 324 patients aged up to 70 yr and the 108 older patients were dealt with as separate groups. Irrespective of the primary chemotherapy regimen, the level of response was not significantly influencing in the OS time, and this was true in both age categories. The median OS of the patients up to 70 yr of age who had any response was 57 months, compared to the 10 months of those with PD (p <0.001). The corresponding figures for the older patients were 40 and 4 months (p <0.001), respectively. The pretreatment prognosticators for the patients with minimal responses were not more favourable than for patients with responses at higher levels. The prolonged primary chemotherapy was favourable in patients having responses less than PR. Accordingly, the primary goal of conventional chemotherapy for multiple myeloma is stabilization of disease, not the level of response.
Clinico‐prognostic implications of simultaneous increased serum levels of soluble CD23 and β2‐microglobulin in B‐cell chronic lymphocytic leukemia
Authors: Molica Stefano , Levato Domenico , Cascavilla Nicola , Levato Luciano , Musto Pellegrino
Published: July 2010

Abstract: Soluble CD23 (sCD23) and beta‐2 microglobulin (β2‐m) are reliable prognostic parameters in B‐cell chronic lymphocytic leukemia (CLL); however, their merit over well‐established clinical variables such as clinical stages, bone marrow (BM) histology and lymphocyte doubling time (LDT) remains to be defined. Furthermore, information dealing with the impact on overall survival of the simultaneous increase of either β2‐m or sCD23 are lacking. In this prospective study based on 106 B‐cell CLL patients, we propose a combination of β2‐m and sCD23 as a strong prognostic system whose statistical significance was mainly due to an excess of deaths in the subgroup displaying increased serum levels of either β2‐m or sCD23. Multivariate survival analysis confirmed the important dominant role of such a finding, thus excluding features with a high degree of codependence (i.e. clinical stages, LDT) and including variables with low association (i.e. BM histology) in the final regression model. The presence of increased serum levels of β2‐m/sCD23 and diffuse BM histology signified high‐risk disease, whereas the absence of any adverse variable was associated with prolonged survival; in between there was a subgroup with only 1 characteristic which displayed an intermediate pattern of survival. Finally, on the basis combined increased serum levels of β2‐m and sCD23, a better stratification of low‐ and intermediate‐risk patients could be obtained, thus allowing the formulation of a clinico‐biological staging for CLL.
Elevated serum concentrations of hepatocyte growth factor in acute myelocytic leukaemia
Authors: Hjorth‐Hansen H. , Seidel C. , Lamvik J. , Börset M. , Sundan A. , Wagge A.
Published: July 2010

Abstract:  Serum concentrations of hepatocyte growth factor (HGF) were measured in 60 patients suffering from acute myelocytic leukaemia (AML). At the time of diagnosis elevated HGF concentrations (>1.25 ng/ml) were found in 28% of the patients. HGF levels correlated with the presence of disseminated intravascular coagulation (DIC), levels of lysozyme, creatinine, peripheral blood blast counts and lactic dehydrogenase. In the group of patients with high HGF (>1.25 ng/ml) we found a tendency towards an increased early mortality; 41 % of them died within 15 d from diagnosis, as opposed to 5% of the patients with normal HGF (log rank test p = 0.07). DIC‐related bleeding or thrombosis contributed to this early mortality. In responders, HGF levels normalized after treatment. HGF levels are low in neutropenia and neutropenic infections.
In vitro treatment with retinoids decreases bcl‐2 protein expression and enhances dexamethasone‐induced cytotoxicity and apoptosis in multiple myeloma cells
Authors: Tosi Patrizia , Pellacani Annalisa , Visani Giuseppe , Ottaviani Emanuela , Ronconi Sonia , Zamagni Elena , Benni Monica , Cavo Michele , Tura Sante
Published: July 2010

Abstract:  All‐trans retinoic acid (ATRA) has been shown to inhibit in vitro growth of multiple myeloma (MM) cells, and this effect can be further potentiated by the addition of Dexamethasone (DEX). We here extended this study by testing the activity of 9‐cis retinoic acid (9‐cis RA) and 13‐cis retinoic acid (13‐cis RA), both alone and in combination with DEX, in two MM cell lines, U266 and RPMI 8226. Furthermore, we aimed at investigating the mechanisms involved in the interactions of retinoids and DEX in this setting. 9‐cis RA appeared to be the most active agent in U266 cell line (IC50 = 1.2 μmol/l vs 10.5 and 9.8 μmol/l obtained with ATRA and 13‐cis RA, respectively) while, in RPMI 8226 cell line, 9‐cis RA and 13‐cis RA were almost equally cytotoxic (IC50 = 1 and 0.8 μmol/l) and ATRA was less effective. Co‐incubation with DEX resulted in a synergistic cytotoxic activity in both the cell lines except for the combinations DEX + 9‐cis RA in U266 cell line and DEX + 13‐cis RA in RPMI 8226 cell line, where the effect was merely additive. A synergistic cytotoxic effect of retinoids and DEX was also observed on fresh MM cells obtained from 7 patients. Both retinoids and DEX are known to be inducers of apoptosis; we verified that the combined inhibitory activity of retinoids and DEX could be attributed to an increased induction of apoptosis. This effect may be mediated by a reduced intracellular expression of BCL‐2 protein, which indeed observed after prolonged in vitro treatment with retinoids. It has been described recently that an enhanced expression of BCL‐2 protein can contribute to the occurrence of early chemoresistance; the downregulation of BCL‐2 protein induced by retinoids could thus be exploited, by means of novel chemotherapy plus retinoids combinations, in order to improve the efficacy of conventional chemotherapy in MM.
Bcr‐abl protein detection in peripheral blood mononuclear cells for follow‐up of chronic myelogenous leukaemia patients
Authors: Schleuning M. , Mittermüller J. , Kolb H.‐.J.
Published: July 2010

Abstract:  We have assessed the value of p210 protein detection in peripheral blood cells for follow‐up of chronic myelogenous leukaemia (CML) patients. Quantification was achieved by comparing the relative intensities of the p210 bands with those of the normal abl protein (p145). Serial dilution of Ph‐positive K562 cells with Ph‐negative HL60 or KG1 cells revealed a linear correlation between the p210/p145 ratio and the number of Ph‐positive cells (r = 0.998; p < 0.001) with a sensitivity of detecting less than 1% Ph‐positive cells in 5 times 106 cells. Ninety‐six consecutive patients were enrolled in the study and a total of 155 Western blot analyses have been performed and compared to chromosomal analyses of bone marrow. Parallel RT–PCR analyses have been performed on 99 occasions. All patients with positive cytogenetic findings also probed positive for p210. In 23 instances p210 was detectable despite negative chromosomal analysis. In 16 samples these results were confirmed by RT–PCR. In patients with partial cytogenetic remission (n = 26) the results of the p210 assay correlated significantly with the percentage of Ph‐positive metaphases (r = 0.69; p < 0.001). In conclusion, monitoring of CML patients by quantification of the bcr‐abl protein is a feasible and sensitive alternative to chromosomal analysis of bone marrow.
Circulating haemopoietic progenitor cells in primary and secondary myelofibrosis: relation to collagen and reticulin fibrosis
Authors: Čolović Milica D. , Wiernik Peter H. , Janković Gradimir M. , Vidović Ana D. , Janošvić Slobodanka , Basara Nadežda M.
Published: July 2010

The relationship between the extent of bone marrow reticulin and collagen fibrosis and the concentration of granulocytic (CFU–GM), erythroid (BFU–E) and megakaryocyte (CFU–Mk) progenitor cells in the peripheral blood of patients with primary agnogenic myeloid metaplasia (AMM) and secondary myelofibrosis (sMF) has not been definitively correlated. We studied 23 patients with established diagnosis of AMM and 12 patients with sMF for the extent of reticulin and collagen bone marrow fibrosis and for the spontaneous colony (sCFU–GM, sBFU–E and sCFU–Mk) formation. The control group consisted of 11 healthy volunteers. Trephine biopsy of the posterior iliac crest was performed in all individuals studied to determine the type and degree of reticulin and collagen fibrosis. Gomori's silver impregnation technique was used. sCFU–GM, sBFU–E and sCFU–Mk colony formation was related positively to spleen size, the white blood cell counts and the degree of collagen fibrosis in AMM (p < 0.01). Stimulated CFU–GM were also significantly correlated with the degree of bone marrow reticulin and collagen fibrosis. There was no correlation between the extent of peripheral blood progenitor concentration and the degree of bone marrow reticulin or collagen fibrosis in sMF and in control individuals. In conclusion, the extent of bone marrow fibrosis is significantly correlated with the peripheral blood progenitor colony formation in AMM but not in sMF.