bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased liposome‐mediated gene transfer into haematopoietic cells grown in adhesion to stromal or fibroblast cell line monolayers
Authors: Marit Gérald , Cao Yunxu , Froussard Patrick , Ripoche Jean , Dupouy Maryse , Elandaloussi Abdel , Lacombe Francis , Mahon François Xavier , Keller Héléne , Pla Marika , Reiffers Josy , Theze Jacques
Published: March 2003

Abstract: We investigated transfection rates of CD34+ hematopoietic progenitor cells (HPC) or haematopoietic cell lines (TF‐1, KG1a and K562) using the LacZ gene as a reporter and cationic liposomes. The transfection efficiency of CD34+ hematopoietic progenitor cells (HPC) or TF‐1, KG1a and K562 grown in suspension is very low (average percentage of 0.013 for HPC and 0,03 for cell lines). Adhesion of HPC or cell lines to plates by immunological or physical methods significantly enhances transfection efficiency; however, the percentage of transfected cells still remained low. We found that adhesion of TF‐1, KG1a and K562 HC to MS‐5 stroma cells or NIH‐3T3 fibroblast cells increased transfection efficiency. Under these conditions transfection is achieved in 11.2–25% (mean 18.3%) for the cell lines and 13.6% (range 8.2%–24.2%) for CD34+ HPC. These results indicate that liposome‐mediated transfection of HC is significantly increased when cells are grown in adherence to stroma or fibroblast monolayers.
Flow cytometric analysis of defensins in blood and marrow neutrophils
Authors: Klut Maria E. , Whalen Beth A. , Hogg James C.
Published: March 2003

Abstract: Polymorphonuclear neutrophils (PMN) are vital in host defense against microbial infections. This study provides a flow cytometric method for the quantitative analysis of microbicidal peptides (defensins) in cells of PMN lineage. Rabbit neutrophil peptides, NP‐2 and NP‐5, were measured in all PMN and in subpopulations of PMN expressing l‐selectin. PMN lineage counts were made on Wright's‐stained blood smears and marrow cytospins. Immunoreactivity for NP‐2, and NP‐5 was detected by using the alkaline phosphatase anti‐alkaline phosphatase technique. The results show that marrow PMN express higher levels of NP‐2 and NP‐5 than blood PMN, p<0.001 and that these levels are associated with elevated numbers of myeloid precursors. In both blood and marrow, NP‐2 occurs in two PMN subpopulations and the mean fluorescence intensity of NP‐2 is consistently higher than that of NP‐5. Increased levels of defensins are observed in circulating PMN depicting the most
l‐selectin p<0.05. Immunocytochemical results indicate that PMN defensins reside in cytoplasmic granules and are not constitutively expressed on the cell surface. Furthermore, defensins are not detected in monocytes, eosinophils, lymphocytes and erythrocytes. The flow cytometric method described here provides a novel means of quantitating host natural defenses, allows the characterization of PMN subpopulations and has clinical applications.
Long‐term proliferation in vitro of hematopoietic progenitor cells from children with congenital bonemarrow failure: effect of rhGM‐CSF and rhEPO
Authors: Martínez‐Jaramillo Guadalupe , Espinoza‐Hernández Laura , Benítez‐Aranda Herminia , Mayani Hector
Published: March 2003

Abstract: We have characterized the proliferation kinetics of hematopoietic cells in long‐term marrow cultures (LTMC) from five normal children and seven children with congenital bone marrow failure (four with Fanconi anemia [FA] and three with congenital pure red cell aplasia [PRCA]). Total nonadherent and adherent cells, as well as nonadherent progenitors, were determined weekly in the presence or in the absence of rhGM‐CSF (10 ng/ml) or rhEPO (3 U/ml). As compared to normal LTMC, hematopoiesis was drastically reduced in cultures from FA patients. Myeloid and erythroid progenitor cells reached undetectable levels after only 3 and 1 weeks of culture, respectively. This was observed even in cultures supplemented with rhGM‐CSF, in which no response to this cytokine occurred. In LTMC from PRCA children, the growth of erythroid and multipotent progenitors was also drastically reduced. Myelopoiesis, on the other hand, showed normal levels during the first three weeks of culture; however, from week 4, there was a significant decrease in the levels of both progenitor and mature cells, reaching undetectable levels several weeks before normal cells did. Response to rhGM‐CSF and rhEPO was transient and deficient. Our results suggest that in FA, alterations at the level of primitive progenitor cells are so severe that myeloid, erythroid and multipotent progenitors are unable to proliferate in LTMC, even in the presence of rhGM‐CSF. In patients with PRCA the erythroid arm of hematopoiesis is preferentially affected and addition of rhGM‐CSF and/or rhEPO to these cultures had little or no effect on erythroid cell production. Interestingly, myelopoiesis in this culture system was deficient as well and response to rhGM‐CSF was defective, suggesting that the myeloid lineage is also altered in congenital PRCA.
Long‐term follow‐up of autologous stem‐cell transplantation for follicular and transformed follicular lymphoma
Authors: Berglund Åke , Enblad Gunilla , Carlson Kristina , Glimelius Bengt , Hagberg Hans
Published: March 2003

Abstract: Despite the fact that follicular lymphomas are both chemo‐ and radiosensitive, the disease is generally non‐curable. These lymphomas often undergo transformation to a more malignant state. In order to improve the prognosis, high‐dose treatment with stem cell support has been tested, but its role in the treatment of this disease is still unclear. Fourteen men and eight women with a median age of 45 yr (34–59) were treated with high‐dose therapy with autologous stem cell transplantation between 1987 and 1996. The patients were selected to undergo intensive therapy because of an estimated short survival (median <3 yr), even though they had chemosensitive disease and adequate performance status. Eleven patients' lymphomas had transformed, and the other eleven patients had one or more unfavourable prognostic signs such as advanced stage, bulky disease, multiple relapses, or short remission duration. The conditioning regimen has varied over the period, but BEAC (Becenum, etoposide, cytarabine, cyclophosphamide) or etoposide/cyclophosphamide with or without total body irradiation (TBI) was used in most patients. Nine patients had their stem cells purged. After a median follow‐up time of 74 months overall survival was 81% and disease‐free survival 72%. One toxic procedure‐related death occured. There was no difference in outcome between patients with a transformed lymphoma compared to those without transformation. The patients treated with TBI had a significantly worse outcome. Toxicity was also much higher in TBI‐treated patients, including four cases of secondary malignancy (three myelodysplastic syndrome (MDS) cases and one patient with breast carcinoma). This retrospective study, with the longest follow‐up time so far reported, shows a promising 6‐yr DFS of 72% in a group of follicular lymphoma patients with a bad prognosis. The outcome of patients with transformed lymphoma compared to historical controls is especially encouraging. The high incidence of MDS is worrying. The role of TBI should be questioned because this and other studies have not shown any advantage of using TBI. In the absence of randomised trials the role of high‐dose treatment for patients with follicular lymphoma is still not defined.
Different alterations in lactate dehydrogenase activity and profile of peripheral blood mononuclear cells in Hodgkin's and non‐Hodgkin's lymphomas
Authors: Jurišić V. , Konjević G. , Banićević B. , Duričić B. , Spužić I.
Published: March 2003

Abstract: The enzyme lactate dehydrogenase (LDH) activity of peripheral blood mononuclear cells (PBMC), LDH isotype H and M pattern and PBMC spontaneous LDH release activity were examined in 55 non‐Hodgkin's lymphoma (NHL) patients, 46 Hodgkin's disease (HD) patients and 47 controls. The intracellular LDH and M isotype activity of PBMC, their spontaneous LDH release activity significantly increase (p<0.01) in NHL with progressing histological grade of malignancy. Contrary to this, all classical HD patients have a significant elevation (p<0.05) of each of these parameters. Furthermore, unlike HD, in NHL clinical stage is associated with significant (p<0.05) increase in the level of spontaneous LDH release activity in each histological form. It is also shown that spontaneous LDH release activity of PBMC for HD and NHL patients demonstrates significant positive correlation (p<0.005) with serum LDH level, although elevation of spontaneous LDH release precedes serum LDH increase in both diseases. The results obtained regarding alterations in intracellular, isotype and spontaneously released LDH activity of circulating PBMC show that these parameters are dependent, in NHL patients, on the grade of malignancy and tumor burden, while they are persistently present in HD patients.
Alterations in binding activity of T cell transcription factor CD28 responsive element binding complex (CD28RC) following allogeneic bone marrow transplantation
Authors: Tanaka Junji , Mori Akio , Ota Shuiti , Yamamoto Yasushi , Kobayashi Sumiko , Hashino Satoshi , Asaka Masahiro , Imamura Masahiro
Published: March 2003

Abstract: The CD28 responsive element binding complex (CD28RC) has an important role in transducing CD28/B7 costimulatory signals. Using electrophoretic mobility‐shift assay (EMSA), we have analyzed the binding activity of CD28RC in the mixed lymphocyte culture (MLC) using peripheral blood mononuclear cells (PBMC) obtained from the patients before and after allogeneic bone marrow transplantation (allo‐BMT). The binding activity of CD28RC was low in MLCs using PBMC from patients without acute GVHD and it was also low in MLCs using PBMC from patients without chronic GVHD (cGVHD). In contrast, this activity in patients with cGVHD was estimated to be high. The relative values of CD28RC in comparison with third party MLCs were significantly higher in MLCs using PBMC from patients with cGVHD than those in MLCs using PBMC from patients without GVHD (0.55±0.31 versus 0.23±0.12, respectively, n=10, p=0.05). IL‐2 concentrations in the MLC medium from patients without GVHD were undetectable; however, a detectable level of IL‐2 was present in MLC medium from a patient with extensive cGVHD. These data were interpreted to suggest that the CD28 costimulatory pathway was specifically activated against recipient antigen in allo‐BMT patients with GVHD. In other words, it was suggested that the CD28 costimulatory pathway was specifically suppressed in allo‐BMT patients without GVHD, and this suppression might contribute immunological tolerance after allo‐BMT.
EORTC QLQ‐C30 and FACT‐BMT for the measurement of quality of life in bone marrow transplant recipients: a comparison
Authors: Kopp Martin , Schweigkofler Hansjörg , Holzner Bernhard , Nachbaur David , Niederwieser Dietger , Fleischhacker W. Wolfgang , Kemmler Georg , Sperner‐Unterweger Barbara
Published: March 2003

Abstract: The purpose of the study was to compare two differentquality‐of‐life self‐rating instruments, namely the EORTC QLQ‐C30, developed by the quality‐of‐life study group of the European Organisation for Research and Treatment of Cancer, and the FACT‐BMT (version 3), the Functional Assessment of Cancer Therapy – Bone Marrow Transplantation scale, which is the FACT‐G(eneral measure) in combination with a module developed specifically for evaluating quality of life of bone marrow transplant (BMT) patients. Fifty‐six BMT recipients completed both the EORTC QLQ‐C30 and the FACT‐BMT (German language version) during the same session. Questionnaire data were analyzed on a subscale basis using correlation analysis and multiple linear regression. Correlations between corresponding subscales of EORTC QLQ‐C30 and the FACT‐BMT ranged from r=0.30 for the emotional domain (poor agreement) to r=0.77 for global QOL (good agreement). This suggests that the instruments, despite considerable overlap, possibly focus on different aspects of QOL, in particular in addressing emotional and social issues of BMT patients. It appears that the FACT‐BMT gives a more comprehensive overview regarding the multidimensional construct of quality of life. The EORTC QLQ‐C30 gives more insight into the physical aspects of quality of life and helps to identify symptoms which effectively decrease quality of life from the patient's perspective. The QLQ‐C30 might be improved by the incorporation of a BMT‐specific module currently under development. We therefore conclude that neither of the two instruments can be replaced by the other in the assessment of QOL of BMT patients and that a direct comparison of results obtained with the two instruments is likely to be misleading.
B‐lymphocytes in CLL and NHL differ in the mRNA splicing pattern of the CD45 molecule
Authors: Yu Y. , Rabinowitz R. , Polliack A. , Ben‐Bassat H. , Schlesinger M.
Published: March 2003

Abstract: In the present study the cell surface expression of CD45 isoforms on normal and neoplastic human B cells was correlated with splice products of the CD45 mRNA, using RT‐PCR technology. In non‐Hodgkin's lymphoma cells in the leukemic phase (NHL) the majority of the cells expressed a high level of CD45RA, while in CLL most of the cells expressed a low level. In the Raji and Daudi Burkitt B‐cell lymphoma lines the main CD45 mRNA product was the largest, unspliced, full‐length isoform (456) and the 56 splice product. Similar results were obtained with B‐cell lymphoma cells isolated from the peripheral blood of patients with NHL in the leukemic phase. In EBV‐transformed B‐cell lines, the 456 and the 56 isoform of CD45 mRNA were predominant, but in addition a low level of the 5‐ and 0‐exon splice products was detected. A strikingly different pattern was obtained with B‐CLL cells. In CLL the level of the 456 and the 56 isoforms was low, while that of the 5‐ and 0‐exon splice products was increased. Thus, in contrast to the heterogeneity in the expression of CD45RO in B‐CLL, the majority of the cells contained the CD45 mRNA splice product coding for CD45RO. Analysis of splice products of the CD45 mRNA may serve as an additional tool to differentiate CLL from the leukemic phase of NHL.
Clonal patterns of X‐chromosome inactivation in female patients withaplastic anaemia studied using a novel reverse transcription polymerase chain reaction method
Authors: Mortazavi Y. , Chopra R. , Gordon‐Smith E. C. , Rutherford T. R.
Published: March 2003

Abstract: Conflicting results have been published on the frequency of clonal patterns of X‐chromosome inactivation in female patients with aplastic anaemia. Previous studies have used DNA methylation to measure X‐inactivation, but aberrant methylation is known to occur in some situations. We have developed a non‐radioactive reversetranscription polymerase chain reaction (RT‐PCR) method to study expression of the polymorphism at nt.1311 of the G6PD gene at the RNA level. Using this, and a similar method for the iduronate‐2‐sulfatase (IDS) gene, we have re‐evaluated X‐inactivation in AA patients. 32/35 normal individuals showed polyclonal haemopoiesis. Patients with presumed clonal diseases showed both monoclonal and polyclonal patterns, consistent with previous reports. Overall, clonal patterns were observed in granulocytes of 10/26 AA patients (38%), a significantly higher proportion than in controls (p<0.01). Two cases showed discordance between lymphocytes and granulocytes, indicating clonality arising within the myeloid lineage. Eight cases showed clonal patterns in both myeloid and lymphoid cells, indicating the involvement of a pluripotent stem cell. Clonal patterns did not correlate with age, but there appeared to be an association with duration of disease. In PNH patients, CD59‐negative cells showed clonal patterns of X‐inactivation. In two cases, however, clonal patterns were also detected in CD59‐positive cells.
Gene frequencies of the HPA‐3 and HPA‐5 platelet antigen alleles among the Amerindians
Authors: Covas D. T. , Bı´scaro T. A. , Nasciutti D. C. , Guerreiro J. F. , Santos S. E. B. , Zago M. A.
Published: March 2003

Abstract: Platelet antigens are of importance in several clinical situations and in population genetics. Data are scarce on allele frequencies in ethnic groups other than whites, Asians and African Americans. The frequencies of the alleles of the systems HPA‐3 and HPA‐5 were determined using the allele‐specific restriction enzyme for five South‐American Amerindian populations and compared with those obtained for Afro‐Americans, Japanese and whites from Brazil. The frequency of the HPA‐3a allele among the Amerindians as a group did not differ from the values obtained for the other populations. However, differences were observed among the Amerindians, varying from 0.27 to 0.75, the highest frequency thus far observed for a population of Asian origin. Only the HPA‐5a allele was found among 130 Amerindian chromosomes. The determination of gene frequencies of the HPA systems in different populations allows inference of gene flows and genetic constitution of populations and the estimation of the risk of platelet‐specificalloimmunization.
Incidence, clinical features and outcome of essential thrombocythaemia in a well defined geographical area
Authors: Jensen Morten Krogh , De Nully Brown Peter , Nielsen Ove Juul , Hasselbalch Hans Carl
Published: March 2003

Abstract: In an attempt to characterise the clinical features, incidence and outcome of essential thrombocythaemia (ET) we report our experience in a large unselected series of patients from a well defined region. All new cases of ET in the County of Copenhagen were registered during the period 1977–98. We identified 96 cases of ET, yielding an age‐ and sex‐adjusted annual incidence rate of 0.59/100.000 and a point‐prevalence at last follow up of 11/100.000. The overall incidence rate was 0.31 and 1.00 per 100.000 population during the consecutive periods 1977–89 and 1990–98, respectively, corresponding to a 3.2‐fold increase. Median age at diagnosis was 67 yr (females 68 yr, males 66 yr, range 18–87 yr), and the female to male (F/M) ratio was 2.6:1. At diagnosis, 52% of the patients displayed no ET‐related symptoms and were discovered fortuitously by a routine platelet count. Forty‐eight percent presented with thrombohaemorrhagic phenomena, of which microvascular disturbances of the central nervous system (CNS), extremities and skin were most frequently observed (23%). Compared to patients diagnosed after 1989, patients diagnosed before 1990 had a significantly higher mean platelet count, white blood cell (WBC) count, lactate dehydrogenase (LDH) value and alkaline phosphatase value. With a median follow up of 70 months, 5‐yr survival was 76%, significantly lower than the expected survival of an age‐ and sex‐matched control group (p=0.0052). Thirty‐seven patients experienced a total of 55 thrombohaemorrhagic events during follow‐up, corresponding to an incidence of thrombosis and microvascular disturbances or haemorrhage of 8.1% per pt‐yr and 2.5% per pt‐yr, respectively. The number of patients experiencing thrombosis or microvascular disturbances was significantly higher among the 29 patients who never received acetylsalicylic acid (ASA) compared to the 67 patients who received ASA during follow up (45% vs. 21%; p=0.017). This study provides population‐based data suggesting the benefit of treatment with low‐dose ASA in a non‐selected population of patients with ET.
Growth factor receptor profile of CD34+ cells in AML and B‐lineage ALL and in their normal bone marrow counterparts
Authors: De Waele M. , Renmans W. , Vander Gucht K. , Jochmans K. , Schots R. , Otten J. , Trullemans F. , Lacor P. , Van Riet I.
Published: March 2003

Abstract: Leukaemic cells show a low clonogenic activity and a heterogeneous proliferative response to growth factors. We investigated whether this could be due to an altered expression of growth factor receptors on the leukaemic precursors. Receptors for G‐CSF, stem cell factor (SCF), IL‐3, IL‐6 and IL‐7 were detected on CD34+ cells in AML and B‐lineage ALL with monoclonal antibodies and flow cytometry. The expression was compared with that on myeloid and B‐lymphoid CD34+ cells in normal bone marrow. Leukaemic CD34+ cells expressed the same receptors as their normal counterparts. AML and B‐lineage ALL could be distinguished by the growth factor receptor profile of their CD34+ cells. SCFR, G‐CSFR and IL‐6Rα were found in AML, IL‐7R in B‐lineage ALL and IL‐3Rα in both. IL‐3Rα was upregulated in AML and B‐lineage ALL CD34+ cells, while samples with low or high expression were present for the other receptors. This variable expression could correlate with the heterogeneous response of leukaemic cells to growth factors. Functional studies on isolated CD34+ cells are needed to investigate this further.
Lack of pathogenic mutations in the 5′‐untranslated region of the thrombopoietin gene in patients with non‐familial essential thrombocythaemia
Authors: Allen Anthony J. R , Gale Rosemary E , Harrison Claire N , Machin Samuel J , Linch David C
Published: March 2003

Abstract: Thrombopoietin (TPO) is thought to be the major physiological regulator of thrombopoiesis, and, in general, circulating levels are inversely proportional to megakaryocyte and platelet mass. However, normal or elevated TPO levels are found in patients with essential thrombocythaemia (ET) and the reason for this is not fully understood. Recent studies have shown that four kindreds with hereditary thrombocythaemia (HT) have point mutations in the 5′‐untranslated region (UTR) of the TPO gene which lead to increased TPO translation. In order to determine whether similar mutations are present in apparently acquired ET, in particular in those patients with polyclonal myelopoiesis, we have studied this region in 50 ET patients using neutrophil DNA. The known HT mutations were investigated using polymerase chain reaction with mismatch primers and restriction enzyme digestion; only wild‐type alleles were detected. Single‐stranded conformation polymorphism (SSCP) analysis of exons 1–4 identified a C→T substitution at nucleotide 3767. However, this appears to be a common polymorphism, as it was present at the same frequency in haematologically normal controls and is unlikely to be of pathological significance. These results demonstrate that mutations in the 5′‐UTR of the TPO gene are not the cause of the normal or elevated TPO levels in acquired ET.
Immunomodulatory effects of moxifloxacin in comparison to ciprofloxacin and G‐CSF in a murine model of cyclophosphamide‐induced leukopenia†
Authors: Shalit Itamar , Kletter Yehudith , Halperin Drora , Waldman Dalia , Vasserman Elad , Nagler Arnon , Fabian Ina
Published: March 2003

Abstract: We analyzed the effect of the two quinolones moxifloxacin and ciprofloxacin on the repopulation of hematopoietic organs and on the production of cytokines by various organs of cyclophosphamide (CP)‐induced leukopenic mice. The effect was compared to that of G‐CSF. Cyclophosphamide injection induced a severe leukopenia, with nadir at day 4 post‐injection. All the quinolone and G‐CSF‐treated animals showed WBC>500/μL at the nadir, compared to 50% of saline‐treated mice. Cyclophosphamide induced a marked decrease in the number of myeloid progenitors (CFU‐C) in bone marrow (BM) and spleen. Quinolone or G‐CSF treatment resulted in a 1.4–4.3‐fold increase in CFU‐C numbers in the BM; no enhancement was observed in the spleen. Treatment with CP resulted in enhanced colony‐stimulating activity (CSA) in bone shaft and spleen and decreased activity in bladder and lung. Treatment of CP‐injected mice with quinolones significantly enhanced CSA in the bone shaft, spleen, lung and bladder on different days.

In normal mice the highest levels of GM‐CSF and IL‐6 were observed in lung‐conditioned medium (compared to bone shaft, spleen and bladder). Injection of CP resulted in a 22.5‐ and 93‐fold decrease in GM‐CSF and IL‐6 levels, respectively, in lung‐conditioned medium, while treatment with quinolones resulted in 2–4‐fold increase in GM‐CSF with no effect on IL‐6 production. G‐CSF treatment had no enhancing effect on GM‐CSF nor on IL‐6 production. We conclude that moxifloxacin and ciprofloxacin administered to CP‐injected mice revert some of the immune suppressive effects of cyclophosphamide.
Myelofibrosis with myeloid metaplasia in adult individuals 30 years old or younger: presenting features, evolution and survival
Authors: Cervantes Francisco , Barosi Giovanni , Hernández‐Boluda Juan‐Carlos , Marchetti Monia , Montserrat Emilio
Published: March 2003

Abstract: Myelofibrosis with myeloid metaplasia (MMM) usually affects older people and is associated with a median survival of 3.5–5 yr. Survival of MMM patients 55 yr old or younger is longer, but there is no information on the youngest subset. The presenting features, evolution and survival were analyzed in 9 patients with MMM aged 30 yr or less, representing 2.8% of two series including 323 cases. Age ranged from 17 to 30 yr; 3 patients were males and 6 females. Five patients were asymptomatic and none had constitutional symptoms. Anemia was observed in 4 patients, being severe in only one. The WBC count was normal in 7 patients and 2 had mild leukocytosis; moderate thrombocytosis was observed in 7 patients. No patient showed blood blast cells or bone marrow cytogenetic abnormalities. With a median follow‐up of 6.8 (range 0.8–28) yr, 2 patients died 10.7 and 9.9 yr from diagnosis, one from complications of bone marrow transplantation performed after progression of the disease and the other from gastrointestinal bleeding secondary to portal hypertension. Among remaining patients, one developed Budd–Chiari syndrome, another was lost to follow‐up at 3 yr, and 5 remain asymptomatic and currently without treatment (one had been splenectomized at diagnosis). These results indicate that in most young adults with MMM the disease presents without adverse prognostic factors and may remain stable for years, which may be of interest when considering the treatment of such patients.
Incidence of hepatitis virus infection and severe liver dysfunction in patients receiving chemotherapy for hematologic malignancies
Authors: Kawatani Toshio , Suou Takeaki , Tajima Fumihito , Ishiga Kiyomi , Omura Hiromi , Endo Akira , Ohmura Hiroshi , Ikuta Yujiro , Idobe Youko , Kawasaki Hironaka
Published: March 2003

Abstract: Hepatitis virus infection through virus reactivation has a high risk of mortality in patients with hematological malignancies receiving chemotherapy. We examined the incidence of both hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and severe liver dysfunction (alanine aminotransferase >ten times the normal upper limit and total bilirubin >5 mg/dl) during chemotherapy in 268 patients with hematological malignancies. Eight patients (3.0%) were infected with HBV and 22 patients (8.2%) were infected with HCV. One patient (0.4%) was infected with both HBV and HCV. HBV‐ or HCV‐infected patients showed severe liver dysfunction at a significantly higher incidence than non‐infected patients (11/31 (35.5%) vs. 0/237 (0%), p<0.0001). Furthermore, the incidence of severe liver dysfunction in HBV‐infected patients was significantly higher than in HCV‐infected patients (6/8 (75.0%) vs. 4/22 (18.2%), p<0.01). Three of eight HBV‐infected patients were initially negative for hepatitis B surface antigen (HBsAg) by latex agglutination and became positive for HBsAg during chemotherapy. Furthermore, all three patients developed severe liver dysfunction and two developed fatal fulminant hepatitis. From an examination of the original stock of serum samples before chemotherapy, two patients were found to be positive for HBV‐DNA by polymerase chain reaction (PCR). Although post‐transfusion HBV infection was suspected in the one remaining patient, the cause of HBV infection could not be clarified due to the impossibility of examination in blood donors. Since HBV‐infected patients develop severe liver dysfunction at a higher incidence than either patients not infected with virus or HCV‐infected patients before chemotherapy for hematological malignancies, it is recommended that HBV‐DNA should be tested by PCR to detect HBV marker‐negative carriers and liver function tests should be carefully monitored.
Cancer‐related anemia in a rat model: α2‐macroglobulin from Yoshida sarcoma shortens erythrocyte survival
Authors: Bhanushali Aparna A. , Raghunathan R. , Kalraiya Rajiv D. , Mehta Narendra G.
Published: March 2003

Abstract: Implantation of Yoshida ascites sarcoma in rats was found to lead to a reduction in the hemoglobin content, the erythrocyte count and the packed cell volume of blood to 30% of normal in 4 d; however, there was no decrease in the mean cell hemoglobin, the mean cell volume and the mean corpuscular hemoglobin concentration, or suppression of erythropoiesis. The red cells from the circulation of tumor‐bearing animals, tagged with 51Cr and injected intravenously in normal rats, showed significantly faster clearance than normal. The erythrocytes contaminating the tumor ascites exhibited extremely short survival, suggesting that one or more secreted tumor product(s) may be responsible for the effect. Incubation of red cells from normal rats in the cell‐free ascites fluid, or with an isoform of α2‐macroglobulin purified from it, also led to reduction in the survival; but the ascites fluid depleted specifically of α2‐macroglobulin was without any effect. The erythrocytes exhibiting reduced survival showed a proportionate decrease in their cellular deformability. The study identifies a tumor product that is directly responsible for the causation of anemia in the host, and the mechanism by which it does so.
Improved outcome in adult acute myeloid leukemia is almost entirely restricted to young patients and associated with stem cell transplantation
Authors: Wahlin Anders , Markevärn Berit , Golovleva Irina , Nilsson Marie
Published: March 2003

Abstract:Prognostic factors were studied in a series of 318 patients with acute myeloid leukemia (AML), 17–90 yr old, treated at a single centre during 1982–98, and representing 79% of the total number of cases registered in the area during this period. Risk group stratification based on cytogenetics, occurrence of antecedent hematological disorder, and leukocyte count could be performed in 93%. Five percent were allocated to the favourable risk group, 40% to standard risk, and 55% to adverse risk. Complete remission (CR) was attained in 52%. The CR rate was higher in the favourable (80%) and standard risk groups (69%) than in the adverse risk group (37%). The CR rate increased from 44% in the 1980s to 60% in the 1990s. The 5‐yr survival rate for all patients was only 12%. Low age, promyelocytic leukaemia, treatment in the 1990s, high induction treatment intensity, and non‐adverse risk group were favourably associated with survival. The median survival time increased from 115 to 349 d between the 1980s and the 1990s, but the 5‐yr survival rate was only 11% for patients over 55 yr of age even in the last decade. For the younger patients, the 5‐yr overall survival rate increased from 9% to 35% in the last decade. The median time in first remission was 365 d. Age below 56 yr, allogeneic and autologous transplants, and non‐adverse risk group were associated with prolonged response duration. The duration of response among all patients increased from 250 d in the 1980s to 451 d in the 1990s, but event‐free survival time did not improve significantly in patients above 55 yr of age. Among patients below 56 yr of age, overall survival and event‐free survival were significantly better for those who received allogeneic or autologous transplants in first remission than for those who were treated with chemotherapy only. Overall survival times did not improve from the 1980s to the 1990s among those patients below 56 yr who were treated with chemotherapy only in first remission, in spite of the use of transplants in second remission.
Comprehensive comparison of FISH, RT‐PCR, and RQ‐PCR for monitoring the BCR‐ABL gene after hematopoietic stem cell transplantation in CML
Authors: Kim Yoo‐Jin , Kim Dong‐Wook , Lee Seok , Kim Hee‐Je , Kim Yoo‐Li , Hwang Ji‐Yeon , Oh Il‐Hoan , Park Yoon‐Hee , Lee You‐Kyoung , Min Chang‐Ki , Kim Tai‐Gyu , Han Tae‐Hee , Min Woo‐Sung , Kim Chun‐Choo
Published: March 2003

Abstract: The reverse transcriptase‐polymerase chain reaction (RT‐PCR) was compared with fluorescence in situ hybridization (FISH) and real‐time quantitative RT‐PCR (RQ‐PCR) for minimal residual disease (MRD) monitoring in 266 post‐transplant bone marrow samples from 78 patients with chronic myelogenous leukemia (CML). The sensitivities of FISH to BCR‐ABL positive samples determined by first‐round (1st) RT‐PCR, second‐round (2nd) RT‐PCR, and RQ‐PCR were 64.2%, 25.8%, and 20.7%, respectively. The BCR‐ABL/ABL ratio by RQ‐PCR had a mean of 0.000 13 in the 1st RT‐PCR‐negative samples and 1.42 in the 1st RT‐PCR‐positive samples (P<0.001), and means of 0.000 39 and 0.51 in the 2nd RT‐PCR‐negative and ‐positive samples (P< 0.001). The mean ratios of BCR‐ABL/ABL by RQ‐PCR were significantly different in N/N (1st/2nd RT‐PCR) or N/P and P/P (P<0.001), but not in N/N and N/P, which showed that the discriminative power of RQ‐PCR is confined to the 1st RT‐PCR level. In this respect, monitoring of the 1st RT‐PCR might be useful for estimating normalized BCR‐ABL levels after transplantation. Nested RT‐PCR was of limited use, as RQ‐PCR quantified the BCR‐ABL transcripts in 60 (91%) of 66 samples determined to be negative by 2nd RT‐PCR. FISH was significantly correlated with RQ‐PCR in FISH‐positive samples (n=24, r=0.79, P=0.001). An increase of FISH preceded that of RQ‐PCR in a few cases with molecular relapse. By analyzing a large number of samples post‐transplant, we found that RQ‐PCR might be the most useful assay for MRD monitoring; however, FISH and RT‐PCR were found to be useful complementary tools.
Expression of the bcl‐2 family of pro‐ and anti‐apoptotic genes in multiple myeloma and normal plasma cells
Authors: Spets Helena , Strömberg Thomas , Georgii‐Hemming Patrik , Siljason Jan , Nilsson Kenneth , Jernberg‐Wiklund Helena
Published: March 2003

Abstract: Aberrant expression of genes regulating apoptosis/survival seems to be essential in the stepwise development of human multiple myeloma (MM). In this paper we have compared the expression of bcl‐2 family pro‐ and anti‐apoptotic genes in MM cell lines, primary MM cells and normal plasma cells. The Bcl‐2, Mcl‐1, Bcl‐xL/S, Bcl‐w, Bax, Bak, and Bad were shown to be expressed in both malignant and non‐neoplastic, normal plasma cells. Quantitative analysis revealed that the malignant phenotype seemed to correlate with an elevated expression of Mcl‐1, a decreased expression of Bax and, to a lesser extent, an increased Bcl‐2/Bax expression ratio. The possible influence of interleukin‐6 (IL‐6) in regulating the expression of the bcl‐2‐related genes was also examined. Using the IL‐6‐dependent MM cell lines U‐1958 and U‐266–1970 it was clearly shown that IL‐6 deprivation induced cell cycle arrest in both cell lines, whereas apoptosis was only detected in the U‐1958 cells. Furthermore, the anti‐apoptotic proteins Bcl‐2, Mcl‐1 and Bcl‐xL were down‐regulated, while the expression of the pro‐apoptotic Bax protein was increased. To conclude, we suggest that the expression pattern of the Bcl‐2 family of proteins separates the malignant phenotype of MM from normal plasma cells, and that the protecting effect of IL‐6 may be conducted via an altered balance between these proteins.
Anthracyclines potentiate activity against murine leukemias L1210 and P388 in vivo and in vitro
Authors: Szmigielska‐Kaplon Anna , Ciesielska Ewa , Szmigiero Leszek , Robak Tadeusz
Published: March 2003

Abstract:  The interactions of 2‐chlorodeoxyadenosine (2‐CdA, cladribine) and three anthracyclines: doxorubicin (DOX), idarubicin (IDA) and mitoxantrone (MIT) were evaluated on murine leukemias P388 and L1210. Prolongation of survival time of animals receiving drugs in combination compared to mice treated with drugs in monotherapy was tested. We have also evaluated interactions of the cytostatics on murine leukemias in vitro by measuring their inhibitory effects on P388 and L1210 cell proliferation. We have observed a synergistic effect of MIT and IDA in combination with 2‐CdA on P388 leukemia resulting in an increase of life span (ILS)=226% in case of MIT+2‐CdA and ILS=126% in the case of IDA+2‐CdA, whereas 2‐CdA used as a sole drug resulted in an ILS=47%. The survival time of animals inoculated with P388 leukemic cells and treated with DOX+ 2‐CdA was similar to ILS gained by DOX monotherapy (178% and 200% respectively). The mice bearing L1210 leukemia receiving combined chemotherapy lived significantly longer than the animals on single agent regimens. The animals treated with schedule 2‐CdA+MIT lived significantly longer (P=0.004) as compared to the groups receiving drugs in monotherapy (ILS of 2‐CdA+MIT group=60%, ILS of MIT group 33%, and 2‐CdA group 33%). Finally, combination of DOX or IDA with 2‐CdA resulted in ILS =73% (2‐CdA+DOX regimen), and ILS=60% in case of 2‐CdA+IDA regimen, which is significantly higher than ILS gained on monotherapy schedules. In vitro tests revealed that all tested anthracyclines enhance the antiproliferative activity of 2‐CdA against L1210 and P388 leukemic cells (P<0.05).

Our study has shown that all anthracyclines potentiate 2‐CdA antileukemic activity, both in vivo and in vitro. It failed however to point the best one to be combined with cladribine. We suggests that further clinical trials with such combinations are needed.
Development and rapid dissemination of Merkel‐cell carcinomatosis following therapy with fludarabine and rituximab for relapsing follicular lymphoma
Authors: Cohen Yossi , Amir Gail , Polliack Aaron
Published: March 2003

Abstract: This report deals with an unusual case of a patient with follicular small cleaved lymphocytic lymphoma who developed Merkel‐cell carcinoma soon after receiving chemoimmunotherapy with a fludarabine‐containing regimen and rituximab. The presentation of the Merkel‐cell carcinoma in this patient was atypical because of the absence of dermal involvement and the very rapid clinical progression. In the light of recent reports which suggest a possible link between the immunocompromised state and the development of Merkel‐cell carcinoma, the atypical presentation seen in this patient may indeed imply a possible link between the therapy given and the development of Merkel‐cell carcinoma. To the best of our knowledge, this is the first documentation of Merkel‐cell carcinoma appearing in a patient soon after treatment with fludarabine and/or rituximab.
Mononucleosis syndrome and acute monocytic leukemia
Authors: Tamayose Kenji , Sugimoto Koichi , Ando Miki , Oshimi Kazuo
Published: March 2003

Abstract: The association of infectious mononucleosis and an immunocompromised host such as occurs in acute leukemia is reported. The most common cause of infectious mononucleosis is Epstein–Barr virus (EBV) and cytomegalovirus (CMV). Patients with mononucleosis syndrome caused by other agents are rare. We report a case of acute monocytic leukemia (AMoL) who developed varicella zoster virus (VZV) mononucleosis syndrome in the bone marrow recovery phase after myelosuppression due to high‐dose cytarabine. Mononuclear leukocytes appearing during the mononucleosis syndrome were very similar to the initial leukemic cells. Varicella zoster virus mononucleosis syndrome was confirmed by delayed herpes zoster rash with dermatomal distribution.
Disappearance of a mucosa‐associated lymphoid tissue (MALT) lymphoma of the urinary bladder after treatment for Helicobacter pylori
Authors: Van Den Bosch J. , Kropman R. F. , Blok P. , Wijermans P. W.
Published: March 2003

Abstract: A 59‐yr old man presented with macroscopic hematuria. A non‐Hodgkin’s lymphoma of the MALT type of the urinary bladder, stage I A‐E, was diagnosed. Radiotherapy was advised, but the patient refused this kind of treatment. Because of the known relationship of a gastric MALT lymphoma and Helicobacter pylori, the patient was treated with HP eradication therapy. Afterwards the lymphoma disappeared. The patient is 3 yr later in a persistent complete remission.

This is the first known case of the disappearance of a MALT lymphoma of the urinary bladder after treatment with HP eradication therapy.
Rituximab (anti‐CD20 monoclonal antibody) as consolidation of first‐line CHOP chemotherapy in patients with follicular lymphoma: a phase II study
Authors: Jäeger Gerald , Neumeister Peter , Brezinschek Ruth , Höfler Gerald , Quehenberger Franz , Linkesch Werner , Sill Heinz
Published: March 2003

Abstract: Advanced stages of follicular lymphoma are deemed incurable by conventional approaches. Immunotherapy with the humanised monoclonal anti‐CD20 antibody rituximab represents a new therapeutic option. The aim of our study was to determine the effectiveness and safety of rituximab in the consolidation setting of first‐line treated patients with follicular lymphomas. Thus the goal was first to reduce tumour burden using the CHOP regimen as induction treatment followed by consolidation with rituximab administered on a standard 4 wk schedule at a dosage of 375 mg m−2 body surface area. Between August 1998 and April 2001, 41 patients were enrolled in the study. All patients were evaluable with regard to tumour response and toxicity. The overall remission rate in the intent‐to‐treat analysis was 100%. On subgroup analysis, 20 [83% (95% CI: 63–95%)] of the 24 patients with grade 1 or 2 histology entered complete remission (CR), in 10 cases (42%) after additional rituximab therapy. Rituximab thus led to CR in 10/14 patients [71% (95% CI: 42–92%)] who had merely achieved partial remission (PR) with CHOP. Of 16 evaluable patients with grade 3 histology (excluding one patient achieving CR on CHOP who refused further treatment with rituximab), 15 [94% (95% CI: 63–97%)] achieved CR, 13 (81%) of these while still receiving CHOP. Two of the three patients achieving only PR on CHOP entered CR following rituximab. Thirty‐four patients (83%) continued to be in remission during a median follow‐up period of 24.3 (9–40) months. Our data suggest that the use of rituximab for consolidation after CHOP may improve CHOP‐induced remission and thus increase the CR rate. Furthermore, it was accompanied by a reduced rate of infusion‐related side‐effects.
Severe respiratory syncytial virus pneumonia complicating fludarabine and cyclophosphamide treatment of chronic lymphocytic leukemia
Authors: Field K. , Slavin M.A. , Seymour J.F.
Published: March 2003

Abstract: The potential for life‐threatening pneumonia due to respiratory syncytial virus (RSV) infection is recognised among patients with acute leukaemia and recipients of allogeneic or autologous bone marrow transplantation. RSV pneumonia has a high mortality rate in these settings. Less intensively treated patients are not usually considered to be at risk for serious RSV pneumonia. We describe the case of a 62‐yr‐old patient with chronic lymphocytic leukaemia (CLL) treated with fludarabine and cyclophosphamide who developed severe RSV pneumonia and recovered following treatment including intravenous ribavirin.
The enigma of the metabolic fate of circulating erythropoietin (Epo) in view of the pharmacokinetics of the recombinant drugs rhEpo and NESP
Authors: Jelkmann Wolfgang
Published: March 2003

Abstract:  Recombinant human erythropoietin (rhEpo) is a mainstay in the treatment of anaemia, primarily in renal failure. Because the half‐life of circulating rhEpo is relatively short (4–8 h), the drug is usually administered 2–3 times weekly. Recently, a novel erythropoiesis‐stimulating protein (NESP) with a longer half‐life (24–26 h) has been approved. NESP possesses two additional N‐glycans compared to endogenous Epo or rhEpo. The pharmacokinetics of rhEpo and NESP in humans have been investigated in detail. The composition of the N‐glycans is clearly important in determining the biological activity and the velocity of the degradation of Epo and its analogues. However, due to the lack of knowledge of the main site and mechanism of the removal of Epo from circulation, the difference in survival of rhEpo and NESP has remained phenomenological. Investigators have implicated the liver, kidneys, and bone marrow as possible sites of the catabolism of Epo. However, while hepatocytes take up desialylated Epo, the liver does not appear to play a major role in the degradation of intact Epo. Likewise, renal Epo clearance is apparently of secondary importance. Studies showing non‐linear pharmacokinetics of Epo suggest that Epo is eliminated by saturable mechanisms. The hormone, as well as the recombinant drugs, can be incorporated by erythrocytic progenitors and other tissues expressing the Epo receptor. The affinity of the Epo receptor for rhEpo is 4.3‐fold higher than for NESP. Taken together, it seems most likely that native Epo, rhEpo and NESP are degraded following Epo receptor‐mediated uptake, mainly in bone marrow.
Recurrent palmar–plantar erythrodysaesthesia following high‐dose cytarabine treatment for acute lymphoblastic leukemia
Authors: Crawford Julie H. , Eikelboom John W , McQuillan Andrew
Published: March 2003

Abstract: Palmar–plantar erythrodysaesthesia (PPE) is an uncommon cutaneous complication of cytotoxic chemotherapy which generally presents as a painful erythema involving the palms and soles. It has been suggested that PPE caused by cytarabine does not recur with subsequent cytarabine re‐challenge. We report a patient with recurrent, increasingly severe episodes of PPE, ultimately complicated by a severe bullous eruption, following successive cycles of high‐dose cytarabine for the treatment of acute lymphoblastic leukaemia. Contrary to previous recommendations, our experience cautions against the further use of high‐dose cytarabine in patients who develop PPE, and is a timely reminder of the potential toxicity of this agent, which is now increasingly being used as first‐line treatment in the management of haematologic malignancies.
Rebound hepatosplenomegaly in type 1 Gaucher disease
Authors: Tóth Judit , Erdős Melinda , Maródi László
Published: March 2003

Abstract: A 19‐yr‐old male patient with type 1 Gaucher disease was put on regular biweekly infusions of alglucerase. After 1 yr of treatment, hepatic and splenic volumes decreased from 38 and 45 mL/kg to 31 and 34 mL/kg, respectively. In addition, hemoglobin concentration, platelet count and white cell count increased, acid phosphatase level decreased, and the patient gained weight and energy. Despite improvement, the patient refused enzyme replacement therapy (ERT) because of muscle rigidity, chest pain, trembling and anxiety, which he attributed to enzyme substitution. Two and 4.5 yr after cessation of therapy, hepatic and splenic volumes increased to 36 and 53 mL/kg and to 53 and 110 mL/kg, respectively. The patient developed non‐tractable hematuria because of compression and dislocation of the left kidney by the enlarged spleen, which necessitated splenectomy. This report suggests that cessation of ERT in Gaucher disease may result in severe and complicated rebound visceromegaly.