bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment of erythropoietin‐resistant anaemia with desferrioxamine in patients on haemofiltration
Authors: Goch J. , Birgegård G. , Danielson B.G. , Wikström B.
Published: July 2010

Abstract: We studied 5 anaemic patients with Hb 74 ± 8 g/l on haemofiltration (HF) treatment. They were iron overloaded. Their serum ferritin was 2667 ± 8 μg/l. All patients' haemoglobin (Hb) levels decreased after an initial response to recombinant human erythropoietin (r‐HuEPO) treatment. An increase in r‐HuEPO dose from 100 to 400 U/kg s.c. thrice weekly with addition of oral and intravenous iron treatment for 8 weeks arrested the fall in Hb level. However, there was no significant increase in Hb during these 8 weeks. Iron was withdrawn at week 24 and desferrioxamine (DFO) treatment (i.v. doses of 2 g thrice weekly) was added to r‐HuEPO from week 26 to 36. Two weeks after DFO initiation the Hb level increased to 110 g/l. Thereafter the r‐HuEPO doses were reduced from 400 to 25 U/kg within 3 months. The Hb remained stable at a level of 110 g/l during the study, i.e., 17 months after the DFO treatment. Serum ferritin levels fell at a more rapid pace during DFO treatment and continued to decrease after DFO cessation for the following 17 months. In accordance with previous observations we found a positive effect of DFO treatment on erythropoiesis in patients with anaemia and iron overload. DFO treatment should be considered in patients with iron overload and r‐HuEPO‐resistant anaemia.
Disseminated intravascular coagulation in acute leukaemias at first diagnosis
Authors: Nur Samina , Anwar Masood , Saleem Muhammad , Ahmad Parveen Ara
Published: July 2010

Abstract: Haemorrhagic diathesis is the commonest cause of morbidity and mortality in acute leukaemias (AL). It is most commonly due to thrombocytopenia resulting from bone marrow failure. However, in a significant number of cases, disseminated intravascular coagulation (DIC) plays an important part. Previously it was thought that this mechanism was mainly confined to acute promyelocytic leukaemia (APL), but recently it has also been reported to occur in other subtypes of acute leukaemia. We report the results of a study carried out to find the incidence of DIC in various types of AL at the time of first diagnosis and in the absence of other recognisable causes. DIC was observed in 14(13.4%) cases out of 104 cases of AL studied. Nine out of 49(18.4%) cases of AML and 5 out of 55(9.1%) cases of acute lymphoblastic leukaemia (ALL) showed coagulation abnormalities consistent with DIC. Out of the 9 cases of AML showing DIC, 63 (66.67%) belonged to APL (FAB ME) subtype. Three (60%) out of 5 cases of ALL with DIC had T‐cell immunophenotype. The results indicate that DIC may also occur in types of AL other than APL, particularly in T‐ALL, and should be looked for.
In vitro drug sensitivity of leukemic progenitors and P‐glycoprotein expression in adult acute myeloid leukemia: correlation with induction treatment outcome
Authors: Basara Nadežda , Radoševic‐Radojković Nina , Čolović Milica , Bošković Darinka , Rolović Zoran
Published: July 2010

Abstract: We have investigated the self‐renewal capacity (PE2) and in vitro sensitivity to cytosine‐arabinoside (ara‐C) and daunorubicine (DNR) of leukemic progenitors (CFU‐AML) to determine the significance of these tests for predicting induction treatment outcome in 75 adult acute myeloid leukemia (AML) patients. In addition, in a part of this group of patients (n = 46) we determined the expression of P‐glycoprotein (P‐gp) immunocytochemically and correlated those results with the therapeutic response. We have evaluated 66 patients who showed the following responses: 28/66 complete remissions (CR), 16/66 resistant leukemias (RL) and 22/66 early deaths (ED). The PE2 value was significantly higher in patients with RL than in patients with CR (p<0.00375). CFU‐AML sensitivity to ara‐C and DNR alone was not different between response groups, but the difference in CFU‐AML sensitivity to the combination of two drugs between patients with CR and RL was not significant, although a trend was noted (p<0.06). P‐gp expression was found in only 1/18 patients who achieved CR but in 9/11 patients with RL and 7/11 patients with ED, which is a highly significant difference (p<0.0006). We concluded that both PE2 and P‐gp expression in AML cells are valuable predictors of therapeutic response in adult AML and should be included in creating the best therapeutic approach to AML patients.
COP‐BLAM regimen combined with granulocyte colony‐stimulating factor and high‐grade non‐Hodgkin's lymphoma
Authors: Niitsu N. , Umeda M.
Published: July 2010

Abstract: The clinical efficacy of COP‐BLAM chemotherapy combined with human recombinant granulocyte colony‐stimulating factor (G‐CSF) was evaluated in 104 previously untreated patients with non‐Hodgkin's lymphoma (NHL). According to the method of Laurence et al., a modified COP‐BLAM regimen was administered every 21 days. G‐CSF was added when the granulocyte count fell below 1000 × 109/l. Ninety‐eight of 104 (94.2%) patients achieved a complete remission; the 4‐year survival rate was 82.4% with a median duration of observation of 26 months. Survival was significantly longer in patients with low serum LDH levels, B‐cell type or low CRP or in earlier clinical stages, than in patients with high serum LDH levels, T‐cell type or higher CRP levels or in advanced clinical stages. The mean duration of administration of G‐CSF was 5.4 days. Twelve patients developed infections during treatment. The adverse effects of G‐CSF included interstitial pneumonia, bone pain and fever. Patients administered COP‐BLAM combined with G‐CSF achieved a high rate of remission and had a low incidence of infection. Nearly all the patients could be treated in 21‐day cycles. The results suggest that G‐CSF combined with COP‐BLAM was effective in treating NHL, because the patients can tolerate a higher dose of the anticancer agents.
Assessment of the value of treatment with granulocyte colony‐stimulating factor in children with acute lymphoblastic leukemia: a randomized clinical trial
Authors: Dibenedetto S. P. , Ragusa R. , Ippolito A. M. , Lo Nigro L. , Di Cataldo A. , D'Amico S. , Miraglia V.
Published: July 2010

Abstract: The present trial was designed to test the effects of G‐CSF on the duration of the second phase of induction chemotherapy in children with newly diagnosed acute lymphoblastic leukemia (ALL). A total of 32 patients were assigned randomly to a group that received (14 patients; group A) or a group that did not receive (18 patients; group B) G‐CSF (10 g/kg/day subcutaneously and daily) throughout of the second phase of induction therapy. One of 14 (7.1%) patients in group A and 2 of 18 (11.1%) patients in group B completed the course of chemotherapy within the planned time. The median length of this phase was 37 days (range, 29 to 65; mean, 40; SD, 8.6) for patients in group A and 36 days (range, from 29 to 55; mean, 38; SD, 7.4) for those in group B, and the difference was not statistically significant. The number of days during which patients had granulocyte counts of less than 2 × 109/l, the number of febrile episodes of unknown origin, the number of bacterial and fungal infections and the number of days of hospitalization did not differ in a statistically significant manner between the two groups. Our data suggest that G‐CSF supportive therapy may be unnecessary in children with neutropenia of short duration, for whom the risk of infection is low.
Study of the role of vitamin B12 and folinic acid supplementation in preventing hematologic toxicity of zidovudine
Authors: Falguera M. , Perez‐Mur J. , Puig T. , Cao G.
Published: July 2010

Abstract: A prospective, randomized study was conducted to evaluate the role of vitamin B12 and folinic acid supplementation in preventing zidovudine (ZDV)‐induced bone marrow suppression. Seventy‐five human immunodeficiency virus (HIV)‐infected patients with CD4 + cell counts < 500/mm3 were randomized to receive either ZDV (500 mg daily) alone (group I, n = 38) or in combination with folinic acid (15 mg daily) and intramascular vitamin B12 (1000 μg monthly) (group II, n = 37). Finally, 15 patients were excluded from the study (noncompliance 14, death 1); thus, 60 patients (31 in group I and 29 in group II) were eligible for analysis. No significant differences between groups were found at enrollment. During the study, vitamin B12 and folate levels were significantly higher in group II patients; however, no differences in hemoglobin, hematocrit, mean corpuscular volume, and white‐cell, neutrophil and platelet counts were observed between groups at 3, 6, 9 and 12 months. Severe hematologic toxicity (neutrophil count < 1000/mm3 and/or hemoglobin < 8 g/dl) occurred in 4 patients assigned to group I and 7 assigned to group II. There was no correlation between vitamin B12 or folate levels and development of myelosuppression. Vitamin B12 and folinic acid supplementation of ZDV therapy does not seem useful in preventing or reducing ZDV‐induced myelotoxicity in the overall treated population, although a beneficial effect in certain subgroups of patients cannot be excluded.
Hereditary anaemias and iron deficiency in a tribal population (the Baiga) of central India
Authors: Reddy P. H. , Petrou M. , Reddy P. A. , Tiwary R.S. , Modell B.
Published: July 2010

Abstract: We have studied the prevalence and molecular nature of hereditary anaemias (abnormal haemoglobins, β‐thalassaemia, α‐thalassaemia, and Glucose 6 phosphate dehydrogenase (G6PD) deficiency) in a primitive central Indian tribe, the Baiga. 43% of the population appear to be iron‐deficient. Hereditary anaemia gene frequencies are, sickle cell 0.0824, G6PD deficiency (in males) 0.0457, β‐thalassaemia 0.0057, and deletional α‐plus thalassaemia 0.65. Both – α3.7 and –α4.2 deletions were observed and non‐deletional α‐thalassaemia was suspected. The overall gene frequency of Xmn I + polymorphism (C→T – 158 cap site; upstream of G gamma region) is 0.35. This polymorphism is preferentially linked to βS genes. It appears that sickle cell disease covers a wide range of severity in the Baiga tribe based on higher mortality in the offspring of AS × AS parents (2.5/couple) compared to AA × AS (0.75/couple) and AA × AA (0.76/couple) parents. This is compatible with the high frequency of genetic modifying factors, i.e., the Xmn I polymorphism and α‐thalassaemia. The results also indicate that “normal” red cell values must be defined for each population where thalassaemias, G6PD deficiency and iron deficiency are common.
Development of a criterion for response to therapy at 6 months in multiple myeloma
Authors: Guillemin F. , Guerci A‐P. , Feugier P. , Péré P. , Pourel J. , Guerci O.
Published: July 2010

Abstract: To investigate the prognostic value of therapy at 6 months on survival in multiple myeloma, to develop a new criterion assessing response to initial therapy at 6 months, and to compare it to a current criterion. The types of initial and 6‐month therapy were considered in a prognostic factor analysis in 70 patients treated in routine practice. Using the response to initial therapy defined by the clinician's decision as grouping variable in this group, a discriminant analysis identified the characteristics of responder patients. The validity of the resulting criterion was tested in another test group. Its prognostic ability was compared to the CLMTF criterion (50% M‐component reduction from baseline). The therapy at 6 months, reflecting the clinician's appraisal of response to initial therapy, predicted survival significantly. A criterion combining two variables (M‐component change and haemoglobin level at 6 months) classified 70% and 72.4% of patients correctly regarding response status in the training and test groups respectively. This criterion was shown to perform better than the CLMTF criterion in predicting survival. Conclusion: A new criterion for response to therapy at 6 months, also presented in a nomogram, combines M‐component change and haemoglobin level at 6 months.
Low serum levels of interleukin‐6 in children with post‐infective acute thrombocytopenic purpura
Authors: Gangarossa Simone , Romano Vincenzo , Munda Silvana E. , Sciotto Antonella , Schiliro‘ Gino
Published: July 2010

Abstract: Interleukin‐6 plays an important role in host defense mechanisms and it appears to be a major mediator of the acute‐phase response. IL‐6 is also an important thrombocytopoietic factor. High serum levels of IL‐6 are present in reactive thrombocytosis. The number and function of circulating platelets are the major factors that affect megakaryocytopoiesis by thrombopoietin. High levels of thrombopoietin have been observed in patients with thrombocytopenic purpura. To evaluate a possible thrombopoietin‐like function of IL‐6, we measured IL‐6 levels in the serum of patients affected by post‐infective acute thrombocytopenic purpura using a sensitive ELISSA assay. As controls, we studied normal subjects and patients with reactive thrombocytosis. No significant difference was observed between thrombocytopenic patients and normal controls. High IL‐6 levels were present in patients with reactive thrombocytosis. In conclusion, we had not observed high levels of IL‐6 in acute thrombocytopenic purpura and, very probably, IL‐6 is not involved in the regulation of platelet mass for the hemostatic function. The thrombocytopoietic activity of IL‐6 is another acute‐phase response and it is consistent with the other functions of this cytokine. This suggests an active participation of platelets in host defense mechanisms.
Interleukin‐4 inhibits the production of interleukin‐1 by adult T‐cell leukemia cells
Authors: Mori Naoki , Shirakawa Fumihiko , Murakami Shuichi , Oda Susumu , Eto Sumiya
Published: July 2010

Abstract: Freshly isolated leukemic cells from patients with adult T‐cell leukemia (ATL) produce high levels of interleukin‐1 (IL‐1), which is believed to play an important role in neutrophilia, elevation of C‐reactive protein, osteolytic bone lesions, hypercalcemia, and fever in ATL. However, relatively little is known regarding the regulatory mechanism of IL‐1 production in ATL. Interleukin‐4 (IL‐4) affects the monocytes‐ and neoplastic cells‐mediated cytokine production. In this study, we investigated the effect of IL‐4 on IL‐1 α and IL‐1 β production by ATL cells in vitro. IL‐4 was found to markedly inhibit the release of IL‐1 α and IL‐1 β into the conditioned medium in a dose‐dependent manner. Northern blot analysis of steady‐state IL‐1 mRNA demonstrated that IL‐4 treatment of ATL cells resulted in a reduction of IL‐1 mRNA. These results support the notion that ATL cells spontaneously produce IL‐1 α and IL‐1 β; however, such production can be inhibited by the immunomodulating agent, IL‐4. IL‐4 may play an important regulatory role in the production of IL‐1 in ATL.
Association of Hb S/Hb lepore and δβ‐thalassemia/Hb lepore in Sicilian patients: Review of the presence of Hb lepore in Sicily
Authors: Mirabile E. , Testa R. , Consalvo C. , Dickerhoff R. , Schilirò G.
Published: July 2010

Abstract: The hemoglobin (Hb) lepore‐Boston is a β‐globin structural variant, produced in a reduced amount and formed from the fusion of N‐terminus δ‐(residues 1–87) and C‐terminus β‐chains (residues 116–146). This type of fusion protein is quite common in Southern Italy (Campania, Calabria, and Sicily). We report here the hematological and hemoglobin data on 96 unrelated Sicilians with Hb lepore trait. Particularly interesting are the subjects where Hb lepore occurs with Hb S or Sicilian type δβ‐thalassemia. In these individuals, striking features are clinical variability and different hematological pictures. These observations underscore the importance of thalassemia screening in these geographic areas, such as Southern Italy, principally Sicily, where the mutations in globin gene clusters are especially prevalent. Moreover, as from the second half of the last century, owing to high migratory flux from Sicily to Northern Europe, North and South America, and Australia, the Hb lepore, as well as other hemoglobin variants, have become prevalent, making the identification of the heterozygotes a problem of general interest.
Effect of cytokines on the toxicity of cytostatic drugs on leukemic cells in vitro and in vivo
Authors: Sundman‐Engberg Britt , Tidefelt Ulf , Paul Christer
Published: July 2010

Abstract: Most cytostatic drugs exert their effect on cells in active cell cycle. To improve the effect of cytostatic drugs we have tried, prior to treatment in vitro, to recruit tumor cells from G0 with growth factors. Leukemic cells from the bone marrows of 26 patients with AML and CML in blast crisis were incubated with G‐CSF, GM‐CSF and IL‐3 for 24 h prior to incubation with cytostatic drugs. The cells were incubated with mitoxantrone, etoposide or daunorubicin for 1 h, or with Ara‐C continuously. Prior to treatment, 4 patients with AML received GM‐CSF for 24 h, after which blast cells from bone marrow were incubated with cytostatic drugs. After incubation with the cytostatic drugs, cells were cultured in a suspension culture for 4 d. The drug effect was determined with a bioluminescence ATP method. Leukemic cells were significantly stimulated by all three cytokines compared to an untreated control. GM‐CSF and IL‐3 increased the amount of cells 3‐ to 4‐fold and G‐CSF increased the amount 3 times compared to untreated cells. G‐CSF significantly enhanced the cytotoxic effect of daunorubicin, mitoxantrone, etoposide and Ara‐C by 20–40%, while GM‐CSF and IL‐3 showed a significantly increased toxicity for Ara‐C only. Although the cytokines induced a higher percentage of cells killed with the cytostatic drugs, proliferation of the remaining cells resulted in an increased total number of cells from 1.5 to 3 times compared to the unstimulated incubations. We conclude that cytokines induce a higher level of toxicity of cytostatic drugs on leukemic cells, but the increased proliferation of the remaining cells may offset the clinical benefit.
A good response rate to recombinant erythropoietin alone may be expected in selected myelodysplastic patients. A preliminary clinical study
Authors: Di Raimondo F. , Longo G. , Cacciola E. , Milone G. , Palumbo G. A. , Cacciola R. R. , Alessi M. , Giustolisi R.
Published: July 2010

Abstract: The use of recombinant erythropoietin for treatment of anemia in myelodysplastic patients has so far produced poorer results than expected. Most clinical studies have been conducted without any selection of patients. In the present study we report our experience with the use of rhEPO in a population of selected MDS subjects. Only patients affected by refractory anemia according to FAB criteria, without significant WBC and platelets reduction, with normal LDH and short history of disease were eligible for the study and were treated with rhEPO at a dosage of 150 mg/kg three times a week for 2 months. Among 12 so treated patients, 7 (58.3%) achieved complete remission, 2 partial remission and 3 failed to respond. This high response rate makes more than acceptable the cost/benefit ratio for rhEPO in RA patients and may identify a subgroup of patients that can be treated successfully with rhEPO alone.
Prolongation of medium exchange is associated with a decrease in function but not expression of the P‐glycoprotein pump in leukaemic cells
Authors: Hegewisch‐Becker S. , Faltz C. , Hossfeld D. K.
Published: July 2010

Abstract: Daunorubicin (DNR) accumulation studies as functional tests of the multidrug resistance (MDR1) gene product P‐glycoprotein have produced diverging results when correlated to response to chemotherapy in acute leukaemia. To investigate possible reasons for this diversity a starvation experiment, based upon prolongation of medium exchange, was set up in the multidrug resistant cell line CEM/VBL100. DNR accumulation (1 μg/ml) was measured flow cytometrically in the presence or absence of Verapamil (10 μmol/1). In cells permanently kept under ideal growth conditions, addition of Verapamil resulted in an average 90% increase in DNR enhancement in five successive experiments. In contrast, DNR accumulation increased by only 26% when the medium exchange was prolonged by 30 h to 42 h. This effect was not accompanied by changes in the MDR1 gene expression at the RNA or protein level. Consequently, 53 leukaemic blast samples of 30 newly diagnosed and 18 relapsed or refractory patients with acute leukaemia (ALL‐18, AML‐37) were processed without any delay and under the most stringent conditions possible. Evidence of the classical MDR phenotype was arbitrarily defined by a greater than 20% enhancement in DNR accumulation in response to Verapamil (10 μmol/1) or Cyclosporin A (3 μmol/1). Using this cutoff point for analysis of newly diagnosed leukaemia we found DNR uptake better correlated to response to treatment (p = 0.002) than P‐gp detection by means of immunocytochemistry, using a panel of monoclonal antibodies (p = 0.03). We conclude that DNR accumulation studies are a sensitive method for predicting therapy outcome in acute leukaemia when performed with necessary precautions.
Influence of selected clinical events on the need for red blood cell transfusion in patients with severe cytopenia following myeloablative chemotherapy
Authors: Friis L. S. , Christensen B. E.
Published: July 2010

Abstract: The transfusional need in patients treated with myeloablative chemotherapy appears to be greater than that resulting from bone marrow suppression alone. The erythrocyte transfusional need may be expressed as Δhb = the daily loss of haemoglobin (mmol/1/day), and multivariate analyses of factors influencing Δhb have been carried out. We have studied 124 patients, mainly with acute myeloid leukaemia, treated with 537 courses of chemotherapy, inducing cytopenia (a white cell count < 1.0 times 109/1) in 476 cases. The transfusional need was 2.5–3 times greater than expected, suggesting the presence of haemolysis and/or occult bleedings. Using multiple regression analysis and repeated measurement analysis the main factor influencing the transfusional need was type/dose of chemotherapy. The higher the dose of cytarabine the greater the transfusional need. Other factors significantly influencing the transfusional need were tumour burden, fever, treatment‐related events such as positive blood cultures, systemic fungal infection, mucosal bleeding, melaena/haematemesis and diarrhoea. Finally, it was found that the transfusional need decreased gradually during the cytopenic period.
Osteocalcin is not a marker of progress in multiple myeloma
Authors: Mejjad O. , Loët X. Le , Basuyau J. P. , Ménard J. F. , Jego P. , Grisot Ch. , Daragon A. , Grosbois B. , Euller‐Ziegler L. , Monconduit M. ,   le Groupe d'Etude et de Recherche sur le Myélome (GERM)
Published: July 2010

Abstract: The aim of this study was to evaluate the usefulness of serum osteocalcin (OC) levels in multiple myeloma (MM) in order to assess its significance and activity, and to predict its course. Serum OC was measurement in 117 patients with MM and 116 healthy controls matched for age and sex. Serum OC levels were weakly correlated with Karnofsky index (r = 0.22; p<0.03). Lowest OC levels were observed when lytic bone lesions increased (p <0.05). There was no relationship between serum OC levels and vertebral crush fractures, serum calcium concentrations or stage of MM, neither was there any relationship between initial serum OC levels and survival. Progression of the disease was associated with a clear fall of serum OC in 61.5% of the “progressive” patients, versus 41% of the persisting “stabilized” cases. Serum OC level was strongly correlated with bone formation (p = 0.005), but not with bone resorption. Serum OC level is a sensitive marker of osteoblast activity, but a poor marker of the severity of MM. We do not consider it as a marker of MM activity or prognosis.
Inhibition of hypercoagulation by antithrombin substitution in E. coli L‐asparaginase‐treated children
Authors: Nowak‐Göttl U. , Kun N. , Wolff J. E. A. , Boos J. , Kehrel B. , Rath B. , Jürgens H.
Published: July 2010

Abstract: Acquired deficiency of antithrombin (AT), which in some patients could lead to thrombosis, has been a serious side effect of protocols which incorporate E. coli L‐asparaginase (ASP) for the treatment of acute lymphoblastic leukaemia (ALL). In a longitudinal, prospective, non‐randomized study children with ALL (n = 27) were treated according to the protocol ALL‐BFM‐90. During the induction phase using prednisone, vincristine, daunorubicin and ASP, AT substitution was performed in 15/27 patients, when their plasma concentration decreased below 60% of normal with a concomitant increase of D‐dimer formation. After the administration of the AT concentrate the patients, plasma concentration of AT increased and remained elevated after 18, 48 and 72 h. In addition, the plasma concentration of enhanced thrombin generation, D‐dimer formation and plasminogen activator inhibitor 1 decreased towards normal levels. Although the observed laboratory findings may serve as evidence for a possible clinical benefit of AT substitution during ASP treatment, further randomized studies are requested to evaluate whether the use of prophylactic AT administration could reduce the incidence of thromboembolic events in childhood acute leukaemia.
Correlation between argyrophilic nucleolar organizer region counts and labelling index in multiple myeloma
Authors: Marmont F. , Pich A. , Chiusa L. , Locatelli F. , Falda M. , Boccadoro M. , Resegotti L.
Published: July 2010

Abstract:The correlation between the bromodeoxiuridine (BrdU)‐labelling index (LI) of plasma cells and a new proliferation marker, the Argyrophilic Nucleolar Organizer Regions (AgNORs), was investigated in 44 myeloma patients at diagnosis. A preliminary analysis was made to verify the reproducibility of the assessment of plasma cell infiltration (PC%) in bone marrow aspirates, used to collect cells for LI determination, and in bone marrow biopsies, used for AgNORs evaluation. Although an overall good correlation was observed between PC% in biopsies and aspirates (r= 0.58,p= 0.001), the ratio between PC% in biopsies and in aspirates ranged from 0.35 to 7.5. Only 17 patients (38.6%) were within the 0.5–1.5 range. A positive correlation between LI and AgNORs was observed in these patients (r= 0.68,p= 0.003), whereas the correlation was lost in patients with higher ratio between PC% in biopsies and in aspirates (r= 0.08,p= 0.69). The prognostic significance of AgNORs was confirmed by survival analysis, showing a reduced survival for patients with high (>4.4) AgNOR counts (14 monthsvs35 months,p= 0.004). The AgNORs analysis therefore allows the simultaneous evaluation of myeloma cell infiltration, degree of differentiation and kinetics of growth in bone marrow biopsies. AgNOR counts deserve to be included in the procedures for diagnosis and prognostic evaluation of myeloma patients.
Angiogenesis spectrum in the stroma of B‐cell non‐Hodgkin's lymphomas. An immunohistochemical and ultrastructural study
Authors: Ribatti D. , Vacca A. , Nico B. , Fanelli M. , Roncali L. , Dammacco F.
Published: July 2010

Abstract: Samples of lymph nodes from 88 patients with B‐cell non‐Hodgkin's lymphoma (B‐NHL) grouped by the Working Formulation (WF) and from 15 patients with benign lymphadenopathies were investigated immunohistochemically and ultrastructurally for changes in angiogenesis and stromal distribution of two subendothelial basement membrane (BM) components, namely laminin and type IV collagen. The microvessel number was usually low in lymphadenopathies, and increased significantly in low‐grade B‐NHL. Intermediate‐grade tumors displayed a further significant increase that was mainly due to their diffuse subtypes rather than to the follicular subtype. High‐grade B‐NHL showed the highest counts. By contrast with the lymphadenopathies studied, the stroma of B‐NHL reacted intensely with both BM components, whose linear co‐expression was significantly associated with low‐grade and follicular intermediate‐grade B‐NHL, while expression of laminin alone in a granular pattern was detected in diffuse intermediate‐grade and high‐grade tumors. Ultrastructural analysis revealed immature vessels more frequently in diffuse intermediate‐grade, and in high‐grade B‐NHL. These in situ data suggest that angiogenesis occurring in B‐NHL increases along their progression path, and emphasize the importance of angiogenesis as an epigenetic phenomenon of B‐NHL progression.
Radioprotective effects of combination broncho‐vaxom, a macrophage activator, and indomethacin, an inhibitor of prostaglandin production: relationship to myelopoiesis
Authors: Fedoročko P. , Macková N. O.
Published: July 2010

Abstract: The effects of the bacterial extract broncho‐vaxom (BV; radioprotective immunomodulator; 500 μg/mouse i.p., 24 h) and indomethacin (INDO; inhibitor of prostaglandin production; 2 times 40 μg/mouse i.m., ‐ 24 h and ‐ 3 h) on the post‐irradiation recovery of hemopoietic functions in mice were investigated. Both agents were administered either alone or in combination. Endogenous spleen colony formation was increased in all treatment groups, with combination‐treated mice exhibiting the greatest effects. Similarly, 24 h after combined administration of BV and INDO (i.e. at the time of presumed irradiation) to the non‐irradiated mice granulocyte‐macrophage colony‐forming cell (GM‐CFC) numbers were greater in the bone marrow and spleen. Also, as determined by hydroxyurea injection, there was an increase in the number of GM‐CFC in the S‐phase of the cell cycle in the bone marrow. However, GM‐CFC in the spleen of combination pretreated mice was not stimulated to significant proliferation as compared to GM‐CFC in the spleen of mice injected with BV alone. Combined modality treatment was also more effective than single agent treatments in accelerating bone marrow cellularity and GM‐CFC regeneration, but not in accelerating GM‐CFC regeneration in the spleen. Combined administration of BV and INDO to mice prior to lethal irradiation exerted an additional radioprotective effect and protected 95% of the C57B1/6 mice.
3‘‐Azido‐ 3’‐deoxythymidine inhibits erythroid‐specific transcription factors in human erythroid K562 leukemia cells
Authors: Bridges E. G. , Trentesaux C. , Lahlil R. , Spiga M.‐G. , Jeannesson P. , Sommadossi J.‐P.
Published: July 2010

Abstract:The present study examines genetic mechanism(s) possibly involved in the observed 3′‐azido‐3′‐deoxythymidine (AZT)‐induced inhibition of globin gene transcription by evaluating the direct phenotypic erythroid effects of AZT on erythroid‐specific transcription factors which regulate globin gene promoters. In vitro binding of GATA‐1 or NFE‐2 to its consensus sequence was decreased in the presence of AZT reaching a maximum inhibition as early as 24 h after AZT treatment. Nuclear extracts from butyric acid‐induced K562 cells treated with an IC50concentration of AZT exhibited a decrease in GATA‐1 and NFE‐2 binding by approximately 30% and 35%. In contrast, 2′,3′‐dideoxycytidine which inhibits cell growth without affecting hemoglobin synthesis, had no effect on binding of GATA‐1 and NFE‐2 factors. Northern blot analysis revealed a 25% decrease by AZT in GATA‐1 mRNA steady‐state levels at 24 h and this inhibitory effect was maintained until 72 h after drug addition. A similar decrease in NFE‐2 mRNA steady‐state levels was observed at 72 h after AZT treatment. This study suggests that AZT inhibition of erythroid differentiation is subsequent to a decrease of nuclear factors gene expression which affect their DNA binding.
NRAS mutations are rare in acute myeloid leukaemias with t(8;21) or inv(16)
Authors: Panagopoulos I. , Åman P. , Johansson B. , Malm C. , Lenhoff S. , Billström R. , Mitelman F.
Published: July 2010

Abstract: Using PCR and direct sequence methodology, 19 haematologic malignancies with trisomy 8, 18 with t(8;21)(q22;q22) and 8 with inv(16)(p13q22) were screened for NRAS mutations. Of the 45 samples analyzed, 4 (9%) had a mutation; both wild‐type and mutated alleles were observed in these 4 cases. Three of the mutations (involving codons 12 and 13) were found in the trisomy 8 group and 1 (codon 61) among the inv(16) samples. No specific clinical similarities were found in the 3 patients with + 8 and NRAS mutation. By analyzing two sequential samples from the patient with inv(16) and NRAS mutation, it was shown that the mutation had occurred after the inversion. Since no NRAS mutations were detected among the t(8;21) samples and only 1 was found in the inv(16) group, we conclude that acute myeloid leukaemias with t(8;21) or inv(16) generally arise and progress without the involvement of NRAS mutations.
Detection of platelet antibodies in chronic idiopathic thrombocytopenic purpura (ITP). A comparative study using flow cytometry, a whole platelet ELISA, and an antigen capture ELISA
Authors: Stockelberg D. , Hou M. , Jacobsson S. , Kutti J. , Wadenvik H.
Published: July 2010

Abstract: Chronic idiopathic thrombocytopenic purpura (ITP) is a consequence of rapid platelet destruction caused by circulating platelet antibodies. In this study we compared three methods for detecting serum platelet antibodies in a population of 65 patients with chronic ITP. In two of the techniques intact platelets were used as the antibody target, i.e. the whole platelet ELISA and the flow cytometric assay; in the third an antigen‐specific modified antigen capture ELISA (MACE) was employed. By using the whole platelet ELISA and the flow cytometric assay 35% and 45% of the patients, respectively, displayed an antiplatelet antibody. In most cases (26 of 29 patients) IgG was the predominant antiplatelet immunoglobulin. As analysed using the MACE‐technique glycoprotein (GP) Ib/IX‐specific antibodies occurred with the same frequency as antibodies specific for GPIIb/IIIa. Moreover, there was a poor correlation between the MACE results on the one hand and results from the intact platelet‐based techniques on the other, i.e. several patients were positive in one assay whereas they were negative in the other. We conclude that all three techniques have their merits and demerits; it appears reasonable that they should be used together in the evaluation of the autoimmune process of chronic ITP.
Prognostic impact of interferon alpha‐induced cytogenetic remission in chronic myelogenous leukaemia: long‐term follow‐up
Authors: Kloke O. , Niederle N. , Opalka B. , Hawig I. , Seeber S. , Becher R.
Published: July 2010

Abstract:To evaluate the long‐term impact of the reduction of Philadelphia chromosome (Ph)‐positive metaphases by treatment of chronic myelogenous leukaemia (CML) with interferon (IFN) alpha, we examined the outcome of 62 patients who had been enrolled between 1984 and 1990 into 2 IFN trials at our institution. As best cytogenetic response, 9 patients had achieved a complete remission and an additional 9 patients a partial remission. The remaining 44 patients had obtained either a minimal (n = 29) or no cytogenetic response (n = 15). Of the total of 62 patients, 9 were still on schedule and responsive to IFN in January 1995, including 7 patients in ongoing complete cytogenetic remission. The overall 5‐year survival rate after a median follow‐up from diagnosis of 51 months (range 3–102 months) was 62% and the median survival was reached at month 87. The effect of cytogenetic remission on survival was examined by “landmark” studies showing a significant survival advantage for patients with karyotype responses. In conclusion, in the patients studied, cytogenetic improvement was found to translate into improved survival expectancy. Long‐term control by IFN alpha of CML, however, was restricted to a small minority of patients, predominantly to those attaining a complete suppression of the leukaemic cell clone as judged by cytogenetic criteria.
Magnetic resonance imaging is superior to computed tomography and ultrasonography in imaging infectious liver foci in acute leukaemia
Authors: Anttila Veli‐Jukka , Lamminen Antti E. , Bondestam Sören , Korhola Ossi , Färkkilä Martti , Sivonen Aulikki , Ruutu Tapani , Ruutu Petri
Published: July 2010

Abstract: We conducted a prospective study in order to compare ultrasonography, computed tomography and magnetic resonance imaging in the detection of liver foci in patients with acute leukaemia and clinical suspicion of hepatic candidiasis. 28 adult patients fulfilling set entry criteria after recovery from neutropenia were studied. Lesions in the liver were detected by at least one imaging modality in 21 patients: by ultrasonography in 7 (33% of detected cases), computed tomography in 12 (57%) and by magnetic resonance imaging in 20 patients (95%). Magnetic resonance imaging was significantly more sensitive than ultrasonography (p<0.001) and computed tomography (p<0.02). The difference between computed tomography and ultrasonography was not statistically significant (p = 0.1). Invasive procedures performed in 10 patients provided definite proof of candidiasis in 5 patients, and nodes on the liver surface, compatible with yeast infection, were seen during laparoscopy in 3 other patients without proof of fungal infection. We confirm that magnetic resonance imaging is superior to ultrasonography and computed tomography in imaging liver foci in leukaemic patients recovering from neutropenia with persistent non‐specific signs of infection or hepatic involvement.
Cell differentiation in acute myeloid leukemia
Authors: Olsson Inge , Bergh Gösta , Ehinger Mats , Gullberg Urban
Published: July 2010

Abstract: Acute myeloid leukemia (AML) is characterized by a differentiation block leading to accumulation of immature cells. Chromosomal translocations in AML affect transcription factors that are involved in regulation of myeloid differentiation. Aberrant expression of these factors interferes with differentiation events and has a role in the pathogenesis of AML through superactivation or (dominant negative) repression of genes regulating proliferation and differentiation or by interference with assembly of the transcription complex for these genes. The maturation arrest can be reversed by certain agents as judged by results from investigations of myeloid leukemic cell lines and from treatment of acute promyelocytic leukemia (APL) patients with all‐trans retinoic acid. Inactivation of the p53 and retinoblastoma (Rb) tumor suppressor genes is also associated with the pathogenesis of leukemia through effects on the cell cycle, and manipulation of these genes can affect differentiation of AML cells. With differentiation therapy, when successful as in APL, the leukemic cell mass is reduced to allow restoration of normal hematopoiesis and clinical remission, but the disease is not cured. However, initial reduction of the cell mass by maturation can increase the probability for cure with chemotherapy. Overexpression of suppressor genes may increase the probability for differentiation. Most probably, particular molecular defects of subgroups of AML have to be explored to find optimal strategies for treatment including both blocking the cell cycle, promoting terminal differentiation, and inducing apoptosis as well as strengthening the immune response.
Monoclonal antibody‐based methods for quantitation of hemoglobins: application to evaluating patients with sickle cell anemia treated with hydroxyurea
Authors: Epstein Nava , Epstein Michael , Boulet Arielle , Fibach Eitan , Rodgers Griffin P.
Published: July 2010

Abstract: High‐titer monoclonal antibodies (mAb) were raised against chromatographically purified human hemoglobin (Hb) species.

These mAb were specific for either Hb A, Hb F, Hb S or Hb C. Based on these antibodies, which were directly conjugated with either fluorochromes or an enzyme (horseradish peroxidase), we developed immunoassays for determining the Hb profile in the peripheral blood; an enzyme‐linked immunosorbent assay (ELISA) for determining the absolute and relative quantities of various Hb species and one‐step immunolabeling for fluorescence microscopic and flow cytometric analyses of the distribution of RBC with respect to their Hb types. We utilized these methods for monitoring the Hb F level and the percentage of Hb F‐containing cells in patients with sickle cell anemia undergoing treatment with hydroxyurea.
Little evidence for clonal evolution of malignant haematopoietic cells following relapse after autologous bone marrow transplantation
Authors: Jørgensen Hanne , Hokland Peter , Jensen Arne Willy , Hokland Marianne
Published: July 2010

Abstract: By screening for immunoglobulin (Ig) and T‐cell receptor (TCR) gene rearrangements in bone marrow samples aspirated at different time points during the course of disease from 43 patients with acute leukaemia we have analysed the extent of clonal evolution after autologous bone marrow transplantation (ABMT) and addressed the issue of whether the Southern Blot method has the power to reveal clonal proliferations representing minimal residual disease (MRD) in the autologous bone marrow grafts. Our results show that no clonal proliferations were detectable in any of the 43 bone marrow grafts analysed, even after we analysed DNA preparations in 5 cases from cells highly enriched for cells of the original malignant immunophenotype. Moreover, as judged by the Ig‐ and TCR gene configurations in 11 patients, relapse arose from the original clone even though minor clonal variations did occur in about half of the relapsing patients. We conclude that while the Southern Blot method can detect gene receptor rearrangements in the majority of patients with acute leukaemias and high‐grade non‐Hodgkins lymphomas, it is not useful for predicting relapse after ABMT. On the other hand, it is possible – by employing it – to evaluate whether or not relapse after ABMT arises from the original malignant clone and to what extent clonal evolution has taken place.
In vivo biological response following low‐dose interleukin‐2 in complete remission B‐cell non‐Hodgkin's lymphoma patients
Authors: Vecchi Angela , Cavanna Luigi , Avanzini Paolo , Callea Vincenzo , Velardi Andrea , Albi Nicola , Tartoni Pierluigi , Bensi Laura , Valentini Paola , Longo Rosina , Sacchi Stefano ,   Italian Lymphoma Study Group(GISL)
Published: July 2010

Abstract: The aim of the present study is to verify whether recombinant interleukin‐2 (rIL‐2) at low doses is well tolerated in aggressive lymphoma in complete remission (CR), and if there may be a biological justification for its use as a remission‐maintenance therapy able to reduce the percentage of relapses. We treated 6 patients with B‐cell non‐Hodgkin's lymphoma (B‐NHL) in CR following PM‐Cytabom with rIL‐2 3 IMU s.c. x 5 d per wk, every other wk for 8 wk. Our results show that this treatment provokes statistically significant changes in the absolute number of lymphocytes, eosinophils, CD25+ and CD122+ cells and soluble IL‐2 receptors; these doses, however, are not sufficient to modify CD3+, CD16+ and CD56+ cell values or natural killer and lymphokine activated killer cell activity. Thus these findings do not appear to constitute a biological rationale for the use of rIL‐2 at this dose and schedule as a remission‐maintenance therapy in B‐cell NHL. Nevertheless, the results are a valid basis for further study of the use of the same rIL‐2 doses for a longer period of time in combination with other cytokines, in the hope that the biological effects can be augmented without increasing the toxic side effects.
Deficiency of P‐selectin in a patient with grey platelet syndrome
Authors: Mazurov Alexey V. , Vinogradov Dimitry V. , Khaspekova Svetlana G. , Krushinsky Anatoly V. , Gerdeva Ludmila V. , Vasiliev Sergei A.
Published: July 2010

Abstract: Patient B.G. is a 29‐yr‐old female with a lifelong bleeding disorder characterized clinically by a highly increased bleeding time, menorrhagias, long‐lasting bleeding after cuts and tooth extractions and large post‐traumatic haematomas. Her coagulation tests were within normal range, platelet count was 140,000–160,000 per μl, but platelet function was impaired as demonstrated by the absence of collagen‐induced aggregation, although no abnormalities were detected in aggregation response to ADP and ristocetin. Morphologically her platelets were characterized by gigantic size – average profile area was about 2.5 times higher than that of control donors, and severe deficiency of α‐granules – only 16% of their number in control donors. These features taken together indicated the diagnosis of grey platelet syndrome. As has been shown by quantitative immunoblotting, patient's platelets contained small amounts of α‐granule membrane protein P‐selectin – about 15% of that in control donors. The content of plasma membrane glycoproteins IIb–IIIa and Ib was not reduced, suggesting the specific deficiency of α‐granule membrane protein. Thus, B.G. is the second patient described in the literature (see also Lages et al, J Clin Invest 1991: 87: 919–929) with combined deficiency of α‐granules and P‐selectin.
Intensive remission induction therapy for chronic myeloid leukemia in blast phase with a goal of post‐remission bone marrow transplant →— a pilot study
Authors: Lipton J. H. , Messner H. A. , Curtis J. E. , Atkins H. L. , Minden M. D.
Published: July 2010

Abstract: An intensive protocol utilizing mitoxantrone, high‐dose cytarabine, vincristine, etoposide and methylprednisolone as induction therapy for chronic myeloid leukemia in blast transformation is described. Fourteen patients were treated, with a remission/second chronic phase achieved in 64%. None of the 3 patients older than 50 yr responded. Complete hematological responses were seen in 9 of the 11 younger patients, 4 of whom also became BCR‐ABL negative by Southern Blot analysis. Four patients went on to allogeneic bone marrow transplant. Median remission durations were 4.5 (1–5) and 8.5 (5–16) months in the non‐transplanted and transplanted cohorts, respectively. Median survival is 1.5 (0.5–3), 9.5 (7–14) and 17 (14–61 +) months in the non‐responding, responding non‐transplanted and transplanted cohorts, respectively. Toxicity, particularly gastrointestinal, was significant. This represents an aggressive protocol that should be reserved for patients who are potential transplant candidates.
Functional abnormalities in granulocytes predict susceptibility to bacterial infections in chronic lymphocytic leukaemia
Authors: Itälä M. , Vainio O. , Remes K.
Published: July 2010

Abstract: Leukocyte functions were studied in 22 patients with chronic lymphocytic leukaemia (CLL) and were related to the patients' susceptibility to infections. In CLL patients with a history of infections, compared with CLL patients without infections or healthy controls, there were significant impairments of most granulocyte functions; random migration, N‐formyl‐methionylleucylphenylalanine (fMLP) and C5a stimulated chemotaxis and chemiluminescence response were decreased. No differences in these functions between CLL patients without infections and healthy controls were observed. Phagocytosis and intracellular killing of granulocytes were intact in all patients with CLL. By univariate analysis, neutrophil count and serum IgG level also predicted susceptibility to infections. By multivariate analyses, granulocyte chemotaxis and chemiluminescence remained as statistically significant predictors of infections. The lymphocyte functions (mitogen‐induced lymphocyte proliferation and immunoglobulin production in vitro) were equally impaired in all patients with CLL and differed significantly from the respective functions in healthy control subjects. We conclude that impairments in granulocyte functions contribute to susceptibility to infections in CLL.