bluuebunny/crossref_metadata_2025 huggingface dataset:
Observations on two members of the Swedish family with congenital dyserythropoietic anaemia, type III
Authors: Wickramasinghe S. N. , Wahlin A. , Anstee D. , Parsons S. F. , Stopps G. , Bergstrom I. , Eriksson M. , Sandstrom H. , Shiels S.
Published: July 2010

Abstract: Two affected individuals of the Swedish family with CDA, type III, in which the disease is transmitted as an autosomal dominant character, were studied. Both cases displayed features hitherto undescribed in this family but described in patients with CDA, type III, in whom the inheritance may have been as an autosomal recessive character. Such features were: (a) haemosiderinuria, (b) grossly disorganised erythroblast nuclei, (c) differences in the ultrastructural appearances of individual nuclei within the same multinucleate erythroblast and (d) intraerythroblastic inclusions resembling precipitated globin chains. In both cases the giant mononucleate erythroblasts and the multinucleate erythroblasts had total DNA contents up to 28c (1c = haploid DNA content) and 48c respectively, and some DNA synthesising bi‐ and multinucleate erythroblasts contained one or more nuclei which were unlabelled with 3H‐thymidine. These findings are similar to those in patients with the autosomal recessive type of disease. Thus no major phenotypic differences are yet apparent between cases of CDA, type III, with different patterns of inheritance. Analysis of the surface erythrocyte proteins of the 2 Swedish CDA, type III, patients with monoclonal antibodies recognising Band 3, glycophorins A, B, C and D, Rh, CD44, CD47, CD55, CD58, CD59, Lutheran, Kell, LW and acetylcholinesterase did not reveal any gross abnormality of expression of these proteins. A slightly altered expression of blood group antigens A and H was revealed by the lectins Dolichos biflorus and Ulex europaeus and the Mr of Band 3 as judged by SDS polyacrylamide gel electrophoresis was also slightly reduced, suggesting that there may be minor alterations in the degree of N‐glycosylation of some red cell membrane constituents.
Glucose phosphate isomerase (GPI) “Morcone”: A new variant from Italy
Authors: Alfinito F. , Ferraro F. , Rocco S. , de Vendittis E. , Piccirillo G. , Sementa A. , Colombo M. B. , Zanella A. , Rotoli B.
Published: July 2010

Abstract: Here we report the 4th Italian case of glucose phosphate isomerase (GPI) deficiency. The propositus is a young man suffering from chronic haemolytic anaemia since birth with occasional transfusion requirement. Biochemical characterization of the defective enzyme revealed increased affinity for F‐6‐P, decreased affinity for G‐6‐P and marked thermoinstability. Electrophoretic mobility appeared normal. GPI from both parents showed similar but less pronounced biochemical alterations. The variant described here seems to be different from those previously reported. Thus, we propose the provisional name of GPI “Morcone”.
Increased intracellular concentrations of doxorubicin in resistant lymphoma cells in vivo by concomitant therapy with verapamil and cyclosporin A
Authors: Tidefelt Ulf , Juliusson Gunnar , Elmhorn‐Rosenborg Annika , Peterson Curt , Paul Christer
Published: July 2010

Abstract: The tumor cell uptake of doxorubicin was studied in vivo in cells from 2 patients with clinically resistant leukemic lymphomas treated with continous infusions of doxorubicin 9 mg/m2 and vincristine with oral dexamethason. After 24 hours, intravenous or oral verapamil was added and in one treatment course intravenous cyclosporin A was given. Plasma and intracellular doxorubicin concentrations and plasma concentrations of verapamil and norverapamil were determined with HPLC. In the 1st patient the intracellular uptake rate of doxorubicin was increased from 0.007 to 0.013 nmol/mg protein/h after the start of verapamil infusion. In the first treatment course of patient number 2, the intracellular concentration of doxorubicin was increased by 280% during a 6‐h infusion of verapamil. When this patient in the next treatment course was given oral verapamil, no significant effect on doxorubicin uptake was seen. However, when 100 mg of cyclosporin A was added in three intravenous injections at 8‐h intervals, the doxorubicin concentration in the tumor cells increased from 0.027 to 0.086 nmol/mg protein. In conclusion, this study shows that the intracellular concentrations of doxorubicin can be increased also in vivo during patient therapy by the addition of verapamil or cyclosporin A.
Incidence of circulating cells positive for GpIb and GpIIIa in infants with Down's syndrome
Authors: Fisher A.M. , Cockwell A. E. , Jacobs P. A. , Parry H. F.
Published: July 2010

Abstract: Blood samples from 43 infants with Down's syndrome and from 67 with a normal karyotype were screened for the presence of cells positive for platelet glycoproteins GpIb and GpIIIa. There was 1 case of fatal leukaemia and 1 case of transient leukaemia among the Down's syndrome patients, both cases had increased numbers of circulating blood cells positive for platelet glycoprotein markers. These cells were present in small numbers in both the Down's and control groups, especially among stillbirths, and there was no difference in the numbers of such cells found in Down's syndrome patients in comparison with the controls.
Pattern of iron excretion in relation to haemoglobin level and iron load in 8 haematological patients following the administration of subcutaneous deferrioxamine
Authors: Di Palma A. , Moratelli S. , Tolomelli P. , Giuberti M. , Tenan R. , Fagioli F. , Landi L. , Toffoli C. , Atti G. , Vullo C.
Published: July 2010

Iron excretion following subcutaneous administration of deferrioxamine (DFO) was measured between two transfusions of packed red cells in 6 patients with beta‐thalassaemia major on the high level Hb transfusion regime; and in a single 3‐day period in 2 other patients, 1 with transfused beta‐thalassaemia major and the other with haemolytic anaemia due to PK deficiency. The pattern of iron excretion did not change significantly during the period between the two transfusions and was found to be related to serum ferritin levels. The proportion of iron excreted in the stools was inversely related to the serum ferritin level. These observations on iron excretion are of practical importance in relation to DFO administration, especially when evaluated in thalassaemics with normal haemoglobin levels and low iron stores.
Comparison of younger versus older B‐cell chronic lymphocytic leukemia patients for clinical presentation and prognosis. A retrospective study of 53 cases
Authors: Molica Stefano , Brugiatelli Maura , Callea Vincenzo , Morabito Fortunato , Levato Domenico , Nobile Francesco , Alberti Antonio
Published: July 2010

Abstract: Fifty‐three patients affected with B‐cell chronic lymphocytic leukemia (CLL) younger than 50 years and observed in two hematological institutions have been retrospectively evaluated in order to verify whether this disease has different clinico‐hematological features at presentation and different prognosis as compared to older cases. In our experience young cases with B‐CLL diagnosis, confirmed by immunophenotype in 90.5% of patients, accounted for 7.1% of the whole CLL population. Sex distribution, mean peripheral lymphocyte count, platelet count, distribution among Rai's and Binet's stages, total tumor mass (TTM) score, histological pattern of bone marrow infiltration and lymphocyte doubling time (LDT) were similar to a series of 201 CLL cases older than 50 years. Only hemoglobin mean level was significantly higher in younger patients (13.1 ± 2.1 vs 12.2 ± 2.6 g/dl; p<0.01). The overall median survival was 7.1 years. Rai and Binet staging classifications and TTM score system retained their prognostic value in this CLL population. In addition, cases fulfilling criteria of “smoldering” CLL, had a very long survival (75% survival probability at 16 years). Life‐expectancy of younger patients was significantly longer than that of older ones (median survival, 7.1 versus 4.1 years; p < 0.05). However, when the background mortality due to non‐CLL related deaths (i.e., cardiovascular complications, epithelial cancers) was removed, survival advantage of young cases disappeared. In conclusion this study confirms that prognosis of young CLL patients can be easily assessed using the current well‐defined criteria. Since age is not by itself a criterion for intensifying treatment, further efforts to identify those young CLL patients who qualify for more aggressive therapy should be made.
Salvage therapy with low‐dose cytosine arabinoside in refractory or relapsed acute non‐lymphocytic leukaemia:
Authors: Jensen M. Krogh , Johansen P. , Stentoft J. , Jensen M. Krogh
Published: July 2010

Abstract: Thirteen patients with de novo acute non‐lymphocytic leukaemia (ANLL) refractory to standard chemotherapy for remission induction and 12 patients with ANLL in relapse were treated with low‐dose cytosine arabinoside (LD ara‐C) 10 mg/m2 subcutaneously every 12 hours for 21 days. Five of 13 patients (38%) and 6 of 12 patients (50%), respectively, obtained a complete remission (CR). Of these, 7 patients subsequently relapsed after 2–76 months, while 4 patients remain in CR after 7–131 months. Compared to standard intensive regimens treatment with LD ara‐C was rather non‐toxic, requiring platelet transfusions and antibiotics in only 6 and 13 cases, respectively. Three patients (12%) died during induction therapy with LD ara‐C; 2 had a cerebral haemorrhage and 1 developed anuria following a staphylococcol septicaemia. In conclusion, therapy with LD ara‐C may be preferable to more intensive and toxic regimens in the treatment of patients with relapsed or refractory ANLL.
Effects of alpha thalassaemia and haemoglobin F (HbF) level on the clinical severity of sickle‐cell anaemia
Authors: Falusi A. G. , Olatunji P. O.
Published: July 2010

Abstract:  Three clinical parameters — average steady‐state haematocrit (ASSH), number of crises per year (Cr/Y), and number of transfusions per year (Tx/Y) — were evaluated in 52 patients with sickle‐cell anaemia in relation to their foetal haemoglobin (HbF) levels. No correlation was observed between HbF and any of these parameters. A comparison of these three clinical parameters and the alpha globin gene status was also made in 28 of these patients. The relationships between (ASSH) or (Cr/Y) and alpha globin gene status were not significantly different (p>0.05) but a significantly different value (p<0.05) was observed between (Tx/Y) and the alpha globin gene status in these patients. It is concluded that, although HbF levels did not affect any of these parameters, alpha thalassaemia deletion significantly reduces the transfusion requirements of these patients.
Induction of apoptosis in childhood acute leukemia by chemotherapeutic agents: Failure to detect evidence of apoptosis in vivo
Authors: Matsubara K. , Kubota M. , Adachi S. , Kuwakado K. , Hirota H. , Wakazono Y. , Akiyama Y. , Mikawa H.
Published: July 2010

Abstract:  This study is designed to investigate whether apoptosis occurs in vivo in pediatric patients with acute leukemia during induction therapy. When patients with common acute lymphoblastic leukemia (cALL) and acute myeloblastic leukemia (AML) were treated with prednisolone (60 mg/m2/day, p.o. or i.v.) and etoposide (150 mg/m2/day, i.v.), respectively, the blast cell counts fell to below 30% and 5%, respectively, in 1 week. However, during this cytoreduction phase, neither morphologically apoptotic cells nor fragmentation of DNA derived from peripheral blast cells were detected at any preparations. On the other hand, cALL but not AML cells spontaneously undergo apoptosis following their culture in vitro. The addition of autologous serum instead of fetal calf serum substantially prevented apoptosis from occurring spontaneously in cALL cells. When cALL and AML cells freshly obtained from patients before therapy were treated in vitro with 10 μmol/1 prednisolone and 20 μg/ml etoposide, respectively, these cells underwent apoptosis within 6 hours, as determined by a morphological and DNA fragmentation assay. These in vivo and in vitro findings suggest that, although anticancer drugs may induce apoptosis in vivo, these apoptotic cells cannot be detected due to their rapid removal from the circulation.
Hereditary elliptocytosis associated with spectrin Le Puy in a Japanese family: Ultrastructural aspect of the red cell skeleton
Authors: Maréchal Joelle , Wada Hideho , Koffa Trigona , Kanzaki Akio , Wilmotte Rick , Ikoma Kimiko , Yawata Ayumi , Inoue Takafumi , Takanashi Kaoru , Miura Akira , Alloisio Nicole , Delaunay Jean , Yawata Yoshihito
Published: July 2010

Abstract:  A dominantly‐inherited hereditary elliptocytosis of intermediate severity was recorded in a Japanese family from Yamagata. The condition was associated with a spectrin truncated β‐chain (MW: 214 kD; 31% of total β‐spectrin), and a defect of mutant spectrin as regards tetramerization and phosphorylation. cDNA analysis revealed skipping of exon X, the third‐to‐last exon of the spectrin β‐gene. At the gene level, a one‐base substitution (A→G) changed position +4 of the 5′ donor splice site consensus sequence of intron X. This mutation has been described before in a French kindred, defining spectrin Le Puy. Electron micrographs following quick‐freeze deep‐etching showed that the skeletal network was disorganized.
Haemoglobin Köln as de novo mutations in Sweden: Diagnosis by PCR and specific enzymatic cleavage
Authors: Landin Britta , Fröstad Björn , Brune Mats , Ljung Rolf
Published: July 2010

Abstract:  Three independent cases of chronic haemolytic anaemia in Sweden have recently been demonstrated to be due to the unstable haemoglobin variant Hb Köln. The patients, all of whom have partially compensated chronic haemolytic anaemia, presented with aggravated haemolysis during acute infections in childhood. In one case, acute B19 parvovirus infection induced an aplastic crisis. The substitutions all seem to have occurred as de novo mutations. Diagnosis was based on haemoglobin instability testing and isoelectric focusing of haemoglobin dimers. The final identification procedure for the substitutions included extraction of DNA from whole blood, polymerase chain reaction (PCR) amplification of parts of the β‐globin gene and nucleotide sequencing of the resulting material, or studies of restriction length polymorphisms (RFLPs) using the restriction endonucleases Mae II or Nla III. The use of PCR‐RFLP is recommended as a valuable tool for diagnosing Hb Köln.
Analysis of the partial nucleotide sequences and deduced primary structures of the protease domains of mammalian blood coagulation factors VII and X
Authors: Murakawa Masahiro , Okamura Takashi , Kamura Takumi , Kuroiwa Mika , Harada Mine , Niho Yoshiyuki
Published: July 2010

Abstract:  In order to obtain sequence data for the blood coagulation factor VII and factor X in several mammalian species, we amplified and sequenced the DNA segments of exon VIII from each gene by means of the polymerase chain reaction (PCR) method. The DNA segments from the following species were successfully amplified: factor VII from the rhesus monkey and dog, and factor X from the rhesus monkey, Syrian hamster and rat. In each factor, the nucleotide sequences and predicted primary structures of the protease domain showed a high degree of homology among species; amino acid identities of approximately 68%‐92% and 80%‐98% were demonstrated among species in factor VII and factor X, respectively. The locations of the active site residues and five Cys residues were evolutionarily conserved in both factors. Interestingly, the amino acids involved in the human genetic variants, both factor VII 304‐Arg and factor X 326‐Arg, were always conserved across species. The data presented here will be helpful for investigating human genetic variants of factor VII or X, and will provide considerable information for constructing in vitro site‐specific mutants of these factors.
Effect of recombinant human erythropoietin on platelets in patients with anemia of renal failure: correlation of platelet count with erythropoietic activity and iron parameters
Authors: Beguin Yves , Loo Martine , R'Zik Samir , Sautois Brieuc , Lejeune Françoise , Rorive George , Fillet Georges
Published: July 2010

Abstract:  We examined the effect of treatment with rHuEpo on platelet counts in 61 hemodialysis patients and correlated them with changes in erythropoietic activity, iron status and inflammation. Platelets (109/l) increased from 220 ±80 to 245 ±102 after 14 days and stabilized at that level up to day 90 (p<0.0001). The increment was similar in complete or partial responders but was not observed in failures. Serum transferrin receptor (sTfR, a measure of total erythropoiesis) and Hct rose much more progressively, but relative platelet increments correlated with relative increases in sTfR and Hct. Relative platelet increments correlated inversely with relative changes of SeFe or transferrin saturation, but not with their absolute values, nor with baseline ferritin or its progressive decrease. Although baseline platelet count was 12% higher in patients with inflammation and correlated with serum haptoglobin, relative increases were similar in patients with or without inflammation. In conclusion, rHuEpo produced a clinically minor but consistent elevation of platelet counts. These modifications were not related primarily to modifications in iron stores, functional iron deficiency, or inflammation, but paralleled the expansion of erythropoietic activity. The results suggest that rHuEpo has a small positive effect on platelet production, but it cannot be ruled out that this could be partially mediated through functional iron deficiency.
Cell function studies in long‐term bone marrow culture after cell concentration and cryopreservation for autologous transplantation
Authors: Gilabert R. , Ayats R.
Published: July 2010

Abstract: Autologous bone marrow transplantation (ABMT) is frequently used in the treatment of neoplastic diseases. It involves several manipulations in vitro that can damage the stem cells responsible for grafting. Long‐term bone marrow cultures (LTBMC) reproduce the bone marrow microenvironment and support haematopoiesis in vitro for several weeks. This technique provides information on the extent of the injury of stem cells during manipulations of bone marrow cells in vitro. We studied the effect of the usual process of bone marrow cells in vitro in autologous transplantation: cell concentration and cell cryopreservation. 28 bone marrows from healthy donors and 45 from patients who had undergone ABMT were assayed in LTBMC. The cultures were initiated after manual or automatic buffy coat cell separation or after density gradient or automatic mononucleated cells suspension. Bone marrows were studied pre‐ and post‐cryopreservation. The results show that several manipulations can disturb normal cell behaviour in LTBMC. Following automatic mononucleated cell separation and after thawing, the development of adherent cell layer in LTBMC is anomalous. These manipulations led to a delay in covering 50% of the flask surface, absence of adypocytes in adherent cell layer and absence of haemopoietic precursors in the supernatant at the 4th week of culture. The results suggest that LTBMC can be used for control of the manipulations in vitro related with ABMT.
Circulating megakaryocytic progenitor cells in patients with primary thrombocythaemia and reactive thrombocytosis: results using a serum‐deprived culture assay and a positive detection technique
Authors: Sawyer B. M. , Westwood N. B. , Pearson T. C.
Published: July 2010

Abstract: A serum‐free culture method was used to study the growth of megakaryocytic progenitor cells (CFU‐Meg) from patients with elevated platelet counts. The culture technique was combined with immunocytochemistry (APAAP, CD61) for the identification of CFU‐Meg derived cells in cytopreparations of cells eluted from the culture dishes. Twenty‐six patients with primary thrombocythaemia (14 untreated patients, UPT, 12 treated patients, TPT), 14 patients with reactive thrombocytosis (RT) and 9 normal individuals were studied. Unstimulated growth of CD61‐positive cells was detected in 8/14 UPT, 8/12 TPT, 12/14 RT and 5/9 normal subjects (with mean CD61‐positive cell counts of 75, 579, 236 and 7 per cytopreparation respectively). Cultures supplemented with interleukin 3 contained CD61‐positive cells in 11/14 UPT, 7/12 TPT, 14/14 RT and 5/9 normal subjects (with mean CD61‐positive cell counts of 157, 589, 250 and 7 per cytopreparation respectively). Thus, this serum‐free culture technique combined with sensitive positive identification of CFU‐Meg derived cells failed to discriminate between PT and RT. These results cast doubt on the usefulness of serum‐free culture assays for the detection of unstimulated CFU‐Meg growth in the differential diagnosis of patients with elevated platelet counts.
Increased peripheral blood normal myeloid progenitor cells (CFU‐GM) in chronic lymphocytic leukemia: A perspective for autologous peripheral blood stem cell transplantation
Authors: Tsatalas Costas , Chalkia Panayota , Athanasiadis George , Venizelos Ioannis , Karakolios Apostolos , Papageorgiou Athanasios , Kakoulidis Ioannis , Bouricas George
Published: July 2010

Abstract: We assayed granulocyte‐macrophage committed progenitor cells (CFU‐GM) in the peripheral blood of 34 patients with chronic lymphocytic leukemia (CLL) and 12 normal individuals. The patients were divided into separate groups on the basis of previous therapy (i.e. analysis performed at diagnosis, during and after chemotherapy) and clinical features of the disease (i.e. disease stage, pattern of bone marrow infiltration, peripheral blood lymphocytosis). The mean CFU‐GM colony count of the patients was 30 times higher than that of the controls (206.4 ± 197.8 (SD) CFU‐GM per 5 times 105 cells plated versus 6.5 ± 3.6). There was no statistical difference in the numbers of circulating CFU‐GM between the patients studied at diagnosis (257 ± 215.4 CFU‐GM/5 times 105 cells) and those studied during (117.6 ± 169.2 CFU‐GM/5 times 105 cells) or after chemotherapy (207.5 ± 105.9 CFU‐GM/5 times 105 cells), although a trend towards a higher recovery of myeloid progenitors was observed as a function of time elapsing from the last treatment. In addition, we found no significant difference in the in vitro CFU‐GM growth of patients grouped according to their disease stage, pattern of bone marrow infiltration and degree of peripheral blood lymphocytosis. In conclusion, our data indicate that intensification with peripheral blood stem cell support may be feasible in CLL, since progenitor cells of myeloid‐monocytic series are markedly increased in the peripheral blood of these patients. Moreover, it is possible to extend this kind of therapy to patients who have undergone previous extensive chemotherapy and who might have persisting bone marrow infiltration.
Effects of granulocyte‐macrophage colony stimulating factor produced in Chinese hamster ovary cells (regramostim), Escherichia coli (molgramostim) and yeast (sargramostim) on priming peripheral blood progenitor cells for use with autologous Done marrow after high‐dose chemotherapy
Authors: Hussein Atif M. , Ross Maureen , Vredenburgh James , Meisenberg Barry , Hars Vera , Gilbert Colleen , Peters William P. , Coniglio David , Kurtzberg Joanne , Rubin Peter , Peters William P.
Published: July 2010

Abstract: Peripheral blood progenitor cells (PBPCs) were collected without prior association with chemotherapy but after the administration of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) produced in Chinese hamster ovary cells (CHO‐GM, regramostim), Escherichia coli (E. coli‐GM, molgramostim), or yeast (Yeast‐GM, sargramostim) and used in conjunction with autologous bone marrow after high‐dose chemotherapy in 69 patients with breast cancer or melanoma. The mean peripheral white blood cell (WBC) counts increased by 2.2 to 2.7‐fold after regramostim, 4.5 to 7.3‐fold after molgramostim and 4.3‐fold after sargramostim. All patients underwent three leukaphereses. The mean (& standard error) total nucleated pheresed cells per kg × 108 were 4.15 & 0.56, 15.10 & 1.77 and 7.24 & 1.00 for patients receiving regramostim, molgramostim or sargramostim respectively. The mean (& standard error) granulocyte‐macrophage colony‐forming units per kg × 104 mobilized into the PB were 8.75 & 3.63, 71.03 & 17.85, and 65.11 & 18.74 for patients receiving regramostim, molgramostim, or sargramostim respectively. The total mean (& standard error) CD34+ cells per kg × 107 collected by three leukaphereses were 3.28 & 1.62, 1.34 & 0.51 and 2.57 & 1.93, for patients receiving regramostim, molgramostim or sargramostim respectively. The use of either molgramostim‐ or sargramostim‐primed PBPCs led to complete elimination of absolute leukopenia with a WBC count under 100/mm3 in 64% and 77% of patients treated, respectively. Patients receiving molgramostim‐primed PBPCs required fewer red blood cells transfusions than patients receiving regramostim‐primed PBPCs (p = 0.0062). Our data indicate that PBPCs collected without prior association with chemotherapy but after either molgramostim or sargramostim with autologous bone marrow support and GM‐CSF shorten the hematopoietic recovery after myeloablative chemotherapy in patients with breast cancer or melanoma.
Intensive treatment of AIDS‐related non‐Hodgkin's lymphomas with the MACOP‐B protocol
Authors: Schürmann D. , Grünewald T. , Weiß R. , Jautzke G. , Pohle H. D. , Ruf B.
Published: July 2010

Abstract: The usefulness of intensive chemotherapy with the MACOP‐B protocol was evaluated in 8 patients with AIDS‐related non‐Hodgkin's lymphoma (NHL). Four patients had a prior AIDS diagnosis. The median CD4+ lymphocyte count was 0.079 cells × 109/1 (range 0.016–0.330). All patients responded to treatment. Four patients finished chemotherapy, all with complete remission, while another 3 patients deteriorated prior to finishing treatment and died. The median survival was 4 months (range 1 to 86 months). Major causes of the poor outcome were AIDS‐related opportunistic infections and meningeal CNS involvement by NHL developing during or after chemotherapy. Patients with AIDS‐related NHL usually do not appear to benefit from treatment with MACOP‐B protocol. Advanced immunodeficiency is associated with poor tolerance to treatment and inability to finish this chemotherapy protocol. MACOP‐B chemotherapy does not prevent meningeal spread of lymphoma in spite of using repeatedly systemic methotrexate crossing the blood‐brain barrier. CNS prophylaxis with repeated application of intrathecal methotrexate may lower the risk of meningeal spread of lymphoma, which developed in 1 of 5 patients given CNS prophylaxis as compared to 2 of 3 patients without CNS prophylaxis.
All trans retinoic acid enhances human LAK activity
Authors: Fegan C. , Bailey‐Wood R. , Coleman S. , Phillips S. A. , Neale L. , Hoy T. , Whittaker J. A.
Published: July 2010

Abstract: All trans retinoic acid has various effects on normal and malignant cells. Lymphokine‐activated killer (LAK) activity can be derived from T lymphocytes and natural killer cells. This study shows that all trans retinoic acid significantly enhances this activity by increasing production of tumour necrosis factor and gamma interferon, which results in enhanced expression of the p55 part of the interleukin 2 receptor. This effect is dependent on the concentration of all trans retinoic acid and the length of time of culture.
Investigation of microcytosis: a comprehensive approach
Authors: Mach‐Pascual Sara , Darbellay Regis , Pilotto Pierre‐Antonio , Beris Photis
Published: July 2010

Abstract: Microcytosis is a highly prevalent finding during blood examination. This study investigates the causes of microcytosis (defined as mean corpuscular volume (MCV)<82 fl) in 466 patients referred to our laboratory for suspected hemoglobinopathy. The following data were obtained: Hb, MCV, serum iron, transferrin, ferritin, HbA2, HbF, isoelectric focusing of the Hb, gene mapping of chromosome 16 with Xba I and Bgl II and hybridization with an α‐ and a ζ‐probe, inflammatory status. Results show that iron deficiency remains the first cause of microcytosis (35.2% of our patients), even in a selected population such as ours. Deletional α‐thalassemia, probably the most frequent hemoglobinopathy throughout the world, represents the second most frequent cause of microcytosis (31.1%), followed by β‐thalassemia heterozygous state (18.9%). Of our patients, 1.3% had microcytosis due to the presence of an abnormal hemoglobin (HbC, Hb S/C, HbE). Three cases (0.6%) had other possible causes of microcytosis. Of the remaining 60 cases, 28 had an inflammatory state. Finally, 32 cases (6.9%) remain unexplained; taking into consideration the origin of these cases, their hematological parameters and their family history, we postulate that these cases are at high risk for non‐deletional α‐thalassemia.
The use of recombinant SCF protein for rapid determination of c‐kit expression in normal and abnormal erythropoiesis
Authors: McGuckin C. P. , Uhr M. R. , Liu W.‐M. , Gordon‐Smith E. C.
Published: July 2010

Abstract: Stem cell factor (SCF) is the ligand for the dimeric c‐kit tyrosine kinase receptor. Binding of SCF to c‐kit is a crucial element in the developmental stimulus of late stem cells and early progenitor cells. In the erythroid lineage the SCF stimulus is important not only for proliferation and differentiation, but is also known to enhance later haemoglobin production. In an earlier report we described a rapid nonradioactive technique using the extended ester‐attached labelled SCF protein itself for detecting c‐kit expression in marrow and peripheral blood mononuclear populations. In the present study we have taken this a step further to analyse c‐kit expression in developing erythroid cells in vitro, principally using normal donor samples. This was designed for use as a foundation for the comparison of haematological disorders. In this case we tested 4 patients with the congenital disorder of erythropoiesis, Diamond‐Blackfan anaemia (DBA), finding that in all cases DBA c‐kit expression was elevated over normal, in 1 case as high as 348% of the normal average. This may be indicative of the reduced state of progenitor development in these patients. These results show that the described technique is beneficial for analysis in the stem and progenitor compartment.
Monocytic differentiation and synthesis of proteins associated with apoptosis in human leukemia U‐937 cells acquiring resistance to Vincristine
Authors: Pantazis Panayotis , Chatterjee Devasis , Han Zhiyong , Wyche James , DeJesus Albert , Giovanella Beppino
Published: July 2010

Abstract: Human leukemia U‐937/WT cells were exposed to stepwise increased concentrations of Vincristine so that Vincristine‐resistant cell sublines (termed U‐937/RV) were developed. Established U‐937/RV cell sublines have continuously propagated over a year, both in absence and presence of VCR, and have demonstrated similar features. In contrast to U‐937/WT cells, U‐937/RV cells have longer doubling time, and are more differentiated as determined by appearance of distinct morphological features and synthesis of mRNA that codes for the monocyte colonystimulating factor‐1 receptor (c‐fms). Both apoptosis‐suppressing Bcl‐2 and Bcl‐XL proteins were undectable in U‐937/WT cells, whereas Bcl‐2 was nearly detectable and Bcl‐XL readily detectable in U‐937/RV cells. The apoptosis‐promoting Bax protein was also absent in U‐937/WT cells and readily detected in U‐937/RV cells. Vincristine‐resistant cells with different levels of resistance synthesize similar levels of c‐fms mRNA and Bax protein. Finally, unlike U‐937/WT cells, U‐937/RV cells have no ability to induce tumors when xenografted in immunodeficient mice. The findings collectively suggest that development of resistance to Vincristine in U‐937/WT cells may correlate with cell differentiation and synthesis of proteins that regulate apoptosis.
Biweekly COP‐BLAM (cyclophosphamide, vincristine, prednisone, bleomycin, doxorubicin and procarbazine) regimen combined with granulocyte colony‐stimulating factor (G‐CSF) for intermediate‐grade non‐Hodgkin's lymphoma
Authors: Niitsu Nozomi , Umeda Masanori
Published: July 2010

Abstract:  We evaluated the efficacy and adverse effects of biweekly COP‐BLAM (cyclophosphamide, vincristine, prednisone, bleomycin, doxorubicin and procarbazine) therapy combined with granulocyte colony‐stimulating factor (G‐CSF) for treating non‐Hodgkin's lymphoma (NHL). A complete remission was achieved in 65 (90.3%) of the 72 patients. The median follow‐up period was 28 months, and 64 patients were alive at the time of writing. Treatment was delivered as scheduled to 61 patients. G‐CSF made it possible to shorten the interval between courses of chemotherapy. One of the 72 patients died of granulocytopenia and pneumonia; no other severe infections were noted. Further studies regarding adverse effects on organs other than the bone marrow are required to improve the long‐term results of combination therapy on NHL.
Biological and clinical significance of in vitro prednisolone resistance in adult acute lymphoblastic leukaemia
Authors: Tosi P. , Visani G. , Ottaviani E. , Manfroi S. , Tura S.
Published: July 2010

Abstract:  It has been reported that in vitro prednisolone (PDN) resistance provides a prognostic value in childhood acute lymphoblastic leukaemia (ALL). This study aimed at investigating the biological and clinical significance of in vitro PDN resistance in adult ALL. Blast cells from 30 patients were exposed to PDN (0.1 μm–35 μm) and cytotoxicity was determined by the soluble tetrazolium formazan 2,3‐bis (2‐methoxy‐4‐nitro‐5‐sulphophenyl)‐5‐[(phenylamino) carbamyl]‐2H‐tetrazolium hydroxyde (XTT) colorimetric assay. The IC50 (defined as the drug concentration that results in 50% growth inhibition) varied greatly among the samples, from 0.3 μm to >35 μm; 15 μm was subsequently chosen as IC50‐cut‐off point between in vitro resistant and sensitive cases. PDN‐induced cytotoxicity was significantly related to apoptosis, as demonstrated by regression analysis; in sensitive cases, however, the percentage of apoptotic cells after in vitro PDN treatment was significantly increased compared with control (p=0.002). Immunofluorescence evaluation of intracellular BCL‐2 protein showed an equal percentage of positive cells in the two groups, but in resistant cells a higher mean fluorescence intensity (p=0.04) was demonstrated. In vitro sensitive and resistant patients did not display differences in clinical characteristics, in cytological, karyotypic and immunophenotypic features and in the outcome of induction therapy. Disease‐free survival (DFS), however, was significantly better in sensitive patients (p=0.02).
Quantitation of resistance to cytosine arabinoside by myeloid leukemic cells expressing bcl‐2
Authors: Guedez Liliana , Suresh Alaparthy , Tung Frank , Zucali James
Published: July 2010

Abstract:  The presence of bcl‐2 in myeloid leukemias has been associated with a decrease in therapy‐induced apoptosis, reduced patient survival and in vitro autonomous growth of leukemic cells. The present study focuses on the quantitation of resistance to increasing doses of 1‐β‐d‐arabinofuranosylcytosine (Ara‐C) by using hematological tumors expressing different levels of bcl‐2. Scanning densitometry of Western blots demonstrated that the myeloid U‐937 cells express low levels of bcl‐2 (RD=0.008), whereas the follicular lymphoma RL‐7 expressed very high levels (RD=3.084). Colony formation was also examined following incubation with Ara‐C and RL‐7 cells demonstrated a higher clonogenic survival (LD50=0.5 μm) when compared with U‐937 cells (LD50=0.005 μm). Similarly, the level of bcl‐2 expression in each cell line was also related to apoptosis with U‐937 cells demonstrating increased DNA fragmentation when compared with RL‐7 cells. To further evaluate the effect of upregulated bcl‐2 on Ara‐C treatment, U‐937 cells were transfected with a retroviral vector carrying the murine bcl‐2 or vector alone. Upregulation of bcl‐2 by myeloid leukemic cells increased the resistance by 3 logs to Ara‐C when comparing LD50 values from clonogenic assays, and decreased apoptosis by at least 3 logs when measuring dUTP positive cells by flow cytometry.
NK cell‐mediated killing of AML blasts: role of histamine, monocytes and reactive oxygen metabolites
Authors: Brune M. , Hansson M. , Mellqvist U.‐H. , Hermodsson S. , Hellstrand K.
Published: July 2010

Abstract:  Blasts recovered from patients with acute myelogenous leukaemia (AML) were lysed by heterologous natural killer (NK) cells treated with NK cell‐activating cytokines such as interleukin‐2 (IL‐2) or interferon‐α (IFN‐α). The cytokine‐induced killing of AML blasts was inhibited by monocytes, recovered from peripheral blood by counterflow centrifugal elutriation. Histamine, at concentrations exceeding 0.1 μM, abrogated the monocyte‐induced inhibition of NK cells; thereby, histamine and IL‐2 or histamine and IFN‐α synergistically induced NK cell‐mediated destruction of AML blasts. The effect of histamine was completely blocked by the histamine H2‐receptor (H2R) antagonist ranitidine but not by its chemical control AH20399AA. Catalase, a scavenger of reactive oxygen metabolites (ROM), reversed the monocyte‐induced inhibition of NK cell‐mediated killing of blast cells, indicating that the inhibitory signal was mediated by products of the respiratory burst of monocytes. It is concluded that (i) monocytes inhibit anti‐leukemic properties of NK cells, (ii) the inhibition is conveyed by monocyte‐derived ROM, and (iii) histamine reverses the inhibitory signal and, thereby, synergizes with NK cell‐activating cytokines to induce killing of AML blasts.
Salvage chemotherapy for relapsed or refractory non‐Hodgkin's lymphoma with a combination of ACES (high‐dose Ara C, carboplatin, etoposide and steroids) therapy
Authors: Niitsu N. , Umeda M.
Published: July 2010

Abstract:  ACES (Ara‐C, carboplatin, etoposide, steroids) therapy using granulocyte‐colony stimulating factor (G–CSF) was designed for relapsed or refractory non‐Hodgkin's lymphoma (NHL), and the therapeutic effects and adverse reactions were studied. The subjects were 40 patients, including 19 relapsed cases and 21 refractory cases, subjected to chemotherapy using anthracycline type agents. The ACES therapy consisted of carboplatin at 100 mg/m2 and etoposide at 80 mg/m2 for 4 d from the first day, Ara‐C at 2 g/m2 on the fifth day, solumedrol at 500 mg for 5 d from the first day and G–CSF at 2 μg/kg from the seventh day. This therapy was performed every 3 weeks, in principle. The doses were reduced to 70% of the above values for patients aged 70 yr or older. Among the 40 patients, complete remission (CR) was achieved in 14 (35%) and partial remission (PR) in 14 (35%) for a response of 70%. The 50% survival period was 526 d, and the 2‐yr disease‐free survival rate was 58.3%. Adverse reactions of grade 3 or higher included granulocytopenia in 62.5%, anemia in 17.5% and thrombocytopenia in 50%, but there was no death related to treatment. Four patients underwent transplantation of hematopoietic stem cells and have survived for long periods. This treatment was effective against relapsed NHL and could be performed safely with few adverse reactions.
Adhesive interactions between tumour cells and bone marrow stromal elements in human multiple myeloma
Authors: Faid L. , Van Riet I. , De Waele M. , Facon T. , Schots R. , Lacor P. , Van Camp B.
Published: July 2010

Abstract:  Long‐term bone marrow cultures (LTBMC) were established from marrow samples obtained from 6 myeloma patients and 5 healthy donors and were examined by in situ immunogold–silver staining. During the culture period, the established stroma in myeloma LTBMC revealed a lower level of confluency compared to the normal LTBMC. In addition, an increasing proportion of macrophages and osteoclasts was observed in the myeloma stroma throughout the culture period. Moreover, plasma cells were detectable by wk 8, mostly organized in small clusters. They strongly expressed VLA‐4 (6/6), H‐CAM (6/6), ICAM‐1 (6/6) and N‐CAM (3/6). In most cases, a weak expression of the other members of β1‐integrins was observed. The expression of β2‐integrins was always absent. Stromal fibroblasts were found to be weakly positive for VLA‐2, VLA‐3 and VLA‐5 and showed strong expression of VCAM‐1, H‐CAM and ICAM‐1. N‐CAM expression could not be detected. By comparing the adhesion molecule profile of the stromal cells in myeloma cultures with normal bone marrow (BM) cultures, no particular defects could be observed. The stroma displayed most of the potential ligands which could interact with adhesion molecules detected on the myeloma cells. Among these ligands we could find fibronectin and VCAM‐1 for VLA‐4, collagen I for VLA‐2 and VLA‐3 and laminin for VLA‐2, 3 and 6. Four myeloma cell lines, i.e. OPM‐1, U266, RPMI 8226 and JJN3, with a representative phenotype, were used to study the adhesive interactions of myeloma cells with the BM microenvironment. All the myeloma cell lines bound strongly to the marrow cell layers and also showed a high binding to purified fibronectin (FN). However, the adhesion of the cell lines to intact stroma could not be significantly inhibited by anti‐FN receptors antibodies. Nor could it be prevented when the latter were combined with anti‐H‐CAM, V‐CAM and ICAM‐1 antibodies, as tested in the JJN3 cell line. This implies that other unknown mechanisms contribute to the myeloma cell binding.
Increased serum levels of macrophage colony‐stimulating factor (M–CSF) in α‐and β‐thalassaemia syndromes: correlation with anaemia and monocyte activation
Authors: Wiener Edith , Wanachiwanawin Wanchai , Chinprasertsuk Sriprapa , Siripanyaphinyo Uamporn , Mawas Fatme , Fucharoen Suthat , Wickramasinghe Sunitha N.
Published: July 2010

Abstract:  Serum levels of M–CSF were determined by an ELISA method in 29 and 34 patients with HbH disease (α1/α2 or α1/HbCS) or β0‐thal/HbE, respectively, in 28 haematologically normal subjects and in five patients with anaemia due to iron deficiency or myelodysplasia. In HbH disease and β0‐thal/HbE, M–CSF concentrations were significantly higher than those in the normal subjects [986 ± 138 and 1385 ± 133, respectively, vs. 500 ± 33 pg/ml (mean ± SEM); p <0.01, and p <0.001, respectively]. By contrast, in patients with anaemia due to iron deficiency, M–CSF levels were within the normal range. In HbH disease and in β0‐thal/HbE, M–CSF levels correlated inversely with mean basal Hb values (r = –0.39, p = 0.05 and r = –0.60, p <0.001, respectively). In addition, in some of the HbH and β0‐thal/HbE patients, monocyte ADCC activities towards red cells were tested and found to be approximately twice as high as those in normal controls [38.3±5.7 and 30.7 ± 4.6 vs. 17.8 ± 1.8 % specific lysis (mean ± SEM), respectively; p <0.01 and p <0.02, respectively]. When thalassaemic patients and normal controls were considered together there was a significant correlation between M–CSF levels and monocyte ADCC activities (r = 0.51, p <0.02). The results suggest that in HbH disease and in β0‐thal/HbE, raised serum M–CSF contributes to the anaemia by enhancing the effector function of mononuclear phagocytes towards red cells.
Aetiology of severe aplastic anaemia and outcome after allogeneic bone marrow transplantation or immunosuppression therapy
Authors:   A. Bacigalupo for the Working Party on Severe Aplastic Anaemia of the European Blood and Marrow Transplantation Group
Published: July 2010

Abstract:  This study is based on 2163 patients with severe aplastic anaemia from the European Bone Marrow Transplantation Working Party Registry on Severe Aplastic Anaemia. It was designed to define patients' characteristics according to different aetiology and to assess the impact of aetiology of the aplasia on treatment outcome. Significant differences were found in age, sex, disease severity and DR typing between idiopathic, posthepatitis and drug‐associated severe aplastic anaemia. These differences did not affect the overall outcome of patients treated with bone marrow transplantation (BMT) or immunosuppression (IS) therapy.
The deoxyuridine suppression test in HIV‐1 positive patients: the role of azydothymidine (AZT)
Authors: García‐Die F. , Remacha A. , Cadafalch J. , Barceló M. J. , Parelladaxy6 M. M. , Arza B.
Published: July 2010

The deoxyuridine suppression test (dUST) was used to evaluate human immunodeficiency virus type 1 positive (HIV‐1) patients with low serum levels of vitamin B12 and/or low red cell folate and to assess any possible interferences of azydothymidine (AZT) in this test. The dUST was studied in 29 HIV‐1 positive patients, 18 without low serum vitamin B12 or low red cell folate and 11 with low serum vitamin B12 (6 patients), low red cell folate (4 patients) and 1 case with both. The role of AZT was studied using different concentrations (0.2, 2.5 and 10 μM/ml) in 2 groups: 1 group of 5 patients with vitamin B12 and/or folate deficiency and another group consisting of 13 healthy subjects. Methotrexate (MTX) (50 μg/ml) was added to induce a folate megaloblastic pattern in the latter group. Results of the dUST in the HIV‐1 group without low levels of serum vitamin B12 fell within the health‐related reference interval values. A vitamin B12 deficiency was only detected in 1 case in the HIV‐1 group with low serum vitamin B12, although a folate deficiency pattern was observed in the 4 patients with low red cell folate. In the healthy subjects AZT induced a dose‐dependent decrease of the MTX‐induced folate megaloblastic pattern. The pattern was also observed in the group of patients with vitamin B12 or folate deficiency, although AZT did not entirely interfere with the dUST. The effect of AZT on the dUST was attributed to a decrease in the incorporation of the isotope in the absence of deoxyuridine. The dUST is useful in differentiating vitamin B12 deficient patients from HIV‐1 infected patients with low levels of serum vitamin B12.
Differential expression of apoptosis, Bcl‐x and c‐Myc in normal and malignant lymphoid tissues
Authors: Hermann M. , Scholman H. J. , Marafioti T. , Stein H. , Schriever F.
Published: July 2010

Abstract: Bcl‐x and c‐Myc have an important role for the immune response by regulating the programmed cell death (apoptosis) of lymphocytes. Dysfunction of these selection processes can lead to the development of malignant lymphoma. The present study aimed at defining the differential expression of apoptosis, Bcl‐x and c‐Myc in normal and in malignant lymphoid tissues. Follicular centre lymphoma (FCL‐F) and mantle cell lymphoma (MCL) contained the lowest apoptotic indices (AIs), whereas Burkitt's lymphoma (BL) had the highest AIs. The AIs correlated significantly with the growth rates of the tumours and with the extent of Bcl‐x expression. Bcl‐x was expressed in almost all BL cells, but in few tumour cells in FCL‐F and in MCL. c‐Myc, in contrast, was found in the majority of the tumour cells in FCL‐F and in MCL, but not in BL. Whereas the extent of Bcl‐x expression correlated positively with the growth rates, an inverse correlation was observed between the percentages of c‐Myc‐positive tumour cells and the growth rates of the tumours. We conclude that normal and malignant lymphoid tissues have a distinct pattern of apoptosis and that expression of Bcl‐x and c‐Myc in B cell lymphoma is differentially regulated.
In vitro study of Seraspenide on HIV‐induced inhibition of granulopoiesis
Authors: Coutton C. , Chermann J. C.
Published: July 2010

Soluble(s) factor(s) produced by HIV‐infected cells have been implicated in bone marrow dysfunction observed in AIDS patients. We have shown previously that infected macrophage but not CBMC‐derived conditioned medium inhibited the differentiation of purified CD34+ cells, suggesting that accessory cells play a crucial role in granulopoiesis inhibition induced indirectly by HIV. In order to prevent this inhibition we tested the potential protective effect of Seraspenide, a reversible inhibitor of hematopoietic progenitors. We showed that 20 h pretreatment of mononuclear cells with Seraspenide at 10−10 mol/1 prevents CFU‐GM inhibition induced by HIV‐infected cell‐derived conditioned medium.