bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transfer of p16inka/CDKN2 gene in leukaemic cell lines inhibits cell proliferation
Authors: Quesnel Bruno , Preudhomme Claude , Lepelley Pascale , Hetuin Dominique , Vanrumbeke Michael , Bauters Francis , Velu Thierry , Fenaux Pierre 
Published: November 2003

The gene encoding for p16ink4a, a negative regulator of transition between G1 and S phase, is homozygously deleted in a large proportion of acute lymphoblastic leukaemias (ALL). Transfer of p16ink4a gene in several solid tumour cell lines with functional pRb and lacking both p16ink4a alleles has resulted in a dramatic reduction of cell proliferation, and the aim of this work was to confirm this effect in leukaemic (especially ALL) cell lines. We tested the proliferation in liquid medium and in soft agar after transfer of p16ink4a gene by a retroviral vector in leukaemic cell lines with homozygous p16ink4a gene deletion (K562, CEM, Jurkat cell lines) or with p16ink4a gene hemizygous deletion and a point mutation inactivating the remaining allele (HL60 cell line). The viral titre obtained after transfection of PA317 amphotropic packaging cell line, which has a p16ink4a gene homozygous deletion, was low, suggesting that p16ink4a gene expression could impair viral production of retroviral packaging cell lines derived from the NIH3T3 cell line.After retroviral transfer of p16ink4a in cell lines and G418 selection in liquid medium, a strong cell proliferation inhibition was observed for K562, CEM and Jurkat, but no inhibition was seen for HL60. A strong growth reduction in soft agar was also observed with p16ink4a‐transduced CEM, Jurkat and K562 cells, with a moderate growth reduction in the HL60 cell line. The growth inhibition in liquid culture, of K562 and Jurkat cell lines, was confirmed by electroporation transfer of the p16ink4a gene.Our findings show that p16ink4a gene transfer has a growth‐inhibitory effect in leukaemic cell lines with p16ink4a gene homozygous deletion. These data suggest that p16 could be a suitable gene for gene therapy in ALL.
Pitfalls in the immunophenotyping of leukaemia and leukaemic lymphomas: survey of 9 years of quality control in The Netherlands
Authors: Kluin‐Nelemans J. C. , Van Wering E. R. , Van 't Veer M. B. , Van Der Schoot C. E. , Adriaansen H. J. , Van Der Burgh F. J. , Gratama J. W. 
Published: November 2003

During the last decade, biannual quality controls were performed in The Netherlands focusing on the immunophenotyping of leukaemic haematological malignancies. All results on 48 specimens obtained by 18–34 laboratories were analysed. The interlaboratory variability and percentages of discordant results from 30 markers were measured by assessing false positive or negative (cut‐off 10%) results in comparison with median results of the group. The quality of the immunophenotypic diagnoses obtained from the interpretation of these markers in relation to clinical data was evaluated by scoring them as ‘correct’, ‘minor fault’, ‘major fault’, ‘not based upon the markers used’, and ‘no diagnosis’. CD3, CD8, CD19, CD61 and Smλ had the lowest percentage discordancy (sum of total negative and positive discordant values 5–7.5% of assays); CD13, CD15, cyCD22, CD33 and TdT scored worst with 14–20% cumulative discordancy. The analysis of each diagnosis yielded 78% acceptable immunophenotypic conclusions (correct 54% and minor fault 24%). It appeared that the major faults in immunophenotyping were caused by suboptimal antibody selection and erroneous interpretation of the results obtained, rather than by technical errors. Large differences per diagnostic category were observed, with the best scores for mature B‐cell leukaemias, AMLs and common‐ALL, and the poorest scores for T‐cell malignancies which were correctly diagnosed in only 24–60% of specimens. Mature T‐NHL and T‐PLL were mistakenly diagnosed as T‐ALL by 40% of the centres. Misinterpretation of TdT immunofluorescence or omitting this marker contributed significantly to these wrong diagnoses. A median of 4% of immunophenotypic diagnoses were not based on a correct panel of antibodies, but upon the morphology of the accompanying blood smear, and was often flawed by overinterpretation.In conclusion, both the technical performance of immunophenotyping of haematological malignancies in The Netherlands and the procedure by which a final diagnosis is obtained needs improvement, especially for T‐cell malignancies.
Clearance of marrow infiltration after 1 week of therapy for childhood lymphoblastic leukaemia: clinical importance and the effect of daunorubicin
Authors: Lilleyman J. S. , Gibson B. E. S. , Stevens R. F. , Will A. M. , Hann I. M. , Richards S. M. , Hill F. G. H. 
Published: November 2003

At the commencement of UKALL XI, a national MRC trial for childhood lymphoblastic leukaemia (ALL), the therapy included a bolus of daunorubicin (DR) on the first 2 d of the protocol. This component of treatment was subsequently withdrawn because of concern about long‐term cardiotoxicity. All children both before and after this change of policy had their marrow status at the end of the first week assessed by central review as part of the trial to examine the clinical importance of the rate of disease clearance. This also afforded an opportunity to observe the effect of DR on gross residual disease at an early stage of therapy.1419 children were studied: 342 received DR (‘recipients’), 1077 did not. 44% of the recipients completely cleared their marrow of blast cells after 8 d compared with 13% of the non‐recipients (χ2=158.2, P<0.0001). The difference in the proportion with massive residual disease (>80% blasts) was less impressive but there was still a difference in favour of DR recipients (DR 9%, no DR 15%; χ2=7.7, P=0.006). The rate of disease clearance correlated with disease‐free survival for both recipients and non‐recipients, but there was no significant difference in outcome when comparing the two groups with each other, either in terms of disease‐free or relapse‐free survival.DR accelerated the rate of blast cell disappearance from the marrow but the difference this made to disease free survival is small or non‐existent. It appears to be the relative speed of response to a given therapeutic regimen that is prognostically important rather than the absolute rate of response when comparing one treatment with another.
Common clonal origin of lymphocytes and plasma cells in splenic lymphoma with villous lymphocytes
Authors: KATAYAMA YOSHIO , KOJIMA KENSUKE , YOSHINO TADASHI , MATSUO YOSHINOBU , ISOKAWA MASAFUMI , YANO TOMOFUMI , OKA HIDEYUKI , YAMAGUCHI MIKA , DEGUCHI SEIGO , TSUCHIYAMA JUNJIRO , HAYASHI KYOICHI , TESHIMA TAKANORI , SHINAGAWA KATSUJI , ISHIMARU FUMIHIKO , OMOTO EIJIRO , HARADA MINE 
Published: November 2003

In two‐thirds of patients with splenic lymphoma with villous lymphocytes (SLVL) a small amount of M‐protein can be detected in association with the presence of plasma cells in the peripheral blood (PB) and/or bone marrow (BM). However, it is not known whether lymphoma cells and plasma cells originate from the same clone. In this report we describe a case of SLVL which was characterized by the presence of marked monoclonal gammopathy (IgG‐κ 90 g/l) and increased plasma cells in the BM. In an attempt to elucidate the origin of lymphoma cells and plasma cells, we performed morphological, cytogenetic and molecular studies on PB mononuclear cells (PBMNC) without plasma cells and BMMNC containing 10% plasma cells from this patient.Immunofluorescence showed that lymphoma cells and plasma cells were positive for cytoplasmic γ heavy and κ light chains. Well‐developed endoplasmic reticulum was observed in the cytoplasmic organelles of PBMNC using an electron microscope. The mean IgG concentration in the 3 d supernatant cultures of PBMNC was 374±24μg/l. More than 50% PBMNC differentiated into plasmacytoid cells in 6 d of liquid culture with IL‐3 and IL‐6. Analysis by two‐colour FISH revealed that karyotypic abnormalities of monosomy X and trisomy 17 existed simultaneously in both lymphoma cells and plasma cells. JH gene rearranged bands from PBMNC and BMMNC by Southern blot hybridization were identical, whereas DNAs from PBMNC failed to hybridize with the Cμ probe.These observations strongly suggest that lymphoma cells and plasma cells originate from the same clone, and that plasma cells, as well as lymphoma cells, which have undergone class switch recombination, could produce IgG type M‐protein in this case.
Treatment of adults with acute lymphoblastic leukaemia in first bone marrow relapse: results of the ALL R‐87 protocol
Authors: Giona Fiorina , Annino Luciana , Rondelli Roberto , Arcese William , Meloni Giovanna , Testi Anna Maria , Moleti Maria Luisa , Amadori Sergio , Resegotti Luigi , Tabilio Antonio , Ladogana Saverio , Fioritoni Giuseppe , Camera Andrea , Liso Vincenzo , Leoni Pietro , Mandelli Franco 
Published: November 2003

Sixty‐one adults aged <55 years with acute lymphoblastic leukaemia (ALL) in first bone marrow relapse were enrolled in an Italian cooperative study (ALL R‐87 protocol) from 12 GIMEMA Institutions. The treatment programme consisted of: (1) an induction phase with intermediate‐dose cytarabine (IDARA‐C 1 g/m2, 6 h daily infusion ×6 d) plus idarubicin (IDA; 5 mg/m2/d × 6 d) and prednisone (40 mg/m2/d × 21 d), (2) a consolidation phase followed by (3) bone marrow transplant (BMT). Median first complete remission (CR) duration was 8.5 months (range 1–54 months). 34/61 patients achieved CR (56%); 24 (39%) failed to respond and three (5%) died during induction. Most responders (24 patients) could not enter the BMT programme; 15 relapsed early (median time to relapse 2 months); nine were withdrawn due to toxicity and one died in CR of infection. Nine of the 34 CRs underwent BMT (five autologous and four allogeneic). Three of the four allotransplanted patients are alive in continuous CR at 22, 43 and 63 months; only one of the five who underwent an autologous BMT is alive in CR at 46 months.The estimated disease‐free survival (DFS ± SE) at 36 months was 0.16 ± 0.08 for all responders. Univariate analysis showed that previous therapy was the only prognostic factor influencing DFS. The estimated probabilities of event‐free survival (EFS ± SE) and survival ± SE at 37 months were 0.09 ± 0.04 and 0.10 ± 0.04, respectively. The EFS was significantly better in patients with a preceding CR >24 months, compared to those with a shorter first remission.Our results confirm the tolerance and efficacy of IDARA‐C plus IDA in inducing CR in poor‐risk adult ALL. Even though the number of transplanted patients was small, allogeneic BMT seems to give a real opportunity of cure in this category of patients.
Types and sources of fuels for platelets in a medium containing minimal added fuels and a low carryover plasma
Authors: Niu X. , Whisson M. E. , Guppy M. 
Published: November 2003

The storage of platelets in synthetic media can result in plasma savings and reduced transfusion reactions. Accordingly, a wide range of storage formulations have been developed with the aim of replacing at least a proportion of the plasma in the storage medium. However, the concentrations and types of fuels in the carryover plasma, and the utilization of these fuels by platelets in storage, has not been investigated. We have developed a system which can measure total ATP turnover, and the contribution to total ATP turnover by the oxidation of various fuels and by lactate production, in a bag of partially purified platelets in a buffered saline with minimal carryover citrate phosphate double dextrose (CP2D) plasma.Carryover plasma was about 1% and the final platelet suspension contained, on average, 0.62 mM glucose, 9.6 mg/l free fatty acids, 32 mg/l triglycerides and 0.23 mM total amino acids. The oxidation of carbohydrate (glucose, glycogen and lactate) accounted for 60% of total ATP turnover. The platelets also produced lactate (<6% of total ATP turnover) and consumed free fatty acids and amino acids/proteins (15.2% of total ATP turnover). Therefore we have identified the fuels that account for about 80% of oxygen consumption and ATP turnover by platelets in a medium with low carryover plasma. The implications of these data for storage strategies are discussed.
Heterogenous expression of bcr‐abl fusion mRNA in a patient with Philadelphia‐chromosome‐positive acute lymphoblastic leukaemia
Authors: Inokuchi Koiti , Futaki Makoto , Hanawa Hideki , Tanosaki Sakae , Yamaguchi Hiroki , Iwakiri Rika , Nomura Takeo , Dan Kazuo 
Published: November 2003

We performed molecular and cytogenetic analysis on a 56‐year‐old woman with Philadelphia chromosome (Ph1)‐positive acute lymphoblastic leukaemia (ALL) having two types of major and minor bcr‐abl (M‐bcr‐abl, m‐bcr‐abl) fusion mRNA at diagnosis. In the course of her disease, unexpected heterogenous bcr‐abl fusion mRNA was detected by sequential analysis using the reverse transcription and polymerase chain reaction (RT‐PCR).The RT‐PCR analysis showed both M‐bcr‐abl (b2‐a2 type) and m‐bcr‐abl at diagnosis. Although b2‐a2 type M‐bcr‐abl disappeared after complete remission (CR) was achieved with intensive induction chemotherapy, expression of both m‐bcr‐abl and a new type of M‐bcr‐abl mRNA (b1‐a2 type), which may have been produced through splicing out of exon b2, was detected in the early stage of CR. The leukaemic cells contained these heterogenous bcr‐abl mRNAs in both the CR and relapsed state, and showed more stable expression of the m‐bcr‐abl type mRNA than that of the b2‐a2 type. These findings of heterogenous bcr‐abl mRNA due to alternative or missplicing during the disease course in the present ALL case may provide important evidence of disease progression.
CD56+ NK lymphomas:clinicopathological features and prognosis
Authors: KWONG Y. L. , CHAN  , LIANG RAYMOND , CHIANG ALAN K. S. , CHIM C. S. , CHAN T. K. , TODD DAVID , HO FAITH C. S. 
Published: November 2003

The surface molecule CD56 marks a category of malignant lymphoma of putative natural killer (NK) cell origin. We conducted a retrospective analysis of 24 cases of CD56+ NK lymphoma/leukaemia to define the clinicopathologic and prognostic features of this specific group of lymphomas. 56 cases of nasal lymphomas and 204 cases with an initial diagnosis of peripheral T‐cell lymphoma were retrospectively analysed. To specifically examine lymphomas of putative NK origin, only those that were negative for surface expression of CD3 but positive for CD56 were analysed. 24 cases were identified. The initial predominant sites of involvement were nasal (n =18), palate (n = 1), nodal (n = 1) and multi‐organ (n =4). Clinically, in patients with disease localized to one anatomical site (n = 20), most had symptoms confined to the nose, with a high percentage in early stage (I: 91%; IV: 9%). The marrow was not involved in any of these cases. However, patients with multi‐organ involvement at presentation (n = 4) behaved differently. All presented acutely with pancytopenia, hepatosplenomegaly, and marrow infiltration with haemophagocytosis. A leukaemic phase was observed in one case. Anthracycline containing combination chemotherapy resulted in complete remission in 75% of patients with localized disease, but only in 25% with multi‐organ involvement. The median survival of patients with localized disease was 12 months, compared with 2 months in the multi‐organ group (P =0.06); the disease‐free survival was significantly better in the former (P <0.01). The overall median survival of all patients was still poor at 11 months.We conclude that CD56+ NK lymphomas could be divided into two main patterns of disease presentations: localized (predominantly nasal), and multi‐organ involvement. Each has different clinicopathologic and prognostic features. Conventional chemotherapy appeared ineffective for the majority of patients, and innovative treatment modalities are needed to improve outcome.
Successive transformation of chronic myelomonocytic leukaemia into acute myeloblastic then lymphoblastic leukaemia, both with minor‐bcr rearrangement
Authors: DAUTEL Marie Michèle , FRANÇOIS Sylvie , BERTHEAS Marie‐FranÇoise , BARANGER Laurence , GARDAIS Jacques , BOASSON Marc , IFRAH Norbert , BLANCHET Odile 
Published: November 2003

We report a case of chronic myelomonocytic leukaemia (CMML), which transformed first into acute myeloblastic leukaemia (AML) and then into acute lymphoblastic leukaemia (ALL). In the AML and ALL phases, chromosome analysis showed a classic Philadelphia chromosome (Ph) t(9;22)(q34;q11). Molecular studies showed breakpoint cluster region rearrangement between exons e1 and a2 compatible with a p190bcr/abl breakpoint as observed in Ph‐positive lymphoblastic acute leukaemia. The minor (m‐bcr) rearrangement was also detected during complete remission.This observation supports a multistep pathogenesis of leukaemias, and that the p190bcr/abl breakpoint may influence the course of the disease.
Do HbSS erythrocytes lose KCl in physiological conditions?
Authors: GODART H. , DORMANDY A. , ELLORY J. C. 
Published: November 2003

KCl cotransporter activity in sickle (HbSS) red blood cells (RBCs) was measured in cells suspended in ‘simple’ physiological saline, saline augmented with inorganic salts, and autologous plasma. Our results showed that the transporter was only functioning at 20% of the level of cells in saline when cells were resuspended in autologous plasma. Kinetic analysis of the data showed that plasma decreased both Vmax and Km for K+ of the transporter. The plasma factor(s) responsible was heat‐stable and dialysable (i.e. size &60; 10 kD).Adding magnesium, calcium, inorganic phosphate or bicarbonate to ‘simple’ saline to mimic the effect of plasma revealed that Mg2+ and Ca2+ had no significant effect at physiological concentrations. Pi was not effective at 1.1 mM , but did inhibit significantly (42±2%) at 5.6 mM . HCO−3   had a major inhibitory effect on K+ influx when added to saline, and was identified as the principal candidate for the plasma effect.We suggest bicarbonate may play a significant role in modifying KCl cotransport, and hence HbSS cell volume in vivo. It acts by altering the set point of the transporter via the signalling systems involved in its regulation.
A prospective study of minimal residual disease in childhood B‐lineage acute lymphoblastic leukaemia: MRD level at the end of induction is a strong predictive factor of relapse
Authors: Jacquy C. , Delepaut B. , Van Daele S. , Vaerman J. L. , Zenebergh A. , Brichard B. , Vermylen C. , Cornu G. , Martiat P. 
Published: November 2003

We prospectively investigated minimal residual disease (MRD) in 51 children with B‐lineage acute lymphoblastic leukaemia (ALL) treated according to the Fralle 93 protocol. PCR follow‐up was performed in children in morphological and cytogenetic complete remission, provided an immunoglobulin (IgH) gene rearrangement could be detected using FR3/JH amplimers. MRD was studied according to our previously described methodology, with a few modifications including the use of a consensus JH probe to control for PCR efficiency variations.Out of the initial 51 patients, 34 were assessable for MRD at the end of induction at the time of analysis. MRD levels were as follows: >1/103 in 26%, 1/103 to 1/104 in 50% and <1/104 or not detectable in 24%. With a median follow‐up of 20 months there were five relapses, all of which occurred in the group of patients with MRD >1/103. To date, none of the patients with MRD 1/103 (good molecular responder) has relapsed. Classification according to molecular response at the end of induction did not correlate with the conventional risks groups: there were no statistically significant differences between good and bad molecular responders. Of particular interest is the absence of correlation between WBC at diagnosis and MRD level at the end of induction. We conclude that classification of patients into good and bad molecular responders using PCR seems to be a better prognostic indicator than conventional risk factors in childhood B‐lineage ALL. Patients with MRD level >1/103 have a particularly poor outcome and should always be considered for alternative therapeutic strategies in the future, whereas in good molecular responders belonging to poor or intermediate risk categories, treatment de‐escalation might be contemplated.
MACULAR HAEMORRHAGE IN ADULT ACUTE LEUKAEMIA PATIENTS AT PRESENTATION AND THE RISK OF SUBSEQUENT INTRACRANIAL HAEMORRHAGE
Authors: Jackson Nicholas , Reddy Sagili Chandrasekhara , Harun Mohd Hishamuddin , Quah Soon Hoe , Low Heng Chin 
Published: November 2003

Retinal changes are common in adult acute leukaemia patients at presentation, but whether they correlate with the risk of subsequent intracranial haemorrhage is unknown. A 4‐year study has been carried out in 82 newly‐diagnosed acute leukaemia patients, aged 12–77 years, who were studied prospectively for the presence of intra‐retinal haemorrhages (IRH), white‐centred haemorrhages (WCH), cotton‐wool spots (CWS) and macular haemorrhages (MH). Groups with and without these features were compared for their risk of intra‐cranial haemorrhage (ICH) within the first 30 d following diagnosis.There was no association between the incidence of ICH and the presence of IRH, WCH or CWS. However, 6/13 of those with MH developed ICH, compared to 6/69 of those without MH (relative risk 5.0, CI 95% [2.03–12.33], P=0.003). The only other identifiable risk factor for ICH was the M3 subtype of AML, but if the four cases of M3‐AML were discounted from analysis, MH remained a highly significant risk factor for ICH. Patients with MH should be monitored intensively for the development of ICH, and receive priority in the allocation of platelets where these are in short supply.
Frequent involvement of chromosomes 1, 3, 7 and 8 in splenic marginal zone B‐cell lymphoma
Authors: Solé Francesc , Woessner Soledad , Florensa Lourdes , Espinet Blanca , Mollejo Manuela , Martín Pedro , Piris Miguel Angel 
Published: November 2003

We have studied 19 cases of splenic marginal zone B‐cell lymphoma (SMZBCL) combining cytological features, conventional cytogenetics, and in situ hybridization (ISH) techniques.A clonal chromosome abnormality was found in 11/19 patients (58%). The more frequent recurrent abnormalities were: del(3), del (7q), and involvement of chromosomes 1, 3, 7 and 8. No patient showed the translocation t(11;14)(q13;q32). An outstanding finding was the low incidence of trisomy 3 (36%) compared to patients with MALT lymphoma. These findings support the interpretation that SMZBCL is a distinct lymphoproliferative disorder.