bluuebunny/crossref_metadata_2025_split huggingface dataset:
Different sensitivity of normal and leukaemic progenitor cells to Ara‐C and IL‐3 combined treatment
Authors: Lista P. , Brizzi M. F. , Rossi M. , Resegotti L. , Clark S. C. , Pegoraro L. 
Published: March 2008

Summary. In the present study the effects of a combined treatment with cytosine‐arabinoside (Ara‐C) and inter‐leukin‐3 (IL‐3) on acute myeloblastic leukaemia clonogenic cells and on normal haemopoietic progenitors was investigated, with the aim of improving the tumoricidal effect of cycle specific drugs.Blast cells from 24 acute myeloblastic leukaemia (AMT,) patients were screened with a short‐term proliferative assay based on 3H‐thymidine (3H‐TdR) uptake for their response to IL‐3. To evaluate the synergism between the growth factor and Ara‐C, the cells were pretreated for 3d in liquid culture in the presence or absence of IL‐3 (10 U/ml) and for the last 24 h with Ara‐C (3 μg/ml). The cells were then washed and seeded in semisolid media to assess their clonogenic ability. The results showed that, in those cases which were good responders to IL‐3 in the 3H‐TdR uptake assay (19 out of a), Ara‐C exposure eliminated a greater proportion of clonogenic cells if pretreated with IL‐3 than if untreated (P < 0.001), while in cases unresponsive to IL‐3 this effect was not significant. Moreover, when the same protocol was applied to bone marrow cells from normal donors, it was found that IL‐3 pretreatment did not significantly enhance the toxic effect of Ara‐C on day 14 granulocyte‐macrophage colony forming units (CFU‐GM) and erythroid burst forming units (BFU‐E). Finally IL‐3 pretreatment was also able to increase the cytotoxic effect of Ara‐C on leukaemic cells co‐cultured, to simulate clinical AML remission, with normal bone marrow cells. The results indicate that IL‐3 may improve the therapeutic index of cycle‐specific drugs in AML therapy.
Factor VIII heavy chain polypeptides in plasma and concentrates
Authors: Kemball‐Cook G. , Bevan S. A. , Barrowcliffe T. W. 
Published: March 2008

Summary. Factor VIII polypeptides in plasma and FVIII concentrates have been analysed by an electrophoretic technique based on that of Weinstein et al (1981). Samples were complexed with radiolabelled anti‐FVIII Fab, and the immunocomplexes visualized by SDS‐polyacrylamide electrophoresis. The technique visualized FVIII heavy chain polypeptides in all types of samples, including plasma, without further purification.Fresh or frozen normal plasma (collected into protease inhibitors) contained a range of polypeptides with the largest dominant band at an apparent Mr of 250–300 kDa, and the smallest at 80–90 kDa: no bands were produced from samples of severe haemophilic plasma.Cryoprecipitate had a similar polypeptide distribution to normal plasma, but intermediate purity FVIII concentrates showed more degraded patterns which varied between products: the 250–300 kDa bands were reduced or absent, the 80–90 kDa bands were more pronounced than in plasma, and in one product a polypeptide was seen at approximately 40–50 kDa. In some products heat treatment for viral inactivation increased the proportion of smaller FVIII polypeptides.Highly‐purified FVIII concentrate derived from plasma was also degraded relative to plasma FVIII, and two products obtained by recombinant DNA technology both showed degraded, though slightly different. profiles.The native structure of FVIII in fresh plasma appears heterogeneous with a predominance of higher M, forms: these are degraded to a greater or lesser extent during concentrate production, dependent on the manufacturing processes used.
Blood findings in generalized mastocytosis: evidence of frequent simultaneous occurrence of myeloproliferative disorders
Authors: Horny H.‐P. , Ruck M. , And M. Wehrmann , Kaiserling E. 
Published: March 2008

Summary Blood findings in 61 cases of generalized mastocytosis (GM) were evaluated. The cases were divided into two major variants: Systemic mastocytosis (SM; n= 34) with urticaria pigmentosa‐like skin lesions, and malignant mastocytosis (MM; n = 27), without skin involvement. The following results were obtained:

Significant differences between MM and SM were found in the main haematological parameters (erythrocyte, platelet and leucocyte counts and haemoglobin level): normal values were found in 16 of the SM cases, but never in MM.
The main pathological findings were: in SM, anaemia (9/34) and leucocytosis (5/34); and in MM, leucocytosis (19/ 27), monocytosis (14/271, eosinophilia (12/27), bicytopenia (12/27, mostly anaemia with thrombocytopenia), basophilia (10/27) and isolated anaemia (7/27).
The major finding was a significant difference between MM and SM in the incidence of myeloproliferative disorders (MPD), myelodysplasia and mast cell leukaemia (MCL): these disorders occurred in 23 (92%) MM patients, but only in two (6%) SM patients (P < 0.001). The four instances of MCI, and two of myelodysplasia all occurred with MM. Of the 19 cases of MPD, six (SM, 1; MM. 5) were acute variants (acute myeloid and myelomonocytic leukaemias) and 13 (SM. 1; MM, 12) were chronic variants. No case of malignant lymphoma was noted.
The blood picture in 10 of the 13 chronic MPD cases represented an atypical chronic myeloid leukaemia for which the preliminary descriptive term‘mastocytosis‐associated MPD’is proposed.
A survey of 103 published cases (SM, 77; MM. 26) yielded similar findings, including a high incidence of MPD and MCL in MM. These findings add further weight to the argument for recognizing SM and MM as two separate entities.
Denaturing gradient gel electrophoresis and direct sequencing of PCR amplified genomic DNA: a rapid and reliable diagnostic approach to beta thalassaemia
Authors: Losekoot M. , Fodde R. , Harteveld C. L. , Heeren H. van , Giordano P. C. , Bernini L. F. 
Published: March 2008

Summary The analysis of polymerase chain reaction (PCR)‐amplified β‐globin DNA with allele‐specific oligonucleotide (ASO) probes reveals a very heterogeneous spectrum of β‐thalassaemia in the Netherlands. However about 20% of the β‐thalassaemia mutations cannot be identified with this approach. The combination of specific amplification of certain regions of the β‐globin gene with denaturing gradient gel electrophoresis (DGGE) allowed us to rapidly localize several of these mutations to specific regions of the gene. which were again amplified and directly sequenced.We believe that the combination of DGGE and the direct sequence determination of PCR amplified genomic DNA represents a valid alternative to the‘ASO probes’approach, especially in countries where a very heterogeneous spectrum of β‐thalassaemia mutations occurs.
Osteoblast stimulation in multiple myeloma lacking lytic bone lesions
Authors: Bataille Régis , Chappard Daniel , Marcelli Christian , Rossi Jean‐Franclois , Dessauw Philippe , Baldet Pierre , And Jacques Sany , Alexandre Christian 
Published: March 2008

Summary The reasons why some patients with multiple myeloma (MM) do not develop severe bone loss, or even develop sclerotic bone lesions, remain unclear. In order to answer this question at the cellular and tissue level, we evaluated the histological bone condition of 10 patients with MM who never developed lytic bone lesions during the course of their disease (including two patients with sclerotic MM).Myeloma‐induced bone changes in the close vicinity of myeloma cells were evaluated by quantitative histology (bone histomorphometry). All 10 patients presented a significantly increased osteoblastic activity. This was associated with an increased bone resorption in seven of the 10 cases. Three patients had a pure osteoblastic presentation. These features were the reverse of the pattern observed in seven patients with lytic bone lesions: increased bone resorption with decreased bone formation. Almost all of these 10 patients showing excessive osteoblastic activity had increased serum bone gla protein levels, a specific marker of bone formation. Finally, 90% of these patients were lambda MM (70% of them were IgG lambda MM), an immunoglobulin subtype previously associated with the sclerotic MM variants.In conclusion, a subset of patients with MM never develop severe bone loss because of the stimulation of osteoblastic activity. These patients belong to the same family as osteosclerotic MM, presenting more frequently the IgG type and lambda subtype.
Irradiation of platelets with UV‐B light exposes fibrinogen binding sites via an intracellular mechanism
Authors: Kooy M. van Marwijk , Borghuis L. , Prooijen H. C. van , Aarts‐Kiemens M. I. , Akkerman J. W. N. 
Published: March 2008

Summary Previous studies have shown that ultraviolet irradiation (UVI) causes platelet aggregation. In the present study we exposed platelet suspensions to a relatively high dose of UV‐B (8 J/cm2) under conditions comparable to those of UVI of platelet concentrates in order to obtain more insight into the UV‐induced aggregation response and to evaluate the significance of this phenomenon for the clinical use of UV‐irradiated platelet concentrates.This study provides evidence that UV‐B induced aggregation is mediated by a Ca2+‐dependent process of fibrinogen binding to an intact glycoprotein IIb—IIIa complex on platelet membranes. Although UV‐induced platelet aggregation is independent of thromboxane A2 formation and ADP secretion, it requires metabolic energy, cytosolic Ca2+ and a low cyclic‐AMP level. Thus, UV‐B irradiation causes platelet aggregation by exposing fibrinogen binding sites via an intracellular mechanism. Since the amount of bound fibrinogen following UVI is relatively low (about 2300 molecules/ platelet) and the binding remains reversible, its effect on platelet behaviour after transfusion may be minor.
Prevention of Gram‐positive and Candida albicans infections using teicoplanin and fluconazole: a randomized study in neutropenic children
Authors: Schaison G. , And A. Baruchel , Arlet G. 
Published: March 2008

Summary We have shown that the combination of teicoplanin (T) and ceftriaxone (C) given once daily as first‐line empirical antimicrobial therapy is very effective (85%) and safe in febrile neutropenic children (ICAAC. 1988). Like others (ICAAC. 1989) we have stressed the new life‐threatening problems with Streptococcus mitis and Strep. sanguis. Considering the regular sensitivity of Gram‐positive organisms to T. it is possible that the introduction of the glycopeptide at the onset of aplasia may be helpful. The high incidence of fungal infection or superinfection supports the routine use of an effective antifungal agent when fever is still present or relapse occurs.Our study was designed to assess the prevention of (1) staphylococcal and streptococcal infections using T when the central catheter is being placed, and (2) candida infection by the simultaneous use of a new anti‐mycotic agent fluconazole (F) administered once‐daily. Patients were randomized into two arms of the study. Arm A received T 6 mg/kg/d and F 3 mg/kg/d starting when the catheter was put in place. If fever occurred. C 50 mg/kg/d and amikacin (A) 1 5 mg/kg/d were added and T increased to 10 mg/kd/d. If a febrile relapse occurred, amphotericin B was added. In arm B the combination of the three agents T 10 mg/kg/d, C 50 mg/kg/d and A 15 mg/kg/d was started only if there was fever. Amphotericin B was used in febrile relapses. Forty‐six patients were eligible (23 in each arm: no statistically significant differences between the two groups). The mean age of the patients was 8 years and mean duration of aplasia 23 and 24 d. Results were as follows: arm A the duration of T + F alone was 10 d, 22 patients had a febrile episode with 1 Gram‐positive, 11 Gram‐negative and one candida strain isolated in eight patients. C +A were effective in 72% of these patients. There were no deaths. In arm B, fever occurred in all patients during the first 6 d, with eight Gram‐positive and six Gram‐negative organisms isolated in nine patients.C + T + A were successful in 83% of patients. A bacterial superinfection was documented in four patients, candida superinfection in seven patients and one patient died from Candida parakrusei septicaemia. When the total amount of C and A used in both arms was compared, there was a reduction in 20% of the two agents in arm A, but T was equivalent in both arms. Treatment was well tolerated in both arms.Teicoplanin given very early is an effective agent in preventing Gram‐positive infection, fluconazole given simultaneously is a very promising agent in the prevention of C. albicans superinfection and the value of ceftriaxone in relapsing fevers is confirmed.
Characterization of the pseudogenic and genic homologous regions of von Willebrand factor
Authors: Marchetti G. , Patracchini P. , Volinia S. , Aiello V. , Schiavoni M. , Ciavarella N. , Calzolari E. , Schwienbacher C. , Bernardi F. 
Published: March 2008

Summary The homologous pseudogenic and genic regions of von Willebrand factor (vWF) were studied in DNA from a patient with homozygous deletion of vWF genes and compared with a normal control. This analysis indicates informative restriction patterns for the investigation of restriction fragment length polymorphisms (RFLPs) and gene lesions, and for molecular cloning. A useful new genic Xbal RFLP was found and characterized.A large BgIII fragment of the pseudogenic region was cloned and mapped, and single sequences (9 kb) were used as probes. Corresponding genic and pseudogenic fragments, which contain exons 23–28, and specific restriction patterns were identified, including a new polymorphic TaqI site that was mapped in the gene. A cloned fragment contains the 5’boundary of the pseudogene and recognizes an additional and unknown homologous sequence in the genome.The chromosomal localization of the vWF pseudogene and of the breakpoint cluster region (BCR) gene were compared by ‘in situ’ hybridization: overlapping patterns were detected. The cloning, characterization and mapping of the pseudogenic region improves the analysis of this portion of chromosome 22 affected by several somatic and constitutional alterations, and also of the corresponding genic region on chromosome 12.
Splenic lymphoma with villous lymphocytes: natural history and response to therapy in 50 cases
Authors: Mulligan Stephen P. , Matutes Estella , Dearden Claire , Catovsky Daniel 
Published: March 2008

Summary We studied the natural history and response to treatment in 50 patients with splenic lymphoma with villous lymphocytes followed for a minimum of 6 months and up to 15 years (median 3·7 years). The disease occurs in the elderly (median 68 years) and affects males more than females (M/F ratio 1·77). The median survival for the group was not reached but 82% were surviving at 3 years and 78% at 5 years. Twelve patients (24%) died, one‐third of deaths were disease related (four patients) and one‐half were due to cardiovascular disease or another malignancy (six patients). The outcome was worse for males (31% died) than females (11% died).Fourteen patients were not treated and 10 remain alive between 1 and 6 years from diagnosis. The remaining patients were treated by chemotherapy, splenic irradiation or splenectomy. The response to chemotherapy was poor and only eight of 22 (36%) patients treated achieved a good response. Splenic irradiation was employed in seven patients and three benefited from it. Splenectomy seems to be the treatment of choice, with significant improvement seen in 19 of 20 patients with one post‐operative death. This response, lasting from 6 months to 7 years (median 4 years), was seen irrespective of whether splenectomy was the initial treatment or used later in management.
Late onset immune pancytopenia following bone marrow transplantation
Authors: Bashey Asad , Owen Irene , Lucas Geoffrey F. , Amphlett Nigel W. , Jones Margaret M. , Lawal Adebayo , McMullin Mary‐Frances , Mahendra Prem , Tyfield Linda A. , Hows Jill M. 
Published: March 2008

Summary A 17‐year‐old boy developed autoimmune pancytopenia in the absence of chronic graft‐versus‐host disease 170 d after allogeneic bone marrow transplantation (BMT) from his HLA identical brother. The anaemia and thrombocytopenia responded to conventional immunosuppressive treatment, but the neutropenia was refractory to this and to splenectomy and subsequent removal of splenic remnant.Following total lymphoid irradiation the neutrophil count rose to low normal levels but thrombocytopenia and anaemia secondary to marrow hypoplasia required transfusion support. Bone marrow function was finally normalized by an additional transfusion of donor marrow without prior immunosuppressive therapy. We conclude that late onset immune pancytopenia post BMT caused by antibodies of probable donor origin may be life threatening in the absence of chronic graft‐versus‐host disease.
Rearrangements of the RAR‐α gene in acute promyelocytic leukaemia:
Authors: Coco Francesco Lo , Avvisati Giuseppe , Diverio Daniela , Biondi Andrea , Pandolfi Pier Paolo , Alcalay Myriam , Rossi Giulio De , Petti Maria C. , Cantù‐Rajnoldi Angelo , Pasqualetti Daniela , Nanni Mauro , Fenu Susanna , Frontani Marina , Mandelli Franco 
Published: March 2008

Summary We have used genomic probes which specifically recognize DNA rearrangements of the RAR‐α locus on chromosome 17q21 in patients with acute promyelocytic leukaemia (APL) and acute myeloid leukaemia (AML) subtypes. Molecular data were examined in comparison with morphological and immunophenotypic characterization at diagnosis in 20 hypergranular FAB M3 cases, five microgranular APL (M3v), 51 non‐M3 AML and 12 myeloid CML blast crises.Rearrangements of the RAR‐α locus were only detected in 23/25 APL cases and in none of the other FAB subtypes analysed. Surface marker characterization showed a consistent immunophenotypic profile—HLADR negative. CD9 and CD13/33 positive—in all M3 and M3v cases. Neither HLADR negativity nor CD9 positivity were associated with RAR‐α rearrangements in non M3 AML.Our data indicate that RAR‐α gene rearrangements are relevant diagnostic features of both M3 and M3v, and may prove useful molecular marker for follow‐up analysis in APL patients.
Heterogenous expression of decay accelerating factor and CD59/membrane attack complex inhibition factor on paroxysmal nocturnal haemoglobinuria (PNH) erythrocytes
Authors: Shichishima Tsutomu , Terasawa Takashi , Hashimoto Chokichi , Ohto Hitoshi , Uchida Tatsumi , Maruyama Yukio 
Published: March 2008

Summary In order to clarify the characterization of pheno‐types of paroxysmal nocturnal haemoglobinuria (PNH) erythrocytes (E), we analysed the expression of decay accelerating factor (DAF) and CD59/membrane attack complex inhibition factor (MACIF) on the membrane surface of PNH‐E by means of the flow cytometric method using anti‐DAF and/or CD59/MACIF monoclonal antibodies in nine PNH‐patients. In two‐colour analysis, this expression on PNH‐E was classified into three fractions; negative, intermediate and positive according to intensity. The negative fraction was classified into two groups; one group an exclusively negative population, and the other a negative population having slightly DAF‐positive E. The intermediate fraction was recognized on PNH‐E of cases with PNH II‐E and extremely heterogenous. In the positive fraction, this expression was almost the same as on normal E except for case 8. In single‐colour analysis for DAF or CD59/MACIF, three fractions were classified as well as the definition in two‐colour analysis. In single‐colour analysis, the expression on PNH‐E was also heterogenous in each fraction and among PNH‐patients. However, the intermediate fraction for CD59/MACIF was not found on PNH‐E of cases without PNH II‐E, although intermediate fraction for DAF was recognized on PNH‐E of some cases with PNH III‐E in addition to those with PNH II‐E.The results suggest that expression of CD59/MACIF and DAF on the membrane surface of PNH‐E phenotypes is heterogenous and varies among PNH patients.
High risk of early resistant relapse for leukaemic patients with presence of multidrug resistance associated P‐glycoprotein positive cells in complete remission
Authors: Musto Pellegrino , Melillo Lorella , Lombardi Gina , Matera Rosella , Giorgio Giuseppedi , Carotenuto Mario 
Published: March 2008

Summary. The immunocytochemical detection of multidrugresistance (MDR) associated P‐glycoprotein (P‐170) was longitudinally performed on bone marrow smears from 32 responsive patients with acute leukaemia in the different phases of the disease (at diagnosis, in complete remission, at relapse) by means of APAAP technique and monoclonal antibody C219.The whole group of eight patients with presence of P‐170 positive cells while in complete remission rapidly relapsed with a high proportion of blasts showing MDR phenotype; they were resistant to further treatments.Twelve out of 24 subjects without cells with MDR phenotype in complete remission remained in this condition, six had a responsive relapse (without significant expression of P‐170 in 5/6 patients) and six a resistant relapse. Four patients of this last group significantly expressed P‐170.Our data indicate that the detection of scattered P‐170 positive cells during complete remission might identify a subset of leukaemic patients with high risk of early and resistant relapse.
Analysis of methylation in the c‐MYC gene in five human myeloma cell lines
Authors: Ohtsuki Takemi , Nishitani Koji , Hatamochi Atsushi , Yavvata Yoshihito , Namba Masayoshi 
Published: March 2008

SummaryGenomic alterations of the human c‐MYC gene were analysed in five human myeloma cell lines established in Kawasaki Medical School and compared with those of normal lymphocytes, Raji cells from Burkitt's lymphoma, and an Epstein‐Barr virus positive lymphoblastoid cell line (LCL). Although no structural chromosome aberrations at 8q24, the c‐MYC locus, were distinct, the mRNA level of c‐MYC in these myeloma cell lines was 30–50‐fold that in normal peripheral blood lymphocytes. Regarding the methylation of c‐MYC, DNAs of the myeloma cell lines were digested with MspI plus EcoRI or HpaII plus EcoRI, and hybridized with three genomic 32P‐labelled probes; the first, second and third exons of the human c‐MYC gene, respectively. The extent of methylation in cytosine at a single CCGG site in the third exon substantially decreased in these myeloma cell lines as compared with that in normal tonsillar B, LCL and Raji cells. No significant differences in hypomethylation between these myeloma, normal B, LCL and Raji cells was detected in the first and second exon of c‐MVC. These results suggest that the hypomethylation in the third exon of c‐MYC might be related to the enhanced expression of c‐MYC in these human myeloma cell lines.
Haemophilia B Leyden arising de novo by point mutation in the putative factor IX promoter region
Authors: Royle G. , Van de Water N. S. , Berry E. , Ockelford P. A. , Browett P. J. 
Published: March 2008

SummaryHaemophilia B Leyden is characterized by severe factor IX deficiency during childhood with partial resolution at puberty or following the administration of anabolic steroids. The disorder has recently been associated with point mutations in the putative factor IX promoter region, which contains an imperfect direct repeat spanning a possible start site of transcription. We have identified a T to C transition at position +8 in the promoter region of a patient with the haemophilia B Leyden phenotype. A mutation at this site has not been previously reported and occurs within the repeat consensus sequence in the transcribed but untranslated portion of the gene. There is no family history of haemophilia and sequence analysis of his mother and other family members indicates that the mutation has arisen de novo in this patient. This observation provides futher support for a causal relationship between point mutations in the presumptive promoter region of the factor IX gene and the Leyden phenotype.