bluuebunny/crossref_metadata_2025 huggingface dataset:
A prognostic classification of myelofibrosis with myeloid metaplasia
Authors: Barosi Giovanni , Berzuini Carlo , Liberato Lucio N. , Costa Attilia , Polino Grazia , Ascari Edoardo
Published: March 2008

The dependence of survival time on a set of prognostic factors was explored by means of Cox's regression model in 137 cases of myelofibrosis with myeloid metaplasia (MMM). The following parameters recorded at diagnosis proved to be important independent indicators of a poor prognosis: a higher value for age, a lower value for Hb concentration, a higher value for immature myeloid cells in peripheral blood (IMC), a lower value for total erythroid iron turnover (TEIT), and a bone marrow red cell aplasia (RCA). A prognostic classification tree was constructed whose terminal nodes (risk groups), described by simple logical conditions upon important indicators, were characterized by significantly different expected survival. The two extreme risk groups lend themselves to a simple, but complete description. The low‐risk group (19.7% of the sample) comprises cases who had the diagnosis of MMM before age 45 and a number of IMC constantly lower than 24%. The actuarial proportion of patients surviving at 15 years was 100%. The high‐risk group (29.9% of cases) comprises patients with age greater than 45 and Hb lower than 13 g/dl, associated with RCA, or with a relatively decreased erythropoiesis (TEIT lower than 2 times the normal) or with IMC greater than 24%. Seven out of the 11 who died within this group developed blastic crisis. Median survival time of the group was 69 months.
Platelet peroxidase‐positive blast cells in transient myeloproliferative disorder with Down's syndrome
Authors: Suda Junko , Eguchi Mitsuoki , Ozawa Takebumi , Furukawa Toshiharu , Hayashi Yasuhide , Kojima Seiji , Maeda Hiroo , Tadokoro Kenji , Sato Yuko , Miura Yasusada , Ohara Akira , Suda Toshio
Published: March 2008

Transient myeloproliferative disorder accompanied by Down's syndrome has been characterized as exhibiting self‐limiting haematological abnormalities. We studied six patients suffering from this disorder in order to clarify the biological nature of their blast cells. Metaphases of leucocytes stimulated with phytohaemagglutinin (PHA) showed trisomy 21 in all patients except one. The exception was constitutionally trisomy 21 mosaic (46, XY = 89/ 47,XY. + 21 = 11). However, metaphases from the peripheral blood cells (blast cells: 70%) without PHA stimulation showed exclusively trisomy 21. Simultaneous examination for morphology and chromosomal analysis on single colonies revealed that granulocyte‐macrophage (GM) colonies and an erythroid colony contained only cells with the trisomy 21 karyotype.

The blast cells showed positive reactions for platelet peroxidase (PPO) and with monoclonal antibodies against platelet‐megakaryocyte antigen (TP 80 and TP 82). In methylcellulose and liquid culture systems, high plating efficiencies were observed, and mainly mature basophils containing histamine developed in the presence of PHA‐stimulated leucocyte conditioned medium (PHA‐LCM). In vivo, mature neutrophils, basophils, eosinophils or megakaryocytes coexisted with PPO‐positive blast cells in the peripheral blood of some patients with this disorder.

These findings suggest that transient myeloproliferative disorder is characterized by a proliferation of PPO‐positive blast cells with trisomy 21, although some heterogeneity may be seen.
Nucleoside transport in acute leukaemia and lymphoma: close relation to proliferative rate
Authors: Wiley J. S. , Snook M. B. , Jamieson G. P.
Published: March 2008

Summary The proliferation of mammalian cells requires nucleosides which are provided either by de novo synthesis or by influx of nucleosides via membrane transporters with subsequent metabolic trapping. In this study the density of nucleoside transporters in freshly‐isolated blast cells from patients with leukaemias and lymphomas was quantitated by equilibrium binding of 3H‐nitrobenzylmercaptopurine riboside (NBMPR). In acute myeloid leukaemia (AML) the density of NBMPR binding sites on blast cells ranged from 3800 to 24 200 sites/cell and this value correlated with the 3H‐thymidine labelling index (1–20%) which was used to measure proliferative rate (r=0.80, P < 0.001). Cells from patients with Burkitt's lymphoma, other B‐cell lymphomas, T‐lymphoblastic lymphoma and large cell lymphoma gave a 20‐fold range of NBMPR site densities (from 3700 to 75 300 sites/cell) and site numbers correlated closely with the labelling index (r=0.87, P < 0.001). Non‐proliferating cells from patients with chronic lymphocytic leukaemia expressed the lowest density of NBMPR binding sites (850–2900 sites/cell). Comparison of bone marrow and peripheral blood blasts confirmed the positive correlation between NBMPR binding sites and labelling index for four individual patients. In contrast, the density of NBMPR binding sites on lymphoblasts from non‐T acute lymphoblastic leukaemia (ALL) was low (2300–7400 sites/cell) and showed little dependence on proliferation over a wide range of labelling indices (1–20%). No correlation was observed between NBMPR site density and cell size measured by the intracellular water space. Thus an increased proliferative rate of AML or lymphoma is associated with higher numbers of nucleoside transporters in the cell membrane.
Factor IX Cardiff: a variant factor IX protein that shows abnormal activation is caused by an arginine to cysteine substitution at position 145
Authors: Liddell M. B. , Peake I. R. , Taylor S. A. M. , Lillicrap D. P. , Giddings J. C. , Bloom A. L.
Published: March 2008

Summary Crude barium chloride eluates prepared from 12 unrelated patients with cross‐reacting material positive (CRM +) haemophilia B were activated with celite eluate, the reaction products resolved after reduction by 13% SDS‐ PAGE, and factor IX antigenic material detected by probing with radiolabelled immunopurified rabbit anti‐factor IX antiserum followed by autoradiography. Out of the 12, one sample showed faulty activation with the production of a stable reaction product with a MW compatible with that of a putative light chain‐activation intermediate. In order to confirm this, two oligonucleotide primers that bracketed exon 6 of the factor IX gene were constructed and used to prime a polymerase chain reaction on DNA isolated from the patient's peripheral blood leucocytes. A single 489 nucleotide DNA fragment was obtained, gel purified, subcloned into M13, and DNA sequencing carried out on both strands. A single C to T transition was discovered that changed the Arg residue at position 145, the first residue of the first bond in the activation peptide, to a Cys, a result that confirmed the inferences drawn from the activation studies.
Reduction of membrane band 7 and activation of volume stimulated (K+, Cl−)‐cotransport in a case of congenital stomatocytosis
Authors: Morlé L. , Pothier B. , Alloisio N. , Féo C. , Garay R. , Bost M. , Delaunay J.
Published: March 2008

We report on a case of congenital stomatocytosis in a French boy presenting with a haemolytic anaemia requiring splenectomy at the age of 6. The red cells included 15‐20% stomatocytes and displayed a marked increase of volume. Their osmotic resistance and density were reduced; however, their deformability was unaltered in isotonicity. Erythrocyte Na+ was high (27 mEq/1) and K+ low (65 mEq/1). The newly described (K+, Cl−)‐cotransporter normally triggered by hypo‐osmotic stress, was activated to maximal capacity. Membrane band 7 was reduced by 72%. From anamnestic data, the condition appears to have been transmitted by the father. The mother proved to be strictly normal on clinical, morphological, osmotic and biochemical bases. We suggest that the partly missing band 7 may play an important role in the genesis of stomatocytosis.
Participation of radioresistant Forssman antigen‐bearing macrophages in the formation of stromal elements of erythroid spleen colonies
Authors: Sadahira Yoshito , Mori Masaharu , Kimoto Tetsuo
Published: March 2008

Summary. Using immunohistological methods with anti‐Forssman glycosphingolipid antiserum and F4/80 monoclonal antibody, we investigated the dynamics of stromal macrophages during the development of haematopoietic spleen colonies in lethally irradiated adult mice which had been transplanted with syngeneic bone marrow cells. After irradiation, the spleen had shrunk and both Forssman +F4/80+ and Forssman‐aggregated F4/80+ macrophages tended to aggregate in the red pulp. On the other hand, after irradiation and transplantation, discriminate erythroid foci developed in the shrunken splenic red pulp and both types of aggregating macrophages, which were involved in the foci, began to extend their cytoplasmic processes. Later, the macrophages distributed themselves within the enlarged erythroid colonies and showed fine cytoplasmic extensions among erythroblasts. We also examined the occurrence of stromal macrophages in the haematopoietic foci during spleen development. While Forssman + macrophages were scarcely present at birth when erythropoietic activity had already occurred, many F4/80+ macrophages were observed. Forssman+macrophages gradually increased in number up to 3 weeks after birth. The results clearly demonstrated heterogenicity of stromal macrophages in haematopoietic foci.
AML associated with previous cytotoxic therapy, MDS or myeloproliferative disorders: results from the MRC's 9th AML trial
Authors: Hoyle Christine F. , Bastos Marcosde , Wheatley Keith , Sherrington P. D. , Fischer P. J. , Rees J. K. H. , Gray Richard , Hayhoe F. G. J.
Published: March 2008

Summary. The outcome of treatment with standard first line therapy of 66 patients with acute myeloid leukaemia (AML) secondary to preceding chemotherapy (Group 1), a myelo‐dysplastic state (Group 2) or a myeloproliferative disorder (Group 3) was analysed in relation to the preceding disorder, the cytogenetic pattern where available, and the cytology and cytochemistry of blood and bone marrow.

The complete remission (CR) rate for the secondary AMLs was 36% (24/66), with 24% (16/66) dying in the induction period and 39% (26/66) having resistant disease. The CR rate was 25% (5/20) for Group 1, 42% (15/36) for Group 2, and 40% (4/10) for Group 3. Even after allowance for the generally older age of the secondary AML patients, they still had a significantly poorer CR rate than the de novo AMLs (P= 0.0004). The lower CR rate was chiefly due to resistant disease. Despite this, overall survival was not significantly worse for the secondary AML patients (P= 0.15). For the 36% that achieved remission, remission duration appeared similar to that of de novo cases.

Of 62 cases with adequate cytology, 38 (61%) had evidence of erythroid and/or megakaryocytic dysplasia with a CR rate of 32% (12/38). The CR rate of these multineage leukaemias was not significantly different from that of the 24 (39%) who showed granulocyte/monocyte precursor involvement only, 42% (10) of whom achieved CR. The presence of features of differentiation within blast cells such as Auer rods or sudanophilia (> 50% positive blasts) was associated with a higher remission rate 47% (18/38) than that of poorly differentiated cases 17% (3/18) (P= 0.04) and thus appeared to be a more important determinant of CR achievement than was lineage involvement.

Cases with a normal karyotype had a 33% (7/21) CR rate, while those with chromosomal abnormalities had a 37% (9/24) CR rate. Only 12 of the 45 cases with adequate cytogenetic analysis showed deletions or monosomies involving chromosomes 5 or 7, and seven of these were in Group 1.
Haemolytic anaemia in analpha‐lipoproteinaemia (Tangier disease): morphological, biochemical, and biophysical properties of the red blood cell
Authors: Reinhart Walter H. , Gössi Urs , Bütikofer Peter , Ott Peter , Sigrist Hans , Schatzmann Hans‐Jürg , Lutz Hans U. , Straub P. Werner
Published: March 2008

Summary. A patient with familial analpha‐lipoproteinaemia (Tangier disease) was found to have stomatocytosis and haemolytic anaemia. The analysis of the red cell membrane constituents revealed a low cholesterol content (90 nmol/ml red cells, control 130 nmol/ml red cells), a decreased cholesterol/phospholipid ratio (0·54, control 0·78), high phosphatidylcholine (41·5%, control 30·6%) and low sphingomyelin (18·8%, contol 27·6%). The electrophoretic membrane protein pattern was normal. Osmotic gradient ektacytometry and osmotic resistance showed a decreased surface/volume ratio, which caused an increased filtration resistance in 3 μm pores. The elasticity of the membrane was unchanged. Functional membrane properties were altered: the anion exchange rate was increased, whereas alkali cation fluxes were normal. The capacity to release vesicles was reduced. This case represented a new type of stomatocytosis. It contributes to the understanding of the role of cholesterol and phospholipids in the red cell membrane and biomembranes in general.
Adult T cell leukaemia cells are of CD4+ CDw29+ T cell origin and secrete a B cell differentiation factor
Authors: Yamada Yasuaki , Ichimaru Michito , Shiku Hiroshi
Published: March 2008

Summary. Cells from six cases of adult T cell leukaemia were studied with respect to phenotypical and functional features. All cells were reactive with anti‐CD4 and anti‐CDw29 monoclonal antibodies (antibody against helper inducer T cells) but were unreactive with anti‐CD45R monoclonal antibody (antibody against suppressor inducer T cells). Functionally, these cells secreted a B cell differentiation factor detected by SKW6‐CL4 cell differentiation to IgM‐producing cells, this secretion being enhanced by culture with recombinant IL‐2. Though these results indicate that adult T cell leukaemia cells are of mature helper inducer T cell origin, these cells strongly suppressed PWM‐induced B cell differentiation in the absence of CD8+ suppressor effector T cells.
The DNA deletion in an Indian δβ‐thalassaemia begins one kilobase from the Aγ globin gene and ends in an LI repetitive sequence
Authors: Mishima N. , Landman H. , Huisman T. H. J. , Gilman J. G.
Published: March 2008

High fetal haemoglobin levels of 5–15% are present in adult heterozygotes for δβ‐thalassaemia as the result of large deletions of DNA. We have cloned DNA spanning the deletion breakpoint for a new Indian δβ‐thalassaemia associated with mild anaemia. The 5’ breakpoint is at 42151 of GenBank file HUMHBB, which is about 1 kb 3’ of the Aγ globin gene poly A site at 41003. On the 3’ side of the breakpoint, the sequence is homologous to LI (KpnI) repetitive DNA located 3.6–10 kb 3’ of the β‐globin gene: Indian δβ‐thalassaemia DNA is 74% homologous to the inverted complement of HUMHBB from 69849 to 70020, followed by a region 78% homologous to the direct sequence of HUMHBB from 70534 to 71010. The precise location of the 3’ endpoint of this deletion has not been determined, but it is within LI sequences located more than 10 kb 3’ of the β‐globin gene.
Favourable outcome of patients older than 40 years of age after transplantation with marrow grafts depleted of lymphocytes by counterflow centrifugation
Authors: Bär B. M. A. M. , Witte T. De , Schattenberg A. , Boezeman J. , Hoogenhout J.
Published: March 2008

Nineteen patients with a median age of 43 years (range 40–48) were transplanted for acute myelogenous leukaemia (AML), refractory anaemia with excess of blasts in transformation (RAEBt), acute lymphoblastic leukaemia (ALL) in first complete remission, multiple myeloma, and chronic myelogenous leukaemia (CML) in chronic or accelerated phase. Their outcome was compared with that of 35 patients with a median age of 34 years (range 30–39), and a group of patients with age younger than 30 years (median 24; range 16–29) transplanted for the same indications. Donors were human leucocyte antigen (HLA)‐identical, mixed lymphocyte culture (MLC) negative siblings. All marrow grafts were depleted of lymphocytes by counterflow centrifugation. The estimated event‐free survival at 3 years after allogeneic bone marrow transplantation was 60.7% for patients with age > 39 years, 57.8% for patients with age <30, and 43.0% for the intermediate age group (P>0.3). The estimated transplant‐related mortality showed no tendency to increase with older age of recipients. The incidence of acute GVHD>grade 1 was 15.7% in patients with age > 39 years, 9.5% in patients younger than 30 years, and 23% in the intermediate age group. The incidence of chronic GVHD was higher in the older patients (39% compared to 24% in patients younger than 30 years, 19% in the intermediate age group). Chronic GVHD resolved completely in five out of seven patients aged 40 years or more.

Reduction of the incidence of acute graft‐versus‐host disease by physical lymphocyte depletion allows allogeneic bone marrow transplantation for patients aged 40–50 years without increase of transplant‐related mortality resulting in similar event‐free survival in patients older than 40 years compared to those younger than 40 years.
Allogeneic bone marrow transplantation for secondary leukaemia and myelodysplastic syndrome: a survey by the Leukaemia Working Party of the European Bone Marrow Transplantation Group (EBMTG)
Authors: Witte T. De , Zwaan F. , Hermans J. , Vernant J. , Kolb H. , Vossen J. , Lönnqvist B. , Beelen D. , Ferrant A. , Gmür J. , Yin J. Liu , Troussard X. , Cahn J. , Lint M. Van , Gratwohl A.
Published: March 2008

This retrospective survey of the EBMT Leukaemia Working Party describes 78 patients with myelodysplasia (MDS) or secondary acute myelogenous leukaemia (sAML) who received an allogeneic bone marrow transplant (BMT). The status of underlying disease at the time of transplantation was prognostic for the 2‐year disease‐free survival. Thirty‐four patients received intensive chemotherapy prior to the conditioning for BMT. The 2‐year disease‐free survival was 60% for the 16 patients transplanted in complete remission. The results were significantly less favourable for those with more advanced disease who only partially responded to prior intensive chemotherapy (2‐year diseasefree survival: 18%) while none of those who either relapsed or were resistant to chemotherapy survived BMT for 2 years. Forty‐four patients had not received any prior intensive chemotherapy. The disease‐free survival at 2 years after BMT was 58 ± 19% when a patient was transplanted for refractory anaemia (RA(S)), 74 ± 14% for refractory anaemia with excess of blasts (RAEB), 50 ± 16% for RAEB in transformation (RAEBt), and 18 ± 11% for secondary AML.

Allogeneic BMT can therefore be considered as curative treatment for patients with MDS. Patients with sAML who have a histocompatible donor should be given chemotherapy intensive enough to induce complete remission. If this is achieved these individuals have a prognosis comparable to those with de novo AML in first remission after BMT.
Interleukin 2 does not promote the in vitro and in vivo proliferation and growth of human acute leukaemia cells of myeloid and lymphoid origin
Authors: Foa R. , Caretto P. , Fierro M. T. , Bonferroni M. , Cardona S. , Guarini A. , Lista P. , Pegoraro L. , Mandelli F. , Forni G. , Gavosto F.
Published: March 2008

The effect of recombinant interleukin 2 (IL2) on the in vitro and in vivo proliferation and growth of human acute leukaemia cells of both myeloid and lymphoid origin was investigated. In none of the 25 primary samples tested could a continuously in vitro growing cell line be obtained by adding IL2 to the culture medium. Although IL2 induced a proliferative signal in three of the 31 acute leukaemias analysed, the overall 3H‐thymidine uptake of the neoplastic cells was significantly reduced (P<0.05) in the presence of IL2. The unlikelihood of an important proliferative signal triggered by IL2 was confirmed in a semisolid clonogenic assay, which failed to document an increased colony growth in the 26 samples studied. Furthermore, using a colorimetric assay as a test for cell proliferation and survival, in seven of the 11 fresh acute leukaemia samples tested a 22–40% reduction in viability was observed in the presence of IL2, while in the remaining four, IL2 was ineffective. In order to investigate the effect of IL2 in an in vivo setting, an experimental model in heavily immunosuppressed nu/nu mice was established. In no case did IL2 promote the in vivo proliferation and growth of human myeloid and lymphoid acute leukaemia cells injected in the mice. On the contrary, with seven of the eight leukaemic cell lines which gave rise spontaneously to leukaemic masses, this could be prevented when the mice received locally 300 U of IL2 three times daily for 90 d. IL2 also blocked the growth in vivo of three fresh acute leukaemia samples (two myeloid and one lymphoid). Co‐culture experiments using leukaemic cell lines and increasing numbers of normal lymphocytes suggest that the inhibitory effect of IL2 is probably exerted via an indirect mechanism. These findings, coupled to the well‐documented ability of IL2 to generate lymphokine activated killer cells cytolytic against leukaemic blasts, further point to the potential role of immunotherapy with IL2 in the management of patients with haematological malignancies.
High serum IL‐2 levels are predictive of prolonged survival in multiple myeloma
Authors: Cimino Giuseppe , Avvisati Giuseppe , Amadori Sergio , Cava Maria Carmela , Giannarelli Diana , Di Nucci Gian Domenico , Magliocca Vittoria , Petrucci Maria Teresa , Poti Giampiero , Sgadari Cecilia , Mandelli Franco
Published: March 2008

Summary.In this study we analysed serum IL‐2 levels in 61 patients with multiple myeloma (MM). Patients serum IL‐2 levels were significantly higher than normal controls. Moreover, higher serum IL‐2 levels were associated with a prolonged actuarial survival. In particular, 87% of the MM patients with IL‐2 ≥ 10 U/ml are still alive at 5 years while only 13% of the remaining patients with IL‐2 < 10 U/ml are alive. The multivariate analysis confirmed these data indicating that high serum IL‐2 levels are the most useful predictor index of longer survival in MM patients. Furthermore, among the 50 patients in whom serum beta‐2‐microglobulin (SB2M) determination was available we observed that all patients with serum IL‐2 levels ≥ 10 U/ml had SB2M < 6 μg/ml, whereas in patients with serum IL‐2 < 10 U/ml SB2M ranged from 1.3 to 15 μg/ml. Using these two parameters we were able to identify three groups of patients with different survival duration. Group A (9 patients) defined by serum IL‐2 ≥ 10 U/ml and SB2M < 6 μg/ml in which all patients are alive; group B (26 patients) characterized by serum IL‐2 < 10 U/ml and SB2M < 6 μg/ml in which 24% of patients are alive and group C (15 patients) characterized by serum IL‐2 levels < 10 U/ml and SB2M ≥ 6 μg/ml in which the actuarial survival curve drops to 0 at 2.5 years. A statistically significant difference was observed between groups A and B (P< 0.05), groups A and C (P< 0.01) and groups B and C (P< 0.01). These data could reflect the existence of an active T cell control on B cell neoplasia and may suggest the opportunity of a more extensive use of recombinant biological modifiers such as IL‐2 in the therapeutic strategy of MM.
The common pathway for alpha‐ and gamma‐thrombin‐induced platelet activation is independent of GPIb: a study of Bernard‐Soulier platelets
Authors: Jandrot‐Perrus Martine , Rendu Francine , Caen Jacques P. , Levy‐Toledano Sylviane , Guillin Marie‐Claude
Published: March 2008

Summary. The responses to alpha‐ and gamma‐thrombin were studied in normal and Bernard‐Soulier platelets labelled with [32P]phosphate, to investigate the relationship between thrombin binding to the platelet membrane glycoprotein Ib (GPIb) and thrombin‐induced platelet activation. For this purpose we conducted parallel studies of the kinetics of platelet aggregation, granule secretion, hydrolysis of poly‐phosphoinositides, formation of phosphatidic acid, phosphorylation of the myosin light chain (p20) and of the 43 kDa protein (p43), and thromboxane B2 formation. Like alphathrombin, gamma‐thrombin activated control platelets via all the above metabolic responses, but only after a prolonged lag. In Bernard‐Soulier platelets, alpha‐thrombin induced polyphosphoinositide hydrolysis and phosphatidic acid formation, p20 and p43 phosphorylation, thromboxane B2 formation, secretion and to a lesser extent aggregation, but only after a prolonged lag. The metabolic responses of Bernard‐Soulier platelets to gamma‐thrombin were very similar to those of control platelets.

We have previously showed that GPIb which is not present in Bernard‐Soulier platelets binds alpha‐ but not gammathrombin. The present results indicate that thrombin binding to GPIb is not directly coupled either with the activation of phospholipase C specific to polyphosphoinositides, or with the activation of protein kinase C and phospholipase A2. However, thrombin binding to GPIb appears to promote an early mechanism which accelerates all the platelet responses.
‘MACHO’ chemotherapy for stage IV B cell lymphoma and B cell acute lymphoblastic leukaemia of childhood
Authors: Hann I. M. , Eden O. B. , Barnes J. , Pinkerton C. R.
Published: March 2008

An intensive 6‐month schedule of drugs was devised with both systemic and central nervous system activity, known by the acronym ‘MACHO’, to treat 24 newly and consecutively diagnosed children, 13 with stage IV B‐cell non‐Hodgkin's lymphoma (B‐NHL) and 11 with B‐cell acute lymphoblastic leukaemia (B‐ALL). There were three deaths from complications of chemotherapy (two infective, one biochemical). Five children with central nervous system disease at diagnosis (CNS +) received planned additional megatherapy/bone marrow transplants. Event‐free survival (EFS) at 1 year for the 11 cases of B‐ALL is 64% (95% confidence intervals [CI] 31–89%) and of 13 stage IV B‐NHL cases is 50% (95% CI 19–75%). Patients with bulky abdominal disease had a 32% EFS at 1 year (CI 13–68%) compared with 76% (CI 39–94%) for those without bulky abdominal disease. Overall EFS for eight CNS + patients is 73% at 1 year (95% CI 34–97%) compared with 48% (95% 24–74%) for those without CNS disease (CNS—). However, only two of the CNS+ cases had bulky abdominal disease (patients 10 and 12) and the difference is not significant (P<0·5).

A score of 1 was given for each of the following potential prognostic features: bulky abdominal disease, pleural effusion and severe renal dysfunction within 48 h of presentation. Patients who scored 0 or I fared significantly better than those who scored 2 or 3 (EFS at 1 year 78% [CI 49–95%] versus 24% [6–65%], P<0·04). Two patients with a score of 2 survived past 6 months and another is currently well, but has not regenerated his marrow following autologous transplantation. This protocol is relatively effective for patients who have B‐ALL, but those patients who have bulky abdominal disease, often associated with severe renal dysfunction, and those with CNS disease, do not fare so well and require new approaches to therapy.
A splicing mutation accounts for the lack of p53 gene expression in a CML blast crisis cell line: a novel mechanism of p53 gene inactivation
Authors: Foti A. , Bar‐Eli M. , Ahuja H. G. , Cline M. J.
Published: March 2008

Summary. Alterations of the p53 anti‐oncogene have recently been found to occur frequently in the blast crisis of chronic myelocytic leukaemia. The p53 gene may be altered by gross structural alterations or by point mutations in the coding sequence. We now report a novel mechanism of gene inactivation in a blast crisis cell line where a mutation in a splice donor site at the 5’end of the fifth intron of the gene interrupts RNA processing and gene expression.
Increased numbers of circulating haematopoietic progenitor cells after treatment with high‐dose interleukin‐2 in cancer patients
Authors: Schaafsma M. R. , Fibre W. E. , Van Der Harst D. , Duinkerken N. , Brand A. , Osanto. S. , Franks C. R. , And R. Willemze , Falkenburg J. H. F.
Published: March 2008

Summary Immunotherapy with recombinant interleukin‐2 (IL‐2) and lymphokine‐activated killer (LAK) cells has been applied to patients with metastatic cancers for its antitumour activity. In the present study we investigated the effects of in‐vivo administration of IL‐2 (3.106 U/m2/d, continuously i.v.) on haematopoiesis. Six patients with disseminated renal cell carcinoma, treated with IL‐2 and LAK cells, were monitored for the numbers of white blood cells and circulating haematopoietic progenitor cells (HPC). During IL‐2 treatment lymphopenia developed, followed by lymphocytosis after discontinuation of IL‐2 infusions. IL‐2 administration also resulted in neutrophilia and eosinophilia. Absolute numbers of circulating HPC declined markedly during IL‐2 treatment. However, after completing IL‐2 infusions, the numbers of circulating erythroid (BFU‐E), inyeloid (CFU‐GM) and multipotential progenitor cells (CFU‐GEMM) strongly increased, reaching a maximum after 5 d (day 10 from the start of IL‐2 treatment). This increase did not result from repeated leucaphereses, since patients treated with IL‐2 alone showed a similar response. In comparison with pretreatment levels the pool of circulating HPC expanded about 20‐fold. This study illustrates that IL‐2 treatment has a biphasic effect on the frequency of circulating BFU‐E. CFU‐GM and CFU‐GEMM. causing a decrease during 11,‐2 infusion, followed by an increase after IL‐2 administration. The total number of progenitor cells harvested by four consecutive leucaphereses is in the range that is commonly used for peripheral blood stem cell autografting.
Clinicopathological features of acute undifferentiated leukaemia with a stem cell phenotype
Authors: Brito‐Babapulle F. , Pullon H. , Layton D. M. , Etches A. , Huxtable A. , Mangi M. , Bellingham A. J. , Mufti G. J.
Published: March 2008

Summary Over a 41/2‐year period, 141 patients with acute leukaemia had morphologic, immunophenotypic and cytochemical studies performed at King's College Hospital. Seven cases were noted to have blast cells which did not express myeloid or lymphoid antigens or cytochemical staining indicative of differentiation but were HLA DR and CD 34 positive. Based on these criteria we have used the term stem cell acute leukaemia to denote these patients. There were five women and two men with a median age of 61 years (16–86). Presentation marrows were heavily infiltrated with blasts (>95% in 6/7) which were usually pleomorphic. Type 2 blasts, Auer rods and dysplastic features were absent. Two of six cases studied showed clonal karyotypic abnormalities. Four patients were treated with high dose chemotherapy. Three of these achieved a complete remission but relapsed at 3, 6 and 7 months respectively. The median survival of the group was 7 months (2–12). We conclude that the stem cell acute leukaemias are a distinct clinicomorphological group which appear to have a poor prognosis with conventional chemotherapy.
Definition of two distinct types of AIDS‐associated non‐Hodgkin lymphoma
Authors: Boyle Michael J. , Swanson Cheryl E. , Turner Jennifer J. , Thompson Ian L. , Roberts John , Penny Ronald , Cooper David A.
Published: March 2008

Summary Non‐Hodgkin lymphoma (NHL) in a human immunodeficiency virus (HIV)‐infected person is an AIDS‐defining condition. There is both an increased incidence of NHL and a poorer prognosis for the disease in these HIV‐infected persons, compared with patients who spontaneously develop NHL.

A series of 41 consecutive patients with HIV‐associated NHL treated at St Vincent's Hospital, Sydney. between 1984 and September 1989 was assessed. Of these, 36 had NHL proven on biopsy and NHL was diagnosed clinically in the other five. On histologic examination 11 had high‐grade malignant lymphoma, small non‐cleaved cell (SNCC) type, similar to Burkitt lymphoma, 18 had high‐grade lymphoma large cell immunoblastic (LCI) type, and seven could not be classified due to inadequate material for assessment.

The clinical features of these two groups were compared. Those patients with LCI had advanced CD4+ immunodeficiency and poor survival (median = 6 weeks). The other group with SNCC had better preserved immune function, and longer survival (median=18 weeks). Three patients had survived longer than 3 years all of whom had SNCC lymphoma as their first AIDS‐defining illness.

The clinical features and prognostic factors demonstrated in this study document heterogeneity of NHL in HIV‐infected patients and should be used as a guide to treatment of these patients, by reserving more intensive treatment for those persons with better prognostic features.
Contrasuppressor T cell leukaemia: clonal proliferation of contrasuppressor T cells in a patient with granular lymphocyte‐proliferative disorder
Authors: Saito Hiroshi , Oshimi Kazuo , Akahoshi Masako , Yamauchi Katsumi , Yamada Osamu , Mizoguchi Hideaki
Published: March 2008

SummaryIn 12 patients with granular lymphocyte‐proliferative disorders (GLPD), we studied the capacity of patient peripheral blood mononuclear cells (PBMC) to promote or suppress polyclonal IgG synthesis by normal non‐T cells in pokeweed mitogen‐containing medium using an enzymelinked immunosorbent assay. During the experiments we found a patient whose PBMC possessed contrasuppressor function. The patient was a 27‐year‐old female with anaemia and lymphocytosis of CD3+CD8+granular lymphocytes (GL). Reconstitution experiments using normal donor non‐T cells and CD4+and CD8+T cells showed that addition of the patient's CD8+cells abrogated the suppressor cell function of normal CD8+T cells. The patient's PBMCs were CD3+, CD8+, Ia+, andVicia villosalectin‐adherent characteristics which are consistent with those of normal blood contrasuppressor T cells. The T cell receptor β and γ genes were found to be monoclonally rearranged. Ultrastructurally, this patient's GLs exhibited clusters of dense cytoplasmic bodies, which were not detected in the GL of other patients with GLPDs. These results indicate that the clonal proliferation of contrasuppressor T lymphocytes had occurred in this patient.
Plasma erythropoietin levels and acquired cystic disease of the kidney in patients receiving regular haemodialysis treatment
Authors: Edmunds M. E. , Devoy M. , Tomson C. R. V. , Krishna U. , Clayworth A. , Durrant S. T. S. , Feehally J. , Walls J.
Published: March 2008

SummaryAcquired cystic disease of the kidney (ACDK) in patients with end‐stage renal failure can be associated with development of polycythaemia. The relationship between plasma erythropoietin levels and ACDK in 17 patients on long‐term haemodialysis treatment was studied. There was a significantly higher level of plasma erythropoietin in patients with multiple renal cysts than in those patients with less than five cysts or no cysts. Haemoglobin tended to be higher in the ACDK group, but the difference was not significant. These results indicate that the development of renal cysts results in increased secretion of erythropoietin.
Identification of two point mutations in the von Willebrand factor gene of three families with the ‘Normandy’variant of von Willebrand disease
Authors: Gaucher Christine , Mercier Bernard , Jorieux Sylvie , Oufkir Driss , Mazurier Claudine
Published: March 2008

Summary Plasma von Willebrand factor (vWf) is a multidomain multimerized glycoprotein which has a dual role in haemostasis: it promotes platelet adhesion to subendothelium and is the carrier of blood coagulation factor VIII (FVIII). We previously characterized a functional defect of vWf, limited to its ability to bind FVIII, in two families whose affected members have the same phenotype that mimics mild haemophilia A and was tentatively named von Willebrand's disease (vWD) ‘Normandy’. A homozygous point mutation C T converting Thr 28 to Met in mature vWf subunit was identified in one of these patients who was born of third‐cousin parents. In the present studies we report two unrelated new cases of vWD ‘Normandy’and characterize, using the analysis of the vWf gene intron 40 region containing a variable number of tandem repeats, the recessive inheritance of the disease in two affected families without known consanguinity. Exons 18–24 of the vWf gene encoding for the first 311 amino acids of mature vWf subunit were amplified by the polymerase chain reaction method and sequenced. Two new missense mutations, both corresponding to a C T transition and predicting respectively an Arg 53 Trp and an Arg 91 Gln substitution, were characterized. The three patients from family 1 were homozygous for the first‐mentioned mutation while the patient from family 3 was homozygous for the second. The patient from family 2 was found a compound heterozygote for the two mutations. None of the two point mutations reported, both destroying a MspI restriction site, could be detected in DNA from 50 normal controls screened by restriction endonuclease analysis. Our data show that different mutations may be found in patients with the ‘Normandy’phenotype. The mutations characterized so far are all localized on the N‐terminal region of mature vWf subunit, within or near the major FVIII binding domain, and some of them occur within the epitope of monoclonal antibodies inhibiting the vWf/FVIII interaction. These observations suggest a causal relationship between these mutations and the vWD ‘Normandy’phenotype.
Resistance to malaria in ankyrin and spectrin deficient mice
Authors: Shear Hannah Lustig , Roth Eugene F. , Ng Chui , Nagel Ronald L.
Published: March 2008

Summary Inbred mice carrying mutations in ankyrin and/or spectrin synthesis and assembly were studied for their ability to support the growth of the rodent malarias, Plasmodium chabaudi adami and P. berghei, in vivo. Mice carrying the nb/nb (normoblastosis) mutation which do not synthesize ankyrin and therefore also have a deficiency in membrane‐bound spectrin, were refractory to P. chabaudi adami, which invades mature erythrocytes and to P. berghei, which invades reticulocytes. Similarly, sph/sph mice which do not synthesize the α chain of spectrin but do synthesize ankyrin, were also resistant to both parasites. The heterozygote for the nb defect (nb/+) exhibited a diminution of parasitaemia. We conclude that the host cell spectrin may be necessary for the invasion and/or growth of rodent malarial parasites.
Antithrombin Vicenza, Ala 384 to Pro (GCA to CCA) mutation, transforming the inhibitor into a substrate
Authors: Caso R. , Lane D. A. , Thompson E. A. , Olds R. J. , Thein S. L. , Panico M. , Blench I. , Morris H. R. , Freyssinet J. M. , Aiach M. , And F. Rodeghiero , Finazzi G.
Published: March 2008

Summary. Antithrombin (AT) Vicenza has been previously identified as a functionally abnormal antithrombin associated with familial thrombosis (Finazzi et al, 1985). It binds normally to heparin, but loses its affinity following interaction with thrombin: it is a poor inhibitor of thrombin. AT Vicenza was isolated from plasma by heparin–Sepharose and thrombin–Sepharose chromatography, fragmented with cyanogen bromide (CNBr) and its tryptic peptides were analysed by fast atom bombardment mass spectrometry mapping. An abnormal peptide mass 1112 was identified. Edman degradation confirmed a substitution of Ala to Pro in the sequence Ala 383–Arg 393. Polymerase chain reaction amplification of exon 6 of the gene followed by genomic sequencing, localized the mutation to codon 384, GCA to CCA. The same mutation has recently been reported in AT Charleville (Mohlo‐Sabatier el al, 1989).

Sodium dodecyl‐sulphate polyacrylamide gel electrophoresis of AT Vicenza (/Charleville) under non‐reducing conditions revealed an apparent increase in mol. wt following interaction with thrombin: under reducing conditions the mol. wt was less than that of normal AT. This indicated cleavage and unfolding of the molecule. The site of cleavage was determined by incubation of AT Vicenza (/Charleville) with thrombin‐Sepharose, reduction and S‐carboxymethylation and reverse phase FPLC. A peptide was identified with the NH2‐terminal sequence beginning Ser–Leu–Asn, demonstrating the cleavage had occurred at the reactive site of the variant. It is concluded that the Ala 384 to Pro substitution transforms AT Vicenza (/Charleville) from an inhibitor into a substrate.
Teniposide, dexamethasone and continuous‐infusion cyclophosphamide in advanced refractory myeloma
Authors: Leoni Franco , Ciolli Stefania , Salti Franco , Teodori Paolo , Ferrini Pierluigi Rossi
Published: March 2008

Twenty‐one patients with advanced refractory myeloma (nine primary unresponsive to melphalan, 12 relapsed and resistant to first‐line salvage therapy) received monthly 7 d courses of cyclophosphamide by continuous infusion, associated with teniposide and dexamethasone. Six patients died within 2 months from start of treatment, before receiving an adequate trial. Among the 15 evaluable patients, there were eight objective responses and three improvements (decrease of monoclonal protein > 50% or > 30%, respectively). After a median follow‐up of 10 months, all responding patients are alive and none of them is still relapsed. In heavily pretreated patients, ineligible for more aggressive approaches, cyclophosphamide by continuous‐infusion exerted a good antineoplastic activity without increasing toxicity.
Detection of soluble IL‐2 receptor in the serum of patients with myelodysplastic syndromes: induction under therapy with GM‐CSF
Authors: Zwierzina Heinz , Herold Manfred , Schöllenberger Susanne , Geissler Dietmar , Schmalzl Franz
Published: March 2008

Sera of 15 healthy controls and 33 patients suffering from myelodysplastic syndromes (MDS) were investigated for soluble interleukin‐2 receptor (sIL‐2R) expression with a cell‐free enzyme‐linked immunosorbent assay (ELISA) system (T‐Cell Sciences; Cambridge, U. S. A.). The upper limit of the assay is indicated with 477 U/ml. According to the FAB classification eight refractory anaemia (RA). 15 refractory anaemia with excess of blasts (RAEB), five refractory anaemia with excess blasts in transformation (RAEBt) and five chronic myelomonocytic leukaemia (CMML) were examined. None of the patients had reported infectious episodes or been under treatment with cytotoxic agents and/or cytokines within the previous 3 months. Significant differences in sIL‐2R levels between RA (median 368 U/ml), RAEB (median 675 U/ml) and RAEBt (median 971 U/ml) and between RA and CMML (median 723 U/ml) were detected.

Six patients, who had been under treatment with rhGM‐CSF for at least 2 weeks, demonstrated a three‐ to sevenfold increase of sIL‐2R expression compared to pretreatment levels. In kinetic evaluation of serum samples for 24 h, the increase of sIL‐2R expression begins within 4 h after subcutaneous application of GM‐CSF and reaches its maximum after 12 h.

Our data cannot suggest whether increased sIL‐2R expression is a primary event due to involvement of lymphocytes in the malignant clone or whether it results from secondary alteration of the cytokine network. Application of GM‐CSF in MDS may result in improvement of altered lymphocyte function. As GM‐CSF induces sIL‐2R expression, a down regulation of the immune response caused by neutralization of free IL‐2 cannot be excluded.
Antiproliferative activity of quercetin on normal bone marrow and leukaemic progenitors
Authors: Larocca L. M. , Teofili L. , Leone G. , Sica S. , Pierelli L. , Menichella G. , Scambia† G. , Panici P. Benedetti , Ricci R. , Piantelli M. , Ranelletti F. O.
Published: March 2008

We used an in vitro clonogenic assay in semi‐solid medium to test the sensitivity of normal bone marrow and acute myeloid and lymphoid leukaemia progenitors to the flavonol quercetin. We have studied 14 acute myeloid (AML) and four acute lymphoid (ALL) leukaemias. All ALL and the vast majority of AML (12/14) had a high sensitivity to quercetin with more than 50% growth inhibition at 2 × 10−6 M quercetin. One M3‐AML was partially quercetin‐sensitive displaying 60% surviving AML‐colony forming units (CFU‐AML) at a quercetin concentration of 10−5 M. One M1‐AML was resistant to the growth inhibitory effect of quercetin at a concentration of 2 × 10−5 M. The clonogenic efficiency of both AML and ALL positively correlated with leukaemic colony‐forming unit (CFU‐L) sensitivity to quercetin suggesting that this parameter can be useful in predicting quercetin responsiveness of leukaemic cells. We have also studied the effect of various quercetin concentrations on colony formation by normal bone marrow cells. At a quercetin concentration of 10−5 M, we observed (in five different experiments) a mean recovery of 53% and 65% of erythroid blast‐forming units (BFU‐E) and granulocyte‐macrophage colony‐forming units (CFU‐GM), respectively. Thus, normal bone marrow appeared partially resistant to quercetin, being inhibited less than 50% by quercetin concentration higher than 2 × 10−5 When normal bone marrow were deprived in CD34 + haematopoietic progenitors the resultant population became highly sensitive to quercetin, with a mean recovery of BFU‐E and CFU‐GM of 5% and 12% of controls respectively in the presence of 2 × 10−5 M quercetin. Furthermore, CD34 progenitors, positively selected, appeared fully resistant to quercetin concentrations as high as 2 × 10−5 M. Thus, CD34 + progenitors are a quercetin‐resistant component in normal bone marrow. In conclusion, our results further provide a biological basis for the therapeutic use of quercetin, considering that this compound could inhibit leukaemic cell growth without suppressing normal haematopoiesis.
Possible mechanism of action of interferon alpha in chronic B‐cell malignancies
Authors: Heslop H. E. , Brenner M. K. , Ganeshaguru K. , Hoffbrand A. V.
Published: March 2008

Recent evidence suggests that tumour necrosis factor alpha (TNF) is an autocrine growth factor for the chronic B‐cell malignancies hairy cell leukaemia (HCL) and some cases of B‐chronic lymphocytic leukaemia (B‐CLL). Incubation with TNF in vitro has been shown to increase viability, DNA synthesis and the expression of the protooncogenes myc, fos and jun in the tumour cells from these patients. TNF in vitro also increases expression of TNF‐mRNA, suggesting the existence of an autocrine growth loop for TNF in these cells. Current experiments are compatible with the hypothesis that interferon alpha (IFN) interferes with this autocrine growth loop in HCL and B‐CLL by stimulating degradation of messenger RNAs (mRNAs) for a number of cytokines including that of TNF. This RNA degradation may be mediated through induction of the enzyme 2, 5 oligo‐A synthetase with consequent increased synthesis of 2, 5 oligo‐A which is known to stimulate the activity of a latent ribonuclease capable of degrading cytokine mRNAs. Circulating tumour‐derived TNF may also contribute to the pancytopenia in HCL and B‐CLL. Whether cytokine autocrine growth loops are important in other B‐cell malignancies, e.g. myeloma and non‐Hodgkin's lymphoma, and subject to IFN‐stimulated breakdown needs further study.