bluuebunny/crossref_metadata_2025 huggingface dataset:
Potential salvage therapy for inadvertent intrathecal administration of vincristine
Authors: Michelagnoli M. P. , Bailey C. C. , Wilson I. , Livingston J. , Kinsey S. E.
Published: November 2003

Vincristine is an anti‐neoplastic agent for intravenous use. Inadvertent intrathecal administration of vincristine tends to follow a predictable clinical sequence of a progressive ascending radiculomyeloencephalopathy, usually fatal in outcome. We report a case of a 10‐year‐old girl who suffered an inadvertent intrathecal vincristine administration. We have outlined the management strategy used and her consequent neurological progress. She has survived but with a significant neurological deficit. Her outcome is, however, less severe than in previous literature reports; this may in part be attributable to early aggressive neurological management. This case also serves to highlight the tragic consequences of human error when handling chemotherapeutic agents of this nature.
Hyperbilirubinaemia in heterozygous β‐thalassaemia is related to co‐inherited Gilbert's syndrome
Authors: Galanello R. , Perseu L. , Melis M. A. , Cipollina L. , Barella S. , Giagu N. , Turco M. P. , Maccioni O. , Cao A.
Published: November 2003

The reasons why heterozygotes for β‐thalassaemia have considerable variation in serum bilirubin levels are unkown. High levels of bilirubin could be related to the co‐inherited Gilbert's syndrome, determined either by mutations of the coding region or by variation in the A(TA)nTAA motif of the promoter of the bilirubin UDP‐glucuronosyltransferase gene (UGT‐1). We sequenced the coding and the promoter region of UGT‐1A or characterized the A(TA)nTAA motif of the promoter by denaturing gel electrophoresis of radioactive amplified products. The results were correlated with bilirubin levels in 49 β‐thalassaemia heterozygotes for codon 39 (CAG → TAG) nonsense mutation. 21 normal individuals and 32 unrelated patients with Gilbert's syndrome served as controls. The coding sequence region of the UGT‐1A was normal. Five β‐thalassaemia heterozygotes, who were homozygous for the extra (TA) bases in the A(TA)nTAA element of the promoter of UGT‐1A, the configuration present in homozygosity in Gilbert's syndrome, had higher bilirubin levels compared to those with the (TA)6/(TA)7 or (TA)6/(TA)6 configurations.

In the group of 32 patients with Gilbert's syndrome, 31 of whom had the (TA)7/(TA)7 configuration, we detected 14 heterozygotes for β‐thalassaemia, a figure much higher than predicted on the basis of the carrier rate. Homozygosity for the (TA)7 motif, the typical promoter configuration of Gilbert's syndrome, is one of the factors determining hyperbilirubinaemia in heterozygous β‐thalassaemia.
Same‐day determination of chimaeric status in the immediate period following allogeneic bone marrow transplantation
Authors: Hancock J. P. , Burgess M. F. , Goulden N. J. , Steward C. G. , Knechtli C. J. C , Pamphilon D. H. , Potter M. N , Oakhill A.
Published: November 2003

The accurate and rapid determination of the origin of haemopoietic cells may provide valuable information as to the aetiology of, and most appropriate therapy for, leucopenia following allogeneic bone marrow or peripheral blood stem cell transplantation. We describe an approach to the analysis of chimaerism post bone marrow transplantation (BMT) based on the immunomagnetic capture of white cells combined with microsatellite polymerase chain reaction (PCR) and resolution of products by polyacrylamide gel electrophoresis (PAGE). This non‐isotopic method enables the chimaeric status to be determined from as little as 1.0 ml of profoundly leucopenic peripheral blood (WBC 0.1 × 109/l) and has been applicable to all donor/recipient pairs tested so far. Results are available within 6 h of blood sampling and lineage‐specific chimaerism is possible. Furthermore, blood transfusions do not interfere with the analysis.
Thrombopoietin levels in patients with primary and reactive thrombocytosis
Authors: Cerutti Alberto , Custodi Pietro , Duranti Mauro , Noris Patrizia , Balduini Carlo L.
Published: November 2003

Human c‐mpl ligand or thrombopoietin (TPO) has been proved to be a critical cytokine in the physiological regulation of thrombopoiesis. Previous evidence suggested that TPO production is constitutive and TPO plasma levels are regulated by the platelet–megakaryocyte mass through c‐mpl receptor‐mediated uptake and metabolism. To evaluate whether this mechanism of TPO level regulation is also operative in subjects with an elevated platelet count, we evaluated serum TPO in 32 patients with thrombocytosis due to essential thrombocythaemia (ET) or polycythaemia vera (PV) and in 70 subjects with reactive thrombocytosis; 32 healthy subjects were also studied. TPO levels were significantly higher in the ET and PV groups (median 246.2 pg/ml, range 93.5–4596) and reactive thrombocytosis patients (median 287 pg/ml, range 82.7–1960.0) than in normal subjects (median 156.7 pg/ml, range 62.2–352.7). No significant difference was found between the two groups of patients, indicating that serum TPO levels cannot differentiate between primary and reactive thrombocytosis. No significant correlation was found between platelet count and TPO levels in either ET‐PV or reactive thrombocytosis, whereas a trend toward a correlation between acute‐phase reactants and TPO was observed in patients with reactive thrombocytosis. These results indicate that TPO clearance by platelets–megakaryocytes is not fully operative or is not the only mechanism of TPO level regulation in primitive and reactive thrombocytosis.
Redox regulation of apoptosis in interleukin‐3‐dependent haemopoietic cells: absence of alteration in both mitochondrial membrane potential (Δψm) and free radical production during apoptosis induced by IL3 withdrawal
Authors: Garland J. M. , Søndergaard K. L. , Jolly J.
Published: November 2003

Apoptosis in haemopoietic cells has been linked to the production of free radicals and reduction in mitochondrial transmembrane potential, Δψm. We have found that apoptosis in murine IL3‐dependent haemopoietic cells induced by IL3 withdrawal lowers a principal source of anti‐oxidant (reduced glutathione) but does not increase free radicals nor affect Δψm. Increased free radicals are, however, produced during IL3 signalling. Artificially increasing intracellular free radicals by depleting internal glutathione mimics IL3 signalling in that cells maintain shape and are transiently stimulated into S‐phase in the absence of IL3. Conversely, depletion of free radicals by anti‐oxidants accelerates apoptosis both in the presence and absence of IL3. Our studies suggest that redox conditions rather than free radicals or mitochondrial membrane potential per se may play a significant role in maintaining cell and replicative integrity, and in determining apoptosis in haemopoietic cells.
Preferential adhesion of fetal liver derived primitive haemopoietic progenitor cells to bone marrow stroma
Authors: Blair A. , Thomas D. B.
Published: November 2003

The orderly transfer of haemopoiesis from the fetal liver to the medullary cavities during the ontogeny may result from a decrease in the capacity of the fetal liver to support haemopoiesis. Alternatively it could be facilitated by the preferential migration of primitive haemopoietic cells to the marrow. In this study we have compared the ability of high proliferative potential colony forming cells (HPP‐CFC) derived from murine fetal liver (FL) on the 15th day of gestation and from murine adult bone marrow (BM) to adhere to either FL or BM derived stromal layers. Adhesion of FL HPP‐CFC to bone marrow stromal layers was significantly greater than all other combinations tested. The remarkable affinity of HPP‐CFC derived from FL for BM stromal layers is consistent with the preferential migration of haemopoietic cells from the fetal liver to the medullary cavities.
The detection of minimal lymphoma by molecular and combined culture–molecular methods
Authors: Wu G. Q. , Sharp J. G. , Wu G. , Vose J. , Greiner T. C. , Chan W. C.
Published: November 2003

Seventy‐four patients from a prospective randomized trial comparing autologous bone marrow (ABM) versus blood stem cell (BSC) transplantation after high‐dose chemotherapy for intermediate and high grade non‐Hodgkin's lymphoma (NHL) were studied for the presence of residual lymphoma prior to transplantation. Pre‐transplant bone marrow (BM), peripheral blood (PB) and the ABM or BSC harvest were studied by molecular assays immediately after collection and at weekly intervals after the initiation of in vitro cultures. B‐NHLs with t(14;18) at the major breakpoint region (mbr) were monitored by detecting cells with the translocation. Other B‐NHLs were monitored with tumour‐specific primers and probes to the immunoglobulin heavy chain (IgH) gene complementary determining region (CDR) III. T‐NHLs were similarly monitored using the T‐cell receptor gamma chain gene V‐J junctional region as the tumour‐specific marker. Of the 74 patients, seven did not have adequate tumour biopsies for molecular characterization. Of the remaining 67 cases, 35 had identifiable markers for follow‐up studies and 20/35 cases (52%) had tumour cells detected in either the pretransplant BM/PB samples or the ABM/BSC harvest. Residual tumours were detected at a high frequency in T‐NHL (100%) and t(14;18)+ B‐NHL (86%) but at a lower frequency in B‐NHLs without t(14,18) (44%). In five cases, one or more of the samples were initially negative for residual lymphoma but became positive after a period of culture; additional studies confirmed that in vitro culture enhanced the sensitivity of tumour detection in about half of these samples. Molecular assay for minimal residual disease can be performed in the setting of multicentre prospective clinical trials. The substantial frequency of failure of obtaining tumour‐specific IgH CDRIII sequences in paraffin‐embedded B‐NHLs argues for the storage of frozen tumour samples for possible molecular studies.
A randomized phase II study of BB‐10010: a variant of human macrophage inflammatory protein‐1α for patients receiving high‐dose etoposide and cyclophosphamide for malignant lymphoma and breast cancer
Authors: Bernstein Steven H. , Eaves Connie J. , Herzig Roger , Fay Joseph , Lynch Joseph , Phillips Gordon L. , Christiansen Neal , Reece Donna , Ericson Solveig , Stephan Margaret , Kovalsky Matthew , Hawkins Kim , Rasmussen Henrik , DeVos Arjen , Herzig Geoffrey P.
Published: November 2003

Macrophage inflammatory protein‐1α (MIP‐1α) is a chemokine that can inhibit the cell cycle progression of both primitive haemopoietic and epidermal progenitor cells. This property could potentially be exploited to attenuate both the myelosuppressive effects of chemotherapy as well as mucositis. We evaluated both the biological and clinical effects of BB‐10010, a genetically engineered variant of MIP‐1α, in patients with malignant lymphoma or breast cancer receiving high‐dose etoposide (VP 3.6 g/m2) and cyclophosphamide (Cy 200 mg/kg). 52 patients were randomized to one of three cohorts. Cohort A received no BB‐10010; cohorts B and C received 10 μg/kg and 100 μg/kg of BB‐10010, respectively. All patients received post‐chemotherapy G‐CSF. BB‐10010 was well tolerated. There were no significant differences between groups in recovery to an ANC > 0.5 × 109/l, 1 × 109/l or 1.5 × 109/l, the number of days with an ANC < 0.5 × 109/l, days to a platelet count >50 × 109/l or 100 × 109/l, or the incidence and severity of mucositis. There was no evidence of any effect of BB‐10010 on colony‐forming cell (CFC) or long‐term culture‐initiating cell (LTC‐IC) mobilization, cycling activity in the marrow or on chemotherapy‐induced changes in CFC or LTC‐IC number both of which were in the normal range by 22 d after completion of the chemotherapy. To our knowledge this is the first report of a myelointensive regimen having no apparent long‐term effect on the LTC‐IC compartment. In summary, BB‐10010 is safe when used in patients receiving high‐dose therapy but has no effect on reducing the toxicity of such therapy.
Amount of mpl on bone marrow haemopoietic precursor cells from healthy volunteers and patients with refractory anaemia
Authors: Takeshita Akihiro , Shinjo Kaori , Naito Kensuke , Nakamura Satoki , Izumi Masakazu , Ling Pan , Ohnishi Kazunori , Ohno Ryuzo
Published: November 2003

Using a non‐isotopic ligand binding assay using multi‐colour flow cytometry, we quantitatively examined the amount of mpl in megakaryocyte‐platelet lineage cells. Firstly, we quantified the amount of mpl on cell lines. Mpl gene‐transfected BaF3 cells expressed a large amount of mpl, whereas original BaF3, K562, HL‐60 and NOMO‐1 cells showed no mpl. In bone marrow cells from healthy volunteers, mpl was expressed on CD34+ cells from the very early stage of differentiation when they had no CD38 antigen. The amount of mpl increased with differentiation to CD34+CD41+ cells, but decreased with further differentiation to CD34−CD41+ cells. In CD34+CD41+ cells the amount of mpl varied according to cell size: abundant in large cells, moderate in medium‐size cells and a little in small cells.

In bone marrow cells from patients with refractory anaemia (RA), the amount of mpl was decreased compared with that in bone marrow cells from healthy volunteers. When analysed by the same CD phenotype and same cell size, the amount of mpl was less in RA patients compared with that in healthy volunteers in all phenotypes and sizes tested. The proportion of large CD34+CD41+ cells was less in RA patients than in normal volunteers.
Selective enrichment in human megakaryocyte progenitors by the B203.13 surface differentiation antigen
Authors: Vitale Marco , Zauli Giorgio , Bassini Alessandra , Visani Giuseppe , Rezzani Rita , Croce Carlo M. , Manzoli Francesco Antonio
Published: November 2003

The B203.13 monoclonal antibody specifically recognizes a subset (8–18.5%) of human CD34+ haemopoietic progenitors, which are significantly (P < 0.05) enriched in megakaryocyte progenitors (CFU‐meg). Moreover, B203.13 expression was progressively lost as maturation proceeded along the megakaryocytic lineage. In fact: (i) CFU‐meg derived from CD34+/B203.13+ showed a greater number of cells/colony with respect to CD34+/B203.13−; (ii) after 10 d of liquid cultures with 100 ng/ml of thrombopoietin, only a minority of CD34‐derived cells positive for CD41 coexpressed B203.13; (iii) in situ immunocytochemical staining revealed that B203.13 was expressed exclusively on immature megakaryoblasts; (iv) circulating platelets did not express B203.13.
A sensitive flow cytometric assay for circulating platelet–leucocyte aggregates
Authors: Li N. , Goodall A. H. , Hjemdahl P.
Published: November 2003

A whole blood flow cytometric method for studies of platelet–leucocyte aggregates (PLAs) in vivo, involving neither fixation nor centrifugation, is described. With this method, PLAs in the total leucocyte population (PLA/L) were 15.3 ± 8.5% in 36 healthy volunteers. Blocking antibodies had little effect on PLAs in the absence of in vitro stimulation, suggesting that the aggregates were preformed in vivo. Fixation with formaldehyde or paraformaldehyde increased PLA/L significantly. Similarly, prefixation of the blood or red blood cell lysis, with repeated washing and centrifugation, caused artefactual 3–5‐fold increases in PLAs (P < 0.05). ADP, thrombin, platelet activating factor (PAF) or N‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP) each increased PLA formation in unfixed whole blood dose‐dependently; additive effects were found when they were combined. Experiments with blocking MAbs suggested that different ligand–receptor systems mediate PLA formation by different agonists. PLA formation by the platelet agonist ADP was inhibited by P‐selectin blockade, but enhanced by GPIIb/IIIa blockade (which inhibits platelet–platelet interactions). PLA formation by the leucocyte agonist fMLP was inhibited by GPIIb/IIIa blockade, suggesting linking via fibrinogen. Platelet–leucocyte aggregate analysis by this whole blood method appears to reflect in vivo conditions, and enables investigations of the mechanisms involved in their formation.
The Arg633His substitution responsible for the private platelet antigen Groa unravelled by SSCP analysis and direct sequencing
Authors: Simsek S. , Folman C. , Van Der Schoot C. E. , Kr. von dem Borne A. E. G.
Published: November 2003

We have previously described the private or family platelet antigen, Groa, which was identified in a case of neonatal alloimmune thrombocytopenia. The Groa antigen was found to be located on the GP IIIa (β3) subunit of the GP IIb/IIIa complex, the most prominent fibrinogen receptor of platelets. Initial experiments to characterize the Groa antigen at the molecular genetic level were unsuccessful. We therefore decided to use a different strategy to unravel the molecular basis of this antigen. Platelet GP IIIa mRNA of a Gro(a+) and a Gro(a−) donor was amplified with suitable primers in a reverse transcriptase–polymerase chain reaction (RT‐PCR) and subjected to single‐strand conformational polymorphism (SSCP) analysis. Three regions of the amplified GP IIIa cDNA derived from the Gro(a+) donor showed a different SSCP pattern when compared to that of the Gro(a−) donor. Direct nucleotide sequence analysis of these three segments revealed that two of them contained silent substitutions, A1163C, A1553G and G1565A. The first and the latter changes were described previously. In the third segment a G1996A mutation was found, predicting an arginine  →  histidine substitution at position 633 of the mature glycoprotein. PCR‐ASRA (allele‐specific restriction enzyme analysis) performed on cDNA as well as on genomic DNA with the restriction enzyme MaeIII showed that the His633 form of GPIIIa is restricted to the Gro(a+) phenotype. The observed mutation is three amino acids upstream of the mutation underlying the HPA‐8/Sr system (Arg636Cys), suggesting this region of GP IIIa to be susceptible for mutations. Moreover, the presence of a silent mutation and two low‐frequency forms of the silent polymorphisms strongly suggests that the G1996A mutation did not occur in a direct ancestral allele.
Expression and modulation of cellular receptors for interferon‐γ, tumour necrosis factor, and Fas on human bone marrow CD34+ cells
Authors: Sato Tadatsugu , Selleri Carmine , Anderson Stacie , Young Neal S. , Maciejewski Jaroslaw P.
Published: November 2003

Interferon‐γ (IFN‐γ), tumour necrosis factor‐α (TNF‐α) and Fas‐ligand can mediate potent inhibitory signals in haemopoietic cells. Clinical and laboratory studies have suggested the involvement of these cytokines in the regulation of normal haemopoiesis and in the pathophysiology of bone marrow (BM) failure syndromes. As the effects of cytokines may also be regulated at the cellular receptor level, we studied the expression and modulation of TNF receptor (TNFR), IFN‐γR and Fas‐R on haemopoietic progenitor cells. In freshly isolated BM, using flow cytometry, TNFR1 (p55), TNFR2 (p75), IFN‐γR, and Fas‐R were detected on 5–12% of mononuclear cells. Two‐colour staining showed comparable receptor expression on a CD34+ population, which includes haemopoietic progenitor and stem cells. Using reverse transcriptase‐PCR (RT‐PCR) transcription of mRNA coding for these receptors was demonstrated in fresh, highly purified CD34+ cells. These findings indicate that the effects of these factors on progenitor cells may be directly mediated. In cultured BM cells, expression of TNFR1 was not influenced by IFN‐γ, TNF‐α or apoptosis‐inducing anti‐Fas monoclonal antibody (mAb). IFN‐γ decreased CD34+ cell TNFR2 expression. CD34+ cell Fas‐R expression was increased by IFN‐γ and TNF‐α. IFN‐γR expression was enhanced by anti‐Fas mAb and to lesser degree with TNF‐α. Similar results were obtained with RT‐PCR analysis in cultured CD34+ cells. Potentiation of anti‐Fas mAb‐mediated inhibition of haemopoietic colony formation by IFN‐γ and TNF‐α was observed. Similarly, anti‐Fas mAb enhanced the inhibitory effects of IFN‐γ. These results suggest that, in addition to interacting at the level of intracellular signalling pathways, IFN‐γ, TNF‐α or Fas‐ligand may potentiate or antagonize their effects through modulation of cytokine receptor expression.
Sustained remission of severe resistant autoimmune neutropenia with Campath‐1H
Authors: Killick S. B. , Marsh J. C. W. , Hale G. , Waldmann H. , Kelly S. J. , Gordon‐Smith E. C.
Published: November 2003

We report the course of a patient with severe autoimmune neutropenia in whom only transient responses occurred with corticosteroids, antilymphocyte globulin and granulocyte‐colony stimulating factor, and who was resistant to treatment with azathioprine, cyclosporin and intravenous immunoglobulin. A 10 d course of intravenous Campath‐1H monoclonal resulted in a sustained haematological response. The long‐lasting effect of Campath‐1H may be due to its remarkable ability to induce a profound and prolonged peripheral blood T lymphopenia.
Minimal residual disease with TEL‐AML1 fusion transcript in childhood acute lymphoblastic leukaemia with t(12;21)
Authors: SATAKE NORIKO , KOBAYASHI HIROFUMI , TSUNEMATSU YUKIKO , KAWASAKI HAJIME , HORIKOSHI YASUO , KOIZUMI SHOICHI , KANEKO YASUHIKO
Published: November 2003

We analysed the TEL‐AML1 transcript using reverse transcription‐polymerase chain reaction (RT‐PCR) in order to detect minimal residual disease (MRD) in seven children with t(12;21)‐associated B‐lineage ALL. Leukaemic cells with the TEL‐AML1 transcript appear to be very sensitive to chemotherapy, and may be eradicated in most patients if adequate chemotherapy is given. However, a small number of patients with t(12;21) ALL may relapse under the currently used chemotherapy, and we believe that RT‐PCR for detecting MRD with the transcript is a suitable tool for monitoring the efficacy of chemotherapy or impending relapse in these patients. We analysed the TEL‐AML1 transcript using reverse transcription‐polymerase chain reaction (RT‐PCR) in order to detect minimal residual disease (MRD) in seven children with t(12;21)‐associated B‐lineage ALL. Two sets of primers, TEL exon 5 and AML1 exon 3 or 4, detected two types of transcript in four patients and two other types in two other patients. The two different translocation breakpoints in the AML1 gene with or without splicing out of AML1 exon 3 seemed to result in these four types of transcript in leukaemia samples.
Fibrinogen degradation products in patients with the Quebec platelet disorder
Authors: Hayward C. P. M. , Welch B. , Bouchard M. , Zheng S. , Rivard G. E.
Published: November 2003

The Quebec platelet disorder is a serious bleeding disorder associated with proteolysis of α‐granular proteins, including fibrinogen. We evaluated fibrinogen degradation product (FDP) assays as screening tests for this disorder. Patients with the Quebec platelet disorder (13/13) had elevated serum FDPs (P <0.01) due to secreted degraded platelet fibrinogen, but normal plasma FDPs and D‐dimers. Unrelated controls with bleeding disorders (32/34) had undetectable FDPs, and controls with FDPs due to disseminated intravascular coagulation (DIC) and other illnesses (11/11) had elevated FDPs in all assays (P < 0.01). Immunoblot analyses indicated plasma fibrinogen was normal in the Quebec patients and platelet FDPs were found in their platelet lysates, releasates and serum samples. Their platelet FDPs were not altered by treatment with fibrinolytic inhibitors and were different from the FDPs in DIC and in plasmin‐digested fibrinogen. Tests for serum FDPs may provide a simple rapid way to screen for the Quebec platelet disorder.
Successful treatment of cerebral aspergillosis with a novel triazole (voriconazole) in a patient with acute leukaemia
Authors: Schwartz Stefan , Milatovic Dana , Thiel Eckhard
Published: November 2003

Invasive aspergillosis is an increasing problem in patients with acute leukaemia, bone marrow transplantation, immunosuppression after solid organ transplantation, or acquired immunodeficiency syndrome. Despite available antifungal treatment, the mortality approaches 100% in patients with dissemination of the infection into the central nervous system (CNS). Using a novel triazole, voriconazole, we successfully treated an Aspergillus brain abscess in a patient with acute leukaemia. Drug levels above the minimal fungicidal concentration for Aspergillus species were detected in cerebrospinal fluid (CSF) specimens, and the treatment achieved an objective response.
Australian Leukaemia Study Group Myeloma II: a randomized trial of intensive combination chemotherapy with or without interferon in patients with myeloma
Authors: Joshua Douglas E. , Penny Ron , Matthews Jane P. , Laidlaw Christine R. , Gibson John , Bradstock Ken , Wolf Max , Goldstein David
Published: November 2003

The Australian Leukaemia Study Group has performed a randomized trial of interferon α‐2A (Roferon‐A) as a co‐induction agent together with intensive combination chemotherapy and as maintenance following completion of 12 cycles of induction treatment. When used as a co‐induction agent, interferon‐α did not improve response rates, time‐to‐treatment failure, or overall survival. Patients who had interferon together with intensive combination therapy (PCAB: prednisone 60 mg/m2 days 1–5, cyclophosphamide 600 mg/m2 day 1, BCNU 30 mg/m2 day 1, doxorubicin 30 mg/m2 day 1, repeated every 28 d for a total of 12 cycles) had more leucocyte and granulocyte toxicity and received a lower dose intensive of cytotoxic drugs than those patients who received PCAB without interferon.  There was a trend towards prolongation of plateau phase which did not reach significance. Interferon, however, did improve the survival of patients who achieved plateau; for those patients interferon was associated with a 33% decrease in the rate of death after adjusting for initial beta‐2 microglobulin level.
Chronic eosinophilic leukaemia (CEL): a distinct myeloproliferative disease
Authors: Weide Rudolf , Rieder Harald , Mehraein Yasemien , Wolf Martin , Kaiser Ulrich , Seifart Ulf , Görg Christian , Havemann Klaus
Published: November 2003

Chronic eosinophilic leukaemia has not yet been clearly defined, mainly due to the fact that it has not been conclusively shown as a monoclonal disease which should be separated from chronic myelogenous leukaemia, acute myelogenous leukaemia with eosinophilia (AML, FAB M4Eo), and the idiopathic hypereosinophilic syndrome.

We report a patient with a white blood cell count of 17.6 × 109/l with 74% eosinophils, normal platelet count and haemoglobin. No blasts were seen in the peripheral blood and the percentage of blasts in the bone marrow was <3%. A diagnosis of chronic eosinophilic leukaemia was made. Chromosome analysis of a bone marrow aspirate disclosed a trisomy 15 together with loss of the Y chromosome. Moreover, FISH analysis on May‐Grünwald‐Giemsa‐stained peripheral blood smears demonstrated trisomy 15 in the eosinophils. 3 months after initial diagnosis the patient went into blast crisis and died. The blast cells exhibited trisomy 15 and loss of the Y chromosome in a complex, aberrant karyotype.

In conclusion, the case shows that chronic eosinophilic leukaemia is a monoclonal, myeloproliferative disease with eosinophils as part of the malignant clone. Clinically, chronic eosinophilic leukaemia can be separated into a chronic phase and a blast crisis.
Long‐term haematological reconstitution following BEAM and autologous transplantation of circulating progenitor cells in non‐Hodgkin's lymphoma
Authors: Rossi Andrea , Cortelazzo Sergio , Bellavita Piero , Viero Piera , Bassan Renato , Comotti Benedetto , Marchioli Roberto , Marfisi Rosa‐Maria , Rambaldi Alessandro , Barbui Tiziano
Published: November 2003

We report on long‐term haematological recovery and clinical outcome after high‐dose chemotherapy supported by circulating progenitor cells (CPC) transplantation in non‐Hodgkin's lymphoma (NHL) patients, and analyse the role of variables which might influence engraftment. 63 consecutive NHL patients were enrolled in this study. Two groups of patients were considered for analysis: the first 34 patients had untreated diffuse large cell lymphoma with unfavourable prognostic factors. A second group of 29 patients underwent transplantation for resistant or relapsing NHL with low, intermediate and high grade histology. All patients received the BEAM conditioning regimen. As already reported in many studies, all patients showed a rapid haematological reconstitution. 43 patients (68%) achieved long‐term complete trilineage engraftment within a median of 107 d from CPC transplantation. The neutrophil count was the first parameter reaching complete normalization, and haemoglobin was the last. Failure to meet the trilineage levels was due to lack of platelet recovery and was more frequent in patients transplanted in the setting of salvage protocols. By Kaplan‐Meier analysis, the probability of a full reconstitution was 80% in patients to whom transplant was offered as part of a front‐line therapy and 50% when transplant was given in the salvage programmes. Multivariate analysis showed that sustained long‐term haematological reconstitution was significantly related to younger age, the time taken to achieve short‐term reconstitution, and bone marrow involvement.
Subcutaneous CAMPATH‐1H in fludarabine‐resistant/relapsed chronic lymphocytic and B‐prolymphocytic leukaemia
Authors: Bowen A. L. , Zomas A. , Emmett E. , Matutes E. , Dyer M. J. S. , Catovsky D.
Published: November 2003

Seven patients with B‐cell leukaemia — six with chronic lymphocytic leukaemia (CLL) and one with B‐prolymphocytic leukaemia (B‐PLL) — were treated with CAMPATH‐1H, a genetically reshaped CD52 monoclonal antibody, administered subcutaneously (s.c.) three times a week for 6–12 weeks. Four were resistant to, and three had had a short partial remission (PR) following, fludarabine chemotherapy. The patient with B‐PLL achieved complete remission and three patients with CLL attained PR; two of the latter were retreated. The three remaining patients were non‐responders. Three patients were transfusion‐dependent before CAMPATH and all three became transfusion‐independent after treatment. The overall median survival from starting CAMPATH‐1H was 11 months. Three patients reactivated cytomegalovirus (CMV) during the course of treatment, and two were treated with, and responded to, ganciclovir.
The alternate iron transport pathway: mobilferrin and integrin in reticulocytes
Authors: Umbreit Jay N. , Conrad Marcel E. , Berry Michael A. , Moore Elizabeth G. , Latour Lydia F. , Tolliver Beth A. , Elkhalifa Mohamed Y.
Published: November 2003

Iron transport in reticulocytes is known to occur via the well‐described transferrin‐receptor–endosome pathway. An alternative pathway for iron transport independent of transferrin has been postulated in reticulocytes and other cells. Transport of iron into reticulocytes from ferric citrate solutions was shown to be saturable and independent of transferrin. During transport of iron from ferric citrate, both cell surface integrins, and a soluble protein, mobilferrin, were labelled. This demonstrated that the reticulocyte transferrin independent pathway for iron transport involved integrins and mobilferrin similar to intestinal absorptive cells. This pathway would be expected to transport iron into cells under conditions of iron overload and was capable of providing iron for haemoglobin synthesis. Mobilferrin was also radiolabelled when radioiron labelled transferrin was incubated with reticulocytes and this occurred with a different time course than was observed following reticulocyte exposure to radiolabelled ferric citrate. This suggested that mobilferrin may serve as an intermediary in both pathways.
The role of CS1 moiety of fibronectin in VLA4‐mediated haemopoietic progenitor trafficking
Authors: Craddock C. F. , Nakamoto B. , Elices M. , Papayannopoulou Th
Published: November 2003

In vitro the integrin VLA4 mediates the adhesion of haemopoietic progenitors to bone marrow stroma through an interaction with its ligands VCAM‐1 and the CS1 moiety of fibronectin. The VLA4/VCAM‐1 pathway has been implicated in haemopoietic trafficking in vivo since antibodies to both VLA4 and VCAM‐1 decrease the homing (lodgement) of transplanted progenitors and mobilize progenitors. However, the role of the CS1 domain of fibronectin in progenitor trafficking in vivo has not been explored. We studied the effect of competitive inhibition of the VLA4/CS1 pathway on progenitor homing and mobilization in mice. Pre‐incubation of bone marrow cells with a CS1 inhibitor did not alter the number of CFU‐C or CFU‐S12 lodged to the bone marrow of lethally irradiated mice 3 h after transplantation. In addition, continuous administration of a CS1 inhibitor did not increase the number of CFU‐C in the peripheral blood. In order to study the role of the VLA4/CS1 pathway in trafficking of more primitive progenitors we studied whether administration of a CS1 inhibitor mobilized radioprotective cells. In contrast to the effect of anti‐VCAM‐1 which mobilized cells capable of rescuing 100% of lethally irradiated mice, administration of a CS1 inhibitor did not increase the number of radioprotective cells in the peripheral blood. Haemopoietic progenitors also bind to the RGD motif of fibronectin through an interaction with VLA5 and we therefore also studied the effect of antibodies to VLA5 on progenitor homing and mobilization. Antibody to VLA5 did not alter bone marrow lodgement at 3 h or increase the number of circulating haemopoietic progenitors. These studies therefore imply that, in contrast to VCAM‐1, the CS1 moiety of fibronectin is not a significant ligand in VLA4 mediated progenitor trafficking in vivo.
Trisomy 8 in a patient who responded to therapy with all‐trans‐retinoic acid and developed paroxysmal nocturnal haemoglobinuria
Authors: Viniou N. , Michali E. , Meletis J. , Andreopoulos A. , Vaiopoulos G. , Stavroyianni N. , Loukopoulos D. , Yataganas X.
Published: November 2003

Trisomy 8 is the most common numerical chromosomal abnormality in myelodysplastic syndromes (MDS). Paroxysmal nocturnal haemoglobinuria (PNH) is an aquired haemolytic anaemia, clonal in nature, due to somatic mutation. PNH may evolve to aplastic anaemia, to MDS or to acute myeloid leukaemia. We present a patient who had trisomy 8 mosaicism at disease presentation who received therapy with all‐trans‐retinoic acid, responded to therapy, and developed PNH in the course of the disease. Cytogenetics at the time of PNH diagnosis showed a normal karyotype.
Spontaneous exodus of high numbers of normal early progenitor cells (Ph‐negative LTC‐IC) in the peripheral blood of patients with chronic myeloid leukaemia at the beginning of the disease
Authors: Podestà Marina , Piaggio Giovanna , Sessarego Mario , Pitto Anna , Benvenuto Federica , Vassallo Franca , Fugazza Giuseppina , Carella Angelo M. , Frassoni Francesco
Published: November 2003

Elevated white blood cell counts are frequently found in patients with chronic myeloid leukaemia (CML). Although some studies have disclosed that bone marrow of CML patients may contain some normal Philadelphia‐negative early progenitor cells, it has been assumed that the dramatic increase of white blood cells was entirely related to the leukaemic cell expansion. In this study we attempted to quantify the number of normal and leukaemic progenitor cells in the bone marrow and peripheral blood of newly diagnosed CML patients. Bone marrow and peripheral blood cells of eight newly diagnosed CML patients were analysed for clonogenic colony‐forming cells (CFC) and very early progenitor cells, i.e. long‐term culture initiating cells (LTC‐IC). The leukaemic (Ph‐positive) or normal (Ph‐negative) origin of progenitor cells was revealed by cytogenetic analysis performed on single colonies arising from in‐vitro assays.

In 6/8 patients the marrow CFC frequency ranged from 400 to 9300/106 mononuclear cells (MNC), 0–50% being Philadelphia chromosome negative; the LTC‐IC frequency ranged from 0 to 11/106 MNC, and were 80–100% Ph‐negative. The corresponding absolute values into peripheral blood were: CFC = 1–35.×103/ml, 0–50% Ph‐negative, and LTC‐IC = 0–2.5×103/ml, 0–100% Ph‐negative. In one patient, no LTC‐IC were detected in either the marrow or the peripheral blood.

In conclusion, in the peripheral blood of some CML patients, the number of normal LTC‐IC is more than 3 times the number of leukaemic progenitor cells, and is much higher (50 times) than the corresponding value found in normal subjects in steady state (2.5/ml v 124/ml). These data support the concept that leukaemic ‘stem cells’, with respect to normal ones, may be considerably fewer than previously thought. In addition it is shown that at the beginning of CML high numbers of normal LTC‐IC are spontaneously mobilized into the blood. Finally, the presence of Ph‐negative early progenitors into the blood may represent a potential source of normal stem cells available for autografting providing they can be separated from leukaemic cells.
Outcome of transplantation for standard‐risk leukaemia with grafts depleted of lymphocytes after conditioning with an intensified regimen
Authors: Schaap N. , Schattenberg A. , Bär B. , Preijers F. , Geurtsvan Kessel A. , Van Der Maazen R. , De Boo T. , De Witte T.
Published: November 2003

One hundred and eighty‐one consecutive patients with standard‐risk leukaemia were transplanted with HLA‐identical sibling grafts depleted of lymphocytes using counterflow centrifugation. In 116 patients, standard conditioning was intensified by the addition of anthracyclines.

Multivariate analysis revealed significantly more acute GVHD  grade 2 and a trend towards more chronic GVHD in patients conditioned with the addition of anthracyclines. For all patients the risk for chronic GVHD, but not for acute GVHD, increased with a higher number of T cells in the graft. The projected 5‐year probability of relapse was significantly lower in the group of patients conditioned with anthracyclines; 26% versus 52% (P = 0.015). In multivariate analysis the addition of anthracyclines to the conditioning regimen was the only significant factor contributing to a lower probability of relapse. The projected 5‐year probability of leukaemia‐free survival [LFS] in the patients conditioned with and without the addition of anthracyclines was 56% and 36%, respectively (P = 0.004). In multivariate analysis the addition of anthracyclines to the conditioning regimen correlated significantly with a lower number of mixed chimaeras in patients at 6 and 12 months after BMT. Mixed chimaerism at 6 months after transplantation did not significantly correlate with a higher incidence of relapse in further follow‐up. In contrast, mixed chimaerism at 12 months after BMT was significantly associated with higher relapse rate.

We conclude that the addition of anthracyclines to the conditioning regimen improves outcome of BMT using T‐cell‐depleted grafts.
Incidence and nature of feeding problems in young children referred to a paediatric HIV service in London: FEAD screening
Authors: Melvin D , Wright C , Goddard S
Published: November 2003

A detailed screening assessment was carried out on two matched groups of young children; one group was HIV‐infected and the other was not. Screening included assessments of growth, development and food intake. Parents were also interviewed about their child’s feeding and mealtime behaviours. Half of the HIV‐infected children were reported with serious feeding problems; significantly higher than in the uninfected group. More of the children in the HIV‐infected group were found to have poorer growth and developmental weaknesses than in the uninfected group. A combination of physical and psychological factors are suggested as contributing to these feeding difficulties. Early monitoring of feeding behaviours, daily routines and food intake, together with systematic growth and developmental measures are suggested as important components in the care and management of HIV‐infected children.
Child Health Surveillance in England and Wales: the bad news
Authors: Butler J R
Published: November 2003

Postal surveys were conducted in 1993 among all, or samples of, six groups of providers and managers of pre‐school child health surveillance (CHS) in England and Wales. Content analyses were also carried out of strategic policy statements for CHS produced by 54 district health authorities in England and Wales. The surveys aimed to document the views and experiences of CHS providers and managers about the impact of recent changes affecting the structure and operation of CHS, including the publication of Health for All Children, the 1990 Contract for General Practitioners (GPs), the implementation of the National Health Service and Community Care Act 1990, and the changing roles of community doctors and health visitors. Four adverse findings from the surveys are discussed: fragmentation in the child health service; the unwanted effects of the NHS internal market; the adverse consequences of the changing role of health visitors; and the concerns voiced by the community doctors about the quality of CHS in general practice.
Characterization of the allergen Der f 7 from house dust mite extracts by species‐specific and crossreactive monoclonal antibodies
Authors: SHEN H.‐D. , LIN W.‐L. , TSAI L.‐C. , TAM M. F. , CHUA K.‐Y. , CHEN H.‐L. , HSIEH K.‐H. , LI C.‐S. , THOMAS W. R.
Published: November 2003

Background
The group 7 mite allergens react with IgE in 50% of sera from allergic patients.

Objective
To determine the molecular and antigenic characteristics and heterogeneity of Der f 7 in mite extracts.

Methods
Monoclonal antibodies (MoAbs) produced from mice immunized with recombinant Der f 7 were examined for crossreactivity to Der p 7 and then used for immunoblotting of 1 and 2‐D gel electrophoresis. Deglycosylation was studied with N‐glycosidase‐F and N‐terminal sequencing by Edman degradation. The epitopes of the monoclonal antibodies were compared by cross‐inhibitory immunoassays.

Results
Immunoblotting of D. farinae extracts with all the anti Der f 7 MoAbs showed major reactivities at 31, 30 and 25 kDa. The strongest immunostaining was at 25 kDa which contrasted with Der p 7 where the 31 and 30 kDa bands were strongest. The relative strength of staining however varied between extracts. The 31 and 30 kDa components were glycosylation products of the 25 kDa form which had the N‐terminal sequence predicted from cDNA analysis. Two MoAbs stained an 18 kDa band consistent with a degradation product. The 2‐D gels showed that different components with pls from 5.6–6.4. Both species‐specific and Der p 7 crossreactive MoAbs were produced and a two‐site ELISA assay for detecting group 7 allergen was developed with MoAbs recognizing different epitopes.

Conclusions
Der f 7 has been defined by its natural N‐terminal sequence and MoAbs. It apparently exists as different glycosylation and degradation products in mite extracts, the relative abundance of which differs with different preparations. A two‐site ELISA to measure the allergen was developed.
Effect of the expression of DRαEβNOD molecule on the development of insulitis and diabetes in the non-obese diabetic (NOD) mouse
Authors: Yamane K , Yamamoto K , Yoshikawa Y , Sasazuki T
Published: November 2003

Previous studies have shown that a transgenic I-Eα gene, the mouse homologue of human DRα gene, prevents the development of insulitis and hence of diabetes in NOD mice. To investigate the mechanism of this prevention, we generated two strains of NOD mice expressing DRαEβ molecule: DRα-24-NOD expressing DRαEβ molecule on thymic epithelial cells (TEC) and bone marrow-derived cells (BDC), and DRα-30-NOD expressing DRαEβ molecule only on the TEC, and these mice were monitored for disease development. Because the DRαEβ molecule reconstituted I-E controlled immune regulation, it would become clear which cell type, TEC or BDC, was responsible for the I-E-mediated disease protection. To our surprise, however, DRα-24-NOD developed insulitis and diabetes comparably to non-transgenic littermates. This suggested that the difference in structure between DRα and Eα molecules contributed to the difference in preventive effect on the development of insulitis and diabetes between DRα-24-NOD and Eα-NOD. In an analysis of the T cell proliferative responses to glutamic acid decarboxylase (GAD) 65-derived peptides which were known to be diabetogenic autoantigens, it was shown that DRα-24-NOD and NOD acquired comparable level of T cell response to GAD 509–528 but 5–10-fold higher response was observed in Eα-NOD. This suggested that I-ANOD and EαEβNOD molecules could present GAD 509–528 peptide to T cells, while DRαEβNOD could not. Furthermore, T cells from DRα transgenic mice showed proliferative response to antigen-presenting cells from Eα transgenic mice in primary mixed lymphocyte reaction. This also suggested that the EαEβ molecule does differ in structure and peptide binding from the DRαEβ molecule. Present data suggested a possibility that the T cell repertoire selection, or the T cell response to GAD 65 and/or other unknown antigens specifically mediated by I-E molecule, may contribute to the prevention of disease development in Eα-NOD.
Isolation of cDNAs encoding immunogenic regions of gp330, the autoantigen involved in Heymann nephritis
Authors: Luca M E , de Geus B , Sahali D , Bruijn J A , Verroust P , de Heer E
Published: November 2003

Active Heymann nephritis is an organ-specific autoimmune disease of the rat kidney, characterized by the formation of immune complexes located subepithelially in the glomerulus. The T cell-mediated humoral immune response is directed to gp330, a large renal epithelial glycoprotein which is expressed both in the proximal tubule and on glomerular podocytes. In this study polyclonal rabbit antibodies raised against affinity-purified rat gp330 were used to screen a λ-gt11 expression library of the rat kidney. One cDNA clone that was recognized by the antibodies coded for a 2.7-kb protein that is not described in the sequence database of GenBank/EMBL. Two other groups of cDNA clones were identified that displayed similarity with several members of the low-density lipoprotein (LDL)-receptor gene family to which gp330 belongs. By comparison with the gp330-cDNA sequence, these two clones could be mapped to two remote areas on the extracellular domain of gp330. The antigenicity of these two areas is in accordance with their location in highly hydrophilic regions on the extracellular domain of gp330. The cDNA clones described in this study may represent two main immunodominant regions on rat gp330.
Circulating immunoglobulin-secreting cells are heterogeneous in their expression of maturation markers and homing receptors
Authors: Kantele J M , Kantele A , Arvilommi H
Published: November 2003

Immunoglobulin-secreting cells (ISC) in the peripheral blood are active effectors of the human immune defence system on their way to their site of action. We combined immunomagnetic cell separation and ELISPOT to study the expression of maturation markers and homing receptors (HR) on these cells in healthy volunteers. The results revealed that although highly differentiated, peripheral blood ISC are remarkably heterogeneous both with respect to their expression of maturation markers and HR. Moreover, significant differences were demonstrated between the various isotypes. Fewer IgA-secreting cells expressed both markers of early maturation (HLA-DR, HLA-DQ, CD20, and CD21) and of more mature B cells or plasma cells (CD28, CD38, and α-syndecan) compared with IgG- and IgM-secreting cells. IgA-secreting cells also showed the lowest proportion of cells positive for the peripheral lymph node HR, L-selectin, or the skin HR, cutaneous lymphocyte antigen (CLA). By contrast, the expression of mucosal HR on IgA-secreting cells did not reveal a more pronounced homing attitude to mucosal tissues than IgG- or IgM-secreting cells. We conclude that peripheral blood ISC are a heterogeneous cell population and that IgA-secreting cells seem to differ from the other isotypes both in respect of expression of HR and the various maturational markers studied.