bluuebunny/crossref_metadata_2025 huggingface dataset:
Reversion of autoimmune lymphoproliferative syndrome with an antimalarial drug: preliminary results of a clinical cohort study and molecular observations
Authors: Van Der Werff ten Bosch Jutte , Schotte Peter , Ferster Alice , Azzi Nadira , Boehler Thomas , Laurey Geneviève , Arola Mikko , Demanet Christian , Beyaert Rudi , Thielemans Kris , Otten Jacques
Published: March 2003

Summary. Autoimmune lymphoproliferative syndrome (ALPS) is a paediatric disease characterized by lymphoproliferation and autoimmunity. Most patients are known to carry heterozygous mutations of the TNFRSF6 gene leading to diminished Fas‐mediated apoptosis and failure of activated lymphocytes to undergo apoptosis. A subgroup of patients without the TNFRSF6 gene mutation has similar defective apoptosis and clinical features. No effective treatment has been reported so far. Glucocorticoids, intravenous immunoglobulin and/or immunosuppressive drugs have usually led to only transient clinical improvement. Seven ALPS patients (two type Ia and five type III) were treated with the antimalarial drug Fansidar. No toxicity was observed. An objective response was seen in six of them and, in two, the treatment was stopped without reappearance of the symptoms. Moreover, a marked decrease in interleukin‐10 levels was observed in two patients during the treatment. We found that the drug induced apoptosis in activated lymphocytes through activation of the mitochondrial apoptotic pathway.
Influence of karyotype on outcome of allogeneic bone marrow transplantation for adults with precursor B‐lineage acute lymphoblastic leukaemia in first or second remission
Authors: Lee Seok , Kim Dong‐Wook , Kim Yoo‐Jin , Park Yoon‐Hee , Min Chang‐Ki , Lee Jong‐Wook , Min Woo‐Sung , Kim Chun‐Choo
Published: March 2003

Summary.  The prognostic relevance of karyotype has been established in adult acute lymphoblastic leukaemia (ALL) patients treated with chemotherapy but not definitively evaluated in an allogeneic bone marrow transplantation (BMT) setting. To determine the factors affecting the outcome of allogeneic BMT for adults with precursor B‐lineage ALL in first or second complete remission (CR), a total of 41 consecutive patients with a successful karyotype were enrolled in this study. There were 21 men and 20 women with a median age of 27 (15–43) years. The distribution of French–American–British (FAB) subtypes was as follows: L1 (n = 26), L2 (n = 15). Unfavourable karyotypes (n = 12) were defined as Ph+ or t(4;11). Disease status at the time of transplant was first CR (n = 35) or second CR (n = 6). With a median follow‐up of 36 months, the 3‐year probabilities of relapse and disease‐free survival (DFS) were 36·3 ± 8·4% and 57·3 ± 8·4% respectively. Potential variables predicting worse relapse and DFS were FAB subtype (L2), extramedullary involvement, pre‐BMT status (second CR), unfavourable karyotype and type of graft‐versus‐host disease (GVHD). Further multivariate analysis showed that karyotype and pre‐BMT status were independently associated with relapse and DFS. In addition, chronic GVHD was found to be significantly associated with a lower relapse rate.
Methylenetetrahydrofolate reductase genotype does not play a role in multiple myeloma pathogenesis
Authors: González‐Fraile Maria I. , García‐Sanz Ramoń , Mateos Maria V. , Balanzategui Ana , González Marcos , Váquez Lourdes , Miguel Jesús F. San
Published: March 2003

Summary. Methylenetetrahydrofolate reductase (MTHFR) plays an important role in carcinogenesis. A decreased incidence of cancer has been reported in the presence of MTHFR 677TT, 1298AC and 1298CC polymorphic variants. We have analysed the MTHFR genotype in 107 multiple myeloma (MM) patients and 86 controls. The MM and control polymorphisms frequencies were: 34% and 48% for 677CC, 53% and 41% for 677CT, 12% and 11% for 677TT; 36% and 43% for 1298AA, 51% and 44% for 1298AC; and 12% and 13% for 1298CC respectively. No statistically significant differences were observed. In addition, no differences were seen according to MM stage, presence of p16 gene hypermethylation or response to treatment.
Two new molecular bases for the Dombrock null phenotype
Authors: Rios Maria , Storry Jill R. , Hue‐Roye Kim , Chung Amy , Reid Marion E.
Published: March 2003

Summary.  Red blood cells (RBCs) with the Donull phenotype lack all antigens in the Dombrock blood group system, i.e. Doa, Dob, Gya, Hy and Joa. Sequence analysis of DNA from one proband with the Donull phenotype revealed a single nucleotide mutation of t to c in the donor splice site of DO (IVS1 + 2t > c), with outsplicing of exon 2. Analysis of a second proband revealed a homozygous nonsense mutation 442 C > T in exon 2 predicting a premature stop codon (Gln148 Stop). The molecular bases described in these two probands provide an explanation for the lack of Do glycoprotein on their RBCs.
The post‐transplant cytogenetic response to interferon is a major determinant of survival after autologous stem cell transplantation for chronic myeloid leukaemia in chronic phase
Authors: Olavarria Eduardo , Reiffers Josy , Boque Concha , Sureda Ana , Meloni Giovanna , Michallet Mauricette , Clark Richard E. , Blaise Didier , Carella Angelo M. , Cahn Jean‐Yves , Jouet Jean P. , Rizzoli Vittorio , Van Biezen Anja , Gratwohl Alois , Goldman John M. , Niederwieser Dietger , Apperley Jane F.
Published: March 2003

Summary. We have analysed the outcome of 581 autologous stem cell transplants (SCT) for chronic myeloid leukaemia (CML) in first chronic phase reported to the European Group for Blood and Marrow Transplantation between 1983 and 1998. Out off 207 patients evaluable for cytogenetics within 6 months of SCT, 36 patients (17%) were in complete cytogenetic remission (CCR), 34 (16%) in major remission (MCR), 74 (36%) in minor remission (mCR) and 63 (31%) had no cytogenetic response (NR). Interferon (IFN) was given post SCT to 267 patients. Results of the cytogenetic analysis within 1–2 years from SCT were available for 117 patients, the majority of whom (n = 101) received IFN post SCT: 17 (15%) were in CCR, 18 (15%) in MCR, 24 (20%) in mCR and 58 (50%) NR. The median survival in this series was 96 months (71–125) from SCT. There was no difference in survival according to cytogenetic status pre‐ and immediately post SCT. However, patients in CCR or MCR at 1–2 years post SCT had a 10‐year survival of 66% compared with 36% for patients in mCR or NR (P = 0·003). The 5‐year survival for patients receiving IFN post SCT was 72% compared with 61% for patients not treated with IFN (P = 0·01). Out of 155 patients refractory to IFN pre SCT, 70% achieved a cytogenetic response post SCT, which was complete or major in 31%. IFN refractory patients who sustained a CCR or MCR for 1–2 years after SCT had an excellent outcome.
Erythroid 5‐aminolevulinate synthase, ferrochelatase and DMT1 expression in erythroid progenitors: differential pathways for erythropoietin and iron‐dependent regulation
Authors: Zoller Heinz , Decristoforo Clemens , Weiss Günter
Published: March 2003

Summary. To determine whether erythropoietin (EPO) affects haem biosynthesis and iron transport, we studied the effects of EPO on the expression of erythroid 5‐aminolevulinate synthase (eALAS), ferrochelatase and divalent metal transporter 1 (DMT‐1) in human erythroid progenitor cells, and in the murine and human erythroid cell lines MEL and K562. Cytoplasmic e‐ALAS mRNA levels were significantly increased after incubation of cells with EPO for at least 24 h, which could be the result of a transcriptional mechanism. In contrast, ferrochelatase or DMT‐1 mRNA expression were not affected. Moreover, EPO also increased e‐ALAS enzyme activity after only 4 h of stimulation, when mRNA levels were unchanged. The underlying mechanism was an effect of EPO on e‐ALAS mRNA translation, which was under the control of iron regulatory proteins (IRP) 1 and 2. Thereby, EPO weakened the binding affinity of IRP‐2 to the iron responsive element (IRE) within e‐ALAS mRNA which resulted in the increased expression of e‐ALAS IRE‐controlled reporter gene constructs, following EPO stimulation. Our results show that EPO directly affected haem biosynthesis by stimulating the transcriptional and post‐transcriptional expression of the key enzyme e‐ALAS. These data provide new insights into the complex biochemical interaction between iron metabolism, haem biosynthesis and EPO biology.
The relative role of peripheral blood and bone marrow for monitoring molecular evidence of disease in follicular lymphoma by quantitative real‐time polymerase chain reaction
Authors: Summers Karin E. , Davies Andrew J. , Matthews Janet , Jenner Michael J. , Cornelius Victoria , Amess John A. , Norton Andrew J. , Rohatiner Ama Z. S. , Fitzgibbon Jude , Lister T. Andrew , Goff Lindsey K.
Published: March 2003

Summary.  Peripheral blood (PB) and bone marrow (BM) are used interchangeably for t(14;18) (IgH/BCL‐2) molecular monitoring in follicular lymphoma (FL) and detection of rearrangement after treatment has been correlated to increased risk of relapse. To determine the relative value of each tissue, MBR t(14;18) was quantified by real‐time polymerase chain reaction in 52 simultaneous paired PB and BM samples from 38 FL patients. In total, 79% of sample pairs taken in remission (n = 19) or when no morphological disease was evident in the BM (n = 29) had t(14;18) copy number within one log difference and the median difference was small. These findings suggest that, in remission, PB may be adequately monitored. In general, however, higher copy number was detected in BM than in the corresponding PB sample.
Protein C inhibitor (plasminogen activator inhibitor‐3) and the risk of venous thrombosis
Authors: Meijers Joost C. M. , Marquart J. Arnoud , Bertina Rogier M. , Bouma Bonno N. , Rosendaal Frits R.
Published: March 2003

Summary. Protein C inhibitor (PCI), also known as plasminogen activator inhibitor‐3, is a serine proteinase inhibitor that can inhibit enzymes in blood coagulation, fibrinolysis and fertility. The role of PCI in regulating the blood coagulation mechanism is not known, as it can inhibit both procoagulant (thrombin, factor Xa, factor XIa) and anticoagulant (activated protein C, thrombin‐thrombomodulin, urokinase) enzymes. To determine the relevance of this inhibitor in thrombosis, PCI levels were assessed in the Leiden Thrombophilia Study, a case–control study of venous thrombosis in 473 patients with a first deep‐vein thrombosis and 474 age‐ and sex‐matched control subjects. PCI levels above the 95th percentile of the controls (136·1%) increased the risk 1·6‐fold compared with PCI levels below the 95th percentile (95% confidence interval 0·9–2·8). There was a gradual increase in risk of thrombosis with further increasing levels of PCI. Adjustment for a number of possible confounders led to a reduction of the risk estimates associated with PCI. However, it is unclear whether adjustment for such factors in the risk models is justified. These results indicate that high levels of PCI may constitute a mild risk factor for venous thrombosis.
X‐linked thrombocytopenia in a girl
Authors: Inoue Hirokazu , Kurosawa Hidemitsu , Nonoyama Shigeaki , Imai Kohsuke , Kumazaki Hisami , Matsunaga Takayuki , Sato Yuya , Sugita Kenichi , Eguchi Mitsuoki
Published: March 2003

Summary. We report X‐linked thrombocytopenia (XLT) in a 6‐year‐old girl with petechiae and thrombocytopenia from the age of 3 months. Her 2‐year‐old brother was also diagnosed with XLT. The Wiskott–Aldrich syndrome protein (WASP) gene was detected as a replacement of +5th G to Aon intron 6 using sequence analysis, and the WASP expression levels in this patient were one‐third those of a healthy control. The X‐inactivation analysis of the patients lymphocytes showed a random pattern of X‐chromosome inactivation. To our knowledge, this is the first confirmed report of XLT in a female.
Influence of post‐transplant recombinant human granulocyte colony‐stimulating factor administration on peritransplant morbidity in patients undergoing autologous stem cell transplantation
Authors: Demirer Taner , Ayli Meltem , Dagli Mehmet , Haznedar Rauf , Genc Yasemın , Fen Turgay , Dincer Suleyman , Unal Ekrem , Gunel Nazan , Seyrek Ertugrul , Ustun Tulay , Ustael Nilufer , Yildiz Mustafa , Sertkaya Durdu , Ozet Gulsum , Muftuoglu Osman
Published: March 2003

Summary. This study evaluated of the effect of post‐transplant recombinant human granulocyte colony‐stimulating factor (rhG‐CSF) administration on the parameters of peritransplant morbidity. Three sequential and consecutive cohorts of 20 patients each received either post‐transplant rhG‐CSF at a dose of 5 µg/kg/d i.v. in the morning, starting on d 0, d 5, or no rhG‐CSF. Patients who received rhG‐CSF starting on d 0 and 5 recovered granulocytes more rapidly than those not receiving rhG‐CSF (P < 0·001 for ANC ≥ 0·5 and 1 × 109/l). RhG‐CSF administration was not significantly associated with more rapid platelet engraftment. RhG‐CSF administration starting on d 0 and 5 was significantly associated with a decreased duration of fever (P = 0·002 and 0·001 respectively), antibiotic administration (P < 0·001 and 0·006 respectively) and shorter hospitalization (P < 0·001 and 0·001 respectively) compared with the reference group. There was no difference between the d 0 and d 5 arms regarding the parameters of peritransplant morbidity. In conclusion, rhG‐CSF administration was associated with a faster granulocyte recovery, shorter hospitalization, and shorter period of fever and non‐prophylactic antibiotic administration. This study also showed that starting rhG‐CSF administration on d 5 may be as effective as d 0 on the clinical outcome and may be an economical approach in routine clinical practice in this cost‐conscious era.
Pure red cell aplasia due to parvovirus following treatment with CHOP and rituximab for B‐cell lymphoma*
Authors: Song Kevin W. , Mollee Peter , Patterson Bruce , Brien William , Crump Michael
Published: March 2003

Summary.  A 26‐year‐old woman, diagnosed with diffuse large B‐cell lymphoma, was treated with CHOP (cyclophosphamide, hydroxydaunomycin, oncovin, prednisone), rituximab and radiotherapy. She developed transfusion‐dependant anaemia, which persisted following chemotherapy. Bone marrow aspirate and biopsy were consistent with pure red cell aplasia and parvovirus infection. Serology was negative for previous or acute infection but parvovirus DNA was detected by polymerase chain reaction. Administration of intravenous immunoglobulin (1 g/kg) resulted in reticulocytosis and recovery of her haemoglobin. We hypothesize that rituximab caused depletion of her normal B cells, resulting in an inability to mount a primary immune response to parvovirus infection.
FetalRHDgenotyping in maternal serum during the first trimester of pregnancy
Authors: Costa Jean‐Marc , Giovangrandi Yves , Ernault Pauline , Lohmann Laurence , Nataf Valérie , El Halali Najua , Gautier Evelyne
Published: March 2003

Summary. FetalRHDgenotype determination is useful in the management of sensitized RhD‐negative pregnant women. It can be ascertained early during pregnancy by chorionic villus sampling (CVS) or amniocentesis. However, these procedures are invasive, resulting both in an increased risk of fetal loss and in an increased severity of immunization due to fetomaternal haemorrhage. A reliable determination ofRHDgenotype by fetal DNA analysis in maternal serum during the first trimester of pregnancy is reported in this study. One hundred and six sera from RhD‐negative pregnant women were obtained during the first trimester of pregnancy. These sera were tested for the presence ofRHDgene using a new real‐time polymerase chain reaction assay and the results compared with those obtained later in pregnancy on amniotic fluid cells and byRHDserology of the new‐born. All sera from women carrying a RhD‐positive fetus (n = 62) gave positive results forRHDgene detection and sera from women carrying a RhD‐negative fetus (n = 40) were negative. The high level of accuracy of fetalRHDgenotyping obtained in this study could enable this technique to be offered on a routine basis for the management of RhD‐negative patients during the first trimester of pregnancy.
Human Siglec‐5: tissue distribution, novel isoforms and domain specificities for sialic acid‐dependent ligand interactions
Authors: Connolly Nicholas P. , Jones Margaret , Watt Suzanne M.
Published: March 2003

Summary. Human Siglec‐5 is a sialic acid binding immunoglobulin (Ig)‐like lectin (Siglec), comprising one N‐terminal IgV‐SET domain followed by three IgC2‐SET domains, and a cytoplasmic domain with ITIM and SAP motifs which regulate cell signalling. We report the differential distribution of hSiglec‐5 on neutrophil and macrophage subsets in tissues using monoclonal antibodies, 1A5 and 2H8, which require the first IgC2‐SET domain for binding. Interestingly, hSiglec‐5 was especially prominent on macrophages in reactive lymph nodes. We have identified four isoforms of hSiglec‐5 possessing three (hSiglec‐5‐3L and ‐3C) or four (hSiglec‐5‐4L and ‐4S) extracellular domains linked to long (hSiglec‐5‐3L and ‐4L) or short (hSiglec‐5‐4S) cytoplasmic tails or existing as a soluble isoform (hSiglec‐5‐3C). hSiglec‐5‐4L has the broadest tissue distribution, being detected in adult spleen, thymus, lymph node, peripheral blood leucocytes and bone marrow, and in fetal lung and liver. A soluble Fc chimaeric protein containing the hSiglec‐5‐4L extracellular domain binds in a sialic acid‐dependent manner to glycophorin A on human erythrocytes and to α2‐3‐ and α2‐6‐sialyllactose moieties. Domain deletion mutants of hSiglec‐5(D1‐4)‐Fc reveal that the first three IgC2‐SET domains are required for optimal binding, with adhesion being abolished if the first IgC2‐SET domain is deleted. This indicates that each hSiglec‐5 isoform will interact with sialic acid ligands and provides the first step towards defining structure–function relationships of hSiglec‐5 isoforms.
Low expression of interferon regulatory factor‐1 and identification of novel exons skipping in patients with chronic myeloid leukaemia
Authors: Tzoanopoulos Dimitrios , Speletas Matthaios , Arvanitidis Konstantinos , Veiopoulou Christina , Kyriaki Sofia , Thyphronitis George , Sideras Paschalis , Kartalis Georgios , Ritis Konstantinos
Published: March 2003

Summary. Chronic myeloid leukaemia (CML) is a malignant clonal disorder of the haematopoietic stem cell. Treatment of CML patients with interferon alpha (IFN‐α) has induced haematological and cytogenetic remission. Interferons transcriptionally activate target genes through the JAK–STAT and interferon regulated factors (IRFs) family pathways. Interferon regulated factor‐1 (IRF‐1) is a transcriptional activator of genes critical for cell growth, differentiation and apoptosis. The skipping of exons 2 or 2 and 3 of IRF‐1 in patients with myelodysplastic syndromes and acute myelogenous leukaemia suggests that this factor may have a critical role in leukaemogenesis. The role of IRF‐1 in CML is currently unknown. Therefore, mutational analysis of IRF‐1 was performed and its expression pattern was also studied in CML patients. We studied IRF‐1 in peripheral blood mononuclear cells of 21 patients in chronic phase CML. No point mutations were identified at the cDNA level. Surprisingly, fourfold reduction of full‐length IRF‐1 mRNA expression was established in 17/21 patients compared with normal individuals. Low expression of full‐length IRF‐1 was observed in conjunction with high levels of aberrantly spliced mRNAs, reported for the first time. In three patients who were also analysed during blastic transformation, further reduction of full‐length IRF‐1 mRNA was observed. These findings demonstrate that, in CML patients, IRF‐1 can produce high levels of aberrant spliced mRNAs with subsequent reduction in the levels of full‐length IRF‐1 mRNA. This observation is consistent with the notion that exon skipping may constitute another mechanism of tumour suppressor gene inactivation in this disease.
Low blood dendritic cells in chronic myeloid leukaemia patients correlates with loss of CD34+/CD38– primitive haematopoietic progenitors
Authors: Mohty Mohamad , Isnardon Daniel , Vey Norbert , Brière Francine , Blaise Didier , Olive Daniel , Gaugler Béatrice
Published: March 2003

Summary. Myeloid and plasmacytoid dendritic cells (MDC, PDC) play a key role in the initiation of immune responses. We found a reduction of DC subsets among 20 chronic myeloid leukaemia (CML) patients in chronic phase (MDC, mean, 0·10% ± 0·10, P = 0·02; PDC, mean, 0·11% ± 0·08, P = 0·006 versus controls). The maintenance of blood DC correlated with the presence of high percentages of circulating CD34+/CD38– progenitors that were able to give rise in vitro to CD1a+ DC. The reduced DC numbers may contribute to leukaemia escape from immune control and restoration of DC may be a goal for CML immunotherapy.
Incidence and nature of CD20‐negative relapses following rituximab therapy in aggressive B‐cell non‐Hodgkin's lymphoma: a retrospective review
Authors: Kennedy Glen A. , Tey Siok‐Keen , Cobcroft Ralph , Marlton Paula , Cull Gavin , Grimmett Karen , Thomson Damien , Gill Devinder
Published: March 2003

Summary. Re‐treatment with rituximab for B‐cell non‐Hodgkin's lymphoma (NHL) relapsing after previous rituximab therapy has recently been shown to be clinically efficacious. Although the mechanism of resistance to rituximab re‐treatment in non‐responding patients is unknown, it is possible that loss of CD20 expression in the relapsed NHL could be important in some patients. We examined the incidence and nature of CD20 negative relapses following rituximab therapy in aggressive B‐cell NHL treated at our institution. Of a total of 18 patients who received rituximab, 13 have relapsed, with 10 patients subsequently undergoing repeat tissue biopsy. Six of these 10 patients (60%) were shown to have lost CD20 expression by either immunohistochemistry and/or flow cytometry. Furthermore, three of the six patients who relapsed with CD20‐negative NHL also suffered relapses at unusual anatomical sites. We conclude that loss of CD20 expression in aggressive B‐cell NHL relapsing post‐rituximab therapy is common. As such, repeat tissue biopsy should be undertaken to document CD20 expression by both flow cytometry and immunohistochemistry prior to considering repeated courses of rituximab in relapsed aggressive lymphomas.
Dose‐reduced conditioning regimen followed by allogeneic stem cell transplantation in patients with myelofibrosis with myeloid metaplasia
Authors: Hessling Jörg , Kröger Nicolaus , Werner Mathias , Zabelina Tatjana , Hansen Arne , Kordes Uwe , Ayuk Francis Ayuketang , Renges Helmut , Panse Jörg , Erttmann Rudolf , Zander Axel Rolf
Published: March 2003

Summary. Three patients with myelofibrosis received allogeneic stem cell transplantation after a dose‐reduced conditioning regimen of busulphan (8 mg/kg), fludarabine (180 mg/m2) and antithymocyte globulin (4 × 10 mg/kg). The median age at transplantation was 51 years (range 44–58). All patients engrafted with a leucocyte count > 1·0 × 109/l after a median of 18 d (range 16–20). Grade II acute skin graft‐versus‐host disease (GvHD) occurred in one patient. One limited and one extensive chronic GvHD was observed. All patients achieved complete haematological remission. In one patient the fibrosis resolved completely 180 d post transplant. All patients are alive 126, 466 and 764 d after transplantation.
Autologous bone marrow transplantation in the treatment of refractory systemic sclerosis: early results from a French multicentre phase I–II study
Authors: Farge Dominique , Marolleau Jean Pierre , Zohar Sarah , Marjanovic Zora , Cabane Jean , Mounier Nicolas , Hachulla Eric , Philippe Pierre , Sibilia Jean , Rabian Claire , Chevret Sylvie , Gluckman Eliane ,   for the Intensification et Autogreffe dans les Maladies Auto Immunes Resistantes (ISAMAIR) Study Group
Published: March 2003

Summary. Haematopoietic stem cell transplantation (HSCT) has been proposed for refractory autoimmune diseases, including systemic sclerosis (SSc). A sequential Bayesian phase I–II clinical trial was conducted in SSc patients to assess the feasibility, the tolerance and the efficacy of autologous HSCT. Peripheral blood stem cells (PBSC) were collected using cyclophosphamide (4 g/m2) and recombinant human granulocyte colony‐stimulating factor (5 µg/kg/d) and reinfused after positive CD34+ selection. Conditioning used cyclophosphamide (200 mg/kg) or melphalan (140 mg/m2) according to cardiac function. The main end‐point was the failure of the procedure, defined by failure of either PBSC mobilization, CD34+ selection or intensification procedure, or by procedure‐related death. Among the 12 enrolled patients, three failures occurred: one PBSC mobilization, one CD34+ selection and one CD34+ intensification. Probability of graft failure was estimated at 0·286 (95% confidence interval: 0·095–0·54). Autologous PBSC (n = 10) or bone marrow (n = 1) transplantation was actually performed in 11 patients with one procedure‐related death. Median time to neutrophil (> 0·5 × 109/l) and platelet (> 25 × 109/l) haematopoietic reconstitution was 12 and 10 d respectively. After 18 months (range 1–26), eight out of 11 patients have shown major or partial response. Non‐myeloablative conditioning, followed by a T cell‐depleted autologous PBSC or bone marrow transplantation, appears feasible with low toxicity in severe SSc with short‐term clinical benefits.
Genetic polymorphism of the major regulatory element HS‐40 upstream of the human α‐globin gene cluster
Authors: Harteveld Cornelis L. , Muglia Maria , Passarino Giuseppe , Kielman Menno F. , Bernini Luigi F.
Published: March 2003

Summary. The highly conserved 350‐bp major regulatory element HS‐40 (or αMRE) upstream of the human α‐globin gene cluster is involved in the regulation of α‐globin gene expression. The study of αMRE differences between human populations and the evolution of αMRE sequences in mammals may lead to a better understanding of the function and importance of this element in the regulation of expression of the downstream α‐cluster. Denaturing gradient gel electrophoresis was used to determine the sequence heterogeneity of the αMRE region in 276 unrelated individuals, representing seven different populations. Furthermore, we analysed the α major regulatory elements of chimpanzee, orang‐utan and rhesus monkeys and compared them with the equivalent human and murine sequences. Six different αMRE haplotypes (labelled A to F) were found in humans. Haplotype frequencies between the seven populations showed a gradual shift to a higher haplotype A distribution from west to east, being the highest in Indonesians. The African sample shows the largest divergence in haplotypes. Five out of six different haplotypes were present, three of which were exclusively found in Africans. The high prevalence of the haplotype A in humans, together with the conservation of this haplotype in apes, suggests that it is the ancestral one. The αMRE fragment appears to be a highly polymorphic marker, which could be used in combination with the regular markers in the α‐cluster to extend the haplotype and to follow segregation of α‐thalassaemia genes in population studies more accurately.
Genetic hyperferritinaemia and reticuloendothelial iron overload associated with a three base pair deletion in the coding region of the ferroportin gene (SLC11A3)
Authors: Cazzola Mario , Cremonesi Laura , Papaioannou Maria , Soriani Nadia , Kioumi Anna , Charalambidou Anastasia , Paroni Rita , Romtsou Katerina , Levi Sonia , Ferrari Maurizio , Arosio Paolo , Christakis John
Published: March 2003

Summary. Iron overload may predominantly involve parenchymal or reticuloendothelial cells, the prototype of parenchymal iron overload being HFE‐related genetic haemochromatosis. We studied a family with autosomal dominant hyperferritinaemia in whom the proband showed selective iron accumulation in the Kupffer cells on liver biopsy. Analysis of L and H ferritin genes excluded mutations responsible for hereditary hyperferritinaemia/cataract syndrome or similar translational disorders. Sequence analysis of the ferroportin gene (SLC11A3) in four individuals with hyperferritinaemia singled out a three base pair deletion in a region that contains four TTG repeats. This mutation removes a TTG unit from 780 to 791, and predicts the loss of one of three sequential valine residues 160–162. Denaturing high performance liquid chromatography can be used for its detection. SLC11A3 polymorphism analysis indicates that this probably represents a recurrent mutation due to slippage mispairing. Affected individuals may show marginally low serum iron and transferrin saturation, and young women may have marginally low haemoglobin concentration levels. Serum ferritin levels are directly related to age, but are 10–20 times higher than normal. Heterozygosity for the ferroportin Val 162 deletion represents the prototype of selective reticuloendothelial iron overload, and should be taken into account in the differential diagnosis of hereditary or congenital hyperferritinaemias.
Hydroxyurea exerts bi‐modal dose‐dependent effects on erythropoiesis in human cultured erythroid cells via distinct pathways
Authors: Wang Min , Tang Delia C. , Liu Wenli , Chin Kyung , Zhu Jianqion G. , Fibach Eitan , Rodgers Griffin P.
Published: March 2003

Summary. Hydroxyurea (HU) has been shown to increase the proportion of fetal haemoglobin (HbF) in most sickle cell patients. A low‐dosage regimen increased total haemoglobin (Hb) levels in some thalassaemia intermedia patients by preferentially increasing β‐globin biosynthesis. To further characterize these apparent dose‐dependent effects of HU, we examined erythroid cells exposed to HU (5–100 µmol/l) in two‐phase liquid culture. Low doses (from 5 to 25 µmol/l) increased Hb levels by up to 2·7‐fold, and a high dose (100 µmol/l) increased Hb levels when added at d 3–6 of phase II, with no significant changes in response to HU during the late stage of phase II culture (≥ 9 d). HU exposure during d 0–3 of phase II culture increased the number of erythroid colonies to a maximum of fivefold at 5 µmol/l HU. GATA‐1 mRNA was downregulated at a high dose and GATA‐2 was dose dependently upregulated over a lower dosage range. Treatment with 100 µmol/l HU dramatically upregulated the death receptor DR‐5, caspase 3, as determined by cDNA microarray analysis. In contrast, 10 µmol/l HU modestly upregulated mRNA levels of the early growth response gene. Our results suggest that HU exerts concentration‐dependent effects on HbF production and erythropoiesis and that these two effects are mediated by distinct molecular mechanisms.
Phenotypic and genetic analysis of a compound heterozygote for dys‐ and hypoprothrombinaemia
Authors: Akhavan Sepideh , Luciani Matteo , Lavoretano Silvia , Mannucci Pier Mannuccio
Published: March 2003

Summary. We studied a 2‐year‐old boy with a phenotype of combined hypo‐ and dysprothrombinaemia. Sequencing of polymerase‐chain‐reaction‐amplified genomic DNA revealed three different mutations in heterozygosity, a G to A transition at position 7312, resulting in the replacement of arginine 271 by histidine, an A to G transition at position 20058, resulting in the replacement of histidine 562 by arginine, and a 2‐bp deletion at 20062–20063, causing a frameshift and a premature stop codon in exon 14. The first two mutations are compatible with the dysprothrombinaemia phenotype, whereas the small deletion is thought to be the cause of hypoprothrombinaemia.
Elderly haematological patients with chemotherapy‐induced febrile neutropenia have similar rates of infection and outcome to younger adults: a prospective study of risk‐adapted therapy
Authors: García‐Suárez Julio , Krsnik Isabel , Reyes Eduardo , De Miguel Dunia , Hernanz Nuria , Barr‐Alí Mohamed , Burgaleta Carmen
Published: March 2003

Summary. We prospectively evaluated 131 consecutive episodes of fever and chemotherapy‐induced neutropenia in 85 adults with haematological malignancies to determine whether older patients (aged < 60 years) have different causes of fever and outcome than younger adults (aged < 60 years). Patients were stratified into high‐risk and low‐risk groups according to previously published criteria. High‐risk patients received ceftazidime plus amikacin and low‐risk patients received ceftazidime alone. All patients were hospitalized until fever and neutropenia resolved. Ninety one high‐risk episodes were documented: 56 occurring in older patients (mean age 69 years) and 35 in younger adults (mean age 45 years). Non‐Hodkgin's lymphoma and acute myeloid leukaemia were the most frequent underlying neoplasias in both age groups. Intensity of chemotherapy was similar in both age groups. Mean neutrophil count at entry, median duration of neutropenia, rate of documented infection, incidence of bacteraemia, response to therapy, overall mortality and infectious mortality were similar in the two high‐risk age subgroups. The elderly subgroup had a trend to have more Gram‐negative infections and the younger patients more Gram‐positive infections. In addition, 40 low‐risk episodes were registered: 29 in elderly patients (mean age 68 years) and 11 in younger patients (mean age 44 years). Elderly low‐risk patients had more concurrent diseases that younger ones (P = 0·124). Mean neutrophil count at entry, median duration of severe neutropenia and rate of response were similar in the two age subgroups. All low‐risk patients survived. In conclusion, elderly haematological cancer patients with febrile neutropenia show similar rates of infection and outcome to younger ones.
A peptide (P2) derived from the variable heavy chain of an anti‐P‐selectin monoclonal antibody (LYP20) inhibits leucocyte adhesion to thrombin‐activated platelets and endothelial cells
Authors: Murphy Joseph F. , McGregor John L.
Published: March 2003

Summary. P‐selectin, a member of the selectin family of adhesion molecules, is present in endothelial Weibel–Palade bodies and platelet α‐granules, and is rapidly expressed on their surface upon activation, resulting in leucocyte adhesion. LYP20 is a functional monoclonal antibody previously generated in our laboratory that binds with high affinity and specificity directed against P‐selectin. This binding is largely imparted by the specific sequence of amino acids present on the hypervariable portions of the IgG chains. We now show that a peptide derived from the heavy chain of mAb LYP20 dose dependently inhibits the adhesion of poly morphonuclear cells to resting and thrombin‐activated endothelial cells (EC) and platelets. The scrambled form of this peptide, identical in amino acid composition to the authentic peptide but with altered sequence, was not inhibitory at corresponding concentrations. Binding studies revealed that this peptide also dose dependently bound to both resting and thrombin‐activated EC and platelets. Our results may prove useful for the development of new therapeutic inhibitors to modulate leucocyte interactions in inflammatory disorders.
Differential expression of CD2 on neoplastic mast cells in patients with systemic mast cell disease with and without an associated clonal haematological disorder
Authors: Pardanani Animesh , Kimlinger Teresa K. , Reeder Terra L. , Li Chin‐Yang , Tefferi Ayalew
Published: March 2003

Summary. Recently, aberrant coexpression of CD2 and CD25 has been reported to reliably distinguish neoplastic mast cells from normal or so‐called reactive mast cells. Such expression is included in the consensus diagnostic criteria for systemic mast cell disease (SMCD). In our study of patients with SMCD, we found CD2 expression to be more prevalent on mast cells from patients without an associated haematological disorder (P = 0·04). Furthermore, no correlation was found between mast cell CD2 expression and other clinicopathological features in these patients.
Residual leukaemia after bone marrow transplant in children with acute lymphoblastic leukaemia after first haematological relapse or with poor initial presenting features
Authors: Bunin Nancy , Johnston Dennis A. , Roberts W. Mark , Ouspenskaia Maia V. , Papusha Victor Z. , Brandt Mark A. , Zipf Theodore F.
Published: March 2003

Summary. Relapse is the major obstacle to cure for children with acute lymphoblastic leukaemia (ALL) after allogeneic bone marrow transplant (BMT). Development of salvage therapy for post‐transplant relapse could be expedited by understanding the post‐transplant behaviour of microscopically undetectable leukaemia and the ability to predict impending relapse. We have used a quantitative polymerase chain reaction method (sensitivity of 5·0 × 10−6) to measure residual leukaemia before the conditioning regimen, and at five time‐points after transplantation. In total, 18 patients with ALL transplanted in first or second remission were studied for 1 year: For the first year post BMT, 12 remained in remission, four had haematological relapses, one had a cutaneous relapse, and one died of severe graft‐versus‐host disease. The post‐engraftment levels of the leukaemia‐specific immunoglobulin heavy (IgH) chain gene rearrangement for patients with haematological relapses were significantly different from those who remained in remission. The levels for the patients who remained in remission decreased with time, although there were occasional increases consistent with the known standard deviation of the measurement assay. In contrast, all clinical relapses were preceded by a rapid increase in levels. Both the rate of this increase and its timing were variable. These results suggest that residual leukaemia measurements can be used to direct post‐transplant interventions and measure their effects.
Prospective evaluation of cell kinetics, yields and donor experiences during a single large‐volume apheresis versus two smaller volume consecutive day collections of allogeneic peripheral blood stem cells
Authors: Bolan Charles D. , Carter Charles S. , Wesley Robert A. , Yau Yu Ying , Barrett A. John , Childs Richard W. , Read Elizabeth J. , Leitman Susan F.
Published: March 2003

Summary. We report cell kinetics, yields and donation experiences of 20 demographically matched allogeneic peripheral blood stem cell (PBSC) donors who were prospectively assigned to undergo either a single 25 l or two consecutive daily 15 l (15 l × 2) apheresis procedures. Procedures were performed using prophylactic intravenous calcium administration after standard granulocyte colony‐stimulating factor (GCSF) mobilization (10 μg/kg/d). Central line placements (two each), initial CD34 cell counts (0·077 vs 0·078 × 109/l) and yields (7·9 vs 8·1 × 108 CD34 cells) were similar in the two groups; however, 25 l donors spent significantly less time both in the clinic (7·5 vs 10·8 h) and with central venous catheters in place (8·5 vs 29·5 h) than 15 l × 2 donors. End‐procedure platelet counts were below 100 × 109/l in one out of 10 25 l donors versus five out of 10 in 15 l × 2 donors (41%vs 53% mean decrease in platelet counts, P = 0·02). PBSC collection efficiency increased by 37% after 15 l of the 25‐l volume had been processed, compared with no significant change during 15 l × 2 procedures. Results similar to these prospective findings were also observed in CD34 yields, symptoms and platelet counts in additional 25 l and 15 l procedures performed during the same period and evaluated retrospectively. This study indicates that a single 25‐l apheresis procedure results in similar yields and symptoms, but less donor thrombocytopenia and inconvenience than two consecutive daily 15‐l procedures.
Mutations of the cationic trypsinogen gene in patients with hereditary pancreatitis
Authors: Creighton J E , Lyall R , Wilson D I , Curtis A , Charnley R M
Published: March 2003

Background

Hereditary pancreatitis has been shown to be caused by one of two mutations (R117H and N21I) of the cationic trypsinogen gene (PRSS1). Families with hereditary pancreatitis in the north of England were investigated for these mutations. The clinical features associated with each mutation were compared.

Methods

In individuals from nine families with hereditary pancreatitis, DNA was screened for the R117H and N21I mutations. All five exons of the cationic trypsinogen gene were also sequenced to search for additional mutations. Haplotype analysis was carried out to identify common ancestors. Clinical data were collected.

Results

The R117H mutation was identified in three families and N21I in a further five. The R117H mutation was associated with a more severe phenotype than N21I in terms of mean(s.d.) age of onset of symptoms (8·4(7·2) versus 16·5(7·1) years; P = 0·007) and requirement for surgical intervention (eight of 12 versus four of 17 patients respectively; P = 0·029). Haplotype analysis suggested that each mutation had arisen more than once.

Conclusion

Two mutations in the cationic trypsinogen gene cause hereditary pancreatitis in eight of nine families originating in this region. The R117H mutation is associated with a more severe form of the disease in terms of age at onset of symptoms and requirement for surgical intervention.
Lymphocyte activation after non-thermal trauma
Authors: Walsh D S , Siritongtaworn P , Pattanapanyasat K , Thavichaigarn P , Kongcharoen P , Jiarakul N , Tongtawe P , Yongvanitchit K , Komoltri C , Dheeradhada C , Pearce F C , Wiesmann W P , Webster H K
Published: March 2003

Background

Severe injury causes immunological changes that may contribute to a poor outcome. Longitudinal characterization of lymphocyte response patterns may provide further insight into the basis of these immunological alterations.

Methods

Venous blood obtained seven times over 2 weeks from 61 patients with injury severity scores above 20 was assessed for lymphocyte phenotypic and activation markers together with serum levels of interleukin (IL) 2, IL-4, soluble IL-2 receptor (sIL-2R), soluble CD4 (sCD4), soluble CD8 (sCD8) and interferon γ.

Results

Severe injury was associated with profound changes in the phenotypic and activation profile of circulating lymphocytes. Activation was indicated by increased numbers of T cells expressing CD25, CD69 and CD71, and raised serum levels of IL-2, sIL-2R, sCD4 and sCD8. Relatively higher levels of sIL-2R and sCD4 were found in patients with sepsis syndrome.

Conclusion

Polytrauma is associated with dramatic alterations in the phenotypic and activation profile of circulating lymphocytes which are generally independent of clinical course. In contrast, several lymphocyte soluble factors, including sCD4 and sIL-2R, paralleled the clinical course. These data provide new insight into lymphocyte responses after injury and suggest that further assessment of soluble factors as clinical correlates, including those related to lymphocyte activation or generalized inflammation, may be warranted.
Xenotransplantation
Authors: Soin B , Vial C M , Friend P J
Published: March 2003

Background

The success of clinical transplantation has led to a large discrepancy between donor organ availability and demand; considerable pressure exists to develop an alternative source of organs. The use of animal organs for donation is a possible solution that is not yet clinically applicable.

Methods and results

A literature review was performed based on a Medline search to find articles on xenotransplantation. Keywords included hyperacute, acute vascular, xenograft rejection combined with concordant and discordant. Additional references cited in these articles from journals not included in Medline were obtained from the British Library. Limited information on unpublished, preliminary work has been included from sources known to the authors, based on their research work in the field. One hundred and forty-six references and four personal communications have been included in this review article.

Conclusion

A greater understanding of the pathogenesis of xenograft rejection is developing rapidly. Strategies to abrogate hyperacute rejection have proved successful, but control of antibody-driven acute vascular rejection has not yet been achieved. The safety and viability of xenotransplantation as a therapeutic modality are still unproven.
Apolipoprotein E genotype is associated with differential expansion rates of small abdominal aortic aneurysms
Authors: Gerdes L U , Lindholt J S , Vammen S , Henneberg E W , Fasting H
Published: March 2003

Background

The common polymorphism of the apolipoprotein E (APOE) gene is associated with differential risk of atherosclerosis; the gene could be a candidate gene in abdominal aortic aneurysms (AAA).

Methods

APOE genotypes were determined in 57 men aged 65–73 years with a small AAA (30–50 mm). The patients were included in a population ultrasonographic screening programme and were followed with at least two examinations during an interval of 2–4·5 years. The AAA expansion rates in patients with four different APOE genotypes were studied, with adjustment for initial AAA size and smoking.

Results

APOE genotype was a significant determinant of AAA expansion rate (P = 0·001). The adjusted mean (95 per cent confidence interval) rate was 2·1 (1·7–2·6) mm/year in 31 men with genotype E3E3, 1·3 (0·7–1·9) mm/year in 17 men with E3E4, 3·1 (2·0–4·1) mm/year in six men with E2E3 and 4·2 (2·7–5·6) mm/year in three men with E2E4. The mean expansion rate was 2·2 (1·5–2·8) mm/year in non-smokers and 3·0 (2·5–3·6) mm/year in smokers (P = 0·014).

Conclusion

APOE genotype seems to influence AAA expansion rate, but the effects of the individual genotypes, in particular E3E3 and E3E4, are contradictory when compared with the effects of the genotypes on risk of atherosclerosis.
Systematic review of antimicrobial agents used for chronic wounds
Authors: O'Meara S M , Cullum N A , Majid M , Sheldon T A
Published: March 2003

Background

This paper is one of a series of eight systematic reviews that aim to identify effective interventions for chronic wounds. Here, antimicrobial agents are evaluated.

Methods

Electronic databases and other relevant sources were accessed to identify published and unpublished material. Studies were eligible for inclusion if they used concurrent controls, recruited participants with chronic wounds, evaluated an intervention designed to prevent or treat chronic wounds, and incorporated an objective assessment of wound healing. All included studies were assessed against a comprehensive checklist for methodological quality. A narrative overview was conducted.

Results

Thirty trials were included, 25 of randomized design. Small sample size and other methodological problems meant that findings were often difficult to interpret. Results do not support the routine use of systemic antibiotics for leg ulcers or diabetic foot ulcers without acute infection, but they may be useful as an adjunct to surgery for pilonidal sinuses. Several topical preparations may be helpful, including dimethyl sulphoxide, silver sulphadiazine, benzoyl peroxide, oxyquinoline and gentamicin.

Conclusion

Most of this research requires replication in larger, well designed studies to establish both clinical and cost effectiveness.