bluuebunny/crossref_metadata_2025 huggingface dataset:
Coagulation factor XIII polymorphisms and the risk of myocardial infarction and ischaemic stroke in young women
Authors: Reiner Alexander P. , Frank Michele B. , Schwartz Stephen M. , Linenberger Michael L. , Longstreth W. T. , Teramura Gayle , Rosendaal Frits R. , Psaty Bruce M. , Siscovick David S.
Published: March 2003

Summary. The inconsistent findings among association studies that have examined the relationship between factor XIIIA Val34Leu and thrombosis may be owing to (1) population differences in the prevalence of other risk factors that modify the association with Val34Leu, or (2) linkage disequilibrium with other functional factor XIIIA polymorphisms. We therefore performed genotyping for factor XIIIA Val34Leu, Tyr204Phe and Pro564Leu in a population‐based study of myocardial infarction (MI) and ischaemic stroke among white women < 45‐years of age and 345 demographically similar controls, and examined potential interactions with other risk factors. The presence of the factor XIIIA Leu34 allele was associated with a slight decreased risk of MI [odds ratio (OR) = 0·80] that was most pronounced among women with traditional cardiovascular risk factors. Paradoxically, women carrying two copies of the Leu34 allele had a nearly fourfold increased risk of ischaemic stroke relative to the Val34/Val34 genotype. Heterozygosity for factor XIIIA Phe204 was associated with a milder increased risk of ischaemic stroke, and analysis of a kindred with congenital dysfibrinogenaemia suggested that co‐inheritance of the factor XIIIA Phe204 allele may increase susceptibility to ischaemic stroke. Our results suggest that the factor XIIIA Val34Leu variant may be associated with a decreased risk of MI among young women with other risk factors. The relationship of factor XIIIA polymorphisms to cerebrovascular disease requires further study.
Magnetic resonance imaging of femoral marrow predicts outcome in adult patients with acute myeloid leukaemia in complete remission
Authors: Takagi Shojiro , Tanaka Osamu
Published: March 2003

Summary.  Accurate assessment of residual disease is important for the prediction of outcome in patients with acute leukaemia in complete remission (CR). To investigate whether abnormalities on magnetic resonance (MR) images of femoral marrow in adult patients with acute myeloid leukaemia (AML) in CR can predict outcome, 28 newly diagnosed patients with AML underwent MR imaging when bone marrow aspiration or biopsy was performed to verify the state of CR after induction therapy. MR abnormalities on short TI (inversion time) inversion recovery (STIR) techniques persisted in all four patients who did not achieve CR. In 13 CR patients abnormalities on STIR images resolved, to result in normal appearance at the time CR was achieved. All 13 patients remained in CR for 3–104 months (median, 73 months). In the other 11 CR patients, STIR abnormalities persisted at the time CR was achieved. Seven of them relapsed between 1 and 28 months (median, 3 months) after MR evaluation. Disease‐free survival of patients with persistent abnormal STIR images was significantly shorter than that of patients with normal STIR images (P < 0·01). MR imaging of femoral marrow may predict outcome in adult patients with AML in CR.
Combined arsenic trioxide and all‐trans retinoic acid treatment for acute promyelocytic leukaemia recurring from previous relapses successfully treated using arsenic trioxide
Authors: Au W. Y. , Chim C. S. , Lie A. K. W. , Liang R. , Kwong Y. L.
Published: March 2003

Summary.  The optimal treatment of acute promyelocytic leukaemia (APL) recurring from relapses successfully treated using arsenic trioxide (As2O3) is undefined. Three APL patients relapsing from As2O3‐induced remission were studied. Re‐treatment with As2O3 failed in one patient in third relapse, and resulted in morphological but not molecular remission in another patient. Combination therapy with As2O3 and all‐trans retinoic acid (ATRA), however, resulted in morphological and molecular remission in all three cases, with a follow‐up time ranging from 6 to 16 months. Our results suggest a synergistic therapeutic effect between As2O3 and ATRA in APL in advanced relapse.
Optimizing dendritic cell‐based immunotherapy in multiple myeloma
Authors: Yi Qing , Desikan, Raman , Barlogie Bart , Munshi Nikhil
Published: March 2003

Summary. Vaccination with idiotype protein‐pulsed dendritic cells (DCs) has been explored in multiple myeloma and the results have been disappointing. These studies used immature DCs, which are less potent at activating T cells and could differentiate to macrophages once the cytokines were withdrawn. After intravenous administration, DCs accumulate in the lungs and liver for up to 48 h, thus reducing their potential to migrate to lymphoid organs and interact with T cells. To improve the efficacy of DC vaccination in myeloma, we investigated the use of idiotype‐pulsed mature DCs administered subcutaneously. Five patients (three IgG and two IgA myeloma) with stable partial remission following high‐dose chemotherapy were enrolled. DC vaccines were administered three times at 2‐week intervals at least 4 months post transplantation. Idiotype‐specific T‐cell responses, detected using enzyme‐linked immunospot (ELISPOT) (four patients) and proliferation (two patients) assays, were elicited in four and anti‐idiotypic B‐cell responses in all five patients. The cytokine‐secretion profile of activated T cells demonstrated a type‐1 response. A 50% reduction in serum M‐component was observed in one immunologically responding patient that persisted for 6 months and stable disease (for 6 months) resulted in the other three patients. The remaining patient without an immune response to the vaccination relapsed. No major side‐effects were noted. Thus, subcutaneous administration of idiotype‐pulsed mature DCs induced idiotype‐specific T‐ and B‐cell responses. Current efforts are geared towards optimizing the conditions of DC generation and administration, and the development of in vitro assays to monitor the cytotoxicity of the T cells.
Factor XIII activity levels in patients with allogeneic haematopoietic stem cell transplantation and acute graft‐versus‐host disease of the gut
Authors: Pihusch Rudolf , Salat Christoph , Göhring Peter , Hentrich Markus , Wegner Holger , Pihusch Markus , Hiller Erhard , Kolb Hans‐Jochem , Ostermann Helmut
Published: March 2003

Summary. Severe graft‐versus‐host disease (GvHD) of the gut clinically resembles Crohn's disease and ulcerative colitis. As low plasma levels of factor XIII (FXIII) have been described in chronic inflammatory bowel disease (CIBD) and as beneficial effects of FXIII concentrates in CIBD have been reported, we studied the FXIII plasma activity levels in patients undergoing allogeneic stem cell transplantation (SCT). In 20 of 22 patients with an uncomplicated course of SCT, FXIII stayed within the normal range (median 102 iu/dl, range 74–122), but was significantly reduced with the lowest FXIII levels on d 0 and 7 (d 0: median 83 iu/dl, range 55–165, d 7: median 83, range 70–101). In 20 of 22 patients with histologically proven GvHD of the gut, FXIII levels far below the normal range were observed (median 50, range 21–87) with a strong correlation between FXIII activity levels and degree of GvHD (r = −0·908; P < 0·001). We conclude that FXIII is consumed in patients with GvHD of the gut. As FXIII plays a a crucial role in haemostasis and wound healing, a study on the potential benefit of FXIII substitution in patients with severe GvHD of the gut might be rewarding.
FGFR3 dysregulation in multiple myeloma: frequency and prognostic relevance
Authors: Rasmussen Thomas , Hudlebusch Heidi Rye , Knudsen Lene Meldgaard , Johnsen Hans Erik
Published: March 2003

Summary. The t(4:14) translocation affects two potential oncogenes, FGFR3 and MMSET, in multiple myeloma (MM). We investigated the frequency of FGFR3 dysregulation and its prognostic value in MM. FGFR3 mRNA levels were determined in 110 diagnostic bone marrow (BM) samples from MM patients. In addition, selected BM samples were screened for elevated MMSET mRNA levels. 14·5% (16/110) of MM BM samples showed dysregulated FGFR3 expression. Follow‐up of 76 MM patients showed no significant difference between FGFR3 dysfunction and survival (P = 0·3) or correlation with known prognostic factors. Further, no linear relation was observed between FGFR3 and MMSET levels.
Mean platelet volume is an independent risk factor for myocardial infarction but not for coronary artery disease
Authors: Endler Georg , Klimesch Alexandra , Sunder‐Plassmann Heike , Schillinger Martin , Exner Markus , Mannhalter Christine , Jordanova Nelli , Christ Günter , Thalhammer Renate , Huber Kurt , Sunder‐Plassmann Raute
Published: March 2003

Summary.  After rupture of an arteriosclerotic plaque in a coronary artery, platelets play a crucial role in the subsequent thrombus formation, leading to myocardial infarction. An increased mean platelet volume (MPV), as an indicator of larger, more reactive platelets, may represent a risk factor for myocardial infarction. However, this hypothesis is still controversial and most studies addressing the role of MPV were performed comparing patients suffering from myocardial infarction with healthy controls. We intended to identify patients at high risk of suffering myocardial infarction in a group of patients with known coronary artery disease. One hundred and eighty‐five consecutive patients with stable coronary artery disease were compared with 188 individuals who had suffered myocardial infarction. Patients within the highest quintile of MPV (≥ 11·6 fl) had a significantly higher risk of experiencing a myocardial infarction compared with patients within the lowest quintile (OR = 2·6, 95% CI 1·3–5·1) in a multivariate analysis that included sex, age, body mass index, hyperlipidaemia, hypertension, smoking and diabetes mellitus. Our results indicate that patients with pre‐existing coronary artery disease and an increased MPV (≥ 11·6 fl) are at higher risk of myocardial infarction. These patients can be easily identified during routine haematological analysis and could possibly benefit from preventive treatment.
Three major glucose‐6‐phosphate dehydrogenase‐deficient polymorphic variants identified in Mazandaran state of Iran
Authors: Mesbah‐Namin Seyed A. , Sanati Mohammad H. , Mowjoodi Alireza , Mason Philip J. , Vulliamy Tom J. , Noori‐Daloii Mohammad R.
Published: March 2003

Summary. We report the first investigation of glucose‐ 6‐phosphate dehydrogenase (G6PD) deficiency among the Mazandaranians in the north of Iran. We analysed the G6PD gene in 74 unrelated G6PD‐deficient men with a history of favism. Molecular analysis revealed three major different polymorphic variants: G6PD Mediterranean 66·2% (49 out of 74), G6PD Chatham 27% (20 out of 74), G6PD Cosenza 6·75% (5 out of 74). These findings indicated a higher prevalence of G6PD Chatham in this Iranian population than anywhere else in the world. In addition, the distribution of these G6PD variants is more similar to that found in an Italian population than in other Middle Eastern countries.
Interferon and ribavirin as frontline treatment for chronic hepatitis C infection in thalassaemia major
Authors: Li Chi Kong , Chan Paul K. S. , Ling Siu Cheung , Ha Shau Yin
Published: March 2003

Summary.  Hepatitis C virus (HCV) infection is common in transfusion‐dependent thalassaemia. The clinical usefulness of 12‐month treatment, using interferon alpha 3 MIU/m2 thrice weekly and oral ribavirin 16 mg/kg/d, was evaluated in 18 previously untreated thalassaemia patients. The median age at start of treatment was 16 years (range 7–29). Fourteen were infected with genotype 1b and 4 with genotype 6a. The sustained biochemical and virological response rates 6 months after stopping treatment were both 72·2%. Blood consumption was temporarily increased by 30% due to ribavirin‐associated haemolysis. This study demonstrated a high, sustained response rate to combination treatment despite infection with genotype 1b.
Prothrombin A19911G and G20210A polymorphisms' role in thrombosis
Authors: Pérez‐Ceballos Elena , Corral Javier , Alberca Ignacio , Vayá Amparo , Llamas Pilar , Montes Ramón , González‐Conejero Rocio , Vicente Vicente
Published: March 2003

Summary. The prothrombin G20210A polymorphism, which correlates with the plasmatic prothombin levels, is the second genetic risk factor for deep venous thrombosis (DVT), although its prothrombotic role is mild. Recently, the prothrombin A19911G polymorphism, also associated with slight variations of the prothrombin level, has been suggested to modulate the thrombotic risk of the G20210A polymorphism in a preliminary study including few patients and controls. Our study evaluated the effect of the A19911G polymorphism in the arterial and venous thrombotic risk of the prothrombin 20210G/A genotype, analysing 204 consecutive DVT patients and 204 matched controls. Moreover, we analysed 213 carriers of the 20210G/A genotype (152 with DVT, 26 with arterial thrombosis and 35 healthy subjects) and 10 homozygous 20210 A/A carriers. We developed a simple method to simultaneously determine the genotype of both polymorphisms. In accordance with our case/control study, the A19911G polymorphism did not play a significant role in the development of DVT. Analysis of 120 20210 A alleles demonstrated a complete linkage disequilibrium with the 19911 A allele. These polymorphisms (alone or combined) did not modify the risk of arterial thrombosis. However, the 19911A/G genotype slightly increased the risk of developing DVT in carriers of the 20210G/A genotype (OR 3·34 vs 5·86), supporting that the prothrombin 19911 polymorphism could modulate the risk of the G20210A polymorphism in developing DVT.
Safety of allogeneic Epstein–Barr virus (EBV)‐specific cytotoxic T lymphocytes for patients with refractory EBV‐related lymphoma
Authors: Sun Qi , Burton Robert , Reddy Vishnu , Lucas Kenneth G.
Published: March 2003

Summary. Epstein–Barr virus (EBV) causes lymphomas in immunocompromised individuals such as recipients of stem cell or organ transplants and patients with acquired immunodeficiency syndrome (AIDS). EBV has also been detected in the Reed–Sternberg cells of approximately 50% of all cases of Hodgkin's disease (HD). The purpose of this study was to examine the safety, and the clinical and immunological effects of infusing allogeneic EBV‐specific cytotoxic T lymphocytes (CTL) for patients with refractory EBV‐positive malignancies. In this pilot study, we have treated four patients with EBV‐related lymphoma using allogeneic EBV‐specific CTL. Two patients received EBV‐specific CTL derived from partially human leucocyte antigen (HLA)‐matched donors and the other two from HLA‐matched siblings. No complications were observed as a result of the CTL infusions and all patients showed increased levels of EBV‐specific CTL precursors (CTLp) post infusion. Of the two organ transplant patients, one had refractory disease and has sustained a complete remission following the T‐cell infusions. The second has also been disease free since T‐cell infusions, although the efficacy cannot be definitively attributed to CTL therapy because this patient received local radiation therapy prior to immunotherapy. A patient with AIDS‐related, EBV‐positive lymphoma had disease progression following CTL infusions. One HD patient received HLA 4/6 matched T cells from an unrelated donor and showed a decrease in the size of affected lymph nodes and resolution of B‐symptoms post infusion. In conclusion, adoptive immunotherapy with allogeneic EBV‐specific CTL is safe and mayhave efficacy in patients with high‐risk or refractory EBV‐related tumours.
Relationship between hypercoagulable state and erythrocyte phosphatidylserine exposure in splenectomized haemoglobin E/β‐thalassaemic patients*
Authors: Atichartakarn Vichai , Angchaisuksiri Pantep , Aryurachai Katcharin , Onpun Supornchai , Chuncharunee Suporn , Thakkinstian Ammarin , Atamasirikul Kanlayanee
Published: March 2003

Summary. Small pulmonary arterial thromboses can occur following splenectomy of patients with haemoglobin E/β‐thalassaemia (Hb E/β‐thal). We compared plasma markers of coagulation activation in vivo and red blood cell (RBC) markers of procoagulant activity in 15 Hb E/β‐thal patients who were not splenectomized (NS), 15 who had been splenectomized (S), and 15 normal controls (NC). Levels of plasma thrombin–antithrombin III complex (TAT) were significantly higher in the S group than in either the NS or the NC groups, and levels of prothrombin fragment 1.2 (F 1.2) were significantly higher in the S than in the NC group. Diluted Russell's viper venom clotting times were significantly shorter when RBCs from group S patients were added to the assay compared with RBCs from the NC group. Phosphatidylserine (PS) expression (% of annexin V‐positive RBCs) on the outer leaflet of RBC membrane of both ‘larger’‐ and ‘smaller’‐sized RBCs was significantly higher for the S than the NC group. The RBC PS expression of the S and the NS groups, respectively, accounted for 25·3% (P = 0·174) and 6·3% (P = 0·675) of the variation in plasma TAT levels. Our findings indicated that, when compared with NC, splenectomized patients with Hb E/β‐thal were in a chronic low‐grade hypercoagulable state associated with increased numbers of circulating PS exposed RBCs. This condition may have a role in the risk of these patients for pulmonary arterial thromboses.
Abnormal WT1 expression in the CD34‐negative compartment in myelodysplastic bone marrow
Authors: Van Dijk Jeroen P. , Knops Gertrudis H. J. N. , Van De Locht Louis T. F. , Menke Aswin L. , Jansen Joop H. , Mensink Ewald J. B. M. , Raymakers Reinier A. P. , De Witte Theo
Published: March 2003

Summary. In normal bone marrow, WT1 expression is restricted to CD34+ cells. We assessed WT1 mRNA expression levels with quantitative, real‐time reverse transcription polymerase chain reaction in normal, myelodysplastic (MDS) and secondary acute myeloid leukaemia (sAML) bone marrow subfractions, based on differentiation status. The highest WT1 expression was observed in the primitive CD34+ rhodamine‐123 (rho) dull cells, both in healthy donors and MDS or sAML patients. In contrast to normal CD34‐negative bone marrow cells, WT1 was present in CD34‐negative bone marrow cells in 12 out of 13 MDS patients and two sAML samples. Further analysis of this aberrant WT1 expression was performed in the CD34‐negative subfractions of three MDS patients. In one of these, WT1 expression was found exclusively in the erythroid cells. This patient was completely transfusion dependent and showed morphological dyserythropoiesis. In another MDS patient, WT1 expression was found in a non‐erythroid compartment. We conclude that abnormal WT1 expression may contribute to the disturbed differentiation of haematopoietic cells in MDS patients.
Hodgkin and Reed–Sternberg cells express functional c‐kit receptors and interact with primary fibroblasts from Hodgkin's disease‐involved lymph nodes through soluble and membrane‐bound stem cell factor
Authors: Aldinucci Donatella , Poletto Dalisa , Nanni Paola , Degan Massimo , Gloghini Annunziata , Di Francia Raffaele , Russo Stefania , Carbone Antonino , Pinto Antonio , Gattei Valter
Published: March 2003

Summary. Classic Hodgkin's disease (cHD) is a lymphoid neoplasia characterized by few malignant Hodgkin and Reed–Sternberg (H‐RS) cells, embedded in an abundant background of non‐tumour cells. We have previously demonstrated the expression in primary H‐RS cells of the receptor tyrosine kinase (RTK) c‐kit; here we describe its functional role in the cross‐talk between H‐RS cells themselves with neighbouring cell populations. In particular, we analysed the expression of c‐kit and its ligand stem cell factor (SCF) in a panel of HD‐derived cell lines and fibroblasts from HD‐involved lymph nodes (HDF). While c‐kit was expressed by HD‐derived cell lines, usually in the absence of SCF, this latter molecule, in its soluble and/or membrane‐bound (mb) form, was in turn expressed at a high level by primary HDF. In vitro adhesion between HD‐derived cell lines and HDF was mainly mediated by c–kit/SCF interactions, and this phenomenon was significantly inhibited by an excess of soluble SCF or by neutralizing anti‐c‐kit monoclonal antibodies. Furthermore, both soluble and mb‐SCF increased growth and colony survival of HD‐derived cell lines; these effects were significantly enhanced upon co‐stimulation of H‐RS cells with interleukin 9. Finally, soluble SCF was able to partially rescue H‐RS cells from apoptosis induced by serum starvation. Taken together, our data indicated the expression of functional c‐kit receptor by H‐RS cells and suggests a role of SCF in the pathobiology of cHD.
The impact of transfusion of leucodepleted platelet concentrates on cytomegalovirus disease after allogeneic stem cell transplantation
Authors: Ronghe Milind D. , Foot Annabel B. M. , Cornish Jaqueline M. , Steward Colin G. , Carrington David , Goulden Nicholas , Marks David I. , Oakhill Anthony , Pamphilon Derwood H
Published: March 2003

Summary. The impact of transfusion of leucodepleted platelet concentrates (PCs) on cytomegalovirus (CMV) disease was assessed in 215 allogeneic (145 unrelated and 70 related donor) transplants over 3 years. In 43%, both donor and patient were CMV seronegative (CMV–/–). All received CMV‐seronegative red cells and random leucodepleted PCs. No CMV disease occurred in any CMV–/– (low risk) transplant. CMV infection occurred in 31 seropositive patients (26%); 13 died and five deaths were attributable to CMV disease. When compared with historical controls, who received CMV‐seronegative PCs, we found no difference in transfusion‐acquired CMV in the current cohort.
Human cytomegalovirus induces a direct inhibitory effect on antigen presentation by monocyte‐derived immature dendritic cells
Authors: Grigoleit Ulrich , Riegler Susanne , Einsele Hermann , Laib Sampaio Kerstin , Jahn Gerhard , Hebart Holger , Brossart Peter , Frank Friderike , Sinzger Christian
Published: March 2003

Summary. The hypothesis that productive infection of monocyte‐derived immature dendritic cells (DCs) by the human cytomegalovirus (HCMV) is associated with decreased immunostimulatory capacity was tested in this study. DCs were infected with 60–80% efficiency by HCMV strain TB40/E. Infected versus uninfected cells were analysed by fluorescence‐activated cell sorting and by immunocytochemistry for surface expression of major histocompatibility complex (MHC) and co‐stimulatory molecules as well as cytokine secretion during the 3 d after infection. The immunostimulatory capacity of these cells was measured by mixed leucocyte reaction. In spite of the fact that HCMV infection of DCs induced an increased release of tumour necrosis factor‐α (TNF‐α) and a decreased interleukin 10 (IL‐10) production, expression of MHC class I and II, as well as CD40 and CD80 molecules, were downregulated on infected DCs. The mixed leucocyte reaction showed significantly reduced immunostimulatory capacity of infected DC cultures. Simultaneous detection of MHC antigens and virus antigens by double immunofluorescence revealed that downregulation occurred only on infected cells, but not on uninfected bystander cells. These findings demonstrate on a single cell level, together with the marked downregulation of MHC and co‐stimulatory molecules in the presence of high TNF‐α and low IL‐10 levels, a direct inhibitory effect of HCMV on antigen presentation by immature DCs independent of soluble mediators.
Reconstitution of human haematopoiesis in non‐obese diabetic/severe combined immunodeficient mice by clonal cells expanded from single CD34+CD38– cells expressing Flk2/Flt3
Authors: Ebihara Yasuhiro , Wada Mika , Ueda Takahiro , Xu Ming‐jiang , Manabe Atsushi , Tanaka Ryuhei , Ito Mamoru , Mugishima Hideo , Asano Shigetaka , Nakahata Tatsutoshi , Tsuji Kohichiro
Published: March 2003

Summary. In the present study, we examined the expression of Flk2/Flt3, a tyrosine kinase receptor, on human cord blood CD34+ haematopoietic progenitor/stem cells. In flow cytometric analysis, Flk2/Flt3 was expressed on 80% of CD34+ cells and their immature subpopulations, CD34+CD33– and CD34+CD38– cells. Methycellulose clonal culture of sorted CD34+Flk2/Flt3+ and CD34+Flk2/Flt3– cells showed that most of myelocytic progenitors expressed Flk2/Flt3, but erythroid and haematopoietic multipotential progenitors were shared by both fractions. When 1 × 104 lineage marker‐negative (Lin–)CD34+Flk2/Flt3– cells were transplanted into non‐obese diabetic/severe combined immunodeficient (NOD/SCID) mice, none of the recipients possessed human CD45+ cells in bone marrow 11–12 weeks after the transplantation. In contrast, all recipients transplanted with 1 × 104 Lin–CD34+Flk2/Flt3+ cells showed successful engraftment. Furthermore, clonal cells expanded from single Lin–CD34+CD38–Flk2/Flt3+ cells in the culture with Flk2/Flt3 ligand, stem cell factor, thrombopoietin, and a complex of interleukin 6/soluble interleukin 6 receptor were individually transplanted into NOD/SCID mice. At 20 to 21 weeks after the transplantation, three out of 10 clones harvested at d 7 of culture, and three out of six clones at d 14 could reconstitute human haematopoiesis in recipient marrow. These results demonstrated that Flk2/Flt3 was expressed on a wide variety of human haematopoietic cells including long‐term‐repopulating haematopoietic stem cells.
Lupus anticoagulant testing: improvements in performance in a UK NEQAS proficiency testing exercise after dissemination of national guidelines on laboratory methods
Authors: Jennings Ian , Greaves Mike , Mackie Ian J. , Kitchen Steve , Woods Timothy A. L. , Preston F. Eric ,   on behalf of the UK National External Quality Assessment Scheme for Blood Coagul ation
Published: March 2003

Summary. Laboratory screening for lupus anticoagulant (LA) has been shown to be suboptimal in several studies. Guidelines have recently been published by an expert group for the British Committee for Standards in Haematology, in an attempt to standardize and improve screening procedures. The value of using screening tests conforming with these guidelines was investigated in a United Kingdom National External Quality Assessment Scheme (UK NEQAS) proficiency testing exercise. The correct diagnosis was achieved by 97% of laboratories for a LA‐negative sample. However, 18·3% of centres reported a false‐negative result for a sample from a LA‐positive subject. A significantly higher proportion of centres that used methods conforming with the published guidelines achieved the correct diagnosis for this sample (P < 0·002, chi‐square test). A wide variety of screening tests were used by laboratories in this study. Within‐method agreement could be improved by the use of a common normal pooled plasma to determine ratios. However, between‐method agreement was not improved by this procedure. We conclude that adoption of methods compliant with national guidelines may improve the diagnosis of LA. There is a need, however, for reference and standardization materials to ensure further improvement in the accuracy of LA methods.
The expression of prion protein by endothelial cells: a source of the plasma form of prion protein?
Authors: Starke Richard , Drummond Olive , MacGregor Ian , Biggerstaff John , Gale Rosemary , Camilleri Raymond , Mackie Ian , Machin Samuel , Harrison Paul
Published: March 2003

Summary. The neuronal prion protein (PrPC) is also expressed within peripheral tissues including human blood. The majority of blood PrPC is found within the plasma fraction. We hypothesized that the vascular endothelium could be a source of this PrPC. Reverse transcription polymerase chain reaction demonstrated that both human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC‐1) expressed PrPC mRNA. Flow cytometry confirmed PrPC expression on HMEC‐1s and HUVECs (120 900 ± 15 058 and 58 327 ± 4577 molecules PrPC/cell respectively), with no upregulation following cellular activation. Confocal immunofluorescence microscopy confirmed that HMEC‐1s and HUVECs were positive for PrPC on the plasma membrane. Time‐resolved dissociation‐enhanced fluoroimmunoassay (DELFIA®) analysis of cell culture medium demonstrated a slow constitutive release of soluble PrPC not associated with activation. In contrast to von Willebrand factor antigen, PrPC plasma levels in vivo decrease following desmopressin therapy in patients with von Willebrand disease. Measurement of PrPC plasma levels in patients with varying blood counts demonstrated no association between cell count and PrPC concentration. However, there was a higher level of PrPC in plasma from patients with end‐stage renal failure. In conclusion, endothelial cells of both macrovascular and microvascular origin expressed high levels of PrPC which can be constitutively released into the cell culture medium.
Molecular histogenesis of plasmablastic lymphoma of the oral cavity
Authors: Gaidano Gianluca , Cerri Michaela , Capello Daniela , Berra Eva , Deambrogi Clara , Rossi Davide , Larocca Luigi Maria , Campo Elias , Gloghini Annunziata , Tirelli Umberto , Carbone Antonino
Published: March 2003

Summary. Plasmablastic lymphoma (PBL) of the oral cavity is an aggressive B‐cell lymphoma associated with human immunodeficiency virus infection. Although the lymphoma phenotype is consistent with late B‐cell maturation, the molecular histogenesis of PBL is unknown. We investigated PBL of the oral cavity (n = 12) for mutations of immunoglobulin variable heavy chain (IgVH) and BCL‐6 genes, which are acquired by B cells at the time of germinal centre (GC) transit, and for expression of BCL‐6, MUM‐1 and CD138, which distinguish GC B cells from post‐GC B cells. Somatic IgVH hypermutation occurred in 4/10 PBL whereas 6/10 PBL displayed germline IgVH genes. Among PBL carrying hypermutated IgVH genes, the pattern of IgVH mutations was consistent with antigen stimulation in two cases. Mutations of the BCL‐6 gene were restricted to 1/12 patients with PBL of the oral cavity. All cases of PBL of the oral cavity displayed the BCL‐6–/MUM‐1+/CD138+ phenotype that is consistent with late stage of B‐cell differentiation. Overall, these data indicate that, despite a common phenotype and an apparently similar degree of differentiation, PBL of the oral cavity are characterized by histogenetic heterogeneity. A subset of PBL of the oral cavity carried the molecular clues of GC transit and conceivably originated from a B‐cell subset corresponding to post‐GC B cells. Conversely, another fraction of these lymphomas were devoid of somatic IgVH mutations and appeared to originate from naive B cells that have undergone preterminal differentiation independent of GC transit.
Anti‐myeloma activity of natural killer lymphocytes
Authors: Frohn Christoph , Höppner Maike , Schlenke Peter , Kirchner Holger , Koritke Petra , Luhm Jürgen
Published: March 2003

Summary. Natural killer (NK) cells are assumed to contribute to a graft‐versus‐leukaemia effect. In vitro experiments have shown that many leukaemic cells are NK‐cell sensitive. Nevertheless, no data concerning the influence of purified NK cells on malignant myeloma (MM) cells exist. We co‐incubated NK cells with three different MM cell lines and fresh bone marrow samples of nine MM patients. The proportion of vital MM cells was determined before and after co‐cultivation by a flow‐cytometry‐based assay. All MM cells tested, with the exception of one cell line (NCI H929), were susceptible to a NK‐cell attack even without exogenous interleukin 2 (IL‐2). The mean killing of the native MM samples was 23·1 ± 5·4% and 34·5 ± 6·5% at 10:1 and 20:1 effector:target ratio respectively, This corresponded to about 2/3 of those values obtained with the highly sensitive line K562. In contrast, CD34‐positive haematopoietic stem cells as well as peripheral mononuclear cells were completely resistant under similar experimental conditions (1·3% killing). To elucidate the underlying triggering mechanisms, we measured human leucocyte antigen (HLA)‐class I expression of the MM cells. No evidence for HLA loss, which could have explained the NK‐cell recognition if it occurred, was demonstrated. These findings may contribute to the understanding of in vivo NK‐cell activation and encourage clinical applications of NK cells for MM patients.
Nitric oxide and cyclic GMP levels in sickle cell patients receiving hydroxyurea
Authors: Nahavandi Masoud , Tavakkoli Fatemeh , Wyche Melville Q. , Perlin Elliott , Winter William P. , Castro Oswaldo
Published: March 2003

Summary. Recent studies suggest that nitric oxide (NO) may partly be responsible for the beneficial effect of hydroxyurea (HU) in sickle cell disease (SCD) patients. NO stimulates cyclic guanosine monophosphate (cGMP) production, which mediates vasodilatation. We investigated the association between NO, cGMP and fetal haemoglobin (HbF) levels after HU administration. Our data showed that chronic HU significantly increased NO, cGMP, and HbF levels in SCD. Recently it was shown that HbF production was stimulated by cGMP‐dependent protein kinase. Our results suggest that NO stimulates cGMP production, which then activates a protein kinase and increases the production of HbF.
Intrasinusoidal bone marrow infiltration: a common growth pattern for different lymphoma subtypes
Authors: Costes Valérie , Duchayne Eliane , Taib Jacques , Delfour Christophe , Rousset Thérèse , Baldet Pierre , Delsol Georges , Brousset Pierre
Published: March 2003

Summary. We report a retrospective immunohistochemical study on bone marrow biopsies of 43 patients with different types of lymphomas showing unusual intrasinusoidal infiltration. Most of these patients presented with splenomegaly (74·4%) and peripheral lymphocytosis (83%). In 20/43 patients, lymphoid infiltrates were not detectable on haematoxylin–eosin sections. After immunohistochemistry on bone marrow biopsies and blood and bone marrow smear examinations, the following diagnoses were made: splenic marginal zone lymphoma with villous lymphocytes (SLVL) in 24 patients, large granular lymphocyte (LGL) leukaemia in 14 patients, hepatosplenic T‐cell lymphoma in two patients, anaplastic large cell lymphoma in two patients and intravascular large B‐cell lymphoma in one patient. In the presence of intrasinusoidal infiltrates of small lymphocytes, a B‐cell phenotype (CD20+, CD76/DBA44+/–) was associated with splenic marginal zone lymphoma whereas intrasinusoidal CD3/CD45RA‐positive T‐cell infiltrates were strongly suggestive of LGL leukaemia. Intrasinusoidal bone marrow infiltration appears to be a common feature of distinct lymphoma subtypes. Immunohistochemical analysis is essential to detect intrasinusoidal medullary infiltrates (which may be minimal) and should be systematically performed in patients with splenomegaly and peripheral lymphocytosis.
Risk factors for adults with Philadelphia‐chromosome‐positive acute lymphoblastic leukaemia in remission treated with allogeneic bone marrow transplantation: the potential of real‐time quantitative reverse‐transcription polymerase chain reaction
Authors: Lee Seok , Kim Dong‐Wook , Cho Bin , Kim Yoo‐Jin , Kim Yoo‐Li , Hwang Ji‐Yeon , Park Yoon‐Hee , Shin Ho‐Jin , Park Chi‐Young , Min Woo‐Sung , Kim Hack‐Ki , Kim Chun‐Choo
Published: March 2003

Summary. The aim of this study was to evaluate the outcomes for Philadelphia‐chromosome‐positive acute lymphoblastic leukaemia (Ph+ ALL) patients in remission treated with allogeneic bone marrow transplantation (BMT). Twenty‐three adults were entered onto this study. The 2‐year probabilities of relapse and disease‐free survival (DFS) were 39·4 ± 11·6% and 43·5 ± 10·3% respectively. The presence of chronic graft‐versus‐host disease (GVHD) was found to be an independent predictive factor affecting lower relapse and DFS. To monitor the BCR‐ABL transcript, we also analysed 48 bone marrow samples of eight patients using real‐time quantitative reverse‐transcription polymerase chain reaction (RT‐PCR). The kinetics of the BCR‐ABL transcript correlated well with the patients' clinical course. In six patients who were in continuous remission after BMT, a rapid decrease in BCR‐ABL copy number to the PCR‐negative status was observed after the development of chronic GVHD. Meanwhile, routine bone marrow examination of two patients showed PCR positivity with a 3 or 4‐log increase of BCR‐ABL copy number and subsequent haematological relapse, which occurred 2 and 4 months later respectively. Although our data should be interpreted cautiously, the presence of chronic GVHD may reduce the risk of relapse in Ph+ ALL. Real‐time quantitative RT‐PCR appears to be a useful test for BCR‐ABL transcript monitoring.
Impact of disordered puberty on bone density in β‐thalassaemia major
Authors: Bielinski Basia K. , Darbyshire Phil J. , Mathers Lynne , Crabtree Nicola J. , Kirk Jeremy M. W. , Stirling Heather F. , Shaw Nick J.
Published: March 2003

Summary.Reduction of bone density and its associated morbidity is recognized in young adults with β‐thalassaemia major, but the aetiology is not clear. This study used dual X‐ray absorptiometry (DXA) to look at bone mineral apparent density (BMAD) in children and young adults with thalassaemia in a predominantly Asian population, in the context of sexual maturation. Fifty‐five patients were scanned (mean age 13·8 years, range 5·9–37·5) and BMAD z‐scores were calculated using normal data from locally recruited control subjects. Eighteen patients had undergone bone marrow transplantation (BMT) and the remainder were on a transfusion/chelation regimen. BMAD z‐scores ranged from–3·3–1·6 with a mean of −0·92. No difference in BMAD was found between those patients treated conventionally and those who had undergone BMT. When comparing mean BMAD z‐score according to sexual maturation, there was a highly significant difference (P < 0·0001) between those whose pubertal maturation was age appropriate (mean z‐score −0·22), when compared with those who had disordered puberty (mean z‐score −1·82). We have shown that failure to progress normally through puberty is highly significant in the failure of adequate bone mineralization and achievement of peak bone mass in thalassaemic patients. The management of these patients should therefore be pro‐active to anticipate problems and facilitate normal sexual maturation.
Oral anticoagulants in the elderly
Authors: Froom Paul , Miron Ehud , Barak Mira
Published: March 2003

Summary. Over a 1‐year period, 34 998 prothrombin time tests were performed on 2379 patients aged 40–89 years with a recommended target International Normalized Ratio (INR) value of 2·5. At least one INR value of ≥ 5 was found in 507/2379 patients (21·3%). The elderly demonstrated higher maximum and lower minimum INR values, lower warfarin doses, and an increased number of tests performed. After adjustment for these factors, the risk of INR values ≥ 5 increased with age by 15% every 10 years (95% CI 4–28%). We conclude that age is a risk factor for more unstable prothrombin time results.
In vivo imaging studies of the effect of recipient conditioning, donor cell phenotype and antigen disparity on homing of haematopoietic cells to the bone marrow
Authors: Askenasy Nadir , Farkas Daniel L.
Published: March 2003

Summary. Homing of transplanted bone marrow cells (BMC) to the host bone marrow (BM) is the first step of engraftment towards durable multilineage haematopoietic reconstitution. We used an in vivo assay to track PKH‐labelled cells in the BM of mice after transplantation, using fluorescence microscopy through an optical window placed over the distal femoral epiphysis. Within hours after intravenous injection, the cells moved in and out the femur, and were mobile within the marrow space. One hour after injection of whole BMC into non‐conditioned syngeneic and allogeneic recipients, the homing efficiencies (HE) were 1·23 ± 0·14% and 0·12 ± 0·02% respectively (P < 0·001). Irrespective of antigen disparity, the number of PKH‐labelled cells in the femur decreased by 30% and 50% after 1 and 3 d respectively (P < 0·001). Similar HE of naïve and irradiated cells suggested that the majority of cells (> 80%) were quiescent in the BM during the first 3 d. HE were twofold higher in busulphan‐myeloablated recipients (P < 0·001 vs non‐conditioned), and allogeneic transplantation resulted in 84 ± 9% donor chimaerism at 4 weeks. The HE of lin– cells was 16‐fold higher than that of lin+ cells (P < 0·001), and the subset of lin– SCA‐1+ cells was 4·6‐fold higher in the BM‐homed cell population (P < 0·001 vs lin– cells). Approximately 1500 of the BM‐homed cells rescued 62–71% secondary syngeneic and allogeneic myeloablated recipients. Strikingly, the HE could be predicted during the first 3 d after transplantation by correcting the measurements performed in vivo for the enrichment of progenitors in donor inoculum, donor‐recipient antigen disparity and myeloablative conditioning.
Relative importance of female‐specific and non‐female‐specific effects on variation in iron stores between women
Authors: Whitfield John B. , Treloar Susan , Zhu Gu , Powell Lawrie W. , Martin Nicholas G.
Published: March 2003

Summary. Women have lower iron stores than men because of iron loss during their reproductive years. However, variation between women could result from differences in iron loss, aspects of iron homeostasis common to men and women, or a combination of both. We compared the effects of age, menopause, menstrual blood loss and the number of pregnancies (sex‐specific factors), and the effects of genetic variation, on markers of iron stores. We assessed how much the same genes or other familial factors influence iron status in both men and women. Data from 2039 female twins who participated in studies of reproductive health and iron status were used to estimate the proportions of variation that could be ascribed to genes, environment and measured factors. Significant effects of age, menopausal status and magnitude of menstrual blood loss were demonstrated, accounting for up to 18% of variance in serum ferritin in this sample, but number of children had no significant effect. Genetic effects were more than twice as great as sex‐specific effects. The within‐pair similarity of ferritin values in dizygotic female twin pairs was greater than for dizygotic opposite‐sex pairs, but this difference was not quite significant, consistent with a minor role for sex‐specific factors; and the opposite‐sex within‐pair differences did not diminish significantly with age. We conclude that the contribution of genetic differences between women to variation in iron stores outweighs the comparatively small effects of interindividual variation in iron loss through variation in menstruation and number of pregnancies.
Pattern of lymph node metastases in papillary thyroid carcinoma
Authors: Gimm O , Rath F W , Dralle H
Published: March 2003

Background

Incidence and pattern of lymph node metastasis (LNM) were evaluated in 35 patients who underwent primary surgery for papillary thyroid carcinoma.

Methods

Total thyroidectomy with systematic cervicocentral lymphadenectomy was performed routinely. Additionally, 22 patients underwent lymphadenectomy of the ipsilateral cervicolateral compartment. Sternotomy was performed in four patients.

Results

Primary tumour status was: pT1, n = 7; pT2, n = 12; pT3, n = 4; pT4, n = 12. Twenty-four patients had LNM in the ipsilateral cervicocentral compartment (pT1, n = 2; pT2, n = 8; pT3, n = 4; pT4, n = 10). Nineteen had LNM in the ipsilateral cervicolateral compartment (pT1, n = 4; pT2, n = 6; pT3, n = 2; pT4, n = 7). Contralateral cervicocentral LNM was restricted to patients with pT3 tumours (n = 2) or pT4 tumours (n = 3). No patient had LNM in the contralateral cervicolateral compartment. Only one patient (pT3 tumour) had mediastinal LNM.

Conclusion

The cervicocentral compartment was most commonly involved with metastases, although the ipsilateral cervicolateral compartment was almost as often affected. In some patients ipsilateral cervicolateral LNM was present in the absence of cervicocentral LNM. Contralateral cervicolateral and upper mediastinal LNM, however, were rare. This study helps to plan radical surgery for papillary thyroid carcinoma.
Influence of a defunctioning stoma on leakage rates after low colorectal anastomosis and colonic J pouch–anal anastomosis
Authors: Dehni N , Schlegel R D , Cunningham C , Guiguet M , Tiret E , Parc R
Published: March 2003

Background

Anal sphincter function is increasingly preserved following rectal excision for cancer and provides a better quality of life for patients than does a permanent colostomy. However, anastomotic complications may cause considerable morbidity and mortality. This retrospective study examined the incidence of anastomotic complications following two forms of reconstruction after resection for mid-rectal cancer: colonic pouch–anal anastomosis (CPAA) and low colorectal anastomosis (LCRA).

Methods

Some 258 consecutive patients with mid-rectal cancers between 6 and 11 cm from the anal verge underwent proctectomy with mesorectal excision and either CPAA or LCRA. The incidence of clinical and radiological leaks was determined in these patients who were considered in three groups: LCRA (defunctioning stoma), LCRA (no defunctioning stoma) and CPAA (all defunctioned).

Results

In the LCRA group without a defunctioning stoma, a clinical leak occurred in 17·0 per cent, compared with two of 30 in the LCRA group with a defunctioning stoma. In the CPAA group a clinical leak occurred in 4·9 per cent of patients, which was not significantly different from the rate in those with a defunctioned LCRA. Patients with a non-defunctioned LCRA were more likely to suffer a clinical anastomotic leak (P=0·01), peritonitis (P=0·001) and require unscheduled reoperation (P=0·006) than those with a defunctioned LCRA and/or CPAA.

Conclusion

The use of a protective defunctioning stoma is advocated in conjunction with LCRAs.
POSSUM and Portsmouth POSSUM for predicting mortality
Authors: Prytherch D R , Whiteley M S , Higgins B , Weaver P C , Prout W G , Powell S J
Published: March 2003

Background

There is a need for an accurate measure of surgical outcomes so that hospitals and surgeons can be compared properly regardless of case mix. POSSUM (Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity) uses a physiological score and an operative severity score to calculate risks of mortality and morbidity. In a previous small study it was found that Portsmouth POSSUM (P-POSSUM; a modification of the POSSUM system) provided a more accurate prediction of mortality.

Methods

Some 10 000 general surgical interventions (excluding paediatric and day cases) were studied prospectively between August 1993 and November 1995. The POSSUM mortality equation was applied to the full 10 000 surgical episodes. The 10 000 patients were arranged in chronological order and the first 2500 were used as a training set to produce the modified P-POSSUM predictor equation. This was then applied prospectively to the remaining 7500 patients arranged chronologically in five groups of 1500.

Results

The original POSSUM logistic regression equation for mortality overpredicts the overall risk of death by more than twofold and the risk of death for patients at lowest risk (5 per cent or less) by more than sevenfold. The P-POSSUM equation produced a very close fit with the observed in-hospital mortality.

Conclusion

P-POSSUM provides an accurate method for comparative surgical audit.
Postoperative intussusception in children
Authors: de Vries S , Sleeboom C , Aronson D C
Published: March 2003

Background

Postoperative intussusception is an uncommon and sometimes forgotten cause of postoperative intestinal obstruction in children.

Methods

Eleven consecutive cases of postoperative intussusception during a period of 16 years (1981–1997) were analysed retrospectively.

Results

Symptoms developed after a median period of 5 days after operation. With one exception, postoperative intussusception was not diagnosed before reoperation. In ten of the 11 cases the intussusception was successfully treated with intraoperative manual reduction.

Conclusion

Postoperative intussusception is a rare but typical complication in the paediatric age group and should be kept in mind.