bluuebunny/crossref_metadata_2025 huggingface dataset:
The stromal composition of malignant lymphoid aggregates in bone marrow: variations in architecture and phenotype in different B‐cell tumours
Authors: Vega Francisco , Medeiros L. Jeffrey , Lang Wen‐Hua , Mansoor Adnan , Bueso‐Ramos Carlos , Jones Dan
Published: March 2003

Summary. We present evidence that different B‐cell tumours, in bone marrow, have different relationships to stroma. Marrow core biopsies from 46 patients with B‐cell tumours were immunostained with antibodies for distinct stromal cells. Cases included follicular lymphoma (FL), chronic lymphocytic leukaemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), lymphoplasmacytic lymphoma (LPL), and nodal, extranodal and splenic marginal zone lymphoma (NMZL, MALT, SMZL). In normal marrow, low‐affinity nerve growth factor receptor (LNGFR) highlighted a fine network of adventitial reticular cells (ARC). The nodular aggregates of CLL/SLL, NMZL, MALT and SMZL were characterized by distortion of the ARC network and downregulation of LNGFR. In contrast, the aggregates of FL, LPL and MCL were composed of linear arrays of ARC in tight association with individual tumour cells. LNFGR+ was upregulated in ARC associated with the aggregates in FL, LPL and focally in MCL. Upregulation of CD35, vascular cell adhesion molecule (VCAM‐1) and CD40 on ARC was noted exclusively in FL. Marrow lymphoid aggregates in CLL/SLL, NMZL, MALT and SMZL probably grow by displacing the pre‐existing marrow stroma, while FL and LPL maintain a close association with the ARC network. In FL, expression of follicular dendritic cell‐associated markers is modulated in pre‐existing marrow stromal cells.
Human leucocyte antigen‐Cw‐specific cytotoxic T lymphocytes generated from naive cord blood used for cord blood stem cell transplantation
Authors: Yazaki Makoto , Takahashi Toshitada , Mizutani Keigo , Ito Yasuhiko , Wakiguchi Hiroshi , Inoue Masami , Kawa Keisei , Kato Koji , Kato Toro , Saito Hidehiko , Togari Hajime
Published: March 2003

Summary.  Human leucocyte antigen (HLA)‐Cw‐reactive cytotoxic T lymphocytes (CTL) were generated from cord blood (CB) lymphocytes of two cases used for cord blood stem cell transplantation (CBSCT). In both cases, the CTL were cytotoxic against the patient's leukaemic cells, as well as the patient's Epstein–Barr virus (EBV)‐lymphoblastoid cell line (EBV‐LCL) and phytohaemagglutinin blasts, and the cytotoxicity was blocked by anti‐HLA‐class I monoclonal antibodies. In the first case, the CTL recognized Cw 3 (Cw 9 and Cw 10)‐positive EBV‐LCL, while in the second case, the CTL recognized Cw1 and/or Cw7. These cases suggest that CB T cells may be competent enough for generating CTL to induce a graft‐versus‐leukaemia effect and/or graft‐versus‐host disease in patients with CBSCT and that the mismatching of Cw antigens between patient and CB may be related to the outcome of CBSCT.
A novel inherited mutation of the transcription factor RUNX1 causes thrombocytopenia and may predispose to acute myeloid leukaemia
Authors: Walker Logan C. , Stevens Jane , Campbell Hamish , Corbett Rob , Spearing Ruth , Heaton David , Macdonald Donald H. , Morris Christine M. , Ganly Peter
Published: March 2003

Summary.  The RUNX1 (AML1, CBFA2) gene is a member of the runt transcription factor family, responsible for DNA binding and heterodimerization of other non‐DNA binding transcription factors. RUNX1 plays an important part in regulating haematopoiesis and it is frequently disrupted by illegitimate somatic recombination in both acute myeloid and lymphoblastic leukaemia. Germline mutations of RUNX1 have also recently been described and are dominantly associated with inherited leukaemic conditions. We have identified a unique point mutation of the RUNX1 gene (A107P) in members of a family with autosomal dominant inheritance of thrombocytopenia. One member has developed acute myeloid leukaemia (AML).
Induction of CD94/NKG2A expression on T cells in mixed lymphocyte culture by CD14+ cells from granulocyte colony‐stimulating factor‐mobilized peripheral blood mononuclear cells
Authors: Tanaka Junji , Tutumi Yutaka , Li Zhang , Katho Naoko , Sugita Junichi , Mori Akio , Toyoshima Nobuyasu , Ota Shuichi , Kobayashi Masanobu , Kasai Masaharu , Asaka Masahiro , Imamura Masahiro
Published: March 2003

Summary.  We report the increased expression of inhibitory natural killer (NK) cell receptor, CD94/NKG2A, for human leucocyte antigen (HLA) class I on CD8+ T cells in granulocyte colony‐stimulating factor‐mobilized peripheral blood mononuclear cells (G‐PBMC) after mixed lymphocyte culture (MLC). The addition of purified CD14+ cells to CD14‐depleted G‐PBMC‐induced CD94/NKG2A expression in a dose‐dependent fashion; however, this enhancing effect was inhibited by the membrane between responder cells and CD14+ cells. Therefore, CD14+ cells in G‐PBMC induce CD94/NKG2A expression on CD8+ T cells, which in turn appear to downregulate alloresponses after allogeneic peripheral blood stem cell transplantation.
Chromosomal aberrations are shared by malignant plasma cells and a small fraction of circulating CD19+ cells in patients with myeloma and monoclonal gammopathy of undetermined significance
Authors: Zojer Niklas , Schuster‐Kolbe Judith , Assmann Irene , Ackermann Jutta , Strasser Kathrin , Hübl Wolfgang , Drach Johannes , Ludwig Heinz
Published: March 2003

Summary.  In the present study, we aimed to identify distinct structural and numerical chromosomal aberrations in peripheral blood B cells of patients with myeloma and monoclonal gammopathy of undetermined significance (MGUS), which reflect changes thought to occur at different stages of the disease process. Peripheral blood from 12 patients with multiple myeloma and three patients with MGUS was investigated for the occurrence of retinoblastoma‐1 gene deletions, p53 gene deletions and numerical aberrations demonstrated previously to be present in the patients' bone marrow CD138+ cells. By combining immunocytochemical staining for light chains and interphase fluorescence in situ hybridization (FISH), aberrant light‐chain +ve cells were detected in the circulating CD19+ cell fraction. Each kind of chromosomal change present in the myeloma tumour cells was found to be shared by a small fraction of CD19+ cells (0·1–1·8%; median 0·36%, n = 6). In one MGUS patient, aberrant cells could be identified with a frequency of 0·34% within the CD19‐sorted cell fraction. Clonotypic cells were detected with a frequency of 0·01–0·07% of peripheral blood nucleated cells by m‐RNA in situ hybridization with patient‐specific probes in three investigated patients. These results provide evidence that the circulating clonotypic B cells are closely related to the malignant plasma cells in myeloma and MGUS.
Reliability of lymphoma classification in bone marrow trephines
Authors: Buhr Thomas , Länger Florian , Schlué Jerome , Von Wasielewski Reinhardt , Lehmann Ulrich , Braumann Dietrich , Kreipe Hans
Published: March 2003

Summary. The aim of this study was to test and establish the accuracy and reliability of lymphoma classification in bone marrow trephines according to the new World Health Organization (WHO) classification by considering predominantly the morphology and immunophenotype. Therefore, we retrospectively compared lymphoma diagnoses, rendered exclusively on bone marrow trephines without knowledge of lymph node diagnosis in 124 patients, with the results of the reference centres that had reviewed lymph node (n = 90) or extranodal biopsies (n = 34). The overall concordance rate was higher than 85% and 91%, respectively, when patients with discordant malignancy grades were excluded. The concordance rate for low‐grade B‐cell lymphomas was 93% and for high‐grade B‐cell lymphomas 84%. The main reasons for discordant diagnoses were divergent immunophenotypes among low‐grade B‐cell lymphomas (6 out of 81, i.e. 7·4%) and discrepant malignancy grades within high‐grade B‐cell lymphomas (6 out of 31, i.e. 19·4%). No relationship between discordant diagnoses and chemotherapy given during the course of the disease with the site of biopsy (i.e. lymph nodes, extranodal sites) was noted. We conclude from our results that bone marrow trephines are a reliable tool, not only for establishing bone marrow infiltration, but also for the subtyping of lymphomas.
Assessment of caspase activity as a possible prognostic factor in acute myeloid leukaemia
Authors: Oliver Lisa , Vavasseur Fabienne , Mahé Beatrice , Perrin Pascale , Harousseau Jean‐Luc , Meflah Khaled , Vallette François‐Marie
Published: March 2003

Summary. Caspases are one of the key effector molecules in apoptosis. Caspase‐3 activity (identified by cleavage of the peptide DEVD) was analysed in bone marrow blasts (minimum 70%) from 45 acute myeloid leukaemia (AML) patients. 12 patients (25%) exhibited high levels of specific DEVDase activity. These blast cells, despite having activated caspase‐3, displayed none of the classical caspase‐dependent morphological characteristics of apoptosis, such as degradation of DNA fragmentation factor 45, DNA fragmentation, and appeared to be more resistant to drug‐induced apoptosis. Our results suggest that in these AML cells, resistance to apoptosis occurred downstream of caspase‐3 activation.
MyD88 is involved in the signalling pathway for Taxol‐induced apoptosis and TNF‐α expression in human myelomonocytic cells
Authors: Wang Jingxin , Kobayashi Masanobu , Han Mingzhe , Choi Sungki , Takano Masatoshi , Hashino Satoshi , Tanaka Junji , Kondoh Takeshi , Kawamura Ken‐ichi , Hosokawa Masuo
Published: March 2003

Summary. Taxol is an effective anti‐tumour drug against a variety of tumour cells. Taxol directly induces apoptosis in addition to a G2/M cell cycle arrest. However, it remains poorly understood how Taxol induces apoptosis in tumour cells. Taxol induces the secretion of inflammatory cytokines in murine macrophages in a toll‐like receptor‐4 (TLR‐4)‐dependent manner in addition to its anti‐tumour effects, but the effect of Taxol on human macrophages is controversial. In this study, we demonstrated that low doses (less than 1000 nmol/l) of Taxol induced the expression of tumour necrosis factor (TNF)‐α in human myelomonocytic cells and that the induction of TNF‐α mRNA was inhibited by dominant‐negative myeloid differentiation protein (dnMyD88). Furthermore, we demonstrated that the same doses of Taxol induced apoptosis of the same myelomonocytic cells and that the Taxol‐induced apoptosis was also inhibited by dnMyD88. In accordance with the previous reports, Taxol induced the expression of TNF‐α and apoptosis in a TLR4‐independent manner. These results suggest that TNF‐α expression and apoptosis, both induced by Taxol in human myelomonocytic cells, share the signal transduction molecule MyD88.
Quantification improves the prognostic value of CD38 expression in B‐cell chronic lymphocytic leukaemia
Authors: Mainou‐Fowler Tryfonia , Dignum Helen , Taylor Penelope R. A. , Dickinson Anne M. , Saunders Peter W. G. , Proctor Stephen J. , Summerfield Geoffrey P.
Published: March 2003

Summary. Recent studies have shown that CD38 expressed as a percentage of the antigen positivity can predict prognosis and disease progression in patients with B‐cell chronic lymphocytic leukaemia (B‐CLL). The present study showed that quantification of CD38 expressed as antibody‐binding capacity (ABC) improves the prognostic value of the percentage of CD38 positivity in B‐CLL. In a cohort of 81 patients with B‐CLL, a level of CD38 expression of ≥ 30% and an ABC value of 250 proved statistically valid cut‐off points to predict disease progression (% CD38: P=0·0027; ABC: P < 0·0001). There was a positive and significant correlation between the percentage of CD38 expression and ABC (r=0·7; P < 0·0001). There was a better discrimination of survival using ABC rather than percentage CD38 positivity (P < 0·0001 compared with P=0·0027). Only ABC predicted for survival in patients under 60 years of age (P=0·0076) or with stage A disease (P=0·0195). Both percentage CD38 and ABC discriminated between time to first treatment for all patients but only ABC predicted time to treatment for stage A patients (P=0·0004). In conclusion, CD38 positivity is an important prognostic factor in B‐CLL. However, quantification of CD38 is superior to the percentage positivity and should be used clinically in conjunction with other variables of predictive value to identify B‐CLL patients that are likely to progress.
A Leu55 to Pro substitution in the integrin αIIb is responsible for a case of Glanzmann's thrombasthenia
Authors: Tanaka Shigenori , Hayashi Tomoya , Hori Yuji , Terada Chikahiro , Han Kyou Sup , Ahn Hyo Seop , Bourre Francois , Tani Yoshihiko
Published: March 2003

Summary. Glanzmann's thrombasthenia (GT) is a hereditary bleeding disorder caused by a quantitative or qualitative defect in the integrin αIIbβ3. A new mutation, a T to C substitution at base 258 in the αIIb gene, leading to the replacement of Leu55 with Pro, was found by sequence analysis of a patient's αIIb cDNA. In transfection experiments using COS7 cells, the cells co‐transfected with the mutated αIIb cDNA containing C258 and wild‐type β3 cDNA scarcely expressed the αIIbβ3 complex. The Leu55 to Pro substitution in the αIIb gene was found to be responsible for this case of Glanzmann's thrombasthenia.
Human natural killer cell development in a xenogeneic culture system
Authors: Barão Isabel , Vaz Fátima , Almeida‐Porada Graça , Srour Edward F. , Zanjani Esmail D. , Ascensão João L.
Published: March 2003

Summary. In vivo and in vitro xenogeneic models have shown the ability of a non‐human environment in supporting human haemopoiesis. In the present study, we evaluated the effect of fetal sheep thymic stroma in the invitro development of natural killer (NK) cells from humanhaemopoietic progenitors. CD34+HLA‐DR+ (CD34+ DR+)Lin– and CD34+DR–Lin– bone marrow (BM) progenitors were cultured for 3 weeks with or without interleukin 2 (IL‐2), in fetal sheep thymic stroma contact and transwell cultures. Both progenitors gave rise to NK cells, defined as CD45+CD56+ cells, in the presence or absence of IL‐2; however, the percentage of NK cells originated in cultures with IL‐2 was significantly higher. Direct contact with stroma seemed to be required for the most immature progenitors, CD34+DR–Lin–, to differentiate along the NK cell lineage. Functional assays revealed that only cells grown in the presence of IL‐2 were cytolytic against K562 targets and, curiously, NK cells derived from CD34+DR–Lin– progenitors were more cytotoxic that NK cells derived from CD34+DR+Lin– progenitors. These studies suggest that the ability of fetal sheep thymic stroma in promoting the generation of human NK cells from haemopoietic progenitors may have relevance in terms of NK cell ontogeny and induction of tolerance in transplantation.
CD40L stimulation enhances the ability of conventional metaphase cytogenetics to detect chromosome aberrations in B‐cell chronic lymphocytic leukaemia cells
Authors: Buhmann Raymund , Kurzeder Christian , Rehklau Jutta , Westhaus Doreen , Bursch Sabina , Hiddemann Wolfgang , Haferlach Torsten , Hallek Michael , Schoch Claudia
Published: March 2003

Summary. Conventional metaphase cytogenetics underestimates the frequency of specific chromosome aberrations in B‐cell chronic lymphocytic leukaemia (B‐CLL) as a result of the very low proliferative activity of these cells in vitro. New molecular approaches, such as fluorescence in situ hybridization (FISH) or comparative genomic hybridization (CGH), may circumvent this problem, at least in part, but these techniques are either strongly dependent on the knowledge of candidate regions or detect only unbalanced aberrations. In the present study, we analysed 27 B‐CLL peripheral blood samples by metaphase cytogenetics after CD40 ligand (CD40L)‐induced cell cycle stimulation. In comparison with the simultaneous use of B‐cell mitogens such as 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA), lipopolysaccharide (LPS) and pokeweed mitogen (PWM), CD40L stimulation of B‐CLL cells induced a threefold increase in metaphases amenable to analysis by conventional cytogenetics. The analysis of these metaphases confirmed all genetic abnormalities detected by FISH. Moreover, CD40L‐enhanced cytogenetics revealed complex karyotypic aberrations in 11 out of 27 patients (41%). In one case, a balanced translocation t(11;16)(p15;p13.1), so far unreported in B‐CLL, was detected. Taken together, the results of our study show the potential of CD40L‐enhanced metaphase cytogenetics to detect more and new chromosome aberrations in B‐CLL.
Gene expression analysis of 1,25(OH)2D3‐dependent differentiation of HL‐60 cells: a cDNA array study
Authors: Savli Hakan , Aalto Yan , Nagy Bálint , Knuutila Sakari , Pakkala Seppo
Published: March 2003

Summary. The alterations in gene expression associated with 1,25(OH)2D3‐induced differentiation of HL‐60 cells were studied in order to identify potential targets for further investigation of the genetic basis of acute myeloid leukaemia. Atlas human haematology filters, including 406 genes (Clontech), were used to study gene expression in response to 1,25(OH)2D3 (concentration, 5 × 10−8 mol/l) for 24 and 72 h. Compared with untreated cells, expression differences were found in 43 genes. Downregulated genes at both time‐points were: IL2RA, CMYC, NPM, DEK, AF4, FLI1, htlf, MNDA, BCR, IKAROS, BPI and NFAT4. Upregulated genes at both time‐points were IL1B, CD14 and MCL1. CD55, CD58, IRF2, CREB1, ATF4, RAC1, TIAR, KIAA0053, BAT2, BTK, RCK, EV12B and EDN were downregulated at 24 h, while SPI1, MKK3, BTG1 and IL8 were upregulated. At 72 h the upregulated genes were IL1RA, IL2RG, CXCR4, SCYA1, SCYA3, SCYA4, SCYA5, SCYA22, ANX2, CD83 and UPAR. cDNA array results were confirmed on randomly selected genes using quantitative real‐time polymerase chain reaction for three upregulated (CXCR4, IL1B and CD14) and three downregulated (DEK, AF4 and FLI1) genes. Gene expression analysis after differentiation induction may provide a tool to study the roles of DEK, AF4 and FLI1 in cell proliferation and differentiation. To demonstrate the genes that initiate differentiation, sequential gene expression analysis has to be performed during the first 24 h of the differentiation process.
Identification and characterization of a novel gene encoding a SEREX antigen in chronic myeloid leukaemia
Authors: Rogers Sarah A. , Bowen Derrick J. , Ling Min , Thomson Peter , Wang Zhiqing , Lim Seah H.
Published: March 2003

Summary. The novel gene encoding Clone 4, previously isolated through the Serological screening of recombinant expression cDNA library (SEREX) in chronic myeloid leukaemia (CML), was characterized. It is localized to chromosome 7 at q22, occupies approximately 4·5 kb of DNA and comprises 4 exons. Three single nucleotide polymorphisms and a 13 base deletion/insertion polymorphism have been identified. The mRNA transcript was approximately 2·4 kb and contained a long 5′‐untranslated region of almost 1 kb before the candidate open reading frame. Northern blot analysis and multiple tissue expression array indicated a restricted normal tissue distribution. Our findings provide a platform for future investigations into the role of the gene product in CML.
Monensin‐mediated growth inhibition in human lymphoma cells through cell cycle arrest and apoptosis
Authors: Park Woo Hyun , Seol Jae Goo , Kim Eun Shil , Kang Won Ki , Im Young Hyuck , Jung Chul Won , Kim Byoung Kook , Lee Young Yiul
Published: March 2003

Summary. Monensin, a Na+ ionophore, regulates many cellular functions, including apoptosis. We investigated the in vitro antiproliferative effect of monensin on nine human lymphoma cell lines. Monensin significantly inhibited the proliferation of all the lymphoma cell lines examined with a 50% inhibition concentration of about 0·5 μmol/l, and induced a G1 and/or a G2‐M phase arrest in these cell lines. To address the antiproliferative mechanism of monensin, we examined the effect of this drug on cell‐cycle‐related proteins in CA46 cells (both G1 and G2 arrest) and Molt‐4 cells (G2 arrest). Treatment with monensin for 72 h decreased CDK4 and cyclin A levels in CA46 cells, and cdc2 levels in Molt‐4 cells. In monensin‐treated CA46 cells, increased p21‐CDK2, p27‐CDK2 and p27‐CDK4 complex forms were observed. And, in monensin‐treated Molt‐4 cells, increased p21–cdc2 complex form was detected. Furthermore, the activities of CDK2‐ and CDK4‐associated kinases were reduced in association with Rb hypophosphorylation in monensin‐treated CA46 cells. The activity of cdc2‐associated kinase was decreased in both cell lines, which was accompanied by induction of Wee1. Also, monensin induced apoptosis in these cell lines, as evidenced by annexin V binding assay and flow cytometric detection of sub‐G1 DNA content. This apoptotic process was associated with loss of mitochondria transmembrane potential (Δψm). Taken together, these results demonstrated for the first time that monensin potently inhibits the proliferation of human lymphoma cell lines via cell cycle arrest and apoptosis.
Exposure of erythrocytes to methylene blue shows the active role of catalase in removing hydrogen peroxide
Authors: Gaetani Gian Franco , Rapezzi Davide , Mangerini Rosa , Racchi Omar , Rolfo Michela , Ferraris Anna Maria
Published: March 2003

Summary. Methylene blue (MB) is a powerful reducing agent that is widely used in clinical practice as well as for metabolic studies of the erythrocyte. We have investigated the role of catalase as a specific enzyme for the removal of hydrogen peroxide by measuring the in vitro effects of MB on human red cells. In the presence of MB, catalase underwent inactivation even with the co‐existence of active generation of NADPH, leaving the glutathione concentration unaffected. The data obtained in the present investigation show, using a different tool (MB), that catalase is the active enzyme in H2O2 detoxification and that its integrity is largely dependent on an adequate generation of NADPH.
Impaired circadian variations of haemostatic and fibrinolytic parameters in tetraplegia
Authors: Iversen Per Ole , Groot Patricia D. E. , Hjeltnes Nils , Andersen Trine Opstad , Mowinckel Marie Christine , Sandset Per Morten
Published: March 2003

Summary. Spinal cord injured patients are at increased risk of developing deep vein thrombosis (DVT). Whether these patients have increased blood levels of prothrombotic markers remains to be clarified. In general, the risk of developing DVT is highest in the morning hours. In healthy humans, several haemostatic and fibrinolytic parameters exhibit circadian variations, but it is not known whether this also applies to those with spinal cord injury. The aim of the present study was to examine possible circadian variations in prothrombotic markers in tetraplegic patients. We studied six patients with complete tetraplegia and eight control subjects with repetitive blood sampling over a 24 h period. While the control subjects showed marked circadian variations in factor VIII activity, prothrombin fragments 1+2 and D‐dimer levels, the tetraplegic patients did not (P < 0·05). Circadian variation in plasminogen activator inhibitor type‐1 was present in both groups, being most marked (P < 0·05) in tetraplegia. We conclude that the circadian variations of several factors of the haemostatic and fibrinolytic systems are impaired in spinal cord injury. This could possibly reflect a deregulated autonomic nervous system, leading to a dysfunctional link between central and peripheral circadian oscillators.
Therapeutic effects of idiotype vaccination can be enhanced by the combination of granulocyte–macrophage colony‐stimulating factor and interleukin 2 in a myeloma model
Authors: Stritzke Jan , Zunkel Tim , Steinmann Jörg , Schmitz Norbert , Uharek Lutz , Zeis Matthias
Published: March 2003

Summary. Idiotype (Id) vaccination provides an interesting immunotherapeutic strategy against B‐cell lymphomas. In multiple myeloma (MM), however, the therapeutic efficacy of Id vaccination has been disappointing. In an attempt to improve the antitumoral potential, we added granulocyte‐macrophage colony stimulating factor (GM‐CSF) and interleukin 2 (IL‐2) to the protocol. Balb/c mice were inoculated i.p. (d 2) with different doses (1–5 × 105) of HOPC myeloma cells secreting the IgHOPC Id protein. Two days later, animals were injected with 10 000 U GM‐CSF i.p. for 6 d consecutively (d 0–5). On d 5 and 11, myeloma‐specific immunoglobulin (IgHOPC) was administered i.p. together with incomplete Freund adjuvans followed by IL‐2 (2 × 10 000 U/d; i.p) for 10 d (d 5–14). In animals inoculated with 105 myeloma cells, treatment with IL‐2 given as a single agent prolonged the median survival time (MST, 67 d) when compared with the tumour control group (MST 48 d), whereas GM‐CSF did not elicit any survival benefit (MST 49 d). Complete tumour rejection could be achieved in 27% (4/15) by the combination of Id vaccination and GM‐CSF. Additional treatment with IL‐2 further increased antimyeloma activity. In this case, 59% of the animals showed no signs of tumour recurrence. In mice with high tumour burden (5 × 105), no treatment modality achieved long‐term survivors. Both natural killer (NK) cells and CD8+ T cells may be involved in the antitumoural immune response. These data provide evidence for the combined use of GM‐CSF and IL‐2 to enhance the therapeutic effectiveness of clinical cancer vaccination protocols.
A nitric oxide synthase inhibitor, NG‐nitro‐l‐arginine‐methyl‐ester, exerts potent antiangiogenic effects on plasmacytoma in a newly established multiple myeloma severe combined immunodeficient mouse model
Authors: Uneda Shima , Hata Hiroyuki , Matsuno Fumihiko , Nagasaki Akitoshi , Harada Naoko , Mitsuya Yumi , Matsuzaki Hiromitsu , Mitsuya Hiroaki
Published: March 2003

Summary. Angiogenesis is one of critical factors in sustaining the growth, invasion and metastasis of certain solid tumours and haematological malignancies such as multiple myeloma (MM). In the present study, we examined the anticancer potential of an inhibitor of nitric oxide synthase (NOS), NG‐nitro‐l‐arginine methyl ester (L‐NAME), in a novel severe combined immunodeficient mouse model (KHM mouse) that harbours the highly sanguineous plasmacytoma cell line KHM‐4, derived from a patient with highly chemoresistant MM. KHM mice were intraperitoneally administered with either L‐NAME, doxorubicin, melphalan, or paclitaxel. A significant reduction in tumour sizes was noted in L‐NAME‐administered KHM mice while no significant reduction was observed in melphalan‐ or doxorubicin‐administered mice. A profound decrease in angiogenesis was observed in tumour tissues from L‐NAME‐ and paclitaxel‐administered KHM mice. A marked decrease in human vascular endothelial cell growth factor (VEGF) levels was identified in tumour tissues from L‐NAME‐administered KHM mice, strongly suggesting that L‐NAME suppressed VEGF production by tumour cells through its inhibition of NOS in tumour cells, resulting in reduced neovasculization in mice leading to the regression of tumour sizes. The present data represent the first observations that certain NOS inhibitors potentially serve as experimental agents for the treatment of chemoresistant MM and plasmacytoma.
Re‐entry of tumour B cells into the cycle of somatic mutation and isotype switching in follicular lymphoma
Authors: Shiokawa Satoshi , Matsushima Takamitsu , Choi Ilseung , Abe Yasunobu , Shiratsuchi Motoaki , Suehiro Youko , Muta Koichiro , Ohshima Koichi , Nishimura Junji
Published: March 2003

Summary. Sequencing of the immunoglobulin (Ig) gene transcripts of the tumour B cells in lymph node (LN) and bone marrow (BM) from a follicular lymphoma (FL) patient associated with multiple myeloma identified two dominant clones. One of the clones, present in both LN and BM, had somatic mutations and extensive clonal diversity. Among the diversified clones, two dominant populations of identical sequences (group I and II) were present. Group II was a descendant population of group I and had nine more somatic mutations. Group I contained µ‐, δ‐, γ‐ and α‐expressing clones. Group II clones contained mainly µ‐ and δ‐expressing clones. These findings showed that somatic mutation and isotype switching occurred repeatedly in this patient.
Diagnosis of thrombotic thrombocytopenic purpura based on modulation by patient plasma of normal platelet adhesion under flow condition
Authors: Shenkman Boris , Inbal Aida , Tamarin Ilia , Lubetsky Aharon , Savion Naphtali , Varon David
Published: March 2003

Summary. We have designed a simple test for the early diagnosis and treatment monitoring of thrombotic thrombocytopenic purpura (TTP). We examined plasma from 24 TTP patients and normal plasma using a cone and plate(let) analyser (CPA). Test plasma was mixed with citrated normal whole blood (group O) and subjected to flow at a shear rate of 1800/s. Mixing normal plasma (12·5, 25, 50 or 75 µl) with heterologous normal whole blood (final volume of 200 µl) resulted in a decrease of surface coverage (SC, maximally by 63%) and, to a lesser extent, of average size (AS, maximally by 37%) due to dilution of the blood sample. In contrast, mixing the same quantities of acute TTP plasma with normal blood yielded an increase in both SC (up to 125%) and AS (up to 130%). Increased SC and/or AS were detected in all 15 patients in acute phase and in three out of 14 patients in remission. Following repeated plasmapheresis, the enhanced platelet deposition in five patients with acute TTP returned to almost normal patterns. Mixing plasma from patients with other thrombocytopenic conditions in this way resulted in a decrease in both SC and AS, and did not differ from control subjects. In conclusion, the CPA is a simple and specific laboratory test that can be used for the diagnosis and monitoring of plasma exchange therapy in TTP.
Effect of clopidogrel administration to healthy volunteers on platelet phosphorylation events triggered by ADP
Authors: Contreres Jean O. , Dupuy Evelyne , Job Bernard , Habib Aïda , Bryckaert Marg̈ke , Rosa Jean P. , Simoneau Guy , Herbert Jean M. , Savi Pierre , Levy‐Toledano Sylviane
Published: March 2003

Summary. The action of clopidogrel on platelet receptors was analysed using platelets obtained from 11 healthy volunteers given 75 mg of clopidogrel daily for 8 d. Samples of blood were taken before treatment and after 8 d of medication. Determination of 2‐methylthioadenosine diphosphate trisodium (2MesADP)‐induced platelet aggregation, serine/threonine and tyrosine phosphorylations were performed in the absence or presence of the P2Y1‐receptor‐specific antagonist: adenosine 3′‐phosphate 5′‐phosphate (A3P5P) or the strong inhibitor of GPIIb/IIIa activation: SR121566. Major conclusions: 1) Serine and threonine phosphorylations of the myosin light chain (P20) and pleckstrin (P47) do not behave similarly, although they are both recognized as the result of phospholipase C pathway stimulation triggered by the P2Y1 receptor. P47 is strongly affected by the A3P5P, and this appears to be highly dependent on P2Y12. However, P20 phosphorylation occurs in the presence of A3P5P, suggesting that the P2Y12 receptor signal contributes to P20 phosphorylation mediated by a calcium‐independent pathway. The results suggest that P2Y1 and P2Y12 receptors interact to modulate the phosphorylation of P20 and P47. 2) The inside‐out signalling dependent on both P2Y12 and P2Y1 is necessary for GPIIb/IIIa activation. 3) Clopidogrel and SR121566 inhibited the increase in tyrosine phosphorylation induced by 2MesADP and concomitantly inhibited platelet aggregation, indicating that most of the phosphorylations are GPIIb/IIIa dependent. However, neither clopidogrel nor SR121566 inhibited the first wave of 80 kDa substrate (cortactin) which is involved in the reorganization of the cytoskeleton necessary for shape change and which appeared to be essentially P2Y1 dependent.
High‐dose cyclophosphamide does not eradicate paroxysmal nocturnal haemoglobinuria haematopoiesis in mice carrying a Piga gene mutation
Authors: Schaefer Anne , Jasinski Marek , Bessler Monica
Published: March 2003

Summary. Recently, high‐dose cyclophosphamide (HD CY) has been used in the treatment of aplastic anaemia. Several reports have suggested that the treatment may either eradicate or suppress mutant clonal haematopoiesis such as paroxysmal nocturnal haemoglobinuria (PNH). We therefore treated mice that have a proportion of blood cells deficient in GPI‐anchor molecules (PIGA–) with HD CY, and monitored their peripheral blood counts during and after treatment. HD CY produced a transient myelosuppression; however, the contribution of PIGA– haematopoiesis to the peripheral blood remained unchanged, suggesting that HD CY is unlikely to eliminate an existing PNH clone in patients treated for aplastic anaemia.
Population-based record linkage study of the incidence of abdominal aortic aneurysm in Western Australia in 1985–1994
Authors: Semmens J B , Norman P E , Lawrence-Brown M M D , Bass A J , Holman C D J
Published: March 2003

Background

The incidence of abdominal aortic aneurysm (AAA) has increased steadily during the past 30 years.

Methods

Trends in the incidence and surgical intervention for AAA in Western Australia were reviewed for the interval 1985–1994. A population-based health database was used to link morbidity and mortality records of all patients aged 55 years or more who died from rupture or were admitted and treated surgically for AAA. Three groups were separated for analysis: patients with a ruptured AAA, those admitted for elective repair and those admitted as an emergency with an acute (non-ruptured) aneurysm.

Results

There was a decline in the incidence of both emergency and elective procedures for AAA after 1992. While the mortality rate from ruptured AAA has also fallen since 1991, the overall case fatality rate for ruptured AAA has fallen by only 1·3 per cent (from 80·7 to 79·3 per cent).

Conclusion

The decline in mortality rate and emergency procedures may result from a fall in the incidence of ruptured AAA, due to an increasing rate of elective surgery before 1992. The decline in elective procedures from 1992 may be due to a fall in the prevalence of AAA owing to high rates of elective surgery, or to a fall in the incidence of the disease itself.
Thermotolerance induces heat shock protein 72 expression and protects against ischaemia–reperfusion-induced lung injury
Authors: Javadpour M , Kelly C J , Chen G , Stokes K , Leahy A , Bouchier-Hayes D J
Published: March 2003

Background

Ischaemia–reperfusion injury is mediated by neutrophil–endothelial interaction. Induction of heat shock proteins attenuates neutrophil–endothelial interactions. The aim of this study was to determine whether thermal preconditioning could have a protective effect on neutrophil-mediated lung injury in an animal model of lower torso ischaemia–reperfusion.

Methods

Sprague-Dawley rats were randomized into: control, ischaemia–reperfusion, and ischaemia–reperfusion preconditioned with hyperthermia groups. Ischaemia–reperfusion injury was induced by infrarenal aortic clamping for 30 min and reperfusion for 120 min. Thermotolerance was induced by raising the core body temperature to 40·5–41·5°C for 15 min, 18 h before ischaemia–reperfusion. Wet:dry lung (W:D) weight ratio, bronchoalveolar lavage protein (BALprot) concentration, tissue myeloperoxidase (MPO) activity and bronchoalveolar lavage polymorphonuclear neutrophil (BAL PMN) count were measured. Heat shock protein 72 (hsp72) expression in lung, intestine and mesentery was measured using Western immunoblotting.

Results

Ischaemia–reperfusion resulted in a significant increase in tissue oedema (W:D weight ratio) and BALprot concentration. In addition there was a marked increase in tissue neutrophil infiltration (MPO activity, BAL PMN concentration). Preconditioning with hyperthemia resulted in increased expression of hsp72 and significantly reduced tissue oedema and neutrophil infiltration.

Conclusion

Thermal preconditioning protects against neutrophil-mediated ischaemia–reperfusion-induced lung injury, possibly by increasing the expression of heat shock proteins.
Experimental study of the effect of adrenaline tolerance on intestinal ischaemia–reperfusion
Authors: Baykal A , Kaynarogˇlu V , Demirpençe E , Kilinç K , Sayek I , Sanaç Y
Published: March 2003

Background

Adrenaline tolerance improves survival in animal models of shock. The purpose of this study was to determine the effects of adrenaline tolerance on intestinal ischaemia–reperfusion in a mouse model.

Methods

Adrenaline tolerance was developed by injecting intravenous adrenaline, gradually increasing the dose to 2 mg/kg over 5 days. In experimental animals the superior mesenteric artery was clamped for 120 min and then released. Evans blue dye was given intravenously to all animals to quantify pulmonary microvascular injury. Some 60 min after clamp release, the animals were killed and the effects of reperfusion assessed on tissue samples.

Results

Evans blue dye concentrations were significantly higher in animals with intestinal ischaemia–reperfusion than in those having sham intestinal ischaemia–reperfusion or in adrenaline-tolerant mice having intestinal ischaemia–reperfusion or sham ischaemia–reperfusion (P<0·01). Malonyldialdehyde levels increased significantly in the lung in the intestinal ischaemia–reperfusion group compared with those in the sham ischaemia–reperfusion group (P<0·001 for liver, lung and small intestine), whereas there was no difference in adrenaline-tolerant animals. There was no significant change induced by adrenaline tolerance in myeloperoxidase levels in any organ.

Conclusion

Adrenaline tolerance reduced the lung permeability caused by intestinal ischaemia–reperfusion. Catecholamines may play a role in free radical generation induced by ischaemia–reperfusion.
Endoluminal abdominal aortic aneurysm surgery
Authors: Woodburn K R , May J , White G H
Published: March 2003

Background

The development of devices designed for the endoluminal repair of abdominal aortic aneurysm has led to the emergence of new endovascular techniques.

Methods

Articles and case reports obtained from a Medline search of the English language literature from 1989 to 1997 are reviewed. This search was carried out using the MeSH heading ‘aortic aneurysm, abdominal’ and the keywords ‘endovascular’ and ‘endoluminal’.

Results

Reported mortality and complication rates for endoluminal aneurysm repair are similar to those following conventional repair, with the exception of continued perfusion of the aneurysm sac which remains a major problem following endoluminal repair.

Conclusion

Successful endoluminal aneurysm exclusion is associated with reduced aneurysm diameter. However, longer term results of endoluminal repair, in particular of sealed endoleaks, are required before randomized controlled trials of endoluminal versus conventional repair can be undertaken.
Active or passive chest drainage after oesophagectomy in 101 patients: a prospective randomized study
Authors: Johansson J , Lindberg C-G , Johnsson F , von Holstein C S , Zilling T , Walther B
Published: March 2003

Background

This study evaluates the efficiency and safety of two methods of chest drainage after uncomplicated oesophagectomy.

Methods

A prospective randomized study between active suction drainage and passive chest drainage was carried out in 101 patients who underwent gastric pull-up oesophagectomy.

Results

No difference in the prevalence of pneumothorax during treatment was noted between the active (nine of 55) and the passive (four of 46) drainage groups (P=0·20). Nor was there any difference in the size (P=0·46) and duration (P=0·53) of the pneumothorax. There was no significant difference in right (P=0·84) and left (P=0·61) basal atelectases and the amounts of right (P=0·10) and left (P=0·24) pleural effusions. There were significantly more basal atelectases (P<0·001) and pleural effusions (P<0·001) in the non-operated left side compared with the operated right side. Postoperative hospital stay was the same in both groups (median 13 days; P=0·86). The hospital mortality rate was two of 101, and was not affected by the type of drainage.

Conclusion

Passive drainage did not reduce hospital stay, but was as safe and effective as the active system in draining the pleural cavity after uncomplicated oesophagectomy.
Evidence that aminoguanidine inhibits endotoxin-induced bacterial translocation
Authors: Kavuklu B , Agalar C , Guc M O , Sayek I
Published: March 2003

Background

The role of inducible nitric oxide synthase (iNOS) in endotoxin-induced bacterial translocation was investigated by using its specific blocker aminoguanidine in 46 albino mice (25–35 g) allocated into four groups.

Methods

The first group received intraperitoneal saline (control; 0·9 per cent w v−1 sodium chloride 1 ml kg−1; n = 6), the second group intraperitoneal endotoxin (Escherichia coli lipopolysaccharide 055:B5 20 mg kg−1; n = 19), the third group intraperitoneal aminoguanidine (20 mg kg−1, 20 min before and 12 h after saline; n = 6) and the fourth group both endotoxin and aminoguanidine intraperitoneally (n = 15). Some 24 h later, the animals were anaesthetized with ether and blood samples were collected by cardiac puncture together with mesenteric lymph node (MLN), spleen and liver specimens under aseptic conditions. Specimens were then cultured to determine the presence of colony-forming units as an index of bacterial translocation.

Results

No bacterial growth was detected in samples from the first and third groups. Colony-forming bacteria were found in ten of 14 MLN samples, eight of 14 spleens, four of 14 livers and three of 14 peripheral blood samples in the second group, with E. coli being the predominant pathogen. In contrast, in the fourth group, colony-forming bacteria were found in only three of 14 MLN samples (P = 0·02 versus the second group), three of 14 spleens and one of 14 liver specimens. None of the values in the fourth group was significantly different from those in the saline control group.

Conclusion

The inhibition of iNOS during endotoxaemia by its specific blocker aminoguanidine attenuates the incidence of bacterial translocation in mice. These results may be exploited clinically for the prophylaxis and treatment of septic states.
Role of octreotide on release of intact 1–84 parathyroid hormone from human parathyroid cells
Authors: Sadler G P , Jones D L , Morgan J M , Neonakis E , Woodhead J S , Wheeler M H
Published: March 2003

Background

Octreotide has been shown to lower urinary calcium in primary hyperparathyroidism although the mechanism remains unclear. This study examined the effect of octreotide on parathyroid hormone (PTH) secretion from human parathyroid cells in culture and as isolated cells. Additionally in situ hybridization was performed for somatostatin receptor messenger RNA (mRNA) and immunocytochemistry for somatostatin in eight parathyroid adenomas.

Methods

Tissue from three hyperplastic glands and three adenomas was studied as dispersed cell suspensions. Incubation was in buffers containing high (2·0 mmol/l) and low (0·5 mmol/l) calcium concentrations, with or without octreotide 200 ng/ml. Cells were also seeded into tissue culture wells for 24 h to allow receptors to regenerate. Supernatant was removed at regular intervals and PTH levels were estimated using a two-site chemiluminescent assay.

Results

Mean(s.e.m.) PTH secretion at 90 min in hyperplastic cells was 445(75) pmol/l in low calcium and 160(42) pmol/l in high calcium (P<0·02), and in adenoma cells was 170(21) pmol/l in low calcium and 137(27) pmol/l in high calcium (P=0·37). There was no significant difference in secretion of PTH from cells incubated with octreotide either in culture or as dispersed cells. In situ hybridization failed to demonstrate any mRNA for the somatostatin receptors and no somatostatin was detected in any cells with immunocytochemistry.

Conclusion

Somatostatin has no direct action on PTH production and release from human parathyroid cells and is unlikely to be of any therapeutic value in the treatment of hyperparathyroidism.
Significance of angiogenesis in cancer therapy
Authors: McNamara D A , Harmey J H , Walsh T N , Redmond H P , Bouchier-Hayes D J
Published: March 2003

Background

For most solid tumours, surgery remains the most effective primary treatment. Despite apparently curative resection, significant numbers of patients develop secondary disease due to growth of undetected micrometastases. The ability of a tumour to metastasize is related to the degree of angiogenesis it induces. In addition, micrometastases rely on new vessel formation to provide the nutrients necessary for growth. A better understanding of how tumours acquire their blood supply may lead to more effective adjuvant therapies and improve survival following surgery.

Methods

A systematic review of the literature on angiogenesis between 1971 and 1997 was performed using the Medline database to ascertain current thinking on angiogenesis and its relevance in oncological surgery.

Results

Angiogenesis is a physiological process subject to autocrine and paracrine regulation which has the potential to become abnormal and play a part in a number of pathological states, including cancer. Increased angiogenic stimuli in the perioperative period, associated with concomitant reduction in tumour-derived antiangiogenic factors following resection of a primary tumour, result in a permissive environment which allows micrometastases to grow.

Conclusion

Recognition of the role of angiogenesis in metastatic tumour growth represents a significant development in our understanding of tumour biology. The development of antiangiogenic agents offers new promise in the treatment of malignancy. Such agents may prevent or control the development and growth of primary and metastatic tumours.
Detection of residual disease following breast-conserving surgery
Authors: Beck N E , Bradburn M J , Vincenti A C , Rainsbury R M
Published: March 2003

Background

Assessment of completeness of tumour excision has become an integral part of breast-conserving surgery, but the accuracy of margin analysis has been questioned. This study compared the results of resection margin analysis with an examination of tumour bed biopsies and of the excised cavity wall.

Methods

One hundred and forty-four patients underwent breast-conserving surgery for T1–2 N0–1 breast cancer. Following wide local excision, four bed biopsies were taken from the cavity wall which was then completely excised. The presence of invasive and in situ disease at the inked resection margin (IRM) and in the adjacent bed biopsies and cavity wall was recorded.

Results

Positive margins and/or residual disease in either the bed biopsies or cavity wall was found in 62 (43 per cent) of 144 cases. Residual disease (invasive or in situ) was present at the IRM in 39 specimens (27 per cent) and was present in 25 bed biopsy (17 per cent) and 39 cavity wall (27 per cent) specimens. These comprised different but overlapping groups of patients.

Conclusion

Margin analysis of wide local excision specimens is a poor predictor of completeness of excision. Routine resection and examination of the entire cavity wall increases the detection of residual disease compared with examination of bed biopsies alone and is a useful adjuvant to conventional margin evaluation.