CD34+ Cells (cd34+ + cell)

Distribution by Scientific Domains
Distribution within Medical Sciences

Kinds of CD34+ Cells

  • blood cd34+ cell
  • human cd34+ cell
  • peripheral blood cd34+ cell

  • Terms modified by CD34+ Cells

  • cd34+ cell concentration
  • cd34+ cell count
  • cd34+ cell dose
  • cd34+ cell number

  • Selected Abstracts


    Efficacy of Bone Marrow Mononuclear Cells to Promote Bone Regeneration Compared With Isolated CD34+ Cells From the Same Volume of Aspirate

    ARTIFICIAL ORGANS, Issue 7 2010
    Shinji Yasuhara
    Abstract Autologous bone marrow mononuclear cell (BMMNC) transplantation is currently an emerging clinical treatment in the orthopedic as well as cardiovascular fields. It is believed that the therapeutic effect of the BMMNCs is due to neovascularization enhanced by the CD34+ cells contained therein, which include endothelial progenitor cells. However, isolation of the CD34+ cell fraction for clinical application has many disadvantages such as cost and invasiveness related to cell mobilization with cytokine. To investigate whether a purification step is in fact necessary for bone regeneration, we separated BMMNCs, CD34+, and CD34 - cells from the same initial volume of rabbit bone marrow aspirates. We then transplanted them back into a femoral bone defect of the same rabbit together with atelocollagen gel and basic fibroblast growth factor (bFGF) and evaluated neovascularization and bone regeneration up to 8 weeks after transplantation. The greatest potential for neovascularization and bone regeneration medicated by cells from the same volume of bone marrow aspirate was found in the BMMNC group. Although purified CD34+ cells might be an ideal cell source, BMMNCs could be a practical and feasible cell source for bone regeneration in present clinical settings with limited cost, availability of materials, and technical issues for transplantation. [source]


    A new model for predicting the timing of leukapheresis on the basis of CD34+ cell and hematopoietic progenitor cell levels

    JOURNAL OF CLINICAL APHERESIS, Issue 4 2007
    Hao-Wei Teng
    Abstract We developed a model (depending on peripheral CD34+ cell count and hematopoietic progenitor cell count) to determine the optimal timing of 3-day leukapheresis in patients pretreated with chemotherapy and G-CSF. Marrow potentials were identified on the basis of three patterns of leukapheretic yield. Pattern 1 predicted good marrow potential. The positive predictive value of a first-day leukapheretic yield of >1 × 106 CD34+ cells/kg (mean 3-day yield = 8.18 × 106 CD34+ cells/kg, n = 11) was 100%. Pattern 2 predicted poor marrow potential. The negative predictive value of a 3-day leukapheretic yield of >1 × 106 CD34+ cells/kg (3-day yield = 0.26 × 106 CD34+ cells/kg, n = 1) was 100%. Pattern 3 met neither of the above criteria (mean 3-day yield = 1.37 × 106 CD34+ cells/kg, n = 19). The marrow potential was borderline and patients could be further divided into two subgroups according to peripheral CD34+ cell counts when WBC reached >10,000/,l. The mean yield differed significantly between pattern 1 and 3 (P < 0.001). For patients with good marrow potential, leukapheresis should begin as soon as the WBC count is >5,000/,l. Patients with borderline marrow potential may benefit from delaying leukapheresis until the WBC level is >10,000/,l and leukapheresis extended more than 3 days. J. Clin. Apheresis 2007. © 2007 Wiley-Liss, Inc. [source]


    The impact of CD34+ cell dose on platelet engraftment in pediatric patients following unmanipulated haploidentical blood and marrow transplantation,

    PEDIATRIC BLOOD & CANCER, Issue 6 2009
    Ying-Jun Chang PhD
    Abstract Objective Unmanipulated haploidentical blood and marrow transplantation has been developed as an alternative transplant strategy for pediatric patients with hematological diseases. The aim of this study was to investigate the effects of donor and recipient characteristics on hematopoietic recovery in pediatric patients following unmanipulated haploidentical transplantation. Methods Factors correlating with hematopoietic recovery in 133 pediatric patients after unmanipulated haploidentical transplantation were analyzed retrospectively. Results All patients reached an absolute neutrophil count of 500/µl in a median of 12 days (range, 9,49 days). One hundred thirty-three patients reached an untransfused platelet count of more than 20,000/µl in a median of 15 days (range, 7,180 days). Univariate analysis showed five factors associated with platelet engraftment. These were time to transplantation after diagnosis (P,=,0.072), infused nuclear cells/kg of recipient weight (P,=,0.028), CD3+ cells/kg of recipient weight (P,=,0.082), CD4+ cells/kg of recipient weight (P,=,0.083), and CD34+ cells/kg of recipient weight (P,=,0.012). Multivariate analysis showed that infused CD34+ cells/kg of recipient weight (CD34+ cells more than 2.42,×,106/kg vs. less than or equal to 2.42,×,106/kg, HR,=,1.733; 95% CI 1.222,2.549; P,=,0.002) were significantly associated with an increased risk of platelet engraftment. Patients receiving a CD34+ cell dose more than 2.42,×,106/kg had a short time [12 days (range, 7,176 days)] to achieve an untransfused platelet engraftment, compared to 18 days (range, 7,180 days) in patients receiving a lower dose (P,<,0.001). Conclusions Our results suggest that low number of CD34+ cells in allografts is a critical factor associated with delayed platelet engraftment after unmanipulated haploidentical transplantation in pediatric patients. Pediatr Blood Cancer 2009;53:1100,1106. © 2009 Wiley-Liss, Inc. [source]


    Bone marrow cells from myelodysplastic syndromes show altered immunophenotypic profiles that may contribute to the diagnosis and prognostic stratification of the disease: A pilot study on a series of 56 patients,

    CYTOMETRY, Issue 3 2010
    Sergio Matarraz
    Abstract A heterogeneous spectrum of immunophenotypic abnormalities have been reported in myelodysplastic syndromes (MDS). However, most studies are restricted to the analysis of CD34+ cells and/or other major subsets of CD34, cells, frequently not exploring the diagnostic and prognostic impact of immunophenotyping. Methods: We propose for the first time an immunophenotypic score (IS) based on the altered distribution and immunophenotypic features of maturing/mature compartments of bone marrow (BM) hematopoietic cells in 56 patients with MDS that could contribute to a refined diagnosis and prognostic evaluation of the disease. Results: Although MDS-associated phenotypes were detected in reactive BM, the overall immunophenotypic profile of BM cells allowed an efficient discrimination between MDS and both normal and reactive BM, once the number and degree of severity of the abnormalities detected per patient were simultaneously considered in the proposed IS. Interestingly, increasingly higher IS were found among patients with MDS showing adverse prognostic factors and in low- versus high-grade cases. The most informative prognostic factors included the number of CD34+ cells, presence of aberrant CD34,/CD117+ precursors, decreased mature neutrophils and CD34, erythroid precursors, and increased numbers of CD36,/lo erythroid precursors; in addition, the IS was an independent prognostic factor for overall survival. Conclusions: Assessment of immunophenotypic abnormalities of maturing/mature BM cells allows an efficient discrimination between MDS and both normal and reactive BM, once the number and degree of severity of the abnormalities detected are simultaneously scored. Interestingly, progressively higher IS were found among patients with MDS with adverse prognostic features and shorter overall survival. © 2010 Clinical Cytometry Society [source]


    Increased immature hematopoietic progenitor cells CD34+/CD38dim in myelodysplasia

    CYTOMETRY, Issue 2 2006
    Mariela B. Monreal
    Abstract Background Myelodysplastic syndromes (MDS) are clonal disorders affecting hematopoietic progenitor cells (HPC). Despite the relevance of clonal CD34+ cells in developing MDS, only few studies analyze the phenotype of this cell population. The aim of this study was to evaluate phenotypic changes on HPC in MDS that could reflect abnormalities in the differentiation process of stem cells. Methods We analyzed the expression of CD38 and HLA-DR on CD34+ cells by flow cytometry in 36 patients with MDS, as well as in healthy donors (n = 12) and patients with other hematological disorders: non-Hodgkin lymphomas and multiple myeloma, both in complete remission (CR) (n = 32); acute lymphoblastic leukemia in CR (n = 17); de novo acute myeloblastic leukemia (AML) at diagnosis (n = 22) and in CR (n = 37); and AML secondary to MDS at diagnosis (n = 19). Cases with available karyotype were grouped according to the International Prognostic Scoring System (IPSS). Results Compared to normal BM, the fraction of immature HPC, characterized as CD34+bright, intermediate FSC/SSC, and CD38dim, was significantly increased in high risk MDS and secondary AML, but not in low risk MDS, (P , 0.001, P = 0.03, and P = 0.7). De novo AML showed decreased immature HPC. High numbers of immature HPC correlated with higher IPSS risk groups (P = 0.05) and showed significant impact on disease progression (P = 0.03). Conclusion Our study confirms that evaluation of CD38 expression pattern on HPC is an easy and reproducible test that allows evaluating the immature subset of progenitor cells. Increased immature HPC in high risk MDS and secondary AML may reflect blocked differentiation of CD34+ cells in these diseases. © 2006 International Society for Analytical Cytology [source]


    C-Kit receptor (CD117) expression on myeloblasts and white blood cell counts in acute myeloid leukemia

    CYTOMETRY, Issue 1 2004
    Jolanta Wo
    Abstract Background The c-Kit receptor is considered to play a crucial role in hematopoiesis. Induction of mobilization of hematopoietic cells in the bone marrow requires cooperative signaling through c-Kit and c-Kit ligand pathway, and these interactions are important in the retention of stem cells within the bone marrow. Therefore, we analyzed c-Kit density on the leukemic myeloblasts of patients with acute myeloid leukemia (AML) in relation to white blood cell count (WBC) in the peripheral blood. Methods Bone marrow aspirates collected from patients with AML and bone marrow aspirates and leukapheresis products after granulocyte colony-stimulating factor blood mobilization from adult volunteers were studied. To determine the level of c-Kit receptor expression, we applied quantitative (relative fluorescence intensity and antibody binding per cell) cytometric methods. Results Our data showed negative correlation between the level of c-Kit expression intensity on myeloblasts and the number of leukocytes in blood of AML patients. The c-Kit receptor density on myeloblasts in patients with low WBC was significantly stronger than that on myeloblasts in patients with high WBC. In the latter patient group, the density c-Kit receptor on myeloblasts was similar to that on CD34+ cells in mobilized peripheral blood. Conclusions The obtained data suggest an involvement of c-Kit receptor in the regulation of leukemic myeloblasts egress to the peripheral blood. © 2004 Wiley-Liss, Inc. [source]


    Inorganic arsenic induces necrosis of human CD34-positive haematopoietic stem cells

    ENVIRONMENTAL TOXICOLOGY, Issue 2 2008
    Laurent Vernhet
    Abstract Inorganic arsenic is a major environmental contaminant known to exert immunosuppressive effects. In this study, we report toxicity of As2O3, a trivalent inorganic form, toward isolated human hematopoietic CD34+ progenitor cells. Our results demonstrate that low concentrations of As2O3 (0.1,5 ,M) inhibit in vitro proliferation of CD34+ cells and their differentiation into various hematological cell lineages. These effects were associated with the induction of a necrotic process independent of caspases and likely related to mitochondrial damage. We conclude that As2O3 can impair in vitro human hematopoiesis by decreasing survival of CD34+ progenitor cells. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008. [source]


    The expression of cytosolic phospholipase A2 and biosynthesis of leukotriene B4 in acute myeloid leukemia cells

    EUROPEAN JOURNAL OF HAEMATOLOGY, Issue 6 2007
    Gudmundur Runarsson
    Abstract Leukotrienes (LT) exert stimulatory effects on myelopoiesis, beside their inflammatory and immunomodulating effects. Here, we have studied the expression and activity of the enzymes involved in the synthesis of leukotriene B4 (LTB4) in acute myeloid leukemia (AML) cells (16 clones) and G-CSF mobilized peripheral blood CD34+ cells. CD34+ cells from patients with non-myeloid malignancies expressed cytosolic phospholipase A2 (cPLA2), 5-lipoxygenase activating protein (FLAP), and leukotriene A4 (LTA4) hydrolase but not 5-lipoxygenase (5-LO). The enzyme cPLA2 was abundantly expressed in AML cells and the activity of the enzyme was high in certain AML clones. The expression of 5-LO, FLAP, and LTA4 hydrolase in AML clones was in general lower than in healthy donor polymorphonuclear leukocytes (PMNL). The calcium ionophore A23187-induced release of [14C] arachidonic acid (AA) in AML cells was low, compared with PMNL, and did not correlate with the expression of cPLA2 protein. Biosynthesis of LTB4, upon calcium ionophore A23187 activation, was only observed in five of the investigated AML clones and only three of the most differentiated clones produced similar amounts of LTB4 as PMNL. The capacity of various cell clones to produce LTs could neither be explained by the difference in [1 , 14C] AA release nor 5-LO expression. Taken together, these results indicate that LT synthesis is under development during early myelopoiesis and the capacity to produce LTs is gained upon maturation. High expression of cPLA2 in AML suggests a putative role of this enzyme in the pathophysiology of this disease. [source]


    Human hematopoietic stem/progenitor-enriched CD34+ cells are mobilized into peripheral blood during stress related to ischemic stroke or acute myocardial infarction

    EUROPEAN JOURNAL OF HAEMATOLOGY, Issue 6 2005
    E. Paczkowska
    Abstract:, The hematopoietic and non-hematopoietic stem/progenitor cells harvested directly from the bone marrow (BM) or G-CSF mobilized peripheral blood were demonstrated to play an important role in regeneration of damaged organs (1, 2). Here, we asked if the stroke- or acute heart infarct-related stress triggers mobilization of stem/progenitor-enriched CD34+cells from the BM into the peripheral blood, which subsequently could contribute to regeneration of damaged tissues. To address this question the peripheral blood samples were harvested from patients with ischemic stroke during the first 24 h of manifestation of symptoms and on the second and sixth day afterwards or during the first 24 h of acute cardiac pain as well as on the second and sixth day of infarct. We measured in these patients (i) percentage of circulating hematopoietic stem/progenitor-enriched CD34+ cells in peripheral blood by employing fluorescence activated cell sorter (FACS) and (ii) number of hematopoietic progenitor cells for the granulocyte-monocytic colony-forming unit (CFU-GM) and erythoid burst-forming unit (BFU-E) lineages circulating in peripheral blood. We concluded that stress related to ischemic stroke or acute myocardial infarction triggers the mobilization of hematopoietic stem/progenitor-enriched CD34+ cells from the BM into peripheral blood. These circulating stem/progenitor-enriched CD34+ cells may contribute to the regeneration of ischemic tissues, however, this possibility requires further studies. [source]


    Constitutive expression of the FK506 binding protein 51 (FKBP51) in bone marrow cells and megakaryocytes derived from idiopathic myelofibrosis and non-neoplastic haematopoiesis

    EUROPEAN JOURNAL OF HAEMATOLOGY, Issue 4 2004
    Oliver Bock
    Abstract: Objectives:, Overexpression of FK506 binding protein 51 (FKBP51) in megakaryocytic progenitor cells generated from purified CD34+ cells in patients with idiopathic myelofibrosis (IMF) has been demonstrated. It has been suggested that FKBP51 is involved in the dysregulation of the apoptotic programme with consecutive prolongation of cell survival. The knowledge of FKBP51 and its expression in bone marrow cells and mature megakaryocytes in non-neoplastic haematopoiesis and IMF is sparse. Methods:, To evaluate a potential overexpression of FKBP51 in patients with IMF (n = 37) compared with non-neoplastic haematopoiesis (n = 31), total bone marrow cells as well as single megakaryocytes, isolated by laser microdissection, were quantitatively analysed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). By applying immunohistochemistry, FKBP51 gene expression was correlated with staining pattern and cellular localisation of the corresponding FKBP51 protein. Results:, We demonstrated that FKBP51 is constitutively expressed in non-neoplastic haematopoiesis. FKBP51 gene expression by total bone marrow cells as well as megakaryocytes was not significantly different in IMF. FKBP51 protein expression could be localised to myeloid progenitor cells as well as megakaryocytes. In particular, megakaryocytes were stained almost exclusively nuclear for FKBP51. No differences in expression patterns between both IMF and control cases could be demonstrated. Conclusions:, For the first time, FKBP51 expression, in particular gene expression and subcellular localization was described in bone marrow cells of non-neoplastic and neoplastic haematopoiesis grown in vivo. We conclude that FKBP51 could be temporarily overexpressed in megakaryocytic progenitors rather than contribute to the accumulation of mature megakaryocytes in IMF. [source]


    Mobilisation of tumour cells along with CD34+ cells to peripheral blood in multiple myeloma

    EUROPEAN JOURNAL OF HAEMATOLOGY, Issue 5-6 2001
    Lene Meldgaard Knudsen
    Abstract:Background: Cells belonging to the malignant clone are found in the peripheral blood in myeloma patients. In order to minimise the content of tumour cells in the stem cell product it is crucial to perform stem cell harvest at a time when tumour cells in the peripheral blood are at a minimum. Objective: The aim of the study was to compare the mobilisation kinetics of normal CD34+ cells and myeloma plasma cells during mobilisation with either G-CSF alone or high-dose cyclophosphamide (HDCy) plus G-CSF. Design and methods: Morning blood samples were drawn each day during mobilisation from start of G-CSF or HDCy and to the end of leukapheresis, and were analysed by flow cytometry for content of CD34+ cells and myeloma plasma cells (CD38+ + CD45,). Tumour cells were also estimated by a patient-specific real-time polymerase chain reaction (PCR) method based on the 5, nuclease TaqMan technology. Results: Flow cytometry data from 16 patients showed concomitant mobilisation of CD34+ cells and myeloma plasma cells. Seven patients were mobilised twice; first with G-CSF alone and then with HDCy plus G-CSF. There was no difference between the two mobilisation regimens regarding tumour cell mobilisation kinetics. Real-time PCR was performed in one patient and confirmed the mobilisation of tumour cells at the time when CD34+ blood cells were at a maximum. Conclusions: Tumour cells are mobilised to the peripheral blood at the same time as CD34+ cells in multiple myeloma patients after priming with both G-CSF alone and HDCy in combination with G-CSF. [source]


    CD34+ cells derived from fetal liver contained a high proportion of immature megakaryocytic progenitor cells

    EUROPEAN JOURNAL OF HAEMATOLOGY, Issue 5 2000
    Dong-Chu Ma
    Abstract: Endoreplication and maturation of the megakaryocyte (MK) may be retarded or delayed during ontogenesis. In this study, CD34+ cells were isolated from both human fetal liver and adult bone marrow and incubated with thrombopoietin (TPO). The cell number, morphological characteristics, platelet-associated antigen phenotype, maturation stage and DNA ploidy of CD41+cells were examined from day 0 to day 12 in culture. 1) TPO stimulated the proliferation of fetal liver (FL)-derived CD34+ cells with a mean 73.14-fold increase of CD41+ cells after 12 d in culture. Adult BM-derived CD34+ cells increased only slightly, with a mean 8.18-fold increase of CD41+ cells. 2) Although the membrane phenotype of both FL CD34+ -derived MKs and BM CD34+ -derived MKs analyzed with CD41a, CD42a, CD61 and CD34 were similar, all FL CD34+ -derived MKs were in maturation stage I and II and in low ploidy (<4N) class. By comparison, BM CD34+ MKs possessed 15% MKs in maturation stage III and IV and with 23% MKs in high ploidy class (>4N). 3) Most of cultured FL-derived CD34+ cells did not have a well developed demarcation system (DM) and numerous ,-granules after 12 d incubation. von Willebrand factor (vWF) appeared earlier on the cultured BM-derived CD34+ cells than on FL-derived CD34+ cells. 4) The expression of both cyclin E and cyclin B1 progressively increased in FL CD34+cells induced by TPO during 12 d in culture. 5) The expression of cyclin D1 gradually decreased in FL CD34+cells induced by TPO over 12 d incubation. 6) Immunocytochemical analysis showed that cyclin D3 was detected only in cytoplasm of cultured FL-derived CD34+ cells, whereas in both cytoplasm and nuclei of cultured BM-derived CD34+ cells. These data suggest that FL-derived CD34+ cells contain a high proportion of immature megakaryocytic progenitor cells. It further suggests that TPO can push these progenitor cells into proliferation by upregulating the expression of cyclins B1 and E, and drive a high proportion of cells into megakaryocytic lineage. [source]


    Functional characterization of T cells differentiated in vitro from bone marrow-derived CD34+ cells of psoriatic patients with family history

    EXPERIMENTAL DERMATOLOGY, Issue 8 2010
    Kaiming Zhang
    Please cite this paper as: Functional characterization of T cells differentiated in vitro from bone marrow-derived CD34+ cells of psoriatic patients with family history. Experimental Dermatology 2010; 19: e128,e135. Abstract Background:, The strong but complex genetic background suggests that inherent and intrinsic rather than exogenous factors have a key role in immunopathogenesis of psoriasis. It is reasonable to speculate that the dysfunctional activity of psoriatic T cells may partly originate from the abnormal haematopoietic cells. Objectives:, To test if T cells originated from haematopoietic progenitor cells in psoriasis patients display functional alternations similar to previously reported abnormalities of circulating T cells. Methods:, Bone marrow CD34+ haematopoietic cells were isolated from psoriatic patients with family history and healthy subjects, and differentiated into T cells in vitro in the thymic stromal co-culture system. These cells were further subjected to functional comparisons such as in vitro proliferation, secretion of cytokines such as IL-4, IL-8 and IFN,,, and inducing the production of C-myc, Bcl-xL, and Ki67 proteins in human keratinocytes. Results:, While bone marrow-derived CD34+ cells from both patients and healthy volunteers developed into mature T cells within weeks in the thymic environment in vitro, the differentiated T cells from psoriatic patients showed higher proliferation and stronger capacity to secret TH1 cytokines in response to streptococcal superantigen. The differentiated T cells from psoriatic patients, but not from normal controls, induced overexpression of C-myc and Ki67, but not Bcl-XL, in keratinocytes. Conclusions:, T cells differentiated from CD34+ cells of psoriatic patients, but not normal controls, are functionally similar to psoriatic circulating T cells, suggesting that the dysfunctional activity of T cells in psoriatic patients can be traced back to the early development of haematopoietic cells. [source]


    Functional characterization of highly adherent CD34+ keratinocytes isolated from human skin

    EXPERIMENTAL DERMATOLOGY, Issue 7 2010
    Araika Gutiérrez-Rivera
    Please cite this paper as: Functional characterization of highly adherent CD34+ keratinocytes isolated from human skin. Experimental Dermatology 2010; 19: 685,688. Abstract:, Compared to murine models, data on cells responsible for the homeostasis of human epidermis are scarce and often contradictory. Given the conflicting results and the availability of clinical grade protocols to purify CD34 cells from a given tissue, we pursued to phenotypically characterize human epidermal CD34+ population. After magnetic separation of whole skin CD34+ and CD34, cell fractions and selection for cells highly adherent to extracellular matrix, both CD34± fractions retained the ability to form a stratified epidermis in organotypic cultures and presented similar in vitro migratory phenotypes. However CD34, cells showed higher clonogenic potential and in vitro proliferative capacity. These results indicated that CD34, cell fraction contains stem/early progenitor cells, while CD34+ cells might be a transit-amplifying precursor for hair follicle (HF) sheath cells. The ability to isolate living cells using differential cell adhesion and surface markers provides an opportunity to study cells from different morphological regions of the HF. [source]


    Feasibility and long-term results of autologous PBSC transplantation in recurrent undifferentiated nasopharyngeal carcinoma

    HEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 9 2001
    Mario Airoldi MD
    Abstract Background Recurrent undifferentiated nasopharyngeal carcinoma (UNPC) is a chemosensitive illness. Here we report long-term results of high-dose chemotherapy (HDC) as late intensification, with autologous peripheral blood stem cell (PBSC) support. Methods Six patients (5 men, 1 woman; median age 41years; median ECOG PS = 0) with recurrent UNPC (local, 2; local + nodal, 2; bone metastasis, 2) have been enrolled. All patients had been previously treated with neoadjuvant chemotherapy and radiotherapy; 3 of 4 local relapses had received a re-irradiation. Every patient received three courses of cisplatin + epirubicin and 1 cycle of epirubicin followed by PBSC collection. A median of 7.2 × 106/kg (range, 4.5,18) CD34+ cells were reinfused. HDC was according ICE scheme: ifosfamide, 2.5 g/m2/d, + carboplatin, 300 mg/m2/d, + VP-16, 300 mg/m2/d days 1 through 4. Results After conventional chemotherapy, we had 1 CR (16%), 3 PR (50%), and 2 NC (34%). After HDC, we had 4 CR (66%) ,1 PR (17%), and 1 MR (17%). Toxicity was manageable. After a median follow-up of 30 months (range, 14,50), two patients are alive without disease (34%), one is alive with bone disease (16%), and three (50%) died of disease at 16, 18, and 24 months. Conclusions HDC has an acceptable toxicity, can convert PR in CR, and seems effective, with long-lasting CRs. © 2001 John Wiley & Sons, Inc. Head Neck 23: 799,803, 2001. [source]


    Patients with bisphosphonates-associated osteonecrosis of the jaw have reduced circulating endothelial cells

    HEMATOLOGICAL ONCOLOGY, Issue 4 2007
    A Allegra
    Abstract Osteonecrosis of the jaws (ONJ) associated with the use of bisphosphonates is a newly described entity. To elucidate the mechanism leading to ONJ and to test the hypothesis that in patients with ONJ the bisphosphonates may interfere with endothelial cell proliferation, using flow cytometric analysis we evaluated the number of circulating endothelial progenitor cells (EPCs) and circulating endothelial cells (CECs) in eight patients with bisphosphonate treatment and osteonecrosis, eight multiple myeloma (MM) patients with bisphosphonates treatment without ONJ and five normal subjects. MM patients showed an increase of CD34+ cells with respect the control subjects and ONJ subjects. EPCs and CECs were higher in MM patients compared to controls and ONJ patients. ONJ patients showed a decrease of EPCs compared to control subjects while CECs were similar to the controls group. Our results seem to show the possibility that bisphosphonates could have a antiangiogenic effect and a suppressive effect on CECs of patients with ONJ. Copyright © 2007 John Wiley & Sons, Ltd. [source]


    Total CD34+ cells per 10 HPF in bone marrow trephines of patients with chronic myeloid leukaemia correlates with probability of complete cytogenetic response following imatinib treatment

    HISTOPATHOLOGY, Issue 6 2007
    V Elliot
    No abstract is available for this article. [source]


    Comparative analysis of G-CSFR and GM-CSFR expressions on CD34+ cells in patients with aplastic anemia and myelodysplastic syndrome

    INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 6 2009
    H. XU
    Summary The aim of this article was to explore the pathogenetic differences, as well as to provide a new way for the differential diagnosis of these two diseases by comparative analysis of CD34+ cells numbers and their surface expression of granulocyte colony-stimulating factor receptor (G-CSFR) and granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) in patients with aplastic anemia (AA) and myelodysplastic syndrome (MDS). Twenty-seven patients with AA, 45 patients with MDS, and 20 normal controls were enrolled in this study. The ratio of CD34+ cells and their surface expression of G-CSFR and GM-CSFR were detected by flow cytometry (FCM). The ratio of CD34+ cells in BMMNC of AA, MDS patients and controls were 0.2438 ± 0.1129%, 2.1677 ± 1.1345% and 1.0792 ± 0.3221%, respectively. Compared with normal controls as well as MDS patients, the ratio of CD34+ cells in BMMNC of AA was significantly reduced (P < 0.05). The ratio of CD34+ cells in MDS was significantly elevated than controls (P < 0.05). The ratio of CD34+ cells in BMMNC of MDS-RA and MDS-RAEB patients were 1.2821 ± 0.4658% and 3.7729 ± 2.3360%, respectively. Compared with normal controls and MDS-RA patients, the ratio of CD34+ cells in MDS-RAEB was significantly elevated (P < 0.05). The ratio of CD34+ cells in MDS-RA was significantly elevated than AA patients (P < 0.05). The surface expression of G-CSFR on CD34+ cells of AA, MDS patients and controls were 34.402 ± 21.8357%, 26.376 ± 15.2895% and 21.443 ± 7.4465%, respectively. The surface expression of G-CSFR on CD34+ cells of MDS-RA and MDS-RAEB patients were 22.788 ± 14.7628% and 30.682 ± 15.5346%. The surface expression of GM-CSFR on CD34+ cells of AA, MDS patients and controls were 6.5961 ± 4.4322%, 18.2737 ± 10.9841% and 4.2753 ± 2.6249%, respectively. Compared with AA and controls, the expression of GM-CSFR in MDS patients was significantly elevated (P < 0.05). The surface expression of GM-CSFR on CD34+ cells of MDS-RA and MDS-RAEB patients were 16.1625 ± 6.9487% and 22.1003 ± 14.2983%. In AA patients, the ratio of CD34+ cells in BMMNC less than 0.1% accounts for 75% (6/8) SAA patients, compared with 10.55% (2/19) in CAA (P < 0.05). The detection of CD34+ cells and their surface expression of granulocyte (macrophage) colony-stimulating factor receptors G (M)-CSFR in AA and MDS are helpful in the differential diagnosis or prognosis of these two disorders. [source]


    Estimation of haemopoietic progenitor cells in peripheral blood by the Advia 120 and BD vantage flow cytometer: a direct comparison for the prediction of adequate collections

    INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 5 2005
    H. M. GREENFIELD
    Summary Peripheral blood stem cells are increasingly used to ensure rapid haematological engraftment after myeloablative chemotherapy. After mobilization, progenitor cells in the blood can be enumerated to predict an adequate collection by leukapheresis. The Advia 120 automated counter has an immature cell channel measuring a parameter known as large undifferentiated cells (LUC's), which were quantified to assess their value in refining the timing of apheresis. Data were available from 102 apheresis sessions. Positive correlation was found for peripheral blood CD34+ cells and apheresis counts (r = 0.82, P < 0.0005) but not for total WCC (r = ,0.15, P = 0.13) or LUC count (r = 0.12, P = 0.23). Our results indicate that the LUC population in peripheral blood has no relevance to the subsequent CD34 content of the apheresis product and CD34 cell enumeration by flow cytometry is advocated. [source]


    Low cost autologous peripheral blood stem cell transplantation performed in a municipal hospital for a patient with plasma cell leukaemia

    INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 3 2002
    K. Ghosh
    Autologous peripheral blood stem cell transplantation (PBSCT) is a costly procedure. In India, the cost varies from US$20 000 to 25 000 and most patients cannot afford it. Using several cost-cutting measures, we were able to treat a patient with plasma cell leukaemia by autologous PBSCT. A 42-year-old-male presented with plasma cell leukaemia. He was treated with VAD therapy, followed by high-dose cyclophosphamide and granulocyte colony-stimulating factor (G-CSF) for mobilization of peripheral blood stem cells. The patient was conditioned with high dose melphalan, followed by autologous PBSCT. The procedure was performed in a municipal hospital in which there was no prior experience with stem cell transplantation. Costs were reduced by: (i) using oral medication whenever possible; (ii) having a relative of the patient prepare his food under medical guidance; (iii) starting G,CSF on day 7 rather than on day 1; (iv) short-term storage of the PBSC in an ordinary refrigerator at 4 °C without cryopreservation; (v) infusing a large number of CD34+ cells, which shortened the time to engraftment; (vi) delegating many of the functions of a marrow transplant nurse to a resident physician. The cost of transplantation was thereby reduced to about US$ 6000, with successful engraftment by day +13. The patient remained in remission for 7 months, after which he relapsed and was treated with chemotherapy and electron beam radiation to the skin. [source]


    Increased VEGFR2 expression during human late endothelial progenitor cells expansion enhances in vitro angiogenesis with up-regulation of integrin ,6

    JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 5 2007
    David M. Smadja
    Abstract In vitro expansion of late endothelial progenitor cells (EPCs) might yield a cell therapy product useful for myocardial and leg ischaemia, but the influence of EPC expansion on the angiogenic properties of these cells is unknown. In the present study, we investigated the effect of in vitro EPC expansion on vascular endothelial growth factor (VEGF) receptor expression. EPCs were obtained from CD34+ cord blood cells and expanded for up to 5 weeks. Real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) showed that VEGFR2 expression, contrary to VEGFR1 and VEGFR3 expression, was significantly higher on expanded EPCs than on freshly isolated CD34+ cells or on human umbilical vein endothelial cells (HUVECs). Quantitative flow cytometry confirmed that VEGFR2 density on EPCs increased during the expansion process and was significantly higher than on HUVECs. The impact of VEGFR2 increase was studied on the three theoretical steps of angiogenesis, i.e., EPC proliferation, migration and differentiation. VEGFR2 up-regulation had no effect on VEGF-induced cell proliferation, but significantly enhanced EPC migration and pseudotubes formation dependent on integrin ,6 subunit overexpression. In vitro expansion of late EPCs increases the expression of VEGFR2, the main VEGF receptor, with possible implications for EPC-based angiogenic therapy. [source]


    Mobilization effects of G-CSF, GM-CSF, and darbepoetin-, for allogeneic peripheral blood stem cell transplantation

    JOURNAL OF CLINICAL APHERESIS, Issue 5 2009
    Shi Nae Kim
    Abstract The effects of GM-/G-CSF and darbepoetin-, on stem cell mobilization were investigated. From February 2005 to March 2007, 30 allogeneic sibling donors were randomly assigned to a G-CSF group (5 ,g/kg/day for 5,7 days) or triple group (GM-CSF 10 ,g/kg/day on 1st and 2nd day, G-CSF 5 ,g/kg/day for 5,7 days, and darbepoetin-, 40 mg on 1st day). The MNCs and CD34+ cells were not different between the two groups, although the doses (×108/kg of recipient body weight) of CD3+ cells (3.64 ± 1.75 vs. 2.63 ± 1.36, P = 0.089) and CD8+ cells (1.07 ± 0.53 vs. 0.60 ± 0.30, P = 0.006) were lower in the triple group. The engraftments, frequency of RBC transfusions, and hemoglobin recovery were not different between the two groups. The cumulative incidence of overall and Grades II,IV aGVHD was 64.3% vs. 61.1% and 25.9% vs. 27.1% in the G-CSF and triple regimen group, respectively, whereas the cumulative incidence of cGVHD was 20.8 ± 1.3% and 24.4 ± 1.7%, respectively. In conclusion, the triple regimen did not seem to be superior to G-CSF alone in terms of the CD34+ cell dose, hemoglobin recovery, and GVHD. However, the CD8+ cell count was significantly lower in the triple regimen group. The role of a lower CD8+ cell count in the graft may need to be elucidated in the future. J. Clin. Apheresis, 2009. © 2009 Wiley-Liss, Inc. [source]


    Peripheral blood stem cell collection in multiple myeloma: A retrospective analysis of 6 years leukapheresis activity in 109 patients treated at the Istituto Nazionale dei Tumori of Milan

    JOURNAL OF CLINICAL APHERESIS, Issue 4 2009
    Paola Coluccia
    Abstract Double autologous stem cell transplantation is the standard treatment in newly diagnosed multiple myeloma (MM) patients younger than 65 years; therefore, optimization of leukapheresis is crucial. We performed a retrospective analysis of 297 leukaphereses comparing semiautomated (V4.7 in 20% of collections) versus automated (V6.0 in 80%) Caridian (COBE) Spectra versions and analyzing the influence of M-protein on the outcome. Both methods gave comparable collection efficiencies (CE%) (53.4% vs. 55.7% in V6.0 and V4.7, respectively) with similar leukapheresis time and processed volume. Harvest volume was higher in V4.7 (P < 0.0001) with similar contamination of red blood cells (RBCs) (P = 0.77) and platelets (P = 0.09) when compared with V6.0. In patients with higher peripheral white blood cells (WBCs), V6.0 with adjusted harvest volume (<700 mL), achieved similar CD34+ CE% (P = 0.39) and better enrichment of nucleated cells (P < 0.0,002) but higher RBCs (P < 0.0,001) and platelets contamination (P = 0.001), when compared with a larger cycle volume in patients with lower WBCs. In hard to mobilize patients, CD34+ CE% was significantly more efficient with V4.7 than V6.0 (P < 0.0,001). CD34+ CE% was unaffected by serologic M-protein, but platelet CE% was higher in the absence of M-protein (P = 0.0,003), without any reduction in peripheral patients platelets. We, therefore, conclude that in the setting of MM patients with a high WBCs count and/or low percentage of peripheral CD34+ cells, collections with V4.7 or adjusted cycle volume V6.0 gave comparable result in CD34+ CE%. RBCs and platelets contamination is higher if low cycle volume is chosen. In hard to mobilize patients, V4.7 is advisable. J. Clin. Apheresis, 2009. © 2009 Wiley-Liss, Inc. [source]


    Multiple myeloma patients receiving large volume leukapheresis efficiently yield enough CD34+ cells to allow double transplants

    JOURNAL OF CLINICAL APHERESIS, Issue 1 2009
    A.C. Zubair
    Abstract Current protocols for myeloma patients require more than one autologous transplant. We performed a retrospective study to determine the cost-effectiveness of large volume leukapheresis (LVL) compared with standard volume leukapheresis (SVL) collection when two transplants are required. We evaluated 87 patients who underwent a cumulative total of 260 LVL and SVL collections. The median product volume per collection was 356 ml for LVL, and this was significantly higher than the median product volume per collection for SVL (median 149.5 ml, P < 0.001). The median total CD34+ cell yield/kg was 6.4 × 106 for LVL and 5.2 × 106 for SVL. This difference was statistically significant (P = 0.005). Because the target CD34+ cell dose for a single transplant was 3 × 106/kg at our institution, overall the LVL yields enough CD34+ cells that could allow for two transplants. Therefore, more patients in the LVL group were able to undergo a potential second transplant. Because of the reserved cells for a second transplant, LVL patients received significantly less CD34+ cell/kg per transplant than the patients in SVL group (P = <0.001). As a result, LVL group had statistically significant but clinically insignificant delay in neutrophil (P = <0.001) and platelet (P = 0.02) engraftments. Additionally, using LVL instead of SVL to collect ,6 × 106/kg CD34+ cells may potentially save $7,497 per patient. We therefore conclude that LVL is the method of choice for collection of multiple myeloma patients when two transplants are anticipated. J. Clin. Apheresis, 2009. © 2009 Wiley-Liss, Inc. [source]


    Predictive parameters for granulocyte colony-stimulating factor-induced peripheral blood stem cell mobilization

    JOURNAL OF CLINICAL APHERESIS, Issue 6 2008
    Akira Okano
    Abstract To improve the selection of donors for allogeneic stem cell transplantation, it is important to identify reliable parameters that predict CD34+-cell yields after granulocyte-colony stimulating factor (G-CSF)-induced peripheral blood stem cell (PBSC) mobilization. We retrospectively investigated the peripheral blood (PB) kinetics of white blood cells (WBCs), CD34+ cells, matrix metalloproteinases (MMP)-9 and -2, and tissue inhibitors of metalloproteinases (TIMP)-1 and -2 in 15 healthy donors during their treatment with G-CSF. All donors received 10 ,g/kg of recombinant human G-CSF once a day subcutaneously. Leukapheresis was initiated after 4 days of G-CSF treatment, and G-CSF treatment continued until the last day of leukapheresis. WBC and CD34+ cell numbers in the PB rose after 2 and 3 or 4 days of G-CSF treatment, respectively. The PB CD34+ cell numbers on day 4 correlated weakly with the increase in WBC counts from day 1 to day 2 (R2 = 0.254, P = 0.056). There were also positive correlations between the CD34+ cell numbers in the PBSC products on day 4 and the CD34+ cells in the PB on days 1 and 4 (R2 = 0.768, P < 0.0001 and R2 = 0.816, P < 0.0005, respectively). The MMP-9 plasma levels on days 1 and 4 also correlated positively with the day 4 circulating CD34+ cell numbers (R2 = 0.393, P < 0.05 and R2 = 0.406, P = 0.01, respectively). In conclusion, the CD34+ cell numbers in the PB steady state may be a useful parameter selecting allogeneic PBSC donors. J. Clin. Apheresis, 2008. © 2008 Wiley-Liss, Inc. [source]


    Performance of a new separator system for routine autologous hematopoietic progenitor cell collection in small children

    JOURNAL OF CLINICAL APHERESIS, Issue 6 2007
    Volker Witt
    Abstract The AMICUSÔ system was recently introduced for peripheral blood stem cell (PBSC) aphereses in adults. We conducted a single center field evaluation to obtain data about the performance of this system in children with a body weight (bw) < 25 kg. Results of blood priming procedures were compared to historical data obtained with the Fenwal CS3000+ (CS 3000). From August, 2001 to February, 2007, 47/178 (26%) PBSC aphereses procedures were performed in our institution with the AMICUSÔ system in 35 small patients (median bw 13.9 kg; range 6.7,24; age 2.78 years; range 0.97,7.06). The patients suffered from various malignant primary diseases or recurrences. We primed the system with packed RBC in case of >30% dilution of the RBC volume (n = 31) or with saline (n = 16). Compared to the CS3000, the AMICUSÔ revealed comparable collection efficiencies (CE) for CD34+ cells (median 67%, range 26,120), lymphocytes (75%, 25,138), monocytes (54%, 23,173), and granulocytes (10%, 1.5,36), MNC (57% 24,125), but a significantly higher erythrocyte and granulocyte, and a lower platelet CE. There was a significant negative correlation between total leukocyte count and CE for MNC (r = ,0.566; P < 0.001) and CD34+ cells (r = ,0.517; P < 0.001). There was no significant statistical or clinical difference between the CE in blood-primed procedures and saline-primed procedures. With the AMICUSÔ we saw statistically less citrate reactions compared to the CS 3000. We conclude that the AMICUSÔ system is safe and efficient to harvest PBSC on a routine basis in pediatric patients, even in children ,10 kg bw. J. Clin. Apheresis, 2007. © 2007 Wiley-Liss, Inc. [source]


    The number of CD34+ cells in peripheral blood as a predictor of the CD34+ yield in patients going to autologous stem cell transplantation

    JOURNAL OF CLINICAL APHERESIS, Issue 2 2006
    A.L. Basquiera
    Abstract The number of CD34+ cells in peripheral blood (PB) is a guide to the optimal timing to harvest peripheral blood progenitor cells (PBPC). The objective was to determine the number of CD34+ cells in PB that allows achieving a final apheresis product containing ,1.5 × 106 CD34+ cells/kg, performing up to three aphereses. Between March 1999 and August 2003, patients with hematological and solid malignancies who underwent leukapheresis for autologous bone marrow transplantation were prospectively evaluated. Seventy-two aphereses in 48 patients were performed (mean 1.45 per patient; range 1,3). PBPC were mobilized with cyclophosphamide plus recombinant human granulocyte-colony stimulating factor (G-CSF) (n = 40), other chemotherapy drugs plus G-CSF (n = 7), or G-CSF alone (n = 1). We found a strong correlation between the CD34+ cells count in peripheral blood and the CD34+ cells yielded (r = 0.903; P < 0.0001). Using receiver-operating characteristic (ROC) curves, the minimum number of CD34+ cells in PB to obtain ,1.5 × 106/kg in the first apheresis was 16.48 cells/,L (sensitivity 100%; specificity 95%). The best cut-off point necessary to obtain the same target in the final harvest was 15.48 cells/,L, performing up to three aphereses (sensitivity 89%; specificity 100%). In our experience, ,15 CD34+ cells/,L is the best predictor to begin the apheresis procedure. Based on this threshold level, it is possible to achieve at least 1.5 × 106/kg CD34+ cells in the graft with ,3 collections. J. Clin. Apheresis 2005. © 2005 Wiley-Liss, Inc. [source]


    Autologous peripheral blood stem cell collections in children weighing less than 10 Kg with solid tumors: Experience of a single center

    JOURNAL OF CLINICAL APHERESIS, Issue 2 2005
    Hyun-Jung Cho
    Abstract There have only been a few reports and limited performance of peripheral blood stem cell (PBSC) collection in very small children weighing less than 10 kg. In this study, we intended to evaluate the safety and yield of PBSC collection, with the efficacy of PBSC transplantation (PBSCT) in the smallest children with solid tumors. From January 1998 to February 2004, 173 children underwent PBSC collection in Samsung Medical Center, Korea. Of these, 15 (8.7%) children weighed less than 10 kg and their clinical diagnoses were neuroblastoma (10 cases), rhabdoid tumor (2 cases), rhabdomyosarcoma (2 cases), and Wilms tumor (1 case). PBSCs were collected following chemotherapy plus G-CSF mobilization. The median age and weight at the time of apheresis were 15 months and 9 kg, respectively. The median number of PBSC collection procedures per case was 4 (range, 2,7). The median cell yield per apheresis product was 0.95 (range, 0.01,33.32) × 106/kg CD34+ cells and 1.96 (range, 0.12,23.39) × 108/kg mononuclear cells. No complications associated with citrate toxicity and other adverse effect were observed during the procedures. After high-dose chemotherapy, 14 patients were reinfused with PBSCs alone and all showed successful hematopoietic recovery. We concluded that PBSC collection would be a safe and practical procedure, even when done in the smallest children, provided that adequate intravascular fluid volume and circulating red cell mass were maintained. Also, the use of PBSCs to support high-dose chemotherapy was well tolerated and might enhance hematological recovery in the smallest children showing the excellent efficacy of PBSCT. J. Clin. Apheresis © 2005 Wiley-Liss, Inc. [source]


    Large-volume leukapheresis using femoral venous access for harvesting peripheral blood stem cells with the Fenwal CS 3000 Plus from normal healthy donors: Predictors of CD34+ cell yield and collection efficiency

    JOURNAL OF CLINICAL APHERESIS, Issue 1 2003
    Sang Kyun Sohn
    Abstract The current paper reports on the predicting factors associated with satisfactory peripheral blood stem cell collection and the efficacy of large-volume leukapheresis (LVL) using femoral vein catheterization to harvest PBSCs with Fenwal CS 3000 Plus from normal healthy donors for allogeneic transplantation. A total of 113 apheresis procedures in 57 patients were performed. The median number of MNCs, CD3+ cells, and CD34+ cells harvested per apheresis was 5.3 × 108/kg (range, 0.3,11.0 × 108/kg), 3.0 × 108/kg (range, 0.2,6.6 × 108/kg), and 7.9 × 106/kg (range, 0.1,188.9 × 106/kg), respectively. The median collection efficiency of MNCs and CD34+ cells was 49.8% and 49.7%, respectively. A highly significant correlation was found between the collected CD34+ cell counts and the pre-apheresis WBC counts in the donors (P = 0.013), and between the collected CD34+ cell counts and the pre-apheresis peripheral blood (PB) CD34+ cell counts (P<0.001). Harvesting at least >4 × 106/kg CD34+ cells from the 1st LVL was achieved in 44 (77.2%) out of 57 donors and in 19 (90.5%) out of 21 donors with a PB-CD34+ cell count of >40/,l. There was no significant difference in the harvested MNC and CD34+ cell counts between the 1st and 2nd apheresis. The catheter-related complications included catheter obstruction (n = 2) and hematoma at the insertion site (n = 3). Accordingly, LVL using femoral venous access for allogeneic PBSC collection from normal healthy donors would appear to be safe and effective. J. Clin. Apheresis 18:10,15, 2003. © 2003 Wiley-Liss, Inc. [source]


    Comparison of CD34+ cell collection efficiency on the COBE Spectra and Fenwal CS-3000 plus

    JOURNAL OF CLINICAL APHERESIS, Issue 1 2002
    C.D. Ford
    Abstract Optimal collections of mobilized CD34+ cells are important in terms of both patient toxicity and cost. The factors that determine CD34+ collection efficiency (CD34eff) of cell separators have not been well studied. In addition, because several cell separators are available, the type of collection device may also be a significant variable. Previous studies comparing the Baxter-Fenwal CS3000 and the COBE Spectra have not yielded consistent conclusions. Therefore, we retrospectively analyzed the collection outcomes of 163 consecutive donors with a peripheral CD34+ cell concentration (pCD34) of ,5 cells/,l on the first collection that had been harvested on one or the other device. The CS3000 was found to yield a significantly higher CD34eff (50% vs. 39%, P = 0.006). However, donors were not balanced for several prognostic factors, which may contribute to CD34eff including mobilization with G-CSF vs. chemotherapy+G-CSF, average flow rate, and total volume of peripheral blood processed. When appropriate variables were included in a stepwise multiple variable analysis, cell separator type emerged as a significant independent predictive factor for CD34eff (P = 0.018). Our data indicates that the CS3000 will, on average, show a higher absolute CDeff of 8%. Furthermore, since the two devices differ in mechanism, prognostic factors may also differ. Comparisons suggest that peripheral blood WBC and hematocrit may be more important predictors for the CS3000. J. Clin. Apheresis 17:17,20, 2002. © 2002 Wiley-Liss, Inc. [source]