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  • American Society of Hematology  (5)
  • Goto, Moritaka  (5)
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  • American Society of Hematology  (5)
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  • 1
    Online Resource
    Online Resource
    Myeloid Blastic Cells with MLL-ELL Translocation Can Convert Their Morphology into the Fibroblastoid Cells.
    American Society of Hematology ; 2005
    In:  Blood Vol. 106, No. 11 ( 2005-11-16), p. 4565-4565
    Details
    In: Blood, American Society of Hematology, Vol. 106, No. 11 ( 2005-11-16), p. 4565-4565
    Abstract: Objective: It has been reported that acute leukemia with the fusion product of MLL-related molecule is determined to be the poor prognosis. To make clear one of its biological characters, we cultured AML blasts with t (11; 19) (q23; p13.1), which produced fusion protein of MLL and ELL, and the morphological changes as well as their DNA, and RNA were examined. Method: Bone marrow cells were obtained from two patients with MLL-ELL fusion gene after obtaining informed consent. There were over 90% blastic cells in bone marrow. Mononuclear cells were obtained with Ficoll (SG. 1077), and were cultured in DMEM with 10% FCS in CO2 incubator. The morphology of the cells was observed for 3 months. Also, DNA was extracted, and MLL-ELL translocation was analyzed with genomic Southern blotting with probe x which contains BamHI-digested 0.9 kb fragment from MLL cDNA. RNA was extracted with GTC method, and 1st strand cDNA was synthesized with oligo-dT primer, and PCR was performed with MLL common primer and ELL antisense primer. The reaction of DNA sequencing was performed with BigDye Terminator Cycle Sequencing kit (Applied Biosystems), and analyzed with ABI Prism 3700 DNA analyzer. Result: After one month of the cultures, fibroblastoid cells were expanded without any blastic cells onto them. Southern analysis, RT-PCR study, and DNA sequence revealed that the fusion cDNA product with MLL and ELL was observed in fibroblastoid cells. Discussion: AML blasts with MLL-ELL fusion gene can convert their morphology into the fibroblastoid cells. The fibroblasts have been reported to be resistant to the chemotherapeutic agents. This morphological conversion may contribute one of the causes of poor prognosis in AML patients with MLL-ELL fusion gene. We now characterize precisely these fibroblastoid cells in point of the expression of the specific proteins, and the functions including the binding and proliferating capability to the non-adherent blastic cells.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V106.11.4565.4565
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2005
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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    Online Resource
  • 2
    Online Resource
    Online Resource
    Isolation and Characterization of Murine Novel cDNA Clone Using Newly Constructed Gene Trap Vector.
    American Society of Hematology ; 2005
    In:  Blood Vol. 106, No. 11 ( 2005-11-16), p. 4229-4229
    Details
    In: Blood, American Society of Hematology, Vol. 106, No. 11 ( 2005-11-16), p. 4229-4229
    Abstract: Objective: To understand better the mechanism of the maintenance of immature state of stem cells, murine novel cDNA clones are to be isolated by the gene trap method. Method: The trap vector was constructed with En-2 intron sequence followed by splicing acceptor sequence, IRES signal sequence and b-galactosidase cDNA. NeoR gene was also contained without poly-A additional signal. After electroporation into Embryonic stem (Es) cell lines (D3, and E14.1 Köln), neomycin-resistant Es clones were picked up, and were selected in which the expression of b-gal was detected in the undifferentiated condition cultured with leukemia inhibitory factor (LIF), and feeder layer cells by X-gal staining, but were not detected in the differentiated condition with ATRA, and without LIF nor feeder layers. Using 5′ RACE method, the transcripts from the selected clones were identified. One clone was further analyzed. Full-length cDNA was isolated from a library constructed from Es D3 cells, and its characterization was determined. Result and Discussion: one unique full-length cDNA clone was isolated for the further characterization. It was constructed with 2225 nucleotides, and putative 466 amino acids were encoded in a single long open reading frame. The transcript was detected in the undifferentiated Es cells but not in differentiated Es cells nor in various kinds of organs by northern blotting analysis. When this clone was over-expressed in murine 10T1/2 cells, RT-PCR analysis demonstrated the down-regulation of Id-1 gene. These data suggested that the isolated cDNA clone was possibly related to keeping Es cells to be immature multipotent state. We now try to isolate the related cDNA clones which are expressed mainly in the hematopoietic stem cells.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V106.11.4229.4229
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2005
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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    Online Resource
  • 3
    Online Resource
    Online Resource
    Harvest of Autologous Peripheral Blood Stem Cells Using Ifosfamide/Etoposide Regimen Combined with Granulocyte Colony-Stimulating Factor.
    American Society of Hematology ; 2004
    In:  Blood Vol. 104, No. 11 ( 2004-11-16), p. 5218-5218
    Details
    In: Blood, American Society of Hematology, Vol. 104, No. 11 ( 2004-11-16), p. 5218-5218
    Abstract: Aim: To make clear the feasibility of Ifosfamide/Etoposide (IE) regimen combined with granulocyte colony-stimulating factor (G-CSF) for the collection of autologous peripheral blood stem cells (APBSC), we evaluated the number of the collected CD34 positive cells, and the regimen-related toxicities. Patients’ characteristics and Methods: 19 Patients (male 12, female 7) were received APBSC harvest in our hospitals during Aug. 2000 to Dec. 2003 with age of 54.3 years on average (30–66). Diagnoses were included HD (n=2), NHL (n=13), MM (n=3), and malignant synovioma (n=1). IE regimen was included with Ifomide 2000 mg/m2 (day 1), and Etoposide 200 mg/m2 (day 1–3) (4cases) or 500 mg/m2 (day 1,2) (15 cases). For the prevention of hemorrhagic cystitis patients were administered with Mesna, and NaHCO3 during and after 1 day of the administration of Ifosfamide. G-CSF was administered at the dose of 10 ug/kg (lenograstim) or 300 ug/m2 (filgrastim) when blood absolute neutrophil count came down to 1000/mm3. When the suppression of bone marrow was recovered and blood WBC count came up to 5000/m3, CD34-positive cells were counted in blood, and APBSC was harvested when blood CD34-positive cells were determined above 0.1 %. APBSC was harvested with CS-3000 plus (Fenwall). Result: The median duration from the start of the administration of G-CSF to the finish of APBSC harvest was 6.5 days (4–11). The median number of CD34-positive cells of the harvested was 4.32 x 106/kg (1.10–13.50). All cases were harvested. The toxicities during from the conditioning to the harvest were included with grade 1 headache (2 cases), grade 2 nausea (6 cases), grade 1 bleeding (1 case), grade 1 constipation (1 case), grade 1 fever (1 case), and grade 1 thrombocytopenia (1 case). Conclusion: IE regimen combined with G-CSF was feasible to harvest APBSC on the yield of the collection of the transplantable stem cells, and on the regimen-related toxicities.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V104.11.5218.5218
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2004
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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    Online Resource
  • 4
    Online Resource
    Online Resource
    Comparison of the Feasibility of Adult Allogeneic Umbilical Cord Blood Transplantation between the Myeloablative Conditioning Regimen and the Non Myeloablative One.
    American Society of Hematology ; 2004
    In:  Blood Vol. 104, No. 11 ( 2004-11-16), p. 5174-5174
    Details
    In: Blood, American Society of Hematology, Vol. 104, No. 11 ( 2004-11-16), p. 5174-5174
    Abstract: Aim: To make clear the feasibility of non-myeloablative conditioning on the adult allogeneic umbilical cord blood transplantation, we compared the result between patients receiving the myeloablative (conventional) transplantation (CT) and the reduced intensity stem cell transplantation (RIST) in point of the engraftment, and the incidence of GVHD. Patients’ characteristics and Methods: Patients were admitted to our hospitals to receive umbilical cord blood transplantation during May 1999 to May 2004. CT group (n=23): Median age was 33 years. Diagnoses were included AML (n=9), ALL(n=7), NHL(n=4), MDS(n=2), CML(n=1). Preparative conditioning regimens were; TBI/Ara-C/CY (n=16), TBI/VP-16/CY (n=4), TBI/CY (n=3). GVHD prophylaxis was; Cyclosporine A (CsA) (n=11), CsA with short term Methotrexate (MTX) (n=10), Tacrolimus with short term MTX (n=2). The transplanted median cell number was 2.40 x 10^7/kg. HLA disparities were; 1AMM (n=5), 2AMM (n=18), 3AMM (n=3). RIST group (n=13): Median age was 54 years. Diagnoses were; AML (n=3), NHL (n=5), HD (n=2), ATL (n=1), CLL (n=1), CMMoL(n=1). Preparative conditioning regimens were; Flu/ L-PAM (n=4), TBI/ Flu/ L-PAM (n=1), TBI/ Flu/ CY (n=3), TBI/ Flu (n=1), TBI/ Flu/ BU (n=4). GVHD prophylaxis was; CsA (n=5), CsA with MMF (n=7), Tacrolimus (n=1). The transplanted median cell number was 2.49 x 10^7/kg. HLA disparities were; identical (n=3), 1AMM (n=2), 2AMM (n=6), 3AMM (n=2). Result: The rate of engraftment failure including conditioning failure, and autologous recovery was 14.8 %in CT and 38.1%in RIST, respectively. The median duration of the engraftment of neutrophils in CT and RIST were 24 days and 19 days, respectively. Patients with Grade II-IV acute GVHD were 15 (65.2%) in CT and 8 (61.5%) in RIST, respectively. The incidence of Grade III-IVacute GvHD was 7 ( 30.4%) and 5(38.4%), respectively. Conclusion: Engraftment in RIST was achieved earlier than CT, however the incidence of engraftment failure was increased in RIST. There were no significant differences on the incidence of acute GVHD between two groups. Further observations, especially long-term disease-free survival are seemed to be very interesting to ascertain RIST on adult umbilical cord blood transplantation.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V104.11.5174.5174
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2004
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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    Online Resource
  • 5
    Online Resource
    Online Resource
    Derivative (1) t(1;1) (p36; q21) Produces the Fusion Protein with Phosphatase 10 and MEL1 (MDS1/EVI1-Like Gene 1) in a AML (M5a) Case.
    American Society of Hematology ; 2005
    In:  Blood Vol. 106, No. 11 ( 2005-11-16), p. 4336-4336
    Details
    In: Blood, American Society of Hematology, Vol. 106, No. 11 ( 2005-11-16), p. 4336-4336
    Abstract: Objective: A 25 years-old Japanese male was diagnosed as AML (M5a) with 46, XY, der (1) t(1; 1) (p36; q21). To determine the leukemogenesis in this case, we analyzed the fusion product of this chromosomal translocation. Method: After obtaining informed consent, RNA was extracted from his bone marrow cells (blastic cells were occupied in almost 100%) with GTC method, and poly-A rich RNA was separated with oligo-dT latex particles. 1st strand cDNA was synthesized with oligo-dT primer. Oligo-dA stretch was added with TdT to make 5′ RACE. 1st PCR was performed with the synthetic antisense-1 primers from several candidates including p73, and MEL1, and with QT primer (5′-TGAGCCAGAGTGACGAGGACTCGAGCTCAAGCT17-3′) as a sense primer. 2nd PCR was performed with internal antisense-2 primers from several candidates, and Q0 primer (5′-CCAGTGAGCAGAGTGACG-3′) as the sense primer. The reaction of DNA sequencing was performed with BigDye Terminator Cycle Sequencing kit (Applied Biosystems), and analyzed with ABI Prism 3700 DNA analyzer. Result and Discussion: The fusion cDNA product was obtained, which was consisted of MEL1 and Phosphatase 10. The breakpoint of MEL1 was exon 4, and PR domain of MEL1 was disrupted. This domain is reported to be the key function of MEL1, and when MEL1 without PR domain is expressed, myeloid differentiation is blocked. In this case the fusion of Phosphatase 10 and MEL1 is seemed to be the main cause of leukemogenesis.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    URL: Article
    DOI: 10.1182/blood.V106.11.4336.4336
    RVK:
    XA 33000
    RVK:
    XA 10000
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2005
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
    Location Call Number Limitation Availability
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    Online Resource
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