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    Detection of Integrated Murine Leukemia Viruses in a Mouse Model of Acute Myeloid Leukemia by Fluorescence in Situ Hybridization Combined With Tyramide Signal Amplification
    (Elsevier Science Inc, 2000) Acar, Hasan; Copeland, Neal G.; Gilbert, Debra J.; Jenkins, Nancy A.; Largaespada, David A.
    This study rt as undertaken to develop a reliable method to enumerate and map somatically acquired, clonal, murine leukemia virus (MuLV) proviral insertions in acute myeloid leukemia (AML) cells from the BXH-2 mouse strain. This was achieved by using fluorescence in situ hybridization combined with tyramide signal amplification (FISH-TSA) and an 8.8 kilobase pair (kb) full-length ecotropic MuLV or 2.0 kb MuLV envelope (env) gene probe. Two-color FISH was utilized combining chromosome-specific probes for regions near the telomere and/or centromere and the MuLV probes. The technique reliably detected germline and somatically acquired, tumor-specific, MuLV proviruses in BXH-2 AML cell lines. It rr as possible to readily verify homozygous insertions at endogenous ecotropic,MuLV loci, Emv1 (chromosome 5), Emv2 (chromosome 8) and a BXH-2 strain-specific locus (chromosome 11). This strategy also verified the presence of molecularly cloned proviral insertions within the mouse Nf1 gene and another locus on distal chromosome II, as well as on chromosome 7 and chromosome 9 in BXH-2 AML cell line B117. The technique rc as also used to detect several new tumor-specific, proviral insertions in BXH-2 AML cell lines.
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    Micronucleus Incidence and Their Chromosomal Origin Related to Therapy in Acute Lymphoblastic Leukemia (ALL) Patients: Detection by Micronucleus and FISH Techniques
    (WILEY-LISS, 2001) Acar, Hasan; Çalışkan, Ümran; Demirel, Şennur; Largaespada, David A.
    Micronucleus assay and dual color-fluorescence in situ hybridization (DC-FISH), using centromere-specific and whole chromosome-specific painting probes, are considered a useful screening test to determine the incidence of micronucleus, their origin and contents. The patients with acute lymphoblastic leukemia (ALL), who had undergone chemotherapy, were analysed before and after treatment with vincristine, methotrexate, daunomycin, prednisone, and asparaginase. The incidence of micronuclei after the antileukemic agent treatment was significantly higher than before the treatment. Application of DC-FISH using a combination of whole chromosome-specific painting probes and the same chromosome-specific a-satellite centromeric probe showed that there were no significant differences in the micronucleus incidence for any specific chromosome (chromosomes 7, 8, 11, 17, X, and Y). There were no significant differences between the incidence of centromere-positive micronuclei and the incidence of centromere-negative micronucleus. We concluded that antileukemic agents induced the somatic genetic damage but this damage is not related to any specific chromosome studied.