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  • 1
    In: Nature Genetics, 2001, Vol.27(3), p.257
    Description: Embryonic stem (ES) cells offer unprecedented opportunities for random or targeted genome alterations in the mouse. We present here an efficient strategy to create chromosome-specific loss of heterozygosity in embryonic stem cells by the insertion of the gene neo. Six targeted neo insertions on 4 different chromosomes Foxa2 (Hnf3b) on MMU2; Kdr (Flk-1) on MMU5; Gjal (Cx43) on MMU10 and 3 insertions at Vegf on MMU17 were performed. In each case, analysis of genomic DNA samples from heterozygous and homozygous cells to confirm the genotype at the targeted locus and assess the presence of the 2 simple sequence length polymorphism alleles for the markers located on the same chromosome was done. All the homozygous cells studied have lost heterozygosity, not only at the targeted locus, but also at distant linked markers, located 16-66 cM from the neo insertion. The karyotype of these cells confirmed that all homozygous lines still carry only 2 copies of the neo-targeted chromosome. The combination of this method with genome-wide mutagenesis in ES cells (using chemical mutagens or gene-trap vectors) opens up the possibility for in vitro or in vivo functional screening of recessive mutations in the mouse.
    Keywords: Chromosomes ; Embryonic Stem Cells ; Genes ; Genetic Markers ; Genetically Engineered Organisms ; Genomes ; Heterozygosity ; Homozygosity ; Insertional Mutagenesis ; Loci ; Mutagenesis ; Transgenic Animals ; Karyotypes ; Genetically Engineered Animals ; Genetically Modified Animals ; Genetically Modified Organisms ; Geos ; Gmos ; Transgenic Organisms ; Mice ; Muridae ; Rodents ; Mammals ; Vertebrates ; Chordata ; Animals ; Eukaryotes;
    ISSN: 1061-4036
    E-ISSN: 1546-1718
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  • 2
    Language: English
    Description: Customer satisfaction is one of the hot topics also for a successfull Czech car producer ŠKODA AUTO a.s. This diploma thesis titled: Identification of best practices for customer satisfaction at ŠKODA AUTO a.s., analyses customer satisfaction management of sales and evaluates the applicability of proposed improvement process measurements.
    Keywords: Customer Satisfaction; Prodej; Osvědčené Postupy; Sales; Škoda Auto; Zákaznícka Spokojenost; Best Practices
    Source: Networked Digital Library of Theses and Dissertations
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  • 3
    Language: English
    In: Blood, 12 April 2012, Vol.119(15), pp.3495-502
    Description: In addition to its proapoptotic function, caspase-8 is also important for several other processes, including suppressing necroptosis, cell migration, and immune cell survival. In the present study, we report that the loss of caspase-8 in B lymphocytes leads to B-cell malignancies and that the risk for these tumors is further enhanced in the absence of p53. We also report that deficiency of caspase-8 results in impaired cytokinesis and that casp8(-/-) lymphomas display remarkably elevated levels of chromosomal aberrations. Our data support an important role for caspase-8 in the maintenance of genomic integrity and highlight its tumor-suppressive function.
    Keywords: Caspase 8 -- Physiology ; Chromosomal Instability -- Genetics ; Lymphoma, B-Cell -- Genetics
    ISSN: 00064971
    E-ISSN: 1528-0020
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  • 4
    Language: English
    In: PLoS Genetics, 2011, Vol.7(11), p.e1002360
    Description: Ubiquitylation is fundamental for the regulation of the stability and function of p53 and c-Myc. The E3 ligase Pirh2 has been reported to polyubiquitylate p53 and to mediate its proteasomal degradation. Here, using Pirh2 deficient mice, we report that Pirh2 is important for the in vivo regulation of p53 stability in response to DNA damage. We also demonstrate that c-Myc is a novel interacting protein for Pirh2 and that Pirh2 mediates its polyubiquitylation and proteolysis. Pirh2 mutant mice display elevated levels of c-Myc and are predisposed for plasma cell hyperplasia and tumorigenesis. Consistent with the role p53 plays in suppressing c-Myc-induced oncogenesis, its deficiency exacerbates tumorigenesis of Pirh2 −/− mice. We also report that low expression of human PIRH2 in lung, ovarian, and breast cancers correlates with decreased patients' survival. Collectively, our data reveal the in vivo roles of Pirh2 in the regulation of p53 and c-Myc stability and support its role as a tumor suppressor. ; Tumor suppressors and oncogenes play critical roles in cancer development. The tumor suppressor p53 and the oncogene c-MYC are among the most frequently deregulated genes in human cancer, and their ubiquitylation mediated by several E3 ligases is critical for their turnover and their functions. P53 has been shown to be ubiquitylated by Pirh2; however, the physiological significance of this modification of p53 remains unknown. In this study we have generated mice deficient for Pirh2 and have observed that loss of Pirh2 results in a higher level of p53 and cell death, especially in response to radiation. Remarkably, we also identified that Pirh2 interacts with c-Myc and mediates its polyubiquitylation and degradation. c-Myc accumulates in the absence of Pirh2 and this accumulation is accompanied by increased tumorigenesis of Pirh2-deficient mice. We also report that dual deficiency of Pirh2 and p53 synergizes cancer development. Examination of the expression level of PIRH2 in human cancers indicated that its lower expression level associates with poor survival of patients with lung, ovarian, or breast cancers. Collectively, these data indentify Pirh2 as a novel tumor suppressor involved in the regulation of both p53 and c-Myc.
    Keywords: Research Article ; Biology
    ISSN: 1553-7390
    E-ISSN: 1553-7404
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  • 5
    Language: English
    In: International Journal of Cancer, 10 March 2003, Vol.104(1), pp.36-43
    Description: Cytotoxic drug treatment of neuroblastoma often leads to the development of drug resistance and may be associated with increased malignancy. To study the effects of long‐term cytotoxic treatment on malignant properties of tumor cells, we established 2 neuroblastoma cell sublines resistant to vincristine (VCR) and doxorubicin (DOX). Both established cell lines (UKF‐NB‐2VCR and UKF‐NB‐2DOX) were highly resistant to VCR, DOX and vice‐versa but retained their sensitivity to cisplatin. UKF‐NB‐2VCR and UKF‐NB‐2DOX expressed significant amounts of P‐glycoprotein, while parental cells were P‐glycoprotein negative. GD2 expression was upregulated, whereas NCAM expression was decreased in both resistant cells. Spectral karyotype (SKY) analysis revealed complex aberrant karyotypes in all cell lines and additional acquired karyotype changes in both resistant cells. All cell lines harbored high levels of N‐myc amplification. Compared to parental cells, UKF‐NB‐2VCR and UKF‐NB‐2DOX exhibited more than 2‐fold increase in clonal growth , accelerated adhesion and transendothelial penetration and higher tumorigenicity . We conclude that development of drug resistance and acquisition of certain karyotypic alterations is associated with an increase of additional malignant properties that may contribute to the poor prognosis in advanced forms of NB. The 2 novel neuroblastoma cell sublines also provide useful models for the study of drug resistance in aggressive forms of neuroblastoma. © 2002 Wiley‐Liss, Inc.
    Keywords: Neuroblastoma ; Drug Resistance ; Mdr‐1 ; Ncam ; Gd2 ; Karyotype
    ISSN: 0020-7136
    E-ISSN: 1097-0215
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  • 6
    Language: Czech
    Keywords: Odplynění; Kogenerace; Alternativní Zdroj
    Source: Networked Digital Library of Theses and Dissertations
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  • 7
    Language: English
    In: Cancer research, 15 June 2002, Vol.62(12), pp.3466-76
    Description: Analysis of ovarian carcinomas has shown that karyotypes are often highly abnormal and cannot be identified with certainty by conventional cytogenetic methods. In this study, 17 tumors derived from 13 patients were analyzed by a combination of spectral karyotyping (SKY), comparative genomic hybridization (CGH), and expression microarrays. Within the study group, a total of 396 chromosomal rearrangements could be identified by SKY and CGH analysis. When the distribution of aberrations was normalized with respect to relative genomic length, chromosomes 3, 8, 11, 17, and 21 had the highest frequencies. Parallel microarray expression studies of 1718 human cDNAs were used to analyze expression profiles and to determine whether correlating gene expression with chromosomal rearrangement would identify smaller subsets of differentially expressed genes. Within the entire set of samples, microarray expression analysis grouped together poorly differentiated tumors irrespective of histological subtype. For three patients, a comparison between genomic alterations and gene expression pattern was performed on samples of primary and metastatic tumors. Their common origin was demonstrated by the close relationship of both the SKY and CGH karyotypes and the observed profiles of gene expression. In agreement with the pattern of genomic imbalance observed for chromosome 3 in ovarian cancer, the relative expression profile with respect to a normal ovary exhibited a contiguous pattern of reduced expression of genes mapping to the 3p25.5-3p21.31 and increased expression of genes from 3q13.33-3q28. This study demonstrates that SKY, CGH, and microarray analysis can in combination identify significantly smaller subsets of differentially expressed genes for future studies.
    Keywords: Chromosome Aberrations ; Ovarian Neoplasms -- Genetics
    ISSN: 0008-5472
    E-ISSN: 15387445
    Source: MEDLINE/PubMed (U.S. National Library of Medicine)
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  • 8
    Language: English
    In: Cancer research, 01 November 2002, Vol.62(21), pp.6194-204
    Description: BRCA2 is a breast cancer susceptibility gene of which the product is thought to be involved in monitoring genome integrity and cell cycle progression. Brca2-null mice have a defect in embryonic cellular proliferation and die in utero. Here we report the generation of T-cell lineage-specific Brca2-deficient (tBrca2(-/-)) mice using the Cre-loxP system. Mice with a flanked by loxP allele of Brca2 were crossed to transgenic mice bearing Cre recombinase driven by the T cell-specific promoter Lck. Thymic cellularity and distribution of subset populations were normal in tBrca2(-/-) mutants. Thymocytes from tBrca2(-/-) mice underwent normal apoptosis in response to a variety of stimuli, and activated tBrca2(-/-) T cells had normal proliferative capacity. tBrca2(-/-) T cells were more likely than wild-type cells to undergo spontaneous apoptosis, but apoptosed normally in response to restimulation or DNA-damaging stress signals. Examination of metaphase spreads of tBrca2(-/-) T cells revealed that the chromosomes often exhibited aberrations such as breaks and tri-radial structures. The level of chromosomal abnormalities was enhanced in T cells from tBrca2(-/-); p53(-/-) double-mutant mice. However, tBrca2(-/-); p53(-/-) T cells did not show the enhanced level of spontaneous apoptosis demonstrated by tBrca2(-/-) T cells, a difference that likely accounts for an increase in cell number and (3)[H]thymidine incorporation of double-mutant T cells in culture compared with either single mutant. Despite this increased T-cell number, the onset of T-cell lymphomas was only marginally accelerated in tBrca2(-/-); p53(-/-) mice compared with p53(-/-) mice. Our results support a role for Brca2 in repairing spontaneous DNA lesions, and suggest that loss of Brca2 enhances the susceptibility of mouse T-lineage cells to chromosomal aberrations and deregulation of apoptosis in the absence of p53.
    Keywords: Apoptosis -- Genetics ; Brca2 Protein -- Genetics ; T-Lymphocytes -- Cytology ; Tumor Suppressor Protein P53 -- Genetics
    ISSN: 0008-5472
    E-ISSN: 15387445
    Source: MEDLINE/PubMed (U.S. National Library of Medicine)
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  • 9
    Language: English
    In: Leukemia research, December 2005, Vol.29(12), pp.1465-7
    Description: Patients with 1q duplication have demonstrated a wide range of multiple congenital abnormalities. Alterations involving this chromosomal region have being described in hematopoietic malignancies and a series of candidate genes that may be associated with neoplasias have been mapped in this region. We describe a case of partial trisomy 1q "syndrome" and acute monocytic leukemia. Cytogenetic study of the bone marrow cells by GTG-banding and spectral karyotyping (SKY) showed dup(1)(q23q44) in all cells analyzed. The dismorphological features with the dup(1q) suggest a constitutional chromosome alteration and the first, in our knowledge, association of a trisomy 1q "syndrome" with AML.
    Keywords: Chromosomes, Human, Pair 1 ; Trisomy ; Abnormalities, Multiple -- Genetics ; Leukemia, Monocytic, Acute -- Genetics
    ISSN: 0145-2126
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  • 10
    Dissertation
    Dissertation
    Univerzita Pardubice
    Language: Czech
    Description: Práce je zaměřena na pacienty jiné kultury, etnika a především na specifika, ke kterým je nutno při ošetřování těchto pacientů přihlížet. Je rozdělena na část teoretickou a praktickou. Teoretická část zahrnuje základní informace o problematice multikulturního ošetřovatelství, seznámení se základními pojmy oboru a specifiky jednotlivých menšin. Praktická část se skládá ze dvou kazuistik a čtyř ošetřovatelských procesů, které jsou řazeny v přílohách. V závěru jsou uvedeny ošetřovatelské diagnózy zaměřené na specifika ošetřování pacientů jiné kultury, etnika a sestaven modelový ošetřovatelský proces.
    Keywords: Multikulturní Ošetřovatelství ; Migrace ; Kulturní Šok ; Ošetřovatelská Péče ; Ošetřovatelský Proces ; Multicultural Nursing ; Migration ; Culture Shock ; Nursing Care ; Nursing Process
    Source: University of Pardubice
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