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  • 1
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using somatic cell hybrids, the gene for beta-2 microglobulin has been assigned to human chromosome 15. We found it of interest to study a number of human lymphoid cell lines in light of this finding, to analyze whether spontaneously occurring loss or reduction of beta-2 microglobulin could be correlated with any aberration in chromosome 15. The Daudi cell line was shown to be devoid of any beta-2 microglobulin in total extracts. Chromosome analysis showed that one of the two chromosomes 15 was deleted in the region q14↔q21 on the long arm; in some metaphases, both chromosomes were deleted in this region. The K562 cell line was found to express very low (if any) membrane-associated beta-2 microglobulin. Chromosome analysis showed that this line was near-triploid, with two normal chromosomes 15 and one translocation chromosome t(15;18) involving the long arm of chromosome 15, whereby the segment proximal to the breakage point in band q15 was lost. The Namalwa cell line showed a reduction in membrane-associated and total beta-2 microglobulin. Chromosome analysis showed this line to contain one chromosome 15 which was shorter than its normal homolog. The deletion could be identified as such in the region q14↔q21 in Daudi cells, but is probably somewhat smaller than the one in Daudi cells. Since analyses of beta-2 microglobulin production and chromosomes 15 on several other human cells failed to reveal any abnormalities in either of these respects, we postulate that genes responsible for beta-2 microglobulin synthesis and membrane expression could be located in the region q14→q21 on the long arm of chromosome 15. Since beta-2 microglobulin associated with the membrane was found to be absent in the K562 line, where total beta-2 microglobulin was nearly as high as in cell lines with “normal” membrane expression, it is suggested that membrane expression of beta-2 microglobulin can be regulated independently
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  • 3
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The Daudi cell line, established from a Burkitt lymphoma, has recently been found to be HLA- andΒ 2-microglobulin-negative, although it expresses B lymphocyte alloantigens. This report is concerned with the reexpression of HLA-A10, B38, and B17 on the Daudi cell, after cell fusion with another human cell line (Raji) or with mouse fibroblasts. In the latter fusion, the same HLA specificities are re-expressed, but not humanΒ 2-microglobulin while mouseΒ 2-microglobulin andH-2 could be detected. No such reexpression was observed when Daudi was fused with the F9 mouse teratocarcinoma, which lacks mouseΒ 2-m andH-2. No HLA activity (alloantigenic and xenogenic activity) was detected in the membrane or cytoplasm of Daudi, using salt extraction and sonication. Therefore we postulate thatΒ 2-microglobulin could be necessary for the expression and possible synthesis of the HLA antigen.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The yeast amber suppressor, SUP5-a, was previously shown to cause the insertion of tyrosine at the sites of UAG nonsense codons. Nucleotide sequencing established that this SUP5-a suppressor specifies a mutant tyrosine transfer RNA (tRNA) which has the anticodon CΨA instead of the ...
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Chromosoma 73 (1979), S. 301-315 
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A structural component of mitotic chromosomes that partially explains the compaction of DNA within mitotic chromosomes is suggested on the basis of the occurrence of long, regular cylindrical structures in preparations of isolated human chromosomes. These structures, “unit fibers”, of a rather constant diameter of about 4,000 Å have been postulated to be formed by coiling of the 250T2–300 Å solenoid chromatin fiber that itself is formed by coiling of the 100 Å string of nucleosome fiber. The human chromatid would thus be composed by a hierarchy of helices with contraction ratios for DNA at each level of coiling of 7 (string of nucleosomes), 5 (solenoid) and 40 (4,000 Å “unit fiber” or ”super-solenoid”) which results in an overall contraction ratio for DNA in the “unit fiber” structures of about 1,400, which is approximately 5-fold less than the final contraction of DNA in intact chromatids of condensed metaphase chromosomes. The present report concerns more detailed studies with respect to the dimensions and cytochemical properties of the “unit fiber” structures observed in preparations of isolated human mitotic chromosomes that provide direct and indirect evidence in support of their “super-solenoid” structure and relate to known properties of human mitotic chromosomes.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Chromosoma 73 (1979), S. 317-326 
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Chromosomal unit fibers consisting of long, regular fibers of about 0.40 μm diameter were obtained from disintegrated, isolated chromosomes of two Drosophila melanogaster cell lines. In one cell line with an essentially normal karyotype, three clearly defined size classes of 15, 13, and 11 μm length were observed corresponding to the three larger chromosomes of Drosophila. In a cell line carrying an additional translocation between the two largest chromosomes a 19 μm fiber derived from the translocation chromosome was observed. Direct determinations of the DNA content per μm length of Drosophila unit fibers show that DNA is contracted by a factor of about 1400x in agreement with calculations based on the length of the unit fibers and the known DNA content of the individual Drosophila chromosomes. These findings support our previously proposed model for the unit fiber sub-structure of chromosomes as being derived by a hierarchy of coiling with the corresponding contraction ratios being 7 (100 Å string of nucleosomes), 5 to 6 (250–300 Å thick nucleohistone fiber), and about 40 (unit fiber), resulting in a total contraction of DNA in unit fibers in the order of 1400x.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Human genetics 〈Berlin〉 27 (1975), S. 275-301 
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die cytologische und chemische Analyse des Heterokaryons, des unmittelbaren Produktes der Zellfusion, erschließt neue Möglichkeiten für das Studium von Faktoren, die für die genetische Regulation in eukaryoten Zellen verantwortlich sind. Im Vergleich zu proliferierenden Zellhybriden führt der Heterokaryon-Zustand den wichtigen Vorteil mit sich, daß er zwei vollständige Genome enthält, da Chromosomen verlust nicht stattfindet. Da aber Segregation und Rekombination fehlen, können Heterokaryone nicht in der gleichen Art wie proliferierende Zellhybriden zur Genlokalisation gebraucht werden. Wenn jedoch zwei Zelltypen mit unterschiedlichem genetischen Defekt fusioniert werden, kann man die Analyse von Heterokaryonen zum Studium der Genkomplementierung anwenden. Die biologische Information, die man durch Heterokaryone erhält, hat die Rolle des Cytoplasmas bei der Kontrolle der Kernaktivität hervorgehoben. Wenn ein G1-Kern mit dem Cytoplasma einer S-Phasen-Zelle in Kontakt gebracht wird, wird der G1-Kern zur DNA-Synthese stimuliert. Bringt man den Kern in eine mitotische Zelle, wird das Chromatin des G1-Kernes gezwungen, in vorzeitig kondensierte Chromosomen zu kondensieren. Inaktive Kerne, wie z. B. der dormante Hühnererythrocytkern, werden dazu stimuliert, die RNA- und DNA-Synthese zu beginnen, wenn sie bei der Zellfusion mit aktivem Cytoplasma in Kontakt gebracht werden. Spezifische Kernproteine, die sich im Cytoplasma von sich aktiv teilenden Zellen finden, haben sich als verantwortlich für diesen Prozeß der Reaktivierung erwiesen. Bei anderen inaktiven Kernen, wie z. B. Makrophagen und Spermakernen, konnte vergleichsweise gezeigt werden, daß sie durch Fusion mit aktiven Zellen reaktiviert wurden. Das Ausmaß der Aktivierung in den erwähnten Fällen scheint vom Zustand der aktiven Zelle bestimmt zu werden. Inaktive Kerne werden bis zum gleichen Niveau wie der aktive Kern aktiviert, aber seltener darüber hinaus. Wenn man differenzierte Zellen mit undifferenzierten Zellen fusioniert, geht der Differenzierungscharakter gewöhnlicherweise schnell nach der Fusion verloren. Diese Beobachtung stimmt mit mehreren Studien an proliferierenden Zellhybriden überein, wobei eine gewisse Art von negativer Kontrolle der differenzierten Eigenschaften angedeutet wird. Bei Heterokaryonen, die man durch Fusion von Zellen ähnlicher histiotypischer Differenzierung erhalten hat, beobachtet man gewöhnlich Coexpression der differenzierten Eigenschaften.
    Notes: Summary Cytological and chemical analysis of heterokaryons, the immediate product of cell fusion, offer new possibilities for studying the factors responsible for genetic regulation in eukaryotic cells. In comparison with proliferating cell hybrids the heterokaryon state offers the important advantage that a heterokaryon contains two complete genomes since chromosome loss does not occur, but since segregation and recombination are absent, heterokaryons cannot be used for gene mapping in the same way as proliferating cell hybrids. However, if two cell types carrying different genetic defects are fused the analysis can be used for studies of gene complementation. The biological information obtained with heterokaryons has emphasized the role of the cytoplasm in the control of nuclear activity. When a G1 nucleus is brought into contact with the cytoplasm of an S phase cell the G1 nucleus is stimulated to synthesize DNA. If the nucleus is brought into a mitotic cell, the chromatin of the G1 nucleus is forced to condense into prematurely condensed chromosomes. Inactive nuclei such as the dormant chick erythrocyte nucleus will be stimulated to initiate RNA and DNA synthesis when brought into contact with an active cytoplasm by cell fusion. Specific nuclear proteins have been shown to be responsible for this process of reactivation. Other inactive nuclei such as the nuclei of macrophages and spermatozoa have likewise been shown to be reactivated by fusion with active cells. The degree of activation in all of these cases appears to be determined by the state of the active cell. Inactive nuclei are activated to the same level as the active nucleus but seldom beyond this level. If differentiated cells are fused with undifferentiated cells, usually the differentiated character is lost rapidly after fusion. This observation is in agreement with several studies on proliferating cell hybrids indicating some type of negative control of differentiated properties. In heterokaryons obtained by fusion of cells of a similar type of histotypic differentiation usually coexpression of the differentiated markers is observed.
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  • 8
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The influence of different genetic environments on the expression of HLA complex-controlled antigens has been investigated using cell lines with various defects in the synthesis of these molecules and a somatic cell hybrid derived from them. A very sensitive bacterial binding assay allowing simultaneous evaluation of the morphology of a given cell and the quantity of a surface molecule has been developed for these studies. The fetal erythroid cell line K562, the Burkitt's lymphoma-derived cell line DAUDI, and their hybrid DUTKO1 have been employed. K562 and the hybrid, but not DAUDI, expressed HLA-A, B, C heavy chains as detected by the monoclonal antibody W6/32.HL, while two monoclonal antibodies (TÜ48 and 2BC4) against the supertypic specificities HLA-Bw4 and Bw6 showed no reactivity. The presence of human Ia-like antigens on the cell surfaces was investigated with a panel of eight monoclonal antibodies. K562 cells were completely unreactive, and DAUDI cells gave the expected positive reaction, but about 1% or less of the cells in the DUTKO1 population appeared to express these antigens as well. We discuss possible reasons for the failure to detect HLA-B antigens with monoclonal antibodies and the lack of complete “dominance” of the K562 genome in the hybrid cell line.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 111 (1971), S. 242-255 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The timing of replication of an F'lac during the cell cycle of Escherichia coli B/r has been investigated at different growth rates to clarify the relationship of F factor replication to cell division and the replication of the bacterial chromosome. Cells of a lacZ — strain carrying an F'lac were separated according to their ages in an exponentially growing population after the culture was pulse labelled with a radioactive precursor of DNA and pulse induced for the synthesis of β-galactosidase. The amount of label incorporated at different cell ages reflects the state of replication of the bacteriial chromosome, while the amount of enzyme synthesized in response to a short period of induction is assumed to reflect the state of replication of the F'lac. The F'lac replicates at a time somewhat more than half way through the cell cycle at all growth rates investigated. This time is clearly distinguishable from the time of initiation of chromosomal replication at some of the growth rates studied, implying the existence of at least some different control elements in the replication of these two replicons. The regulation of F'lac replication has been further studied by following F'lac replication in temperature sensitive mutants, which are defective in the initiation of chromosomal replication at elevated temperatures.
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  • 10
    ISSN: 1573-4986
    Keywords: SCLC ; FucGM1 ; sera ; immunoassay ; ELISA ; scintillation proximity assay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract We here report an enzyme linked immunosorbent assay (ELISA) and a scintillation proximity assay (SPA) for detection of the ganglioside FucGM1 in sera from small cell lung cancer (SCLC) patients. The SPA was more sensitive and reproducible than the ELISA. In this assay, monoclonal antibodies specific for FucGM1 were bound to SPA particles and incubated with labelled FucGM1 and 100 µl test-serum overnight, and counted in a β-counter. The sensitivity was 0.2 ng. Seven out of twenty sera from SCLC patients were positive, whereas none of twenty sera from healthy individuals were positive for FucGM1. The SPA was more sensitive than the previously reported HPTLC as well as a direct ELISA.
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