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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    International journal of earth sciences 45 (1956), S. 194-202 
    ISSN: 1437-3262
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences
    Notes: Zusammenfassung Argand, Preiswerk und neuerdings auchNabholz halten die granitoiden Kerngesteine der penninischen Decken der Simplon-Tessin Region (schweizerische und italienische Alpen) für herzynische Eruptivgesteine, die alpin mehr oder weniger vergneist wurden.Nabholz gliedert das penninische Kristallin in einen vorherzynischen und einen herzynischen vortriadischen Gesteinsanteil. Neuere Untersuchungen belegen jedoch das alpine Alter der penninischen Deckengesteine der genannten Region und lassen lediglich eine Gliederung in alpin ältere und alpin jüngere Deckengesteine zu. Die älteren werden Hüllgesteine genannt; aus ihnen sind durch alpin syn-bis posttektonische Granitisation s. 1. die jüngeren Kerngesteine hervorgegangen.
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  • 2
    ISSN: 1573-904X
    Keywords: Caco-2 ; in vitro intestinal epithelial model ; confocal laser scanning microscopy ; tight junctions ; cytoarchitecture ; variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To introduce confocal laser scanning microscopy (CLSM)combined with digital image restoration to characterise Caco-2 cellsunder different culture conditions, and thus to define additional validcriteria for the optimisation of culture models. Methods. Growth curves were established and transepithelial electricalresistance (TEER) measured for cells grown in EMEM or DMEMmedium on Cyclopore™ membranes. Cytoskeleton, cell nuclei and tightjunctions (TJ) were investigated by CLSM. Results. Cultures reached a plateau of ∼4.5 × 105 cells/cm2 after∼ 10 days. At the same time TEER reached 750 Ω cm2. An irregular,fairly complete network of TJ was present at confluence (∼2 d).Between 15 and 30 days a regular TJ network was established. Cellsformed mixed mono- and multilayers under most conditions with twoexceptions: flat monolayers were observed on polycarbonate filterswith EMEM and with the Biocoat™ intestinal epithelium differentiationenvironment system. In multilayers TJ were found in the upper aswell as in the lower cell layers although the regular vertical polaritywas disturbed. Conclusions. CLSM represents an important tool to investigate thecytoarchitecture of Caco-2 cells. 3D-analysis of confocal data givesimportant clues on the characteristics of cell layers and thus helps tovalidate optimisation strategies.
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  • 3
    ISSN: 1573-904X
    Keywords: MDCK cells ; confocal laser scanning microscopy ; cytoskeleton ; tight junctions ; in vitro epithelial cell model ; age-related changes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. Madin Darby Canine Kidney (MDCK) cells were grown in culture, and age-related morphological changes in the cytoskeleton and tight junction (TJ) network were used to define stages in view of establishing an optimal in vitro model for the epithelial barrier. Methods. Growth curves and transepithelial electrical resistance (TEER) were determined, and the cytoskeleton (actin, α-tubulin, vimentin) and TJ (Zonula occludens proteins ZO1, ZO2) were investigated with immunofluorescent methods by confocal laser scanning microscopy (CLSM) and digital image restoration. Results. TEER measurements indicated that TJ were functional after one day. Values then remained constant. Four morphological stages could be distinguished. Stage I (0−1 day): Sub confluent cultures with flat cells; TJ established after cell-to-cell contacts are made. Stage II (2−6 days): Confluent monolayers with a complete TJ network, which remains intact throughout the later stages. Stage III (7−14 days): Rearrangement in the cytoskeleton; constant cell number; volume and surface area of cells reduced (cobble-stone appearance). Stage IV (≥ 15 days): Dome formation, i.e. thickening and spontaneous uplifting of the cell monolayer. Conclusions. Based on the structural characteristics of stage III cell cultures, which are closest to the in vivo situation, we expect them to represent an optimal in vitromodel to study drug transport and/or interactions with drugs and excipients.
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  • 4
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptosis is a morphologically distinct form of programmed cell death that plays an important role in the growth regulation of a variety of tissues and also in the elimination of self-reacting immunocompetent cells. Several techniques for the qualitative and quantitative detection of this process have been established; recently, an in situ nick end-labelling technique based on the detection of DNA fragmentation, which is a molecular characteristic of apoptotic cell death, was described. Applying this method to paraffin sections of human tissues, sensitivity was observed to be inconsistently low with regard to the expected number of apoptotic cells. In the present study we show that irradiation of the tissue sections in 10 mM citrate buffer, pH 6.0, by microwaves at 750 W considerably enhances the sensitivity of this nick end-labelling technique.
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  • 5
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract CD44 is a transmembrane glycoprotein, which can exist in a multitude of isoforms due to alternative splicing of the pre-mRNA. We have generated monoclonal antibodies to several of these variant regions, which are encoded by 10 additional exons in the extracellular part of the molecule. CD44 variant isoforms have been reported to be involved in the malignant progression of rat and human tumours. The precise localization of CD44 variant isoforms in normal developmental and morphogenetic processes is essential for diagnostic studies of human tumorigenesis. Therefore, we have analysed a large number of different human tissues by immunohistochemistry for the expression of CD44 isoforms containing either exons 4v, 6v or 9v. Expression of exon 9v-isoforms was detected in almost all epithelia analysed, with a few exceptions. Exon 6v isoforms are expressed only in squamous and glandular epithelia, e.g. skin epidermis, sweat and sebaceous glands, oesophagus, ducts of the mammary gland, salivary and prostate glands. Detection of exon 4v-encoded isoforms was restricted to the epidermis and the oesophagus. Similar tissue distributions of CD44 variant isoforms were observed in 10-week-old fetal tissues. Since one of the ligands of CD44 is hyaluronic acid (HA), we also analysed the tissue distribution of HA synthetase. HA synthetase was detected in all tissues analysed, showing good correlation with the expression of the standard form of CD44, CD44s.
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  • 6
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract CD44 isoforms have been implicated in tumor progression and metastasis formation. This study presents a thorough immunohistochemical analysis of CD44 standard ann isoform expression in normal human skin appendages and epidermis applying monoclonal antibodies against CD44s, CD44v3,-v4,-v5,-v6, and-v9. An improved immunohistochemical protocol with microwave-based antigen retrieval in paraffin section and heavy metal amplificatio of the diaminobenzidine reaction product provided enhanced resolution and sensitivity as compared to studies on frozen sections. The hair follile, the seborrheic and eccrine sweat glands were strongly positive for all CD44 isoforms studied. In the latter, the clear cells but not the dark (intercalated) cells were positive. The sudoriferous ducts adjacent to the glands were weakly positive for all CD44 isoforms and strongly positive near the skin surface. In the apocrine glands, the basal cells showed only a moderate positivity. The myoepithelial cells expressed only CD44s. In the epidermis, all CD44 isoforms were detectable, with strongest CD44 immunostaining in the lower third of the stratum spinosum and weaker staining in the stratum basale and the upper two-thirds of the stratum granulosum. The stratum granulosum and corneum were unreactive. Thus, a regional and cell type-specific CD44 expression was revealed.
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  • 7
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  CD44 isoforms have been implicated in tumor progression and metastasis formation. This study presents a thorough immunohistochemical analysis of CD44 standard and isoform expression in normal human skin appendages and epidermis applying monoclonal antibodies against CD44s, CD44v3, -v4, -v5, -v6, and -v9. An improved immunohistochemical protocol with microwave-based antigen retrieval in paraffin sections and heavy metal amplification of the diaminobenzidine reaction product provided enhanced resolution and sensitivity as compared to studies on frozen sections. The hair follicle, the seborrheic and eccrine sweat glands were strongly positive for all CD44 isoforms studied. In the latter, the clear cells but not the dark (intercalated) cells were positive. The sudoriferous ducts adjacent to the glands were weakly positive for all CD44 isoforms and strongly positive near the skin surface. In the apocrine glands, the basal cells showed only a moderate positivity. The myoepithelial cells expressed only CD44s. In the epidermis, all CD44 isoforms were detectable, with strongest CD44 immunostaining in the lower third of the stratum spinosum and weaker staining in the stratum basale and the upper two-thirds of the stratum granulosum. The stratum granulosum and corneum were unreactive. Thus, a regional and cell type-specific CD44 expression was revealed.
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  • 8
    ISSN: 1573-8671
    Keywords: Deconvolution ; colocalization ; confocal laser scanning microscopy ; tight junctions ; adherens junctions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Computer Science , Natural Sciences in General
    Notes: Abstract The relative spatial distribution of proteins was investigated with immunofluorescent methods by confocal laser scanning microscopy and digital image restoration. For confocal data sets recorded with a voxel dimension of 50 × 50 × 150 nm noise and blur can be decreased and the resolution in the z-axis increased by applying the maximum likelihood estimation algorithm of the Huygens software. This approach was successfully applied to the study of tight and adherens junctions in relation to the actin cytoskeleton in Madin Darby Canine Kidney cells. Colocalization analysis was done for pairs of probes using a histogram-based method. F-actin, occludin, zonula occludens 1, and E-cadherin were included in the study. Double-labeled preparations were used. The combination of deconvolution with the colocalization of confocal data sets offers a powerful tool to investigate the spatial arrangement of proteins.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 142 (1975), S. 185-191 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The content of 5-methylcytosine (5MC) and 6-methyladenine (6MA) in modified and nonmodified DNAs from B. subtilis and B. subtilis phage SPP1 were determined. Nonmodified SPP1 · O DNA contains about 15 5MC residues/molecule. Each modified SPP1 ·R DNA molecule carries 190 modification specific methyl groups. This number is sufficient to account for modification of the 80 restriction sites in SPP1 DNA (Bron and Murray, 1975) against endo R · Bsu R, assuming each modified site contains two 5MC residues. Resistance of SP01 DNA against endo R · Bsu R restriction both in vivo and in vitro is probably not due to methylation of endo R·Bsu R recognition sites.
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  • 10
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A gene coding for a modifying DNA-methyltransferase which methylates the central C in the BsuR recognition sequence 5′GGCC was identified in the genome of the temperate Bacillus subtilis phage SPβ. This gene is expressed only after induction of the prophage by either mitomycin C or UV. The presence of active methyltransferase in induced cells leads to modification of BsuR recognition sites in SPβ DNA as well as in heterologous DNA.
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