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  • 1
    Electronic Resource
    Electronic Resource
    Influence of Fixation, Antibody Clones, and Signal Amplification on Steroid Receptor Analysis (1998)
    Oxford, UK : Blackwell Science Inc
    The @breast journal 4 (1998), S. 0 
    Details
    ISSN: 1524-4741
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Various factors influence analysis of steroid receptors, including tissue processing, fixation, and antibody clones used. Furthermore, the detection methods, that is, use of epitope retrieval by microwave heating and signal amplification, may affect the reliability of the results. Eleven different fixation/tissue-processing methods as well as different commercial antibodies for the estrogen-and progesterone-receptors and the tyramine amplification technique were evaluated for optimizing the staining procedures.Moreover, the results from the optimal immunohistochemical procedure were compared with results obtained using a biochemical dextran-coated charcoal assay in 88 breast carcinomas from postmenopausal patients. The four antibody clones tested (1D5, 6F11, 1A6, and antiserum) revealed different sensibility to formalin, either favoring shorter or longer periods of fixation. Overall, a delay in the onset of fixation showed the worst results, whereas overfixation for up to 4 days or pH of formalin had little influence. Using the new tyramine technique it was possible to reduce costs per slide considerably by increasing antibody dilution.Comparison of the biochemical assay with optimized immunohistochemistry in the 88 cases investigated showed an overall concordance of 98.9%. Our study suggests an optimized immunohistochemical procedure that can replace the biochemical “gold standard” if standardization of tissue processing is maintained.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1524-4741.1998.410033.x
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  • 2
    Electronic Resource
    Electronic Resource
    Genetic heterogeneity in a prostatic carcinoma and associated prostatic intraepithelial neoplasia as demonstrated by combined use of laser-microdissection, degenerate oligonucleotide primed PCR and comparative genomic hybridization (1998)
    Springer
    Virchows Archiv 433 (1998), S. 297-304 
    Details
    ISSN: 1432-2307
    Keywords: Key words Comparative genomic hybridization ; Laser-assisted microdissection ; DOP-PCR ; Prostatic carcinoma ; Prostatic intraepithelial neoplasia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  We combined laser-assisted microdissection from H&E-stained paraffin sections, degenerated oligonucleotide-primed polymerase chain reaction (DOP-PCR), and comparative genomic hybridization (CGH) to analyse chromosomal imbalances in small tumour areas consisting of 50–100 cells. This approach was used to investigate intratumour genetic heterogeneity in a case of metastatic prostatic adenocarcinoma and chromosomal changes in areas of prostatic intraepithelial neoplasia (PIN) adjacent to the invasive tumour. In four microdissected invasive tumour areas with different histological patterns (acinar, cribriform, papillary and solid) marked intratumour heterogeneity was found by CGH. Recurrent chromosomal imbalances detected in at least two microdissected tumour areas were gains on 1p32→p36, 2p22, 3q21, 7, 8q21→q24, 11q12→q13, 16p12→p13, 17, 19 and loss on 16q23. Additional chromosomal changes were found in only one of the microdissected areas (gains on 16q21→q23, 20q22 and losses on 8p21→p23, 12p11→q12, 12q21→q26, 13q21→q34, 16q12, and 18q22). In PIN, gains on chromosomes 8q21→q24 and 17 were found in both samples investigated (low and high grade PIN), while gains on chromosomes 7, 11q, 12q, 16p, and 20q and losses on 2p, 8p21→p23, 12q were found only in one PIN area. Controls to ensure reliable CGH results consisted in CGH analyses of (i) approximately 80 microdissected normal epithelial cells, which showed no aberrations after DOP-PCR and (ii) larger cell numbers (approximately 105 or 107 cells) of the primary tumour investigated without DOP-PCR and partially displaying the chromosomal imbalances (gain on 16p12→p13, losses on 2p25, 8p21→p23, 12p11→p12, 12q21→q26, 18q22) found in the small microdissected areas. Microsatellite and FISH analyses further confirmed our CGH results from microdissected cells. The combined approach of laser-assisted microdissection, DOP-PCR and CGH is suitable to identify early genetic changes in PIN and chromosomal imbalances associated with the particular histological patterns of invasive prostatic adenocarcinoma.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004280050252
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  • 3
    Electronic Resource
    Electronic Resource
    Antigen retrieval, signal amplification and intensification in immunohistochemistry (1996)
    Springer
    Histochemistry and cell biology 105 (1996), S. 253-260 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this overview we emphasize new methods of improving immunohistochemical results in formaldehyde-fixed tissue samples. The benefit of heat-induced antigen retrieval in demasking of concealed epitopes is demonstrated. We provide guidance on the influence of heat-induced antigen retrieval in commonly applied monoclonal and polyconal antibodies. Moreover, we show the promising methods of signal amplification using biotinylated tyramine and signal intensification of diaminobenzidine reaction products by metallic ions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01463928
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  • 4
    Electronic Resource
    Electronic Resource
    Comparative FISH analysis of numerical chromosome 7 abnormalities in 5-μm and 15-μm paraffin-embedded tissue sections from prostatic carcinoma (1997)
    Springer
    Histochemistry and cell biology 107 (1997), S. 121-126 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Interphase fluorescence in situ hybridization (FISH) was performed on 15-μm-thick paraffin sections from prostatic carcinomas using a chromosome 7-specific α-satellite DNA probe. A confocal laser scanning microscope (CLSM) was used for optical sectioning of the thick sections and reconstruction of 3D images. The number of FISH signals was determined by a gallery of optical sections evaluating only complete nuclei. To investiate the influence of section thickness and truncation and nuclei on scoring results, we compared the FISH data from 15-μm sections with signal counts obtained from 5-μm sections. The latter were evaluated by conventional fluorescence microscopy in the same tumor regions previously defined and marked on the slides. After statistical analysis of spot frequencies in tumor and non-tumorous cells (χ2 test), we transferred the signal frequencies into a cytogenetic classification (−7, +7, polysomy 7). Based on this classification, most cases showed more than one chromosome 7 aberration type. Trisomy 7 (+7) became apparent in 15-μm-thick sections in all 19 tumors, polysomy 7 (〉3 spots) in 18/19 cases, and monosomy 7 (−7) in 13/19 cases. In 5-μm sections, however, trisomy 7 and polysomy 7 were found in only 7/19 and 13/19 cases, respectively, and monosomy 7 in 7/19 cases. When comparing the classification results of tumor cells of the same tumor regions originating either from 5-μm or 15-μm sections, the following discrepancies were noted: in 15-μm sections exclusively, in 12/19 tumors, trisomy 7 was found; in 6/19 cases, polysomy 7; in 8/19 cases, monosomy 7. The high proportion of cases with tumor nuclei expressing only one hybridization signal of chromosome 7 in 15-μm sections could be confirmed as monosomy 7 in five selected cases by double-hybridization using centromere-specific probes for chromosomes 7 and 12. These results demonstrate that numerical chromosome 7 aberrations are more frequently observed in thick (15-μm) paraffin-embedded tissue sections by evaluating only complete nuclei. The use of routine sections (5-μm) for interphase cytogenetic analyses is compromised by a remarkable underestimation of the real chromosome copy numbers.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004180050096
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  • 5
    Electronic Resource
    Electronic Resource
    Detection of numerical karyotype changes in the giant cells of Hodgkin's lymphomas by a combination of FISH and immunohistochemistry applied to paraffin sections (1996)
    Springer
    Histochemistry and cell biology 105 (1996), S. 401-404 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Conventional karyotyping of Hodgkin's disease (HD) has until now yielded only limited insight into karyotypic characteristics of this discase. For this reason, fluorescence in situ hybridization (FISH) using alphasatellite chromosome-specific probes was applied to paraffin sections of HD tumors in order to verify numerical aberrations suggested to be specific for HD in the literature. The FISH technique was combined with immunohistochemical detection of the CD30 antigen to allow easier identification of the Reed-Sternberg and Hodgkin (RS&H) cells. The number of specific FISH signals per nucleus was determined both in CD30-positive RS&H cells as well as in non-malignant “bystander” cells in order to assess differences in the signal distribution. Contrasted with normal lymphoid cells, the tumor cells in HD were found to be polysomic for at least one of the chromosomes analyzed (1,2,4, and 8). The technique described is a reliable method for confirmation of results obtained from conventional cytogenetics, which is especially suited for archival material or samples not containing dividing cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01463661
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  • 6
    Electronic Resource
    Electronic Resource
    Target and signal amplification: approaches to increase the sensitivity of in situ hybridization (1997)
    Springer
    Histochemistry and cell biology 108 (1997), S. 325-333 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In situ hybridization (ISH) has proven to be a very important molecular tool in research and diagnosis. However, its applicability can be limited by its restricted detection sensitivity. During the last few years, several strategies have been developed to improve the threshold levels for ISH detection by amplification of either target nucleic acid sequences prior to ISH or the detection signals after hybridization procedures. In this overview, we outline and analyze the principles, applications, and limitations of in situ polymerase chain reaction, in situ self-sustained sequence replication, primed in situ labeling (PRINS), and in situ transcription as examples of target amplification methods, and catalyzed reporter deposition using biotinylated tyramine as an approach to signal amplification in ISH. We also provide a detailed, 1-day protocol for non-radioactive oligonucleotide ISH including signal amplification, which is suitable for diagnostic purposes. Furthermore, future directions of ISH including combined strategies of target and signal amplification as well as automation are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004180050173
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  • 7
    Electronic Resource
    Electronic Resource
    Comparative genomic hybridisation for the analysis of chromosomal imbalances in solid tumours and haematological malignancies (1997)
    Springer
    Histochemistry and cell biology 108 (1997), S. 403-417 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Comparative genomic hybridisation (CGH) is based on a two-colour, competitive fluorescence in situ hybridisation of differentially labelled tumour and reference DNA to normal metaphase chromosomes. This new technology has made a great impact in molecular tumour pathology due to its possible application to archival specimens and the ability to create copy number karyotypes throughout the whole genome from very small amounts of DNA. If chromosomal imbalances can be correlated with a etiological and clinical features of tumours, CGH could be able to provide new prognostic and diagnostic criteria. CGH findings further provide starting points for the molecular genetic characterisation of altered chromosomal regions harbouring yet unidentified genes involved in tumorigenesis and tumour progression. An overview of the results of published CGH studies on solid tumours and haematological malignancies is presented. Methodological limitations of the CGH technology are reported, as well as future developments which will improve its use in routine analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004180050181
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  • 8
    Electronic Resource
    Electronic Resource
    Concentration Dependent and Adverse Effects in Immunohistochemistry using the Tyramine Amplification Technique (1999)
    Springer
    Journal of molecular histology 31 (1999), S. 195-200 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Although the tyramine amplification technique to enhance sensitivity in immunohistochemistry has been described in numerous methodological papers, it has not yet gained access to diagnostic immunohistochemistry. This is mainly due to problems and pitfalls occurring in adaptation of this method to routine application. In this study a monoclonal antibody and a polyclonal antiserum (pan-cytokeratin and anti-myoglobin) were tested in tissues with different amounts of epitopes, using a checkerboard table and testing a total of 133 different dilution combinations of both the tyramide solution and the primary antibodies. The specific tissue investigated, i.e. the amount of accessable epitope to be detected and the applied concentration of the tyramide solution mainly influenced the staining reaction. Several pitfalls such as an uneven distribution of the staining or dramatic overstaining (paradoxical overstaining) must be considered to achieve optimal results. In conclusion, our data confirm methodological studies that the tyramine amplification technique is a powerful method to enhance immunohistochemical sensitivity. However, for reliable daily practice several pitfalls of the technique have to be circumvented.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003554217994
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  • 9
    Electronic Resource
    Electronic Resource
    Phase diagram and crystallization of polycarbonate/poly-∊-caprolactone blends (1996)
    Weinheim : Wiley-Blackwell
    Angewandte Makromolekulare Chemie 243 (1996), S. 177-187 
    Details
    ISSN: 0003-3146
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Physics
    Description / Table of Contents: Es wird über die Gleichgewichts-Schmelzpunktsdepression von Polycarbonat aus Bisphenol A (PC) in Mischungen mit Poly-∊-caprolacton (PCL) berichtet. Hierzu wird ein Modell entwickelt, das den gekrümmten Verlauf der gemessenen Hoffman-Weeks-Plots zu erklären vermag. Es berücksichtigt die Konzentrationsinhomogenitäten, die mit fortschreitender Kristallisation an der Kristallwachstumsfront entstehen. Die gelegentlich beobachtete Schmelzpunktsdepression der PCL-Komponente wird diskutiert. Der Flory-Hugginssche Wechselwirkungsparameter wird zu -0,09 berechnet; aus diesem kleinen Wert muß geschlossen werden, daß die Mischungspartner enthalpisch nur schwach wechselwirken. Der merkliche Abfall der Glastemperatur des PC nach Einmischen von PCL bewirkt eine beträchtliche Kristallisation aus der Schmelze; es können PC-Kristallinitätsgrade bis zu 25% erreicht werden.
    Notes: It is reported on the equilibrium melting point depression of polycarbonate from bisphenol A (PC) in its blend with poly-∊-caprolactone (PCL). To this end, a model which explains the bended course of the measured Hoffman-Weeks plots is developed which takes into account the composition changes at the crystal growth front upon progressive crystallization. The occasional melting point depression of the PCL component is discussed. The calculated Flory-Huggins parameter of χ12 ≍ -0.09 indicates only weak enthalpic interactions. The remarkable drop in the glass transition temperature of PC with blending allows PC degrees of crystallinity up to 25% upon crystallization from the melt.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/apmc.1996.052430115
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  • 10
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    Variations of oxygen-18 in West Siberian precipitation during the last 50 years (2014)
    COPERNICUS GESELLSCHAFT MBH
    In:  EPIC3Atmospheric Chemistry and Physics, COPERNICUS GESELLSCHAFT MBH, 14(11), pp. 5853-5869, ISSN: 1680-7316
    Details
    Publication Date: 2014-06-23
    Description: Global warming is associated with large increases in surface air temperature in Siberia. Here, we apply the isotope-enabled atmospheric general circulation model ECHAM5-wiso to explore the potential of water isotope measurements at a recently opened monitoring station in Kourovka (57.04° N, 59.55° E) in order to successfully trace climate change in western Siberia. Our model is constrained to atmospheric reanalysis fields for the period 1957–2013 to facilitate the comparison with observations of δD in total column water vapour from the GOSAT satellite, and with precipitation δ18O measurements from 15 Russian stations of the Global Network of Isotopes in Precipitation. The model captures the observed Russian climate within reasonable error margins, and displays the observed isotopic gradients associated with increasing continentality and decreasing meridional temperatures. The model also reproduces the observed seasonal cycle of δ18O, which parallels the seasonal cycle of temperature and ranges from −25 ‰ in winter to −5 ‰ in summer. Investigating West Siberian climate and precipitation δ18O variability during the last 50 years, we find long-term increasing trends in temperature and δ18O, while precipitation trends are uncertain. During the last 50 years, winter temperatures have increased by 1.7 °C. The simulated long-term increase of precipitation δ18O is at the detection limit (〈1 ‰ per 50 years) but significant. West Siberian climate is characterized by strong interannual variability, which in winter is strongly related to the North Atlantic Oscillation. In winter, regional temperature is the predominant factor controlling δ18O variations on interannual to decadal timescales with a slope of about 0.5 ‰ / °C. In summer, the interannual variability of δ18O can be attributed to short-term, regional-scale processes such as evaporation and convective precipitation. This finding suggests that precipitation δ18O has the potential to reveal hydrometeorological regime shifts in western Siberia which are otherwise difficult to identify. Focusing on Kourovka, the simulated evolution of temperature, δ18O and, to a smaller extent, precipitation during the last 50 years is synchronous with model results averaged over all of western Siberia, suggesting that this site will be representative to monitor future isotopic changes in the entire region.
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , isiRev
    Format: application/pdf
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