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  • 1
    ISSN: 1432-2307
    Keywords: Ovarian carcinoma ; DNA ploidy ; MYC amplification ; Multiplex PCR ; Prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract There is increasing evidence that DNA ploidy is a prognostic factor in ovarian carcinomas, but it is uncertain whether MYC DNA amplification is an epiphenomenon of DNA nondiploidy or a distinct biological change with an impact on the clinical course of the disease. To clarify these issues we analysed DNA ploidy by flow and image cytometry and MYC copy number by polymerase chain reaction in archival material from ovarian carcinomas with known follow up. The results were compared with proliferative activity (Ki67 index) and p53 and bcl-2 expression. DNA cytometry revealed nondiploidy in 84 of 144 cases (58.3%). Nondiploidy was statistically significantly correlated with histological tumour type, histological grade, Ki67 index 〉10%, FIGO stage, presence of residual tumour after debulking surgery and adverse postoperative outcome. Furthermore, DNA nondiploidy was associated with p53 accumulation. We found that 84.9% of the p53-positive cases were nondiploid. This points to the paramount importance of wild type p53 for the maintenance of genome integrity in this tumour type. MYC DNA amplification was seen in 33.8% (26/77 cases) of ovarian carcinoma. There was no correlation between MYC DNA amplification and histological tumour type, histological grade, FIGO stage, DNA ploidy, proliferative activity or prognosis. However, when p53 and bcl-2 expression was taken into account, a statistically significant correlation between gene alteration or expression patterns and histological tumour type was revealed. The group of mucinous carcinomas demonstrated both MYC DNA amplification and strong bcl-2 expression in 50% and contained the largest fraction of cases without aberration (37.5%). Endometrioid carcinomas were characterized by strong bcl-2 expression in 85%, whereas serous and undifferentiated carcinomas predominantly exhibited p53 alterations, frequently accompanied by bcl-2 overexpression or MYC DNA amplification. Thus, in interaction with other genes MYC DNA amplification may play a role in the determination of the varying differentiation patterns of ovarian carcinomas.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Nierentransplantation ; Chronische Abstoßungsreaktion ; Nichtradioaktive ; In-situ-Hybridisierung ; Key words Kidney transplantation ; Chronic allograft rejection ; Non-isotopic in situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Chronic rejection is a major problem in contemporary kidney transplantation. The purpose of this study was to determine whether renal cells are repopulated by extra-renal cells over time or whether the graft remains permanently allogenic. We studied nine explanted allografted kidneys of sex-mismatched donors by means of non-isotopic in situ hybridization (NISH). We used biotinylated centromer-specific DNA probes of the human chromosomes Y and X. In a further step, monoclonal and polyclonal antibodies against CD45, CD3, CD20, CD31, CD1a, S100, α-actin, factor VIII and UEA were used to analyse the various infiltrating cell types and the cells involved in allograft arteriopathy. In several cases NISH and immunohistochemistry were combined to facilitate the typing of cells. Our study showed that up to several years after transplantation the glomerular, tubular and endothelial cells retained donor origin. The only cells of recipient origin were the inflammatory cells, predominantly macrophages and T lymphocytes.
    Notes: Zusammenfassung Die chronische Abstoßungsreaktion stellt heute das wichtigste noch ungelöste Problem der Nierentransplantation dar. Die Zielsetzung der vorliegenden Studie war es zu klären, ob es nach Transplantation zu einer Reendothelialisierung oder Reepithelialisierung durch extrarenale Zellen kommt oder ob das Transplantat allogen bleibt. Vor diesem Hintergrund untersuchten wir 9 explantierte Transplantatnieren gegengeschlechtlicher Spender mit Hilfe der nichtradioaktiven In-situ-Hybridisierung (NISH). Wir benutzten biotinilierte zentromerspezifische DNA-Sonden für das humane Chromosom Y und X. In einem weiteren Schritt wurden mono- bzw. polyklonale Antikörper gegen CD45, CD3, CD20, CD31, CD1a, S100, α-Aktin, Faktor VIII und UEA eingesetzt, um die infiltrierenden Zellen und die an der Transplantatvaskulopathie beteiligten Zellen zu charakterisieren. In mehreren Fällen wurde NISH und Immunhistochemie kombiniert. Unsere Studie konnte zeigen, daß auch mehrere Jahre nach Transplantation die glomerulären, tubulären und endothelialen Zellen das Geschlecht des Spenders beibehielten. Die einzigen im Transplantat nachweisbaren Empfängerzellen waren die Entzündungszellen, die sich vornehmlich aus T-Lymphozyten und Makrophagen zusammensetzen.
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  • 3
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Aborte ; Numerische Chromosomenaberrationen ; Placentapathologie ; Interphasezytogenetik ; In-situ-Hybridisierung ; Key words Abortions ; Numerical chromosomal aberrations ; Placenta pathology ; Interphase cytogenetics ; In situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Chromosomal aberrations are an important cause of spontaneous abortions. In order to detect numerical aberrations, paraffin-embedded tissue from 26 abortions with known conventional cytogenetic findings (CCG; 25 numerical aberrations and one partial trisomy 7p) was analyzed by means of interphase cytogenetics (ICG) using centromer-specific DNA probes for chromosomes #X, #Y, #10, #18, and #13/#21. Limit-values for the diagnosis of aneusomy in tissue sections were established by classifying the distribution of hybridization signals by CCG data (for gain ≥15% of nuclei with +1 signal; for deletion 〉40% of nuclei with −1 signal). Signal distribution in tissue sections and nuclear suspensions from paraffin blocks analyzed in parallel showed statistically a highly significant correlation (P〈0.0001). ICG and CCG diagnoses corresponded in 18 of 20 cases suitable for evaluation (90%; no false-positive result). No correlation between cytogenetic and histologic findings could be found. ICG proved to be a reliable tool for the detection of numerical chromosomal aberrations in paraffin-embedded tissue of abortions (sections and nuclear supensions). Thus, data for genetic counselling of the parents can be provided. The limit values for diagnosis of aneusomy could also be important for the application of ICG in tumor cytogenetics.
    Notes: Zusammenfassung Chromosomale Aberrationen sind eine wichtige Ursache von Spontanaborten. Zum Nachweis genetischer Störungen im Paraffin-eingebetteten Gewebe wurden 26 Aborte mit bekanntem zytogenetischem Befund an Metaphasen (konventionelle Zytogenetik/KZG; 25 numerische und eine strukturelle Aberration) retrospektiv mit der sog. Interphasezytogenetik (IZG) mit Zentromer-spezifischen DNA-Sonden (für #X, #Y, #10, #18 und #13/#21) analysiert. Durch Zuordnung der IZG-Signalverteilungen (Chromosomen #X, #Y und #18) zu den KZG-Befunden wurden Grenzwerte für die IZG-Diagnose von Aneusomien ermittelt (für Zugewinn ≥15% Kerne mit +1 Signal; für Deletion ≥40% der Kerne mit −1 Signal). Die Ergebnisse in Schnittpräparaten (6 µm dick) und parallel untersuchten Kernsuspensionen korrelierten statistisch hoch signifikant (p〈0,0001). KZG- und IZG-Diagnosen stimmten in 18 von 20 auswertbaren Fällen überein (90%; kein falsch-positiver Befund). Korrelationen zwischen zytogenetischen und histomorphologischen Befunden fanden sich nicht. Mit der IZG lassen sich numerische Chromosomenaberrationen an Schnitten und Kernsuspensionen von Paraffin-eingebettetem Abortmaterial zuverlässig nachweisen, was die Möglichkeit zur genetischen Beratung der Eltern eröffnet. Die ermittelten Grenzwerte für den Nachweis von Aneusomien sind auch für tumorzytogenetische Untersuchungen mit der IZG-Technik relevant.
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  • 4
    ISSN: 1365-2559
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Aims : To determine the pattern of macrophage infiltration in colon cancers and its correlation with clinicopathological characteristics.Methods and results : Colon cancers from 100 patients were arrayed into a tissue microarray (TMA). Four cores per tumour were taken: three from the invasion front (IF) and one from the tumour surface (TS). Macrophages were quantified by immunohistochemistry with antibodies to the PG-M1, KP-1, MRP8, MRP14 and MRP8/14 antigens. The number of macrophages was significantly higher in the TS cores than in the IF cores and both tumour sites showed a higher number of macrophages than the normal mucosa. The number of macrophages decreased in higher stage tumours. The different tumour-associated macrophage (TAM) subpopulations were positively correlated with each other.Conclusions : The increased number of macrophages in cancers compared with normal colon mucosa indicates that macrophages are attracted to the tumour site. However, decreasing macrophages in higher stage colon cancers suggest that this attraction decreases with tumour progression.
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  • 5
    Publication Date: 2018-03-16
    Description: Purpose: The aim of this study was to identify and independently validate a novel gene signature predicting locoregional tumor control (LRC) for treatment individualization of patients with locally advanced HPV-negative head and neck squamous cell carcinomas (HNSCC) who are treated with postoperative radio(chemo)therapy (PORT-C). Experimental Design: Gene expression analyses were performed using NanoString technology on a multicenter training cohort of 130 patients and an independent validation cohort of 121 patients. The analyzed gene set was composed of genes with a previously reported association with radio(chemo)sensitivity or resistance to radio(chemo)therapy. Gene selection and model building were performed comparing several machine-learning algorithms. Results: We identified a 7-gene signature consisting of the three individual genes HILPDA, CD24, TCF3 , and one metagene combining the highly correlated genes SERPINE1, INHBA, P4HA2 , and ACTN1 . The 7-gene signature was used, in combination with clinical parameters, to fit a multivariable Cox model to the training data (concordance index, ci = 0.82), which was successfully validated (ci = 0.71). The signature showed improved performance compared with clinical parameters alone (ci = 0.66) and with a previously published model including hypoxia-associated genes and cancer stem cell markers (ci = 0.65). It was used to stratify patients into groups with low and high risk of recurrence, leading to significant differences in LRC in training and validation ( P 〈 0.001). Conclusions: We have identified and validated the first hypothesis-based gene signature for HPV-negative HNSCC treated by PORT-C including genes related to several radiobiological aspects. A prospective validation is planned in an ongoing prospective clinical trial before potential application in clinical trials for patient stratification. Clin Cancer Res; 24(6); 1364–74. ©2018 AACR .
    Print ISSN: 1078-0432
    Electronic ISSN: 1557-3265
    Topics: Medicine
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