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  • 1
    ISSN: 1432-2307
    Keywords: Automated laser fluorescence sequencer ; Partial digestion ; Restriction enzyme fragment length analysis ; Asymmetrically labelled fluorescence primer ; Mycobacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract An automated method for the restriction fragment length polymorphism (RFLP) analysis for the differentiation of mycobacteria to the species level is described. After polymerase chain reaction (PCR) amplification of a sequence of the gene encoding the 65-kDa surface antigen common to all mycobacteria the product was investigated by RFLP analysis. For accurate determination of fragment sizes the asymmetrically fluorescein-labelled PCR product was partially digested with restriction site enzymes BstEII and HaeIII. The fragments obtained were analysed electrophoretically using an automated laser fluorescence DNA sequencer. Determination of fragment sizes revealed a deviation of ±1 base pair (bp; 0.6%) when compared to expected sizes. The validity of this approach was confirmed by analysing mycobacterial DNA obtained from pure cultures of Mycobacterium (M.) tuberculosis and alcohol-fixed smears as well as paraffin-embedded sputa of patients with culture-proven tuberculosis. Additionally a diagnostic algorithm was established by investigation of cultured M. bovis, M. bovis bacille Calmette-Guérin, M. avium, M. intracellulare and M. fortuitum. The method allows the identification of restriction enzyme sites which are only 40 bp apart. Partial restriction enzyme digestion of asymmetrically fluorescence-labelled PCR products will presumably lead to the discovery of new restriction enzyme sites.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2307
    Keywords: Gastric cancer ; p53 tumour-suppressor gene ; Mutation spectrum ; Dietary mutagens ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The p53 tumour-suppressor gene plays an important role in gastric carcinogenesis. In an analysis of the spectrum of mutations of the p53 gene seen in 56 primary gastric carcinomas of various types and grades of differentiation, the entire coding sequence (exons 2–11) of the p53 gene was screened by single-strand conformation polymorphism analysis and direct genomic sequencing of polymerase chain reaction products. Intragenic restriction site polymorphisms and the probe YNZ22 were used for the detection of loss of heterozygosity (LOH) of the p53 gene locus on chromosome 17p. p53 overexpression was studied with the anti-p53 antibody CM-1. A total of 21 somatic alterations of the p53 gene were found. Twenty were base-pair substitutions, and one was an eight base-pair deletion. Six tumours with p53 mutations revealed LOH. Abnormalities in p53 expression were found in 17 tumour samples, of which 16 had gene mutations. The spectrum of mutations observed was consistent with the predicted spectrum for dietary mutagens associated with the metabolism of nitrogenous compounds, resulting in deamination of nucleic acids. Our findings suggest that p53 could be a primary target for mutations associated with dietary carcinogens in gastric carcinogenesis.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1963
    Keywords: Schlüsselwörter Mastitis ; Tuberkulose ; Lupus vulgaris ; PCR ; Key words Mastitis ; Tuberculosis ; Lupus vulgaris ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary We report the rare case of granulomatous tuberculous mastitis based on lupus vulgaris in a 73-year-old female patient. The most important diagnostic procedure was the detection of mycobacterial DNA-sequences in paraffin-embedded tissues using the polymerase-chain-reaction (PCR). A combined antituberculous therapy lead to complete remission within 14 months after the diagnosis was established. The incidence of tuberculous mastitis among surgically treated lesions of the breast is less than 0.025 percent. Because of the frequent localisation of this disease in the upper outer quadrant of the breast and suspicious findings in mammography, tuberculous mastitis is one, even if rare, differential diagnosis for breast cancer.
    Notes: Zusammenfassung Berichtet wird über einen seltenen Fall einer auf dem Boden eines Lupus vulgaris entstandenen granulomatösen tuberkulösen Mastitis bei einer 73jährigen Patientin. Wegweisend für die richtige Diagnose war der Nachweis mykobakterieller Gensequenzen mit Hilfe der Polymerase-Ketten-Reaktion (PCR) in Paraffineingebettetem Gewebe. Eine antituberkulöse Kombinationstherapie führte innerhalb 14 Monaten nach Diagnosefindung zur Remission. Die Inzidenz der tuberkulösen Mastitis unter den chirurgisch behandelten Erkrankungen der Mamma liegt heute bei 0,025 %. Aufgrund der häufigen Lokalisation dieser Erkrankung im oberen äußeren Quadranten der Mamma und des suspekten mammographischen Befundes ist die tuberkulöse Mastitis eine, wenn auch seltene, Differentialdiagnose zum Mammakarzinom.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1963
    Keywords: Schlüsselwörter p53 ; Molekularpathologie- Kanzerogenese ; Key words p53 ; Molecular pathology ; Carcinogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Mutations of the p53 tumor-suppressor gene are the most common genetic alterations in human cancer, found in approximately 50 % of all tumors. The importance of p53 in human cancer attracts attention in molecular studies dealing with the pathogenesis, diagnosis and prognosis in tumor pathology. This review summarizes the current understanding of p53 both on the genetic and protein level. Frequency and spectrum of somatic p53 mutations in the carcinogenesis of breast cancer, colorectal cancer, gastric cancer, hepatocellular carcinoma, squamous-cell carcinoma of the skin and malignant melanoma are discussed including our own investigations and studies published in the literature.
    Notes: Zusammenfassung Mutationen des p53-Tumorsuppressorgens lassen sich in etwa 50 % aller menschlichen Tumoren nachweisen. Seine Bedeutung für die Entstehung menschlicher Krebserkrankungen macht dieses Gen zu einem zentralen Interesse molekularpathologischer Untersuchungen, die die Erfassung pathogenetischer, diagnostischer und prognostischer Parameter in der Tumorpathologie zum Inhalt haben. In dieser Übersichtsarbeit werden wesentliche Aspekte der Funktion des p53-Tumorsuppressorgens und des von ihm kodierten Proteins dargestellt. Unter Berücksichtigung eigener Untersuchungen und der Literatur werden Häufigkeit und Spektrum somatischer p53-Mutationen in der Kanzerogenese des Mammakarzinoms, des kolorektalen Karzinoms, des Magenkarzinoms, des hepatozellulären Karzinoms, des Plattenepithelkarzinoms der Haut sowie des malignen Melanoms diskutiert.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Der Onkologe 4 (1998), S. 671-681 
    ISSN: 1433-0415
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassend zeigen die aufgeführten Methoden und Beispiele, daß die Molekularbiologie einen diagnostisch und klinisch relevanten Bereich in der Onkologie darstellt. Die Erfassung Tumor-assoziierter molekularer Veränderungen wie Mutationen, Deletionen, Amplifikationen und Translokationen läßt sich für einige Tumorentitäten diagnostisch, prognostisch und therapierelevant einsetzen. Dem Pathologen kommt hier eine ganz besondere und verantwortungsvolle Aufgabe zu, diese Veränderungen in der Zusammenschau mit dem phänotypischen Erscheinungsbild eines Tumors in Verbindung zu bringen. Literatur
    Notes: Die maligne Transformation einer Zelle ist ein komplexer mehrschichtiger Prozeß, der zu einer Anhäufung von genetischen Veränderungen im Genom der betroffenen Zelle führt, die durch verschiedene Alterationen hervorgerufen werden können. Zelluläre Mechanismen, die das biologische Verhalten einer Zelle wie Proliferation, Differenzierung, Motilität und Absterben regulieren, sind besonders betroffen. Die rasche Entwicklung in der molekularen Genetik und Zellbiologie leitet eine neue Phase der Krebsforschung ein, an die große Erwartungen für die Erkennung und Behandlung von Krebserkrankungen geknüpft werden. Die bisherigen Erkenntnisse, insbesondere über Onkogene und Tumorsuppressorgene, sind u. a. Gegenstand dieses Artikels.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Der Onkologe 5 (1999), S. 847-854 
    ISSN: 1433-0415
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: In den westlichen Industrieländern stellen Tumorerkrankungen z. Z. die zweithäufigste Todesursache nach Erkrankungen des Kreislaufsystems dar. Im Laufe des letzten Jahrzehnts ist vor allem durch die Entwicklung neuer molekularbiologischer Analysemethoden deutlich geworden, daß„Krebs” eine Krankheit ist, die durch genetische Veränderungen bedingt wird. Die Entstehung von malignen Tumoren ist ein komplexer mehrschichtiger Prozeß, der durch eine Anhäufung von Mutationen im Genom der betroffenen Zelle charakterisiert ist. Mutationen können durch physikalische Noxen (z. B. UV-Licht), chemische Karzinogene (z. B. Tabakrauch) oder onkogene Viren verursacht werden [11, 33, 50]. Für die Kanzerogenese und Progression von Tumoren sind vor allem Mutationen dreier bestimmter Gengruppen entscheidend, die als Onkogene, Tumorsuppressorgene und Mutatorgene bezeichnet werden.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1569-8041
    Keywords: Ewing tumor ; primary metastases ; survival
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: In the multicenter European Intergroup Cooperative Ewing's Sarcoma Studies, localized Ewing tumors of bone were treated by combination chemotherapy with surgery and/or radiotherapy. Patients with primary metastases (pm-pts) were treated in high risk protocols. Patients and methods: One hundred seventy-seven pm-pts were registered from January 1990 to December 1995, 171 were evaluable for survival analyses. Thirty-six pm-pts received myeloablative megatherapy with stem cell rescue following conventional treatment. Bilateral whole lung irradiation (WLI) was administered in 57 pm-pts with pulmonary involvement. Event-free survival (EFS) rates were estimated by Kaplan–Meier analysis. Prognostic factors were identified by log-rank statistics, Cox procedures and logistic regression. Results: Eighty-nine deaths were recorded by 1 February 1997, EFS four years after diagnosis for all 171 pm-pts was 0.27. EFS for isolated lung metastases was 0.34, for bone/bone marrow (BM) metastases, 0.28, and for combined lung plus bone/BM metastases, 0.14 (P 〈 0.005). WLI improved outcome in case of isolated pulmonary involvement (0.40 vs. 0.19, P 〈 0.05). In pm-pts with combined pulmonary/skeletal metastases, intensification by megatherapy and/or WLI improved EFS from 0.00 to 0.27 (P = 0.0001). Conclusions: EFS four years after diagnosis in patients with disseminated Ewing tumors is 0.27. Whole lung irradiation and megatherapy improve outcome in subgroups of patients with disseminated Ewing disease.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1569-8041
    Keywords: neuroblastoma ; prognosis ; telomerase activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Treatment of neuroblastoma has remained a major challenge in pediatric oncology because the assessment of the individual prognosis, particularly in disseminated disease is still obscure. Previous studies have correlated clinical outcome with activity levels of telomerase, a cellular reverse transcriptase which has been detected in the majority of human malignant tumors. Patients and methods: In this blind-trial study, a non-radioactive telomeric repeat amplification protocol (TRAP) with an internal telomerase-assay standard was used on an automated laser fluorescence sequencer for the detection and semiquantitative analysis of telomerase activity (TA) in 67 neuroblastomas of all clinical stages from the German Neuroblastoma Trial and 2 ganglioneuromas. TA levels were correlated with event-free and overall survival rates and established prognostic markers such as MYCN. Results: TA was present in 14 of 69 (20%) samples, including 3 of 22 stage IVS, 8 of 14 stage IV, 1 of 10 stage III, 1 of 7 stage II and 1 of 14 stage I neuroblastomas and 0 of 2 ganglioneuromas. We found a strong statistical correlation between the presence of TA and poor clinical prognosis with regard to all tumor stages. Multivariate analysis revealed TA as an independent prognostic marker. In particular, the analysis of TA in IVS neuroblastomas distinguished two different prognostic groups. Conclusions: Our data suggest that TA is an independent prognostic marker in neuroblastoma which, in combination with other markers such as MYCN, may proof useful in assessing the individual patient's prognosis.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1437-9813
    Keywords: Key words Ovarian tumor ; Children ; Hypercalcemia ; Small-Cell carcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Tumors of the ovary in girls represent about 80% of pediatric genital tumors; approximately 30% of these tumors are malignant. The risk of malignancy increases with decreasing age. The most frequent finding is a teratoma; other tumors are rare. Small-cell carcinoma (SCCO) of the ovary is extremely rare, occurring mostly in young women. We present an 8-year-old girl with a SCCO of the hypercalcemic type. The findings and treatment are discussed with emphasis on the poor prognosis in these patients, even in stage 1 disease. The current literature is reviewed.
    Type of Medium: Electronic Resource
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