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  • 1
    Electronic Resource
    Electronic Resource
    The use of fluorescent in situ hybridization for detection of the t(2;5)(p23;q35) translocation in anaplastic large-cell lymphoma (1997)
    Springer
    Annals of oncology 8 (1997), S. 65-69 
    Details
    ISSN: 1569-8041
    Keywords: anaplastic large-cell lymphoma ; fluorescent in situ hybridization ; Hodgkin's disease ; Ki-1 lymphoma ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Anaplastic large-cell lymphoma (ALCL) is a recently recognized, distinctive type of non-Hodgkin's lymphoma characterized by anaplastic large-cell cytology and expression of a member of the TNF-receptor family CD30. A characteristic chromosomal translocation has been identified in ALCL of T- or null-cell lineage which juxtaposes a novel tyrosine kinase (anaplastic lymphoma kinase, ALK) located at 2p23 with the nucleophosmin gene (NPM) at 5q35. A chimeric mRNA transcript is produced, and the translocation results in constitutive expression of a truncated form of the ALK protein, p80. There is controversy concerning whether or not the translocation occurs in Hodgkin's disease. The aim of this study was to develop a methodology for fluorescent in situ hybridization (FISH) to detect the t(2;5)(p23;q35), and to compare the results with conventional cytogenetics, reverse-transcriptase PCR and immunostaining for the p80protein. Patients and methods: Twenty-five cases of malignant lymphoma (11 ALCL and 14 HD) were studied. Immunohistochemistry was performed to confirm the diagnosis and for analysis of p80 expression. Conventional cytogenetics were analyzed on G-banded metaphase spreads. FISH was performed using whole chromosome paints for chromosomes 2 and 5 on metaphase spreads and YAC probes for inter phase nuclei. Reverse-transcriptase PCR using primers for ALK and NPM was used to amplify the translocation breakpoint in extracted mRNA. Results: Among 11 cases of ALCL examined by FISH, the translocation was detected in 4. Two of these cases also had RT-PCR and p80 staining performed, with positive results. Among 7 cases where the t(2;5) was not detected by FISH, 3 cases were examined by RT-PCR with negative results and4 cases by p80 staining, also negative. The RT-PCR was negative in all 14cases of Hodgkin's disease, 4 of which were also examined by FISH and found to be negative. Conclusion: Fluorescent in situ hybridization is useful method for detection of the t(2;5)(p23;q35) in anaplastic large-cell lymphoma. The results concur with those of RT-PCR for the chimeric transcript and immunostaining for the p80 protein. The frequency with which the translocation was found was 36% in this small series, and no evidence of the translocation was found in cases of Hodgkin's disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008234301024
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  • 2
    Electronic Resource
    Electronic Resource
    The use of fluorescent in situ hybridization for detection of the t(2;5)(p23;q35) translocation in anaplastic large-cell lymphoma (1997)
    Springer
    Annals of oncology 8 (1997), S. 65-69 
    Details
    ISSN: 1569-8041
    Keywords: anaplastic large-cell lymphoma ; fluorescent in situ hybridization ; Hodgkin's disease ; Ki-1 lymphoma ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Anaplastic large-cell lymphoma (ALCL) is a recentlyrecognized, distinctive type of non-Hodgkin's lymphoma characterized byanaplastic large-cell cytology and expression of a member of theTNF-receptor family CD30. A characteristic chromosomal translocation hasbeen identified in ALCL of T- or null-cell lineage which juxtaposes a noveltyrosine kinase (anaplastic lymphoma kinase, ALK) located at 2p23 with thenucleophosmin gene (NPM) at 5q35. A chimeric mRNA transcript is produced,and the translocation results in constitutive expression of a truncated formof the ALK protein, p80. There is controversy concerning whether or not thetranslocation occurs in Hodgkin's disease. The aim of this study was todevelop a methodology for fluorescent in situ hybridization (FISH) to detectthe t(2;5)(p23;q35), and to compare the results with conventionalcytogenetics, reverse-transcriptase PCR and immunostaining for the p80protein. Patients and methods: Twenty-five cases of malignant lymphoma (11 ALCLand 14 HD) were studied. Immunohistochemistry was performed to confirm thediagnosis and for analysis of p80 expression. Conventional cytogenetics wereanalyzed on G-banded metaphase spreads. FISH was performed using wholechromosome paints for chromosomes 2 and 5 on metaphase spreads and YACprobes for interphase nuclei. Reverse-transcriptase PCR using primers forALK and NPM was used to amplify the translocation breakpoint in extractedmRNA. Results: Among 11 cases of ALCL examined by FISH, the translocation wasdetected in 4. Two of these cases also had RT-PCR and p80 stainingperformed, with positive results. Among 7 cases where the t(2;5) was notdetected by FISH, 3 cases were examined by RT-PCR with negative results and4 cases by p80 staining, also negative. The RT-PCR was negative in all 14cases of Hodgkin's disease, 4 of which were also examined by FISH and foundto be negative. Conclusion: Fluorescent in situ hybridization is useful methodfor detection of the t(2;5)(p23;q35) in anaplastic large-cell lymphoma. Theresults concur with those of RT-PCR for the chimeric transcript andimmunostaining for the p80 protein. The frequency with which the translocationwas found was 36% in this small series, and no evidence of thetranslocation was found in cases of Hodgkin's disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008234301024
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Treatment of mantle-cell lymphoma with Rituximab (chimeric monoclonal anti-CD20 antibody): Analysis of factors associated with response (2000)
    Springer
    Annals of oncology 11 (2000), S. 117-121 
    Details
    ISSN: 1569-8041
    Keywords: anti-CD20 ; chimeric monoclonal antibody ; mantle-cell lymphoma ; R.E.A.L. Classification ; Rituximab
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background:A retrospective analysis was performed to delineatethe factors associated with response, and to determine the duration ofresponse, in 87 patients with CD20–positive mantle-cell lymphoma (MCL) treatedwith Rituximab (chimeric monoclonal anti-CD20 antibody) in two prior studies. Patients and methods:Patients with newly-diagnosed MCL (MCL1,n = 37), and previously-treated MCL (MCL2, n = 50), receivedsingle-agent Rituximab, in the context of two multicentre clinical studiesusing different schedules and doses, conducted in 1996 and 1997. A follow-upanalysis was performed at the end of 1998, including all 81 patients whocompleted therapy. Statistical modeling of factors associated with responsewas performed using ordered logistic regression. The duration of complete (CR)and partial response (PR), and the time to disease progression (TTP), werealso derived. Results:The overall response rate (RR) was 34% (30 of 87)(81 evaluable patients, RR 37%; CR 14%), and was equivalent forMCL1 and MCL2. On univariate analysis, elevated LDH (P = 0.004);prior therapy with alkylating agents (P = 0.01) or fludarabinephosphate (P = 0.04); WHO performance status = 2 (P = 0.02);MCL2 refractory to last prior therapy (P = 0.04); and splenomegaly(P = 0.04), each at the time of treatment with Rituximab, weresignificantly associated with a lower RR. On multivariate analysis, only LDH(P = 0.007) and prior alkylating agents (P = 0.03) retainedstatistical significance. At a median follow-up of 1.4 years, the median TTP was 7 months. The medianduration of response was one year, and was significantly longer for patientsachieving CR vs. PR (P = 0.04). Conclusions:Rituximab is active in MCL, and can induce completeresponses in a minority of patients. Elevated LDH at the time of therapy, andprior therapy with alkylating agents, are associated with a significantlylower RR. The duration of response of one year is similar to that previouslyreported in follicular lymphoma.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008309405678
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    Variability of polymerase chain reaction detection of the bcl-2–IgH translocation in an international multicentre study (1999)
    Springer
    Annals of oncology 10 (1999), S. 1349-1354 
    Details
    ISSN: 1569-8041
    Keywords: lymphoma ; minimal residual disease ; molecular diagnosis ; polymerase chain reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: The capacity of the polymerase chain reaction (PCR) to detect very low numbers of cells bearing a t(14;18) translocation has led to its application in assessment of the results of treatment for follicular lymphoma, and suggestions that therapy might be guided by molecular studies. To test the reliability of PCR a collaborative study was undertaken to compare results from different laboratories in Europe and North America. Methods: Twenty laboratories with records of publication in molecular diagnostics were sent blood from normal donors with varying numbers of t(14;18)-bearing cells added from a cell line with a translocation in the major breakpoint region (MBR) of the bcl-2 gene. Samples contained 1000, 100, 10, 1 or 0 cells per ml of whole blood and were sent blinded in duplicate. PCR methodology varied widely, with the total number of amplification cycles between 30 and 70, and 13 different primers used for the MBR region. Twelve laboratories used nested PCR and eight single round amplification. Results: The sensitivity of nested and single round PCR was similar at 100 cells/ml but below this the nested method proved significantly more sensitive. The false positive rate was 28%, with 11 samples from 9 laboratories reported as positive when no t(14;18) cells were added. PCR product size and sequence analysis showed that false positives were due to contamination from cell-line DNA rather than background translocations in the donors. There was no significant difference in false positive rates between nested and single round techniques. Conclusion: The polymerase chain reaction to detectbcl-2–IgH rearrangements is presently carried out with widely disparate results. Further effort is required to bring forward a standard PCR protocol which can be re-tested in different laboratories to improve accuracy and reproducibility. The application of quantitative techniques such as real-time PCR may resolve many of the problems presently encountered.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008385924543
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    Paper (German National Licenses)
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