Clinical molecular pathology最新文献

{"title":"Cyclin D1 amplification and expression in human breast carcinoma: correlation with histological prognostic markers and oestrogen receptor expression.","authors":"S D Worsley,&nbsp;B A Jennings,&nbsp;K H Khalil,&nbsp;M Mole,&nbsp;A C Girling","doi":"10.1136/mp.49.1.m46","DOIUrl":"https://doi.org/10.1136/mp.49.1.m46","url":null,"abstract":"<p><p>Aims-To study the amplification of the Cyclin D1 gene (CCND1) in human breast carcinoma; to relate this to Cyclin D1 protein expression; to relate these parameters to recognised pathological prognostic factors, including oestrogen receptor (ER) status.Methods-DNA extracted from frozen sections of breast tumours (n = 36) was used for Southern blotting. Probes for CCND1, c-myc and the immunoglobulin heavy chain locus (IgH) were hybridised to tumour DNA. Immunocytochemical expression of Cyclin D1 protein and ER was studied in paraffin wax sections from the same tumours.Results-Amplification of CCND1 was observed in 11% (four of 36) of tumours studied. Over expression of Cyclin D1 protein was observed in 73% (30/41) of tumours. There was no correlation between recognised histological prognostic markers and either gene amplification or expression. However, a weak association was seen between Cyclin D1 expression and ER status.Conclusions-A disparity exists between locus amplification and over expression of Cyclin D1, suggesting the existence of another mechanism for raised protein expression. No significant correlation was detected between either Cyclin D1 amplification or over expression and established prognostic markers.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1996-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.49.1.m46","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26021481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insulin-like growth factors and IGF binding proteins in cyst fluid from patients with craniopharyngioma prior to intracavitary irradiation with Yttrium and thereafter.","authors":"W Zumkeller,&nbsp;M Sääf,&nbsp;T Rähn","doi":"10.1136/mp.49.1.m51","DOIUrl":"https://doi.org/10.1136/mp.49.1.m51","url":null,"abstract":"<p><p>Aim-To examine a series of cyst fluid samples from patients with craniopharyngioma at various stages of treatment in order to evaluate the use of insulin-like growth factors (IGFs) and IGF binding proteins as tumour markers or indicators of successful treatment, or both.Methods-Cyst fluid samples were obtained by stereotactic puncture prior to the intracavitary application of (90)Yttrium and at subsequent occasions. Analysis was performed by gel chromatography, radio-immunoassays, binding studies, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) with subsequent western blotting.Results-IGF-I, -II and IGF binding protein-1 concentrations were measured in three craniopharyngioma cyst fluid samples. Immunoreactive IGF-I and IGF binding protein-1 concentrations in these three samples were between 6 and 29 ng/ml, and 17 and 48 ng/ml, respectively. In contrast, the IGF-II concentrations measured in 19 cyst fluid samples from seven patients with craniopharyngioma at various stages of treatment were much higher at 25-671 ng/ml. SDS-PAGE and subsequent western blotting using [(125)I]IGF-II as the ligand gave bands with estimated molecular weights of 330, 220, 135, 96, 46, 43, 34, 29, and 13.5 kDa in one adult, and identical bands at 220, 41.5, 37.5, 32, and 19 kDa in three cyst fluid samples from three children with craniopharyngioma.Conclusions-These results suggest that IGFs and IGF binding proteins are secreted by craniopharyngiomas and that they may alter the growth characteristics of these tumours. Furthermore, the distinct pattern of IGF binding protein sizes might be used as a tool for the differential diagnosis of tumours of the central nervous system.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1996-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.49.1.m51","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26021482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MDM-2 protein expression in nasopharyngeal carcinomas. Comparative study with p53 protein expression.","authors":"P Kanavaros,&nbsp;C Kouvidou,&nbsp;Y Dai,&nbsp;M Tzardi,&nbsp;G Datseris,&nbsp;K Darivianaki,&nbsp;D Rontogianni,&nbsp;G Delides","doi":"10.1136/mp.48.6.m322","DOIUrl":"https://doi.org/10.1136/mp.48.6.m322","url":null,"abstract":"<p><p>Aims-To investigate the immunohistochemical expression of MDM-2 protein in comparison with that of p53 protein in nasopharyngeal carcinomas.Methods-Formalin fixed, paraffin wax embedded tissue from 59 cases of nasopharyngeal carcinoma was stained by immunohistochemistry for MDM-2 and p53 proteins.Results-The tumours were divided histologically into seven cases of keratinising nasopharyngeal carcinoma (KNPC), 14 cases of non-keratinising nasopharyngeal carcinoma (NKNPC), and 38 cases of undifferentiated nasopharyngeal carcinoma (UNPC). MDM-2 nuclear expression was observed in 0/7 KNPC, 1/14 NKNPC, and 11/38 UNPC. p53 nuclear expression was observed in 1/7 KNPC, 2/14 NKNPC, and 15/38 UNPC. Parallel MDM-2 and p53 expression was found in 12 cases (11 UNPC and one NKNPC). Discordant MDM-2-/p53 + expression was found in six cases (four UNPC, one NKNPC, and one KNPC), and absence of expression of both proteins in the remaining 41 cases.Conclusions-Expression of MDM-2 and p53 proteins may be associated with the level of tumour cell differentiation in nasopharyngeal carcinoma. Simultaneous expression of MDM-2/p53 in a proportion of UNPC suggests that MDM-2 protein may be responsible for stabilisation of p53 protein in these cases, in view of the previous demonstration of the p53 gene in germ line configuration. This could be important in the pathogenesis of these cases, since MDM-2 may deregulate the p53 dependent growth suppressive pathway. Discordant MDM-2-/p53 + expression in a few cases of nasopharyngeal carcinoma may reflect stabilisation of p53 protein by other proteins, or p53 mutations unable to activate MDM-2.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m322","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26020569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Our first year.","authors":"","doi":"10.1136/mp.48.6.m291","DOIUrl":"https://doi.org/10.1136/mp.48.6.m291","url":null,"abstract":"","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m291","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26020562","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The mammalian mitogen activated protein kinase network.","authors":"P Lenormand","doi":"10.1136/mp.48.6.m292","DOIUrl":"https://doi.org/10.1136/mp.48.6.m292","url":null,"abstract":"","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m292","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26020564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ras and p53 in the prediction of survival in Dukes' stage B colorectal carcinoma.","authors":"M A Bennett,&nbsp;E W Kay,&nbsp;H Mulcahy,&nbsp;L O'flaherty,&nbsp;D P O'donoghue,&nbsp;M Leader,&nbsp;D T Croke","doi":"10.1136/mp.48.6.m310","DOIUrl":"https://doi.org/10.1136/mp.48.6.m310","url":null,"abstract":"<p><p>Aims-To determine possible associations between p53 allelic deletion, c-Ki-ras mutational activation, immunohistochemical detection of p53 and ras proteins, various clinicopathological variables, and patient outcome in 168 Dukes' stage B colorectal carcinomas.Methods-Allelic deletion at the p53 tumour suppressor gene locus was detected using polymerase chain reaction (PCR) based loss of heterozygosity (LOH) assays. Overexpressed proteins were detected using the CM1 polyclonal antibody. A PCR based assay was used to detect the presence of activating mutations at codon 12 of c-Ki-ras. Immunostaining was carried out using a monoclonal antibody to p21(ras).Results-p53 LOH, CM1 immunostaining, c-Ki-ras mutational activation, and p21(ras) immunostaining were not predictive of survival by logrank analysis. Multivariate analysis using Cox regression did not predict survival in this group of tumours.Conclusions-Aberrations in ras and p53 are unlikely to play an important role in the subdivision of patients with Dukes' stage B colorectal carcinoma into more accurate prognostic strata. It is possible that later genetic events are more important in conferring a specific phenotype on the resultant Dukes' stage B tumour.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m310","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26020566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of human leucocyte antigen genes in the development of malignant disease.","authors":"W M Howell,&nbsp;D B Jones","doi":"10.1136/mp.48.6.m302","DOIUrl":"https://doi.org/10.1136/mp.48.6.m302","url":null,"abstract":"","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m302","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26020563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Demonstration of CD44 gene expression in cells from fine needle aspirates of breast lesions by the polymerase chain reaction.","authors":"J Bolodeoku,&nbsp;K Yoshida,&nbsp;P Yeomans,&nbsp;C A Wells,&nbsp;D Tarin","doi":"10.1136/mp.48.6.m307","DOIUrl":"https://doi.org/10.1136/mp.48.6.m307","url":null,"abstract":"<p><p>Aim-To demonstrate the feasibility of studying specific gene expression in fine needle aspirates from clinical lesions. The reverse transcription/polymerase chain reaction (RT-PCR) technique was used to demonstrate CD44 gene expression in cells from diagnostic fine needle aspirates taken from patients attending the outpatient clinic for breast diseases.Methods-Polyadenylated RNA was extracted from the cells remaining in the syringe barrel after fine needle aspirate cytological diagnosis of 41 patients with breast lesions. Analysis of CD44 gene expression was performed by RT-PCR using primers flanking the site for insertion of the variant exons. The resulting products were separated on 1.2% agarose gels, transferred to nylon membranes using Southern blotting and hybridised with specific probes for standard (constitutive) and variant exons of this gene.Results-On hybridisation with the CD44 standard exon probe, the expected amplified product of approximately 482 base pairs was visualised in 22 of 41 samples examined. Further hybridisation with the \"variant\" exon probes (exons 7 (v2), 8 (v3), 9b (v4), 12 (v7), and 15 (v10)) on 12 of these samples showed the presence of large molecular variants in all of these samples. However, the expression pattern detected with the probes for exons 7 (v2), 8 (v3) and 9b (v4) differed among the patients.Conclusions-Expression of the standard and variant regions of the CD44 gene in cells remaining in the syringe after fine needle aspiration was demonstrated using RT-PCR. The 5' variant exon probes seemed to show different patterns of expression among the patients. Further studies are currently being conducted to determine whether there is any correlation between expression of the various components of this gene and cytological diagnosis. Using this method, it would be possible to study the expression of other candidate marker genes in breast cancer using fine needle aspirates.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m307","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26020565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"IGF-II dependent autocrine growth in cell lines derived from renal tumours of childhood.","authors":"W Zumkeller,&nbsp;A Mahmood,&nbsp;R Dellow,&nbsp;P N Schofield","doi":"10.1136/mp.48.6.m333","DOIUrl":"https://doi.org/10.1136/mp.48.6.m333","url":null,"abstract":"<p><p>Aims-To determine the role of insulin-like growth factors (IGF) in the proliferation of tumour cells, by studying the mitogenic response to IGFs of three cell lines of differing phenotype established from both malignant rhabdoid and Wilms tumour, representing a range of cell types (GOS 4, G401, and T3/73).Methods-Production of IGF-II and IGF-I was measured by radioimmunoassay, and the presence of IGF binding protein complexes was observed by gel exclusion chromatography. Following growth analyses in serum-free media to ascertain the dependence of the cell lines on exogenous IGFs, the generation of autocrine growth was measured by a density dependence assay of proliferation in culture. Receptors were measured by radioligand cross linking and autocrine growth through these receptors assayed by the use of blocking antibodies.Results-While GOS 4 and G401 were able to proliferate in serum-free medium over a period of 5 d, T3/73 showed an absolute dependence on IGFs added daily at 1-10 ng/ml. Plating at clonal density showed that cell growth was directly density dependent in serum-free medium. The serum independent proliferation of G401 and GOS 4 was blocked by the addition of an antibody to the type 1 IGF receptor (alpha-IR3) suggesting that the effects of autocrine factors are mediated through type 1 IGF receptors. S1 nuclease protection analysis indicated that all three cell lines produced significant amounts of mRNA derived mainly from the P3 IGF-II promoter, but transcripts for IGF-I were undetectable. Radioimmunoassay of IGFs from conditioned media showed that all the lines made assayable IGF-II (8.6, 8.4, and 6.1 ng/ml/24 h/10(6) cells for GOS 4, G401, and T3/73 respectively). The presence of species consistent with both type 1 and type II IGF receptors was demonstrated using radioligand binding to cell membranes followed by cross linking.Conclusions-Autocrine IGF-II may contribute to the serum independence of GOS 4 and G401 cells, whereas T3/73 may depend on exogenous IGF-II for proliferation.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m333","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26018981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of a nested polymerase chain reaction in the diagnosis of Pneumocystis carinii pneumonia.","authors":"R Evans,&nbsp;A W Joss,&nbsp;D Parratt,&nbsp;T H Pennington,&nbsp;D O Ho-Yen","doi":"10.1136/mp.48.6.m347","DOIUrl":"https://doi.org/10.1136/mp.48.6.m347","url":null,"abstract":"<p><p>Aim-To compare the techniques and results of a nested PCR and an immunofluorescence assay (IFA) for the detection of Pneumocystis carinii infection; to consider the role of the nested PCR in the diagnosis of P carinii pneumonia (PCP).Methods-Serial dilutions of two known P carinii positive samples were tested by IFA and PCR to determine their relative sensitivities. Seventy eight respiratory samples (15 from 11 patients with HIV infection/acquired immunodeficiency syndrome (AIDS) and 63 from 42 patients with other forms of immunodeficiency) were tested using both assays, and the costs and technical requirements of each assay were assessed.Results-The PCR had a greater relative sensitivity over the IFA of 2 x 10(1) to 2 x 10(3) fold in a postmortem lung sample and 2 x 10(5) to 2 x 10(6) fold in a bronchoalveolar lavage sample from a patient with PCP. P carinii was detected in all 15 samples from the patients with HIV/AIDS by both IFA and PCR. Of the 63 samples from the patients with immunodeficiencies other than HIV/AIDS, the PCR was more sensitive than IFA.Conclusions-The nested PCR is a more sensitive assay than the IFA. It may be useful in the diagnosis of PCP in patients with immunodeficiencies other than HIV/AIDS. Similarly, PCR may be of benefit for this patient group as less invasive specimens are needed. PCR has an increasing role to play in the diagnosis of PCP in the routine laboratory.</p>","PeriodicalId":87395,"journal":{"name":"Clinical molecular pathology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1136/mp.48.6.m347","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26018983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}