Analytical Cellular Pathology - Volume 22, issue 4

Authors: Hannen, Egied J.M. | van der Laak, Jeroen A.W.M. | Kerstens, Harold M.J. | Cuijpers, Vincent M.J.I. | Hanselaar, Antonius G.J.M. | Manni, Johannes J. | de Wilde, Peter C.M.

Article Type: Research Article

Abstract: The aims of this study of head and neck tissue samples were to develop an immunohistochemical protocol based on the catalysed reporter deposition (CARD) technique to enhance staining results for use in automated true colour image analysis, to assess the reproducibility of systematic tissue sampling in the angiogenic hot spot selection, and quantification of microvessel density (MVD) and other vessel characteristics. The latter data were compared between six metastasised tongue squamous cell carcinomas, vs. four non‐metastasised. In comparison to the standard immunohistochemical protocol with anti‐CD34 antibodies, CARD amplification resulted in both more intensely stained and larger numbers of vessels. Averaging …the 10 most vascularised fields of the 40 to 60 systematically sampled fields in a tissue section resulted in an overall acceptable interobserver reproducibility for most assessed vessel parameters (r≥0.76 and p≤0.01). The percentage vessels with diameter <5 μm was significantly higher in the non‐metastasised tongue carcinomas (p=0.02). However, for a number of tumours the effect of tissue sampling was significant. We conclude that CARD amplification is needed for reliable segmentation of vessels by image analysis systems, and that tumour heterogeneity is a limiting factor for all procedures in which tumour vascularity is assessed in a single tissue section. Figures on http://www.esacp.org/acp/2001/22‐4/hannen.htm. Show more

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 183-192, 2001

Authors: Tomobe, Mitsuro | Shimazui, Toru | Uchida, Katsunori | Akaza, Hideyuki

Article Type: Research Article

Abstract: Purpose: We have previously demonstrated that the AgNOR count in proliferating cells is a predictor of tumor recurrence in superficial bladder tumor (J. Urol. 162 (1999), 63–68). In the present study, we evaluate the type of AgNOR associated with cell cycles as a prognostic factor in invasive bladder tumor using a double staining technique employing both AgNOR and MIB‐1 labelling. Materials and methods: Forty‐four paraffin sections of invasive bladder tumors were stained simultaneously with AgNOR and MIB‐1. The number of AgNORs in proliferating (MIB‐1 positive) or resting (MIB‐1 negative) cells were counted from a total of 100 nuclei. Correlations …between MIB‐1 associated AgNOR count and clinicopathological parameters were statistically analyzed. Results: The AgNOR count in proliferating cells (proliferating NOR) was significantly higher than that in resting cells (resting NOR) (p<0.01). The resting NOR in tumors with distant metastases was significantly higher than that in tumors without metastases (p<0.05). Patients with a low resting NOR tumor had a better prognosis than those with a high resting NOR tumor, whereas the proliferating NOR was not associated with survival. Survival analysis revealed that the resting NOR was the most powerful prognostic marker in patients with invasive bladder tumor (p<0.05). Conclusions: Resting NOR had a predictive value in the prognosis of patients with invasive bladder tumor. Show more

Keywords: Transitional cell carcinoma, invasive, resting cell, AgNORs, MIB‐1

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 193-199, 2001

Authors: Lenander, C. | Habermann, J.K. | Öst, Å. | Nilsson, B. | Schimmelpenning, H. | Tryggvason, K. | Auer, G.

Article Type: Research Article

Abstract: Expression of the γ2 chain at the invasive front of different tumors has indicated an important role for laminin‐5 in cell migration during tumor invasion and tissue remodeling. As there is considerable need for reliable invasion and prognostic markers we evaluated the correlation of laminin‐5 γ2 chain expression with clinicopathologic parameters and patient survival in 93 primary colon carcinomas. Epithelial cells of normal mucosa were consistently negative for staining. In contrast, positive cytoplasmic staining was observed in 89 tumors (96%). Twenty‐four (26%) cases were scored as sparse, 34 (37%) as moderate, and 31 (33%) as frequent γ2 chain expression. There …was a significant association of laminin‐5 γ2 chain expression and local invasiveness of colon carcinomas according to Dukes stage (A‐C) (p=0.001) and tumor budding (p<0.001). A statistical significance could also be noted in decreasing tumor differentiation (p<0.001) and correlation to tumor size (p=0.032). No correlation was observed to tumor site. Univariate analysis identified laminin‐5 (p=0.010), tumor differentiation (p=0.006) and Dukes grade (p<0.001) as significant variables in predicting prognosis. However, by multivariate analyses, this study could not demonstrate that laminin‐5 γ2 chain expression is an independent predictive factor for survival. The results indicate that laminin‐5 γ2 chain expression is up‐regulated during the progression of human colon cancer and that it plays a role in the aggressiveness of these tumors. Demonstration of laminin‐5 γ2 chain positivity also facilitates detection of individual cells or minor cell clusters invading the surrounding stroma. Figures on http://www.esacp.org/acp/2001/22‐4/lenander.htm. Show more

Keywords: Laminin‐5 γ2 chain, colon carcinoma, immunohistochemistry

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 201-209, 2001

Authors: Remmerbach, Torsten W. | Weidenbach, Horst | Pomjanski, Natalja | Knops, Kristiane | Mathes, Stefanie | Hemprich, Alexander | Böcking, Alfred

Article Type: Research Article

Abstract: Objective. The aim of this prospective study was to report on the diagnostic accuracy of conventional oral exfoliative cytology taken from white‐spotted, ulcerated or other suspicious oral lesions in our clinic. In addition we checked DNA‐image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA‐aneuploidy is a sensitive and specific marker for the early identification of tumor cells in oral brushings. Study design. 251 cytological diagnoses obtained from exfoliative smears of 181 patients from macroscopically suspicious lesions of the oral mucosa and from clinically seemingly benign oral lesions which were exisiced for establishing histological diagnoses were compared with …histological and/or clinical follow‐ups of the respective patients. Additionally nuclear DNA‐contents were measured after Feulgen restaining using a TV image analysis system. Results. Sensitivity of our cytological diagnosis on oral smears for the detection of cancer cells was 94.6%, specificity 99.5%, positive predictive value 98.1% and negative predictive value 98.5%. DNA‐aneuploidy was assumed if abnormal DNA‐stemlines or cells with DNA‐content greater 9c were observed. On this basis the prevalence of DNA‐aneuploidy in smears of oral squamous cell carcinomas in situ or invasive carcinomas was 96.4%. Sensitivity of DNA‐aneuploidy in oral smears for the detection of cancer cells was 96.4%, specificity 100%, positive predictive value 100% and negative 99.0%. The combination of both techniques increased the sensivity to 98.2%, specificity to 100%, positive predictive value to 100% and negative to 99.5%. Conclusions. Brush cytology of all visible oral lesions, if they are clinically considered as suspicious for cancer, are an easily practicable, cheap, non‐invasive, painless, safe and accurate screening method for detection of oral precancerous lesions, carcinoma in situ or invasive squamous cell carcinoma in all stages. We conclude that DNA‐image cytometry is a very sensitive, highly specific and objective adjuvant tool for the early identification of neoplastic epithelial cells in oral smears. Show more

Keywords: Oral cancer, oral exfoliative cytology, DNA‐aneuploidy, DNA‐image cytometry, cancer screening, diagnostic accuracy

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 211-221, 2001

Authors: Tzavelas, C. | Smith, P. | Horefti, E. | Rickwood, D.

Article Type: Research Article

Abstract: Immunoporation is a novel method of cell transfection based upon the use of a new type of beads, Immunofect beads, that can be targeted to make holes in different types of cells depending on the type of bead used. It is known that the efficiency of transfection of cells by some techniques can be affected by the presence of serum and another important factor that appears to affect transfection efficiency and cell viability is the osmolarity of the transfection medium. This report presents studies on the effects of serum and varying osmolarity on the efficiency of transfection using immunoporation. The …results clearly indicate that in hypertonic media the presence of serum decreases the efficiency of transfection. In the case of osmolarity, increasing the osmolarity of the immunoporation medium increases the efficiency of transfection but above about 650 mOsm this increasing efficiency is offset by the much lower viability of the cells. Show more

Keywords: Immunoporation, transfection, osmolarity, serum, HL‐60

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 223-227, 2001

Authors: Heiskanen, Mervi | Kononen, Juha | Bärlund, Maarit | Torhorst, Joachim | Sauter, Guido | Kallioniemi, Anne | Kallioniemi, Olli

Article Type: Research Article

Abstract: Multiple regions of the genome are often amplified during breast cancer development and progression, as evidenced in a number of published studies by comparative genomic hybridization (CGH). However, only relatively few target genes for such amplifications have been identified. Here, we indicate how small‐scale commercially available cDNA and CGH microarray formats combined with the tissue microarray technology enable rapid identification of putative amplification target genes as well as analysis of their clinical significance. According to CGH, the SUM‐52 breast cancer cell line harbors several high‐level DNA amplification sites, including the 10q26 chromosomal region where the fibroblast growth factor receptor 2 …(FGFR2) gene has been localized. High level amplification of FGFR2 in SUM‐52 was identified using CGH analysis on a microarray of BAC clones. A cDNA microarray survey of 588 genes showed >40‐fold overexpression of FGFR2. Finally, a tissue microarray based FISH analysis of 750 uncultured primary breast cancers demonstrated in vivo amplification of the FGFR2 gene in about 1% of the tumors. In conclusion, three consecutive microarray (CGH, cDNA and tissue) experiments revealed high‐level amplification and overexpression of the FGFR2 in a breast cancer cell line, but only a low frequency of involvement in primary breast tumors. Applied to a genomic scale with larger arrays, this strategy should facilitate identification of the most important target genes for cytogenetic rearrangements, such as DNA amplification sites detected by conventional CGH. Figures on http://www.esacp.org/acp/2001/22‐4/heiskanen.htm Show more

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 229-234, 2001

Article Type: Other

Citation: Analytical Cellular Pathology, vol. 22, no. 4, pp. 235-237, 2001