Prognostic significance of new immunohistochemistry scoring of p53 protein expression in cutaneous squamous cell carcinoma of mice.

This study was conducted to investigate the prognostic value and therapeutic response of treatment modalities on p53 protein expression and AgNOR index in squamous cell carcinoma (SCC). Furthermore, based on data, we proposed a new p53 immunohistochemistry (IHC) scoring system. Sixty albino mice were given 7,12-dimethylbenz[a]anthracine (DMBA) and 12-O-tetradecanoly Phorbol-13-acetate (TPA) to produce skin tumors. The retinoids were given after the development of tumors. p53 immunohistochemical and AgNOR staining was performed on the sections taken before and after the retinoid administration. p53 protein was expressed in 31 of the lesions (60.8%). AgNOR index was high in all 51 (100%) of the pretreated lesions. There was a marked decrease in the expression of p53 protein in 16/51 (31.4%) and AgNOR index in 36/51 (70.6%) in post-treated mice. There was no decrease in the expression of both markers in mice harboring malignant neoplasms. p53 IHC scores were 0, I, and II in epidermal hyperplasia, papilloma, and dysplasia, respectively, while they were II, III, IV, and V in SCC in situ, well-differentiated, moderately differentiated, and poorly differentiated SCCs, respectively. Alteration of p53 and AgNOR index occured during the development of SCC. The p53 IHC scores are directly related to the grades of malignancy. Both markers might be used as a supportive tool with routinely performed Hematoxylin and Eosin staining and may help in the diagnosis of SCC. The newly proposed p53 IHC scoring system will help histopathologists in making their differential diagnosis among benign, premalignant, and malignant lesions. It will also help the oncologists to assess the prognosis and effectiveness of their chemotherapy.

Modified method of AgNOR staining for tissue and interpretation in histopathology.

This study was conducted in the department of Pathology King Edward Medical University, from June to December 2002 to introduce the new method of AgNOR staining and its interpretation to increase its reliability. A total of 60 brain specimens were stained with modified AgNOR technique. The diagnosis of malignancy was made on H & E staining. AgNOR counts, variation in size and dispersion of AgNOR dots in cells were graded and compared in malignant and non-malignant lesions. Modified method of AgNOR staining and interpretation was an easy, reliable and reproducible alternative to traditional AgNOR techniques for evaluating proliferation activity of cells in malignant and benign brain lesions. mAgNOR counts of different grades of astrocytoma (2.97+/-0.96, 3.97+/-0.43, 6.01+/-2.74 and 8.01+/-3.56) were significantly (P<0.01) greater when compared with counts of normal brain (0.40+/-0.01), and reactive gliosis (0.60+/-0.01). AgNOR size and dispersion were of higher grade in a significantly greater proportion of malignancy when compared with benign conditions (P<0.05). The AgNOR dots were brighter and more clear with modified staining when compared with previous studies. We conclude that modified AgNOR staining technique is simple, quick and reliable to evaluate cell proliferation by detecting AgNORs size and dispersion. In future, AgNOR size and dispersion should be considered rather than the count only. We recommend the use of morphometry for AgNOR size in future. We also recommend the use of modified AgNOR staining for obtaining sound and confidant results in routine paraffin sections.

Patterns of argyrophilic nucleolar organiser regions in pleural and peritoneal effusions.

OBJECTIVE: To evaluate AgNOR size and dispersion as alternate methods to AgNOR counts in order to differentiate malignant from non-malignant effusions. DESIGN: Comparative study. PLACE AND DURATION OF STUDY: Department of Pathology, Postgraduate Medical Institute, Lahore, from January 2003 to June 2004. MATERIALS AND METHODS: A total of 240 samples of pleural and peritoneal effusions were centrifuged, deposits smeared on slides and stained with H and E and AgNOR stain. The diagnosis of malignancy or otherwise was made on H and E staining. AgNOR counts, variation in size and dispersion of AgNOR dots in smears were graded and compared in malignant and non-malignant effusions. RESULTS: Mean AgNOR counts of 11.47+/-3.60 and 11.04+/-3.89 in malignant pleural and peritoneal effusions, respectively, were significantly (p<0.0001) greater as compared with counts of 3.36+/-0.69 and 3.35+/-0.66 in non-malignant effusions. AgNOR size and dispersion were of higher grade in significantly greater proportion of malignant as compared with non-malignant effusions (p<0.0001). CONCLUSION: Typing of AgNOR size and dispersion was found to be an easy and reproducible alternative to traditional AgNOR counts for differentiating malignant from non-malignant effusions. These parameters should be correlated with already established but expensive techniques of AgNOR area and size imaging by electron microscopy and flow cytometry, as an economical alternative.