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Background: Sentinel lymph node biopsy is a widely accepted method for staging melanoma and breast cancer in indicated cases. However, the use of the method in colorectal cancer is under clinical investigation. The aim of the pilot study was to introduce the technique into the surgical practice in colon carcinoma, to determine the feasibility and potential problems and to evaluate the first experience., Methods: Twenty patients with colon cancer underwent lymphatic mapping and sentinel node biopsy using blue dye, fluorescein or lymphoscintigraphy followed by standard surgical resection. The acquired sentinel nodes were investigated with both standard hematoxylin-eosin staining and immunohistochemical staining for cytokeratin., Results: Lymphatic mapping adequately identified at least one sentinel node (SN) intraoperatively or by a modified ex vivo technique in 20 patients (100%). The average number of SN was 1.5 (range 1-3), non-SN 13.6 (range 1-38) per patient. SN correctly predicted the regional lymphatic basin status in 14 cases (70%). The false negative rate was 40%. No patient has been upstaged on the basis of immunohistochemical staining., Conclusions: Lymphatic mapping and sentinel node biopsy in colon cancer is feasible and safe method with a high SN identification rate. The role and significance of sentinel node biopsy in colon cancer is not as clear as its role in other tumors and remains controversial. Further large prospective studies with standardized techniques are needed to evaluate the potential benefit of this new method.

p53 status is an important predictive factor in breast cancer, but the results of many studies are ambiguous. We tested p53 by functional analysis of separated alleles in yeast (FASAY) as well as by immunohistochemistry (IHC) and evaluated correlations with main prognostic factors, proliferation, and Bcl-2. Thirty-two tumors were tested with antibodies BP53-12, DO1, DO11, DO12, and by FASAY. Spearman rank correlations were tested separately with age, tumor type, pT, grade, pN, NPI, Ki-67, S-phase, proliferation index, Bcl-2, and steroid receptor status determined by ER, PR, and pS2. FASAY showed significant correlations with ductal type, grade and proliferation, and an inverse correlation with functional estrogen receptor and Bcl-2. FASAY provided better correlations compared to p53 IHC. We conclude that FASAY shows significant correlations with main prognostic/predictive factors and provides more reliable biological information compared to p53 IHC. Apoptosis is positively linked to proliferation and is not under the control of p53, which is frequently mutated in highly proliferating carcinomas. FASAY seems to be very important in assessing the predictive significance of p53 for a specific therapy of breast cancer.

The surgeon is frequently faced with the task of bioptic sampling of tissues from patients with tumourous diseases. The importance of such frequently minor procedures is incorrectly underrated. The importance of biopsy is in the foreground in particular at present at a time of individualization of treatment of malignant tumours, based on the development of new diagnostic-therapeutic methods. The bioptic specimen is the fundamental link in the basic decision--benignity or malignity--and also a unique source for assessment of prognostic and predictive factors on the basis of which we select the optimal therapeutic procedure. Contrary to current histopathological examination where the principles of correct collection of tissues specimens are generally known, in recent years the importance of molecular examinations is increasing where the surgeon must respect certain different principles to make the sampling successful. The authors working in a department of surgical oncology along with authors from specialized laboratories formulate rules of correct implementation of biopsies and transport of biological material in conjunction with recent laboratory methods, and based on examples of their own practice, they demonstrate how the initial approach of the surgeon can influence in a decisive way the correct diagnosis and therapeutic procedure in oncological patients.

The immunological dysfunction associated with human cancer is well known phenomenon. It comprises number of pathological changes within immune network including imbalance in cytokines of Th1/Th2 origin. The objectives of our study were (i) to evaluate the abnormalities in serum levels of selected cytokines in malignant melanoma patients with regional lymph node metastases as compared to normal values, (ii) to examine the relationship between postoperative cytokine levels and disease progression and (iii) to correlate cytokine profile changes during IFN-alpha therapy with the disease progression and potential therapeutical response. Nine Th1/Th2 related cytokines and sIL-2R were determined in 26 malignant melanoma patients at clinical stage III prior and during adjuvant immunotherapy. Control group consisted of 26 healthy persons. Patients were treated with rIFN-alpha according to EORTC Melanoma group protocol 18952. Cytokines were quantified in patients sera using commercial ELISA kits. Majority of melanoma patients showed significantly lower values of IL-2 and IFN-gamma and pathologically elevated levels of IL-4, IL-6, IL-10 as compared to healthy subjects what indicates disease associated Th1/Th2 imbalance. In addition increased IL-12 and IL-15 values were noted in some patients (54% and 27%, respectively). All patients who manifested early relapse during immunotherapy had significantly lower pretreatment levels of IL-2 and IL-12 than those remaining without progression and probably benefiting from the treatment. Cytokine changes during immunotherapy disclosed that decreases in IL-2 and IL-12 and raises in IL-6 and IL-10 values occurred at least one month prior to relapse. Moreover, the continuous elevation of TNF-alpha and sIL-2R could be observed in patients who remained without progression during 10 months lasting immunotherapy. Our data illustrate that malignant melanoma associates with Th1/Th2 perturbances which are directed towards extended Th2 responses and that measurement of selected cytokines of Th1/Th2 category may be used as an early signal of disease deterioration and for evaluation of immunotherapy response.

Telomerase plays an important role in maintaining the stability of chromosomes. This ribonucleoprotein prevents chromosome ends (telomeres) from gradual loss with each cell division. It enables tumor cells to maintain telomere length, allowing indefinite replicative capacity. Telomerase activity has been detected in the majority of tumor and germ cells and in immortalized cell lines. Quantitative telomerase PCR-ELISA (TeloTAGGG Telomerase PCR ELISA(PLUS)) was evaluated for distinguishing benign and malignant breast tissue. Activity of telomerase was determined in 27 samples of fibrocystic and dysplastic tissues, 28 fibroadenomas and phylloid tumors, and 154 breast cancer tissues; 59 specimens were analyzed retrospectively. Analytical precision and linearity of the assay was tested using breast carcinoma cell line ZR-75-1 and breast tumor tissue extracts. About 4% of tumor samples were excluded from analysis due to interferences in the PCR reaction. Relative telomerase activity differed significantly in the groups of dysplastic tissues, fibroadenomas and carcinomas. The highest activity was found in breast cancer tissue. This method can identify breast cancer tissue with 73% clinical sensitivity and 93% specificity as compared to benign breast tumors. We did not find a correlation between telomerase activity and the tissue levels of estrogen and progesterone receptors, HER-2/neu oncoprotein concentration, tumor size, and lymph node positivity. Probability of disease-free survival was significantly lower for patients with telomerase activity higher than median value. As the assay for telomerase activity has very high analytical sensitivity and high specificity for cancer cells, this routinely used method may prove useful for distinguishing malignant phenotype of breast tissues.

Objective: To compare histologic detection of apoptotic (AP) figures and terminal transferase UTP nick end labeling (TUNEL) detection in colorectal carcinomas and lymphomas., Study Design: We evaluated the percentage of AP cells by AP figure counting and by the TUNEL technique in formalin-paraffin material from 39 tumors--29 colorectal carcinomas and 10 non-Hodgkin's lymphomas. The Lucia G image analysis system (Laboratory Imaging, Prague, Czech Republic) was used for cellularity evaluation and AP counting., Results: On average, 0.81 +/- 0.5% AP cells were detected by figure counting; 0.91 +/- 0.35% were identified by TUNEL. A statistically high correlation between these techniques was found using Pearson's correlation coefficient (r = .56, P < .001). TUNEL, although probably more sensitive, was difficult to standardize., Conclusion: AP figure counting seems to be the method of choice for routine work, particularly because of its cost effectiveness and reproducibility.

The prognostic and predictive value of p53 mutation in breast cancer is still conflicting. The choice of the p53 status detection method may account for some discrepancies. In this pilot study we compared two differently-based methods for detection of p53 alteration in 32 breast carcinoma samples: the immunohistochemical method using Bp53, DO1 and DO11 monoclonal antibodies for analysis of the p53 protein accumulation in cell nuclei and the functional method FASAY. FASAY - functional analysis of the separated alleles in yeast - tests the capability of the human p53 to transactivate a reporter with a p53 binding site RGC driving the ADE2 gene in yeast. In our group the percentage of breast cancers with accumulated p53 protein was 50%, as well as percentage of mutant p53 scored by FASAY was 50%. Although the agreement of both methods, when comparing the results of individual patients was high (94%), our results show that immunohistochemistry does not reflect the p53 status quite exactly.