Telomerase activity detected by quantitative assay in bladder carcinoma and exfoliated cells in urine.

Early diagnosis is one of the most determining factors for patient survival. The detection of telomerase activity is a potentially promising tool in the diagnosis of bladder and other types of cancer due to the high expression of this enzyme in tumor cells. We carried out a quantitative evaluation of telomerase activity in urine samples in an attempt to determine a cut-off capable of identifying cancer patients. Telomerase activity was quantified by fluorescence TRAP assay in urine from 50 healthy volunteers and in urine and bioptic tumor samples from 56 previously untreated bladder cancer patients and expressed in arbitrary enzymatic units (AEU). Telomerase activity in urine ranged from 0 to 106 AEU (median 0) in healthy donors and from 0 to 282 AEU (median 87) in patients with cancer. A telomerase expression higher than the cut off value determined by receiver operating characteristic (ROC) analysis was observed in 78% of cases, regardless of tumor grade and in 71% (15/21) of cases of nonassessable or negative cytology. The quantitative analysis of telomerase activity in urine enabled us to define cut-off values characterized by different sensitivity and specificity. Cytologic and telomerase determination, used sequentially, enabled us to detect about 90% of tumors.

Relation between food habits and p53 mutational spectrum in gastric cancer patients.

p53 tumour suppressor gene mutations were analysed in gastric cancer in relation to food habits and social class in 56 patients from a high risk region of Italy. Exons 5-8 were analysed with DGGE method on amplified DNA from formalin-fixed paraffin-embedded samples. All p53 mutations were observed in patients belonging to low social class and the majority of mutations were found in intestinal type cancers. A positive association was also found with low raw vegetables, fresh, dried and preserved fruits, and ascorbic acid intake. Moreover, specific types of mutations were significantly associated with particular factors, thus suggesting the presence of specific molecular etiologic process in stomach carcinogenesis.

Correlation between p53 gene mutations and p53 protein accumulation evaluated by different methodologies.

Mutations in the p53 gene are the most common genetic alterations in many tumour histotypes. Many of these mutations induce conformational changes resulting in p53 protein stabilisation and consequently an accumulation detectable with immunochemical methods. Available data on the correlation between p53 gene alterations and p53 overexpression widely vary. In this study we analysed the correlation between p53 gene alterations detected by DGGE, SSCP and sequencing and protein expression detected by flow cytometric and immunohistochemical approaches by using PAb 1801 antibody. The study was performed on 21 bladder tumours and 10 cell lines derived from different tumour histotypes as representative of different methodologic problems which can be met starting from different types of biological material. The best correlation (81%) was observed between p53 mutations and FCM results, using a double evaluation criterion for the latter which includes the percentage of positive cells and "delta values", evaluated as the difference between the mean values of Pab 1801 stained cells and isotypic control. The high correlation obtained between results from this FCM double criterion and p53 gene mutations is a good starting point for the analysis on large series of tumours and for a multiparameter FCM analysis including p53 protein levels.

A phase II study of advanced colorectal cancer patients treated with combination 5-fluorouracil plus leucovorin and subcutaneous interleukin-2 plus alpha interferon.

Twenty-one patients with advanced, pretreated colorectal cancer in disease progression were entered in a phase II study to investigate the use of 5-fluorouracil (5FU) + leucovorin with subcutaneous Interleukin-2 + alpha interferon (alpha-IFN). Eighteen of these patients were evaluable for response to treatment: 1 partial response (PR) (duration 8 months), 9 stable disease (SD) (median duration of 6.5 months, range 2-15) and 8 progressive disease (PD). The PR patient survived for 15 months, the SD patients for a median of 11 months and 8 months for PD patients. Toxicity evaluated in the 21 patients reached grade 4 for mucositis in two cases. Grade 3 toxicity was observed more frequently for fever (52.3%) and diarrhea (33.3%) and was most probably the result of the combined side-effect of chemotherapy and the biological response modifiers (BRMs). Treatment was, for the most part, carried out on an out-patient basis as originally planned. In 15 patients tests were carried out to verify whether any immuno-activation had taken place. Significant increases were found during the course of therapy regarding cluster of differentiation activation (HLA-DR, CD71, CS25). Different curves were observed during the course of treatment with respect to the CD8 value, which proved higher in SD patients than in PD patients. Our study would seem to suggest that the addition of BRMs to 5FU + leucovorin could increase survival. The next step, however, must be to determine lower doses of IL-2 for subcutaneous administration in order to reduce toxicity but maintain the same immunostimulation.

Liver metastases from gastric carcinoma: report of a patient treated with adoptive immunotherapy (tumor-infiltrating lymphocytes plus interleukin-2 and subsequently local-regional lymphokine-activated killer cells plus interleukin-2).

A 37-year-old patient with liver metastases from gastric cancer was treated with a double adoptive immunotherapy regimen comprising tumor-infiltrating lymphocytes plus interleukin-2 and subsequently local-regional lymphokine-activated killer cells plus interleukin-2 because of an extremely high in vitro cytotoxic specific activity on established gastric cancer cell lines. The necrosis verified in the center of the hepatic metastasis would appear to demonstrate treatment efficacy, but no clinical response was seen. In vitro cytotoxicity data alone are insufficient to predict the clinical efficacy of adoptive immunotherapy.

[Applicative aspects of liquid-based cytology in cervical cancer screening]. / Aspetti applicativi della citologia in fase liquida nello screening del cervico-carcinoma.

The quality of cytological services is the very heart of the prevention of cervical pathologies. Indeed, various studies have demonstrated that inadequate sampling, mistakes made in the organisational and management methods of the screening programme, and incorrect diagnoses result in unnecessarily high incidence and mortality rates. The aim of this work is to compare the effectiveness of two different methods, i.e. a conventional smear test and a liquid based ThinPrep (TP) test. Said methods were tested on a sample 453 cases diagnosed as being "Atypical Squamous Cells of Undetermined Significance"/"Atypical Glandular Cells of Undetermined Significance" according to the 1991 Bethesda System. All the women with an "Atypical Squamous Cells of Undetermined Significance "/"Atypical Glandular Cells of Undetermined Significance" cytological diagnosis were called back within 3 months for a ThinPrep test, as part of the Level 2 diagnostic controls of a cervical cancer screening programme. Of the initial diagnoses of "Atypical Squamous Cells of Undetermined Significance"/"Atypical Glandular Cells of Undetermined Significance" with a conventional smear test, 124 cases (27.4%) were classified as being adequate, while 329 (72.6%) were satisfactory, although they did have limited indicators of quality. Upon repetition of the cytology with a ThinPrep test, 322 cases (71.1%) were found to be adequate, 129 (28.4%) "suboptimal" and 2 inadequate (p < 0.0001). The main reasons for insufficient results in conventional smear tests are: bad preservation (40.2%), the presence of granulocytes (36.4%), intense phlogosis (12.1%) and erythrocytes (5.5%). In liquid based smear tests, the main indicator of quality is the absence of endocervical glandular cells (56.7%). As for the cytological diagnosis, the use of ThinPrep supplied the following results: of the 453 cases diagnosed initially as being "Atypical Squamous Cells of Undetermined Significance"/"Atypical Glandular Cells of Undetermined Significance", 371 (84.1%) were negative, 54 (11.9%) "Atypical Squamous Cells of Undetermined Significance "/"Atypical Glandular Cells of Undetermined Significance" and 18 (4%) L-SIL (p < 0.0001). Histological follow-up of the 18 cases with L-SIL confirmed the presence of a dysplastic lesion in 8 out of 12 cases (66.7%); in 4 cases there was no consistency between the cytological and histological diagnoses, and in 6 patients no biopsy had been taken. The preliminary experience of this study, although indeed carried out on a limited number of cases, appears to show that suitable training for the collection of samples in a liquid solution could improve the adequacy of the sample and thus the precision of the cytological diagnosis.