P53 and bcl-2 in colorectal cancer arising in patients under 40 years of age: distribution and prognostic relevance.

Young people (40 years of age) with colorectal cancer (CRC) represent a distinct subgroup with more aggressive disease behaviour compared to older patients. We evaluate whether p53 and bcl-2 could be useful in identifying young patients at higher risk of tumour progression. We reviewed 1340 CRC patients with 58 patients 40 years (4.2%). They had more frequent moderately or poorly differentiated mucinous adenocarcinomas (26% versus 12.3%, p=0.03); higher advanced stage at diagnosis; shorter 5-year overall survival (49.8% versus 71%; p=0.02); more frequent p53 positive (89.8% versus 72.6%, p<0.05) and bcl-2 negative (88.0% versus 66.2%, p<0.05) tumours; no difference in DNA content or proliferation indexes. Moreover, p53+ and bcl-2- resulted in being independent predictors of survival with shorter survival for the p53+/bcl-2- patients. Combining p53 and bcl-2, we could identify young CRC patients at higher risk of progression, who probably require development of a more sophisticated therapeutic approach based on identification of predictive factors.

Nuclear and cytoplasmic cellular distribution of survivin as survival predictor in resected non-small-cell lung cancer.

AIM: Survivin is a member of the inhibitors of apoptosis (IAP) gene family that acts through pathways different from those involving the bcl-2 family. Largely undetectable in normal adult tissues, survivin is deregulated in most human cancers including non-small-cell lung cancer (NSCLC) and may represent a tumor marker with prognostic and therapeutic implications. Aim of our study was to determine the prognostic role of survivin as an apoptosis-related biomarker in a series of resected NSCLC patients. METHODS: A retrospective series of resected NSCLC patients were retrieved from the files of the Regina Elena National Cancer Institute. Survivin was detected by immunohistochemistry (IHC) using a polyclonal antibody. Survivin displayed two kinds of immunoreactivity: (i) a diffuse cytoplasmic staining and (ii) a distinct nuclear staining. A score-scale to distinguish positive (score 1-2) vs. negative (score 0) pattern was applied. Clinical and biological (nuclear and cytoplasmic survivin staining) covariables were screened for a prognostic relationship with overall survival (OS) and disease-free survival (DFS) into the univariate and multivariate analyses. RESULTS: Data referring to 116 NSCLC patients who underwent surgery for stage I-IIIA NSCLC were collected. Multivariate analyses identified tumor size, nodal status and nuclear, but not cytoplasmic, expression of survivin as significant independent predictors of OS, with a hazard ratio of 2.40 (95% CI 1.44, 3.99, p=0.001), 2.03 (95% CI 1.26, 3.26, p=0.003) and 1.83 (95% CI 1.01, 3.30, p=0.044), respectively. Median OS for nuclear survivin positive (score 1-2) and negative (score 0) patients were 23 months (95% CI 15, 31) and 36 months (95% CI 1, 76), respectively (p=0.01); five-year survival for score 1-2 and score 0 patients were 20% and 44.5%, respectively. Conversely, no significant impact on survival is found when patients are stratified according to cytoplasmic survivin expression. CONCLUSIONS: Data presented herein open the issue that prognosis of stage I-IIIA NSCLC can be linked to the cellular pattern of distribution of survivin.

HPV genotypes concordance between sex partners.

The HPV genotype concordance in the sexual couples could support the sexual viral transmission of HPV infection. The present study contains a case-report of a stable Italian sex couple harbouring the same five HPV genotypes in their genital samples. The female partner, affected by vulvar condilomatosis, evidenced positivity in her cervicovaginal scraping with high risk HPV DNA Hybrid Capture 2 test and was negative at liquid-based performed Pap Test and at colposcopic examination. The male partner was clinically healthy regarding his external genitalia. In both male and female genital scrapings, the following HPV genotypes were detected by means of a PCR-based assay: 6, 16, 53, 73 and 84. This considerably high genotype concordance does not appear to be casual and supports, in our opinion, the hypothesis that genital HPV types are sexually transmitted agents

Incidence of human papillomavirus infection in oral leukoplakia. Indications for a viral aetiology.

Epidemiological and experimental studies have indicated a possible role of human papillomavirus (HPV) in the etiopathogenesis of oral premalignant lesions and tumors. The aim of this study was to establish the incidence of HPV infection and the typing of genotypes in some patients with nonmalignant oral cavity pathology and healthy subjects. We selected 80 subjects affected by the following pathologies: oral leukoplakia (20), squamous cell papilloma (6), various forms of stomatitis (30), lichen planus (15), burning mouth syndrome (BMS, 9). Ten healthy subjects were used as control. The patients underwent a brushing directly on the lesion or on the lower gums and on the dorsal surface of the tongue and a new, sensitive method of in situ hybridization (ISH) with colorimetric signal amplification was used for HPV detecting. The samples were tested with a Mix probe, which identifies most of the HPV genotypes and, in positive cases, the specific probe for the 6-11, 31-33-51 and 16-18 genotypes, was used. Leukoplakia and papilloma were , therefore, associated with the HPV infection, differently from the other pathologies and healthy oral mucosa (chi-squared < 0.005). In conclusion, according to our findings, HPV is a specific risk factor for the development of oral premalignant lesions.

Presence and incidence of DNA sequences of human polyomaviruses BKV and JCV in colorectal tumor tissues.

The human polyomaviruses JCV and BKV are widespread within population, as shown by serological studies. However, exposure to these viruses does not seem to have pathological consequences in immunocompetent individuals, while in immunocompromised or immunosuppressed patients, polyomaviruses can be activated, giving rise to serious pathologies. Viral DNA sequences were also found in cells from a number of human tumors of mesothelial origin, suggesting that activation of BKV and JCV, taking place in genetically predisposed and/or in immunodepressed individuals, might be involved in the mechanisms of tumor transformation. In this study, samples obtained from 18 patients with colon rectal carcinoma were probed for the presence of JCV and BKV by three different techniques: Southern blot, PCR and in situ hybridization. Our results demonstrate that viral DNA sequences were present in 16 out of the 18 cases considered (88.9%). In the large majority of cases, viruses were detected both in the tumor mass and in the surrounding healthy tissues. Lymphocytes in the investigated areas were also found to be infected by polyomaviruses. These data indicate, for the first time, a possible involvement of polyomaviruses in the pathogenesis of tumors of endothelial origin, like the human colon rectal carcinoma.

Role of P53 and Bcl-2 in advanced rectal carcinomas treated with adjuvant therapy.

Rectal adenocarcinomas is usually associated to a poorer outcome than colon cancers. In this study we analyzed the impact on overall survival of p53 and Bcl-2, evaluated by immunohistochemical techniques, in 126 advanced rectal cancer patients submitted to 5 fluorouracil based adjuvant therapy. Shorter overall survival was observed in patients bearing p53 positive and Bcl-2 negative tumors, although in multivariate analysis only p53 emerged as independent predictor of a worse outcome. These results seem to indicate that, in stage III-IV rectal cancer, p53 alterations may identify high risk patients to be enrolled in more aggressive and/or innovative adjuvant/neoadjuvant treatments.

[Benign metastasizing pulmonary leiomyoma: description of a case and review of the literature]. / Leiomioma metastatizzante benigno nel polmone: descrizione di un caso e revisione della letteratura.

We report the case of a 48-year-old woman with multiple benign metastasizing leiomyomas in both lungs and a single retroperitoneal leiomyoma. The patient underwent surgical resection of the lesions 15 years after total hysterectomy for uterine myomas. Immunohistochemical studies performed on the retroperitoneal and pulmonary neoplasms showed them to be of mesenchymal derivation with smooth muscle differentiation; the samples were negative for HMB-45. Some histogenetic hypotheses for the multiple leiomyomas are presented.

Colourimetric signal amplification of in situ hybridization assay for human papillomavirus DNA detection in cytological samples.

Signal amplification of In Situ Hybridization (ISH) can be obtained by Catalyzed Reporter Deposition (CARD), the catalyzed deposition of biotinylated tyramide at the location of the labelled probe. We analyzed 156 cervico/vaginal samples in order to evaluate whether ISH-CARD improves conventional ISH technique in detecting Human Papillomavirus (HPV) on cytological smears and can be used in clinical practice. 126 patients were clinically suspected to have a squamous intraepithelial lesions (SIL) and 30 were negative control-patients. We performed a HPV test screening on these 156 patients with both methods. The screening positive cases were tested for the presence of subtypes of HPV: 6/11, 31/33/51 and 16/18, using only the ISH-CARD. We found a significant increase of positivity with the amplified system, ISH: positive = 50 cases, ISH-CARD: positive = 120 cases, even in many cases cytologically negative. Many cases showed coinfections with different types of HPV. We observed overlapping results in some cell lines harbouring specific HPV subtypes (SiHa, CaSki and HeLa cells), whereas there was no reaction in the C33A cells, negative for HPV presence. This method enables the detection and the typing of HPV in cytological smears, represents a useful tool for clinical purposes and even identifies occult HPV infections.

p53 nuclear accumulation and multiploidy are adverse prognostic factors in surgically resected stage II colorectal cancers independent of fluorouracil-based adjuvant therapy.

To identify the prognostically highest risk patients, DNA content and p53 nuclear or cytoplasmic accumulation, evaluated by monoclonal antibody DO7 and polyclonal antibody CM1, were determined in 94 surgically resected stage II (Dukes B2) colorectal cancers, treated or not with adjuvant 5-fluorouracil-based chemotherapy. Sixty-one (65%) of the tumors were aneuploid, 16 (17%) of which had a multiploid DNA content; 50 (53%) displayed DO7 nuclear p53 accumulation, and 44 (47%) showed cytoplasmic CM1 positivity. In multivariate analysis, only multiploidy and p53 nuclear positivity emerged as independent prognostic indicators of a poorer outcome. Positivity for p53 was associated with shorter survival in 5-fluorouracil-treated and untreated patients. Therefore, in patients with Dukes B2 colorectal cancer, a biologic profile based on the combined evaluation of DNA multiploidy and p53 status can provide valuable prognostic information, identifying patients to be enrolled in alternative, more aggressive therapeutic trials.

[The value of the Pap test in the diagnostic approach to HPV infection of the cervix uteri]. / Valore del Pap-test nell'approccio diagnostico dell'infezione da HPV della cervice uterina.

HPV infection of the low genital tract is the most diffuse STD world wide. For this reason it's necessary to follow a simple flow chart to reach a correct diagnosis and to practice an adequate therapy. The authors relate on their experience in the management of a group of patients they have observed in a STD clinic. In this group it was executed a diagnostic protocol including Pap test, in situ hybridization, dot blot and histologic examination on tissue biopsies. From the analysis of the obtained data, the authors state that the Pap test is very useful to start in the diagnostic protocol, for it is precise, inexpensive and allows to identify the patients to follow with further diagnostic procedures such as colposcopy and histologic examination, in order to reach an exhaustive diagnosis and to assess the proper therapy.

Selected monoclonal antibodies can increase the accuracy of cytodiagnosis of neoplastic effusions of cryptic origin expanded in a short term culture.

The use of a selected panel of monoclonal antibodies (MoAb) to tumor associated antigens (TAA) in immunocytochemical (IIC) tests has been shown, in a preliminary study, to be a powerful diagnostic tool for the identification of the primary solid tumor causing metastatic effusion. Despite this improvement in a minority of neoplastic fluids a number of different causes may still determine false negative (FN) immunocytochemical diagnoses. The aim of the present study was to confirm the diagnostic accuracy of this panel of MoAb. This was done by analyzing in IIC tests a larger number of effusions and by evaluating whether the expansion in short term culture of those fluids with an uncertain malignant morphology could provide an adequate cellular substrate for immunocytodiagnosis. The analysis of 314 effusions confirmed the results of the pilot study and demonstrated that the combination of short term culture and immunocytochemical assays can further increase the sensitivity of this novel diagnostic procedure from 84.3% to 95.3%.

Use of MoAb D612 in combination with a panel of MoAb for the immunocytochemical identification of metastases from colon-rectum carcinoma.

During the course of colon-rectum tumours a number of clinical events may occur in which conventional cytopathology can provide only a partial contribution to the definition of a differential diagnosis, i.e. effusions, distant recurrences and second neoplasias. In the present study we have evaluated whether monoclonal antibody (MoAb) D612, recognising a colon-rectum associated antigen, can be used in this context. To this end, MoAb D612 was employed in combination with a panel of MoAb of well defined tumour specificity in immunocytochemical tests. The immunocytochemical findings obtained were compared with the histological and clinical diagnosis. Of 62 effusions and 40 fine needle aspirates studied, MoAb D612 reactivity correlated with the correct diagnosis in 92.8% of the instances. These results indicate that this reagent may help to improve the current cytopathological diagnosis of colon-rectum tumours by identifying the colonic origin of metastases in patients with unknown primary tumour, differentiating ovarian carcinoma from colon metastases to the ovaries and establishing the presence of a second neoplasia in patients with a previous history other than colon carcinoma.

Morphometric analysis of hyperthermia-induced nuclear and nucleolar size modifications in a human melanoma cell line.

Although no prominent morphologic alterations were observed by light microscopy in a human melanoma cell line (M-14) exposed at 42 degrees C for 4 hr, the computerized image analysis demonstrated a significant enlargement of both nuclei and nucleoli. The mean nuclear and nucleolar areas were found to be 32% and 94% increased, respectively, in comparison with normal controls. The gaussian distribution of absolute frequency of the measures demonstrated that this nuclear and nucleolar enlargement was distributed among the whole cell population, taking place in clonogenic cells and in "sterilized" cells, ultimately doomed to die. This finding, not previously described in the context of the heat shock response, may be regarded as morphological evidence of an increase in the protein content of the nuclei of heat-treated cells, as determined by biochemical methods.

Morphometric evaluation of histological sections of the thyroid gland in benign and malignant follicular lesions.

A morphometric analysis was made on histological sections of thyroid tissue in various aspects of follicular cell pathologies (goiter, adenoma, and carcinoma) and in normal follicular cells. Three cases of each aspect and 150 cells of each case were measured. The results obtained demonstrate the importance of the nuclear area in the differential diagnosis of normal thyroid, goiter, adenoma, and carcinoma. The other parameters considered, i.e., perimeter, maximum diameter, and nuclear form factor, proved to be able to exclusively distinguish between benign lesions (normal thyroid, goiter and, adenoma) and malignant ones (carcinoma).

Role of percutaneous fine needle aspiration in the diagnosis of lung cancer: our experience with 140 patients.

The authors have performed percutaneous fine needle aspiration (FNA) biopsy in 140 patients with pulmonary lesions of suspected neoplastic nature. In 63% of the cases histologic diagnosis could be made on the bioptic sample. Comparison of these findings with those obtained with conventional exfoliative cytology suggest that the latter technique gives higher percentage of histological characterization, it is free of complications and easier to perform. Thus, FNA appears to be an additional, irreplaceable tool when conventional methods fail to provide clear-cut information regarding the histologic nature of pulmonary lesions.