o Muzio L

Cyclooxygenase isozymes in oral squamous cell carcinoma:a real-time RT-PCR study with clinic pathological correlations.

COX-2 expression in tumour cells has been associated with carcinogenesis in many human neoplasms, including head and neck cancer, while the COX-1 isoform of the cyclooxygenase enzyme is constitutively expressed in normal tissues. We measured COX-1 and COX-2 m-RNA expression in samples of both oral cancer and matched oral mucosa from 22 patients by RealTime RT-PCR; clinic pathological data (grading, TNM staging, inflammation, follow-up) of all patients were available for statistical evaluation. Most of the tumor samples in our study expressed at least one cyclooxygenase enzyme (COX-1 or COX-2 mRNA) more than their matched normal oral mucosa (p<0.05), with no correlation with the entity of inflammation, and a significant inverse relationship was found between COX-1 and COX-2 in each sample. Higher levels of COX-2 expression were associated with poor disease-free survival (p<0.05), but not with overall survival and higher tumor stage and grade. Our results suggest that COX-1 may play a role in oral carcinogenesis, and could be regarded as a potential therapeutic target by chemo preventive drugs; moreover, COX-2 expression might be addressed as a new prognostic tool in the clinical management of OSCC.

Immune response in patients with oral lichen planus and HCV infection.

In recent years an association between oral lichen planus (OLP) and HCV infection has been reported, but the frequency of this association seems to differ in the various geographic areas. It is clear, instead, that some abnormalities occur in the immune-regulation mechanisms of patients with OLP and it is thought to be due to the chronic antigenic stimulus of HCV that causes functional disorders of the immune system in infected patients. Possible immunologic difference between 17 patients with OLP and HCV+ and 17 patients with OLP and HCV- were investigated using standard immunofluorescence and flow cytometry techniques. The distribution of T and B cells was normal in all patients examined, while NK CD56+ cells were increased, above all in HCV- patients. About 65% of T CD4+ lymphocytes coexpressed the CD45RO isoform (p=0.002), while approximately 32% expressed CD45RA, without significant differences in comparison to HCV+ subjects (p>0.05). Moreover, almost all the CD4+CD45RO+ subpopulation coexpressed CD29 in all patients examined. No significant differences between the two groups of patients were detected as to the increase of cytotoxic T CD8+CD57+ lymphocytes. The B cells CD19+CD5+ responsible for the production of “natural” antibodies were detectable in both the examined groups, even if not in all HCV+ subjects (30% ± 10.1 in HCV- and 27% ± 19.4 in HCV+ patients; p=0.47). These findings suggest the existence of differences in lymphocyte subpopulations between OLP-HCV+ subjects and OLP-HCV- patients.

Direct visualization of oral-cavity tissue fluorescence as novel aid for early oral cancer diagnosis and potentially malignant disorders monitoring

Direct visualization of the oral tissue autofluorescence has been recently reviewed in several studies as a possible adjunctive tool for early recognition and diagnosis of potentially malignant and malignant oral disorders. The aims of this study were to assess: a) the value of a simple handheld device for tissue auto-fluorescence visualization of potentially malignant oral lesions; and b) the sensitivity, specificity and diagnostic accuracy of tested device, using histological examination as the gold standard. 175 consecutive patients, with at least one clinical oral lesion, were enrolled in the study. Clinical conventional inspections were performed for each patient by two blind operators. Then, oral biopsy and histological examination were performed. Pathologist was blind with respect to the autofluorescence results. The 175 histological assessments revealed no dysplasia, mild dysplasia, moderate/severe dysplasia and OSCC, in the 67.4%, 8.6%, 8%, 16% of cases, respectively. Oral lesions diagnosed as OSCC were found as positive under fluorescent light in the 96.4% of cases. Statistically significant correlation was observed between oral dysplastic lesions and the loss of tissue fluorescence (p-value=0.001). Low sensitivity values (60% and 71%) were recorded about the ability of the device in differentiating mild dysplasia vs. lack of dysplasia and moderate/severe dysplasia vs absence of dysplasia, respectively. The device tested in our study was found to not replace the histopathology procedure. However, we assessed its usefulness for oral tissue examination, especially within an oral medicine secondary care facility, before performing a biopsy and in monitoring oral lesions.

Expression of sexual hormones receptors in oral squamous cell carcinoma.

Sexual hormones play an important role in expression of genes involved in a wide variety of biological and neoplastic processes. The information on Estrogen Receptors (ER) expression in non-target tissues is very few and, in particular, the studies in head and neck tumors are still controversial. Recent studies analyzed the role of Tamoxifen (TAM) on Oral Squamous Cell Carcinoma (OSCC) lines in relation to the presence/absence of ER. The purpose of the present study was to evaluate the expression of sexual hormones receptors mRNAs, in particular Estrogen Receptor alpha (ERα) and Androgen Receptor (AR) mRNA in OSCC tissues. The study group comprised 20 samples of OSCC, harvested from 20 otherwise healthy subjects (14 males and 6 females, mean age 58.2y, range 38–74). The control group was formed by 20 samples of normal mucosa harvested around the margins of the specimens (at least 1 cm from the lesion margins). Estrogens Receptor alpha (Era) and Androgen Receptor (AR) mRNA expressions were analyzed by RT-PCR carried out on total RNAs extracted from both cancerous and healthy tissues. Obtained data were evaluated by Shapiro-Walk normality test and compared by Students t test. Results with p<0.05 were considered statistically significant. AR transcripts were less expressed in OSCC specimens than in healthy tissues, while levels of ERα transcripts significantly increased in tumor samples. These preliminary data show different expression patterns of AR and ERa mRNAs in malignant tissues of oral mucosa and could suggest an involvement of these sexual hormones in oral cancer.

Saliva as a diagnostic matrix for drug abuse.

Scientific interest in saliva as a diagnostic matrix has greatly increased over the last decade. The Triage® screening test (Biosite Diagnostics), a rapid immunological test used to detect recreational drugs in the urine, was used to compare two biological matrixes: a non-conventional one, saliva, and a traditional one, urine. Twenty-one drug abusers collected one urine and one saliva specimen, both of which were tested with the Triage kit. Data were validated by gas-chromatography-mass-spectrometry (GC-MS). Results were positive for methadone in 9 saliva and 14 urine specimens, for opiates in 2 and 10, respectively, and for barbiturates in 2 specimens. Saliva specimens were negative for cannabis, THC, benzodiazepines and tricyclic antidepressants, although the GC-MS analysis revealed low concentrations of these drugs in the saliva. The study demonstrates the possibility of using saliva as a diagnostic matrix to test for drug-taking; however, the Triage kit must be improved before being used with saliva.

Oral pathology in untreated coeliac [corrected] disease.

Background Many coeliac disease patients with atypical symptoms remain undiagnosed.

Beta-catenin and survivin expression in keratocystic odontogenic tumor (KCOT). A comparative immunohistochemical study in primary, recurrent and nevoid basal cell carcinoma syndrome (NBCCS)-associated lesions.

AIM To determine the epithelial expression of β-catenin and survivin in sporadic (primary, and recurrent) and nevoid basal cell carcinoma syndrome (NBCCS) keratocystic odontogenic tumour (KCOT) in order to assess activation of the β-catenin pathway and evidence of apoptotic inhibition, processes that may contribute to the known differences in their biological behaviour. MATERIALS AND METHODS Sections from 40 cases of KCOT (19 sporadic/primary; 9 sporadic/recurrent and 12 NBCCS-associated) were immunohistochemically stained for β-catenin and survivin. The extent and intensity of immunoreactivity within the lining epithelium was assessed, using semi-quantitative scales, independently by two pathologists who were blinded to the clinical-pathological data. Data were analysed using Kruskal-Wallis test and, for pair-wise comparisons, Mann-Whitney test with Bonferroni correction. RESULTS All cystic epithelial linings stained for β-catenin and survivin but there were differences in the pattern and intensity of staining among KCOT types. Sporadic primary KCOT showed weaker staining for β-catenin (P=0.0003) and survivin (P<0.0048) that was restricted to the basal and para-basal layers only, compared to sporadic recurrent and NBCCS-associated KCOT, which showed expression throughout all epithelial layers. There were no differences in β-catenin expression among recurrent and NBCCS-associated KCOT, whereas the intensity of survivin staining was higher in NBCCS-KCOT (P=0.0003). Nuclear staining for β-catenin was found exclusively in recurrent (5/9 cases) and NBCCS-associated (4/12 cases) KCOT. CONCLUSION The data demonstrate β-catenin delocalization and survivin over-expression in recurrent sporadic and NBCCS-associated KCOT suggesting that these pathways related to apoptotic inhibition have a role in KCOT growth and recurrence.

Carbamazepine transbuccal delivery: the histo-morphological features of reconstituted human oral epithelium and buccal porcine mucosae in the transmucosal permeation.

Transbuccal drug delivery is an attractive way of administration since several well-known advantages are provided, especially with respect to peroral management. Carbamazepine (CBZ) is an anticonvulsant which is useful in controlling neuropathic pain, and it is currently administered by peroral route, although its absorption and bioavailability is limited due to various factors. The oral cavity could be an interesting site for transbuccal CBZ delivery due to two properties: slow administration of constant low drug doses and less dose-related side effects. However, in transbuccal absorption a major limitation could be the low permeability of the mucosa which results in low drug bioavailability; thus the aptitude of the drug to penetrate the buccal mucosa has to be assessed by using tissue models resembling human normal mucosa. In our experience, CBZ well permeates mucosal membranes. In order to assess the efficacy of CBZ transbuccal delivery and to verify the reliability of these tissues in permeability testing before and after the passage of CBZ, the histo-morphological features of reconstituted human oral (RHO) epithelium (E) and buccal porcine mucosae were investigated. Significant histological changes due to CBZ passage were observed both in RHO-E and porcine mucosa. The main findings detected in RHO samples were cellular swellings with a signet ring-like appearance, nuclear swelling, prominent nucleoli lined against the nuclear membrane and the presence of keratohyalin granules. The most striking finding regarding porcine buccal mucosa was a cytoplasmic vacuolization, mainly involving the basal layer.

Hepatitis E virus co-infection in HIV-infected patients in Foggia and Naples in southern Italy.

Abstract Background: Hepatitis E virus (HEV) infection represents an emerging infection in developed countries and is thought to be a zoonotic infection. It has recently been described as a new causative agent of acute and chronic hepatitis in immunosuppressed subjects, including HIV-infected patients. The aim of this study was to assess the sero-virological prevalence of HEV in HIV patients and in the general population as control group. Methods: A prospective and observational cohort study was carried out in two hospitals in southern Italy. The seroprevalence of HEV was determined in a cohort of 959 subjects, 509 (53%) of whom were HIV-positive patients and 450 were from the general population. Serum samples were tested for anti-HEV antibodies; repeatedly positive results were confirmed by a Western blot assay. In positive patients HEV RNA and genotypes were also determined. Results: A total of 46 (4.8%) of the 959 serum samples examined were reactive to anti-HEV Ig and confirmed by Western blotting. The prevalence of HEV antibodies (IgG and/or IgM) was 2.7% in the control group and 6.7% in HIV-infected patients. Anti-HEV IgM was found in 6/46 (13.0%) of the anti-HEV Ig-positive serum samples, in 5/34 HIV patients and in 1/12 of the general population. No HIV-infected patient presented chronic hepatitis with HEV infection alone. Conclusions: This study indicates a higher circulation of HEV in HIV-infected patients, whereas a low prevalence of HEV antibodies in the general Italian population was shown. Chronic hepatitis with HEV alone was absent, while it was present in subjects with HIV-HEV, co-infected with hepatitis B virus (HBV) and/or hepatitis C virus (HCV).

Detection Level of Salivary Survivin in Patients with OSCC

Abstract Introduction: Survivin is a member of the Inhibitor of Apoptosis (IAP) proteins family, broadly expressed in most solid and haematological malignancies, but almost undetectable in normal adult tissues. Regarding Oral Squamous Cell Carcinoma (OSCC), Survivin expression seems to be correlated with aggressive phenotype and unfavorable outcome. In this work, the effectiveness of Survivin detection in saliva samples as diagnostic tool was investigated. Methods: 55 saliva samples from patients with histological diagnosis of OSCC and 30 samples from healthy controls matched for age and gender were collected. All samples from OSCC were collected before therapy. Saliva was analyzed using Assay Designs human Total Survivin TiterZyme® Enzyme Immunometric Assay (EIA) kit for Survivin detection and quantification. Survivin concentration (pg/ml) was studied in relationship to clinical data. Univariate statistical analysis was performed. Results: 35/55 (63.6%) among OSCC patients were found to be positive for surviving expression in saliva vs 12/30 (40%) among health controls; but the mean value was found to be higher in OSCC group (8,69 pg/ml ± 10,15 vs 2.44 pg/ml ± 4.22) reaching statistical significance (p<0.05). Moreover, samples from patients at an early stage (I+II) displayed a lower content of salivary Survivin than those at advanced stage (III+IV) (7.09 pg/ml ± 8.59 vs 8.13 pg/ml ± 10.94). In addition, cases characterized by lymph node metastasis showed a higher surviving concentration (9.38 pg/ml ± 11.26) than cases without metastasis (5.77 pg/ml ± 8.21). Discussion: Salivary Survivin seems to be an interesting biomarker for OSCC, but its use as a single marker may be not sufficient for the early diagnosis of OSCC