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Cystic masses of the lateral neck are mostly benign. However, the incidence of metastatic squamous cell carcinoma (SCC) in cervical cystic masses initially diagnosed as benign is quite high in patients older than 40 years. The aim of this study was to compare the diagnostic accuracy of preoperative cytology and intraoperative frozen section (FS) in detecting malignancy in cystic masses of the neck. We reviewed 61 patients who underwent preoperative ultrasound-guided fine-needle aspiration cytology (FNAC) and neck biopsy of a cystic neck mass, and analysed the concordance between FNAC and intraoperative FS with respect to definitive histology. HPV status was also tested. Of 49 eligible cases, the accuracy of preoperative FNAC was 70.5% (weighted kappa 0.53), meaning moderate agreement between cytology and final diagnosis. Intraoperative FS consultations detected 16 cases of SCC metastasis while the remaining 33 cases were negative for SCC, showing perfect agreement with histology. Since FS results were useful in evaluating cystic neck masses, despite a moderate accuracy of cytology, we suggest intraoperative FS analysis for all cystic neck masses. This technique can allow us to switch to therapeutic neck dissection, multiple upper aerodigestive tract biopsies, tongue base mucosectomy, and bilateral tonsillectomy in the same surgical setting.

Despite the advances in anticancer therapies, their effectiveness for many human tumors is still far from being optimal. Significant improvements in treatment efficacy can come from the enhancement of drug specificity. This goal may be achieved by combining the use of therapeutic molecules with tumor specific effects and delivery carriers with tumor targeting ability. In this regard, nucleic acid-based drug (NABD) and particularly small interfering RNAs (siRNAs), are attractive molecules due to the possibility to be engineered to target specific tumor genes. On the other hand, polymeric-based delivery systems are emerging as versatile carriers to generate tumor-targeted delivery systems. Here we will focus on the most recent findings in the selection of siRNA/polymeric targeted delivery systems for hepatocellular carcinoma (HCC), a human tumor for which currently available therapeutic approaches are poorly effective. In addition, we will discuss the most attracting and, in our opinion, promising siRNA-polymer combinations for HCC in relation to the biological features of HCC tissue. Attention will be also put on the mathematical description of the mechanisms ruling siRNA-carrier delivery, this being an important aspect to improve effectiveness reducing the experimental work.

Objective. The inflammation is known to be essential in the neoplastic progression, nevertheless this host-tumor interaction has not been yet clarified. Whilst neoangiogenesis might stimulate the tumor progression, simultaneously this process could interfere with tumor growth. The aim of this study was to evaluate the prognostic value of intra- and peri- tumor infiltrating lymphocites (TILs) and correlate this parameter with the overall survival. Material and Methods. We retrospectively collected 50 cases of epithelioid pleural mesthelioma analyzed from 2005 until 2016 by UCO di Anatomia e Istologia Patologica di Trieste. Immunohistochemistry staining of CD3 was performed to define intra-tumor lymphocites count. Intraepithelial, stromal and peritumoral TILs was assessed using the following scoring system: score 1 when TILs percentage ranges between 6 and 25%; score 2 when TILs percentage ranges between 26 and 50% and score 3 between 51% and 75%. Data were statistically analyzed by Kaplan-Meier method using a dedicated software (Statistical Package for Social Sciences, SPSS Inc., Chicago, IL, USA). The level of significance was p < 0.05. Results. In this sample the average and median age were 70 and 71 respectively. The 4-years OS was 16% (n=8) with 42 deaths due to mesothelioma and the event occurred after an average period 14.97 months long. Neither intraepithelial nor stromal TILs scores correlated with OS (p>0.05). On the contrary, high scores (2+3 score) of peritumoral TILs significantly correlates with improved OS (p=0.02). Conclusions. In our experience, peritumoral TILs was a positive prognostic factor in epithelioid pleural mesothelioma. This result suggests to include an immunological analysis in the therapeutical approach to mesothelioma because it has to be clarify how immune elements take part in the tumor progression. Aiming to stimulate a specific antitumor immune response,further studies are needed to optimize the immunotherapy.

Objective. Human Papillomavirus (HPV)-induced oropharyngeal squamous cell carcinoma (OPSCC) represents an increasing tumor entity with typical clinical and histopathological features, associated with a rising incidence and improved prognosis (1). Detection of HPV-DNA is the most widely used method to diagnose HPV infection in clinical samples. Commercially available diagnostic assays show variable sensitivity and specificity and no international standards have been proposed yet (2). The main purpose of this preliminary study was to evaluate the reliability of EasyPap Direct HPV DNA Test (Ulisse BioMed) on formalin-fixed paraffin-embedded samples (FFPE), despite this test has been designed to work in vaginal mucus, where preliminary data showed a sensitivity of 97%. The test is real-time PCR based, it does not require DNA extraction and it can perform high-risk HPV genotyping. Kit performances have been compared with a commercial kit which requires DNA extraction. Materials and Methods. This pilot study included 38 cases of OPSCC that were surgically treated in our institute. Tumor samples were fixed in buffered formalin, embedded in paraffin and stored at room temperature for 1-13 years (mean storage period 4 ± 4 years). In order to confirm histology and lesion grade, all samples were reviewed by hematoxylin-eosin staining. DNA extraction has been performed starting from FFPE samples through QIAamp® DNA FFPE Tissue Kit (Qiagen, Hilden, Germany) and HPV has been typed through Real Time PCR HPV sign® Pyro Mark System Q96 IDTM (CE-IVD Diatech Pharmacogentetics Iesi Italy). We found 16 cases positive for HPV16, 1 case positive for HPV35 and 21 HPV negative samples. In order to perform HPV detection through EasyPap Direct HPV DNA test, samples underwent deparaffinization with xylene and rehydration with ethanol. Genotyping data were compared to those obtained with Qiagen kit. Results. EasyPap Direct HPV DNA test identified 16 HPVpositive cases and 22 HPV negative cases. Genotyping data showed that 15 samples were HPV16 and 1 sample was HPV35 positive. Concordance analysis showed an agreement- coefficient Kappa Cohen K=0.95 IC95% [0.84-1.00] (excellent agreement, Fleiss classification). The sensibility and specificity of the EasyPap HPV DNA test was 94.1% (95%, confidence interval 80.1%-98.8%) and 100% (95%, confidence interval 88.6%-100%), respectively. The positive predictive value was 100% (95%, confidence interval 88.6%-100%) and the negative predictive value was 95.5% (95% confidence interval, 81.9%-99.3%). Conclusions. Hitherto it was well known that DNA extraction contributes to increase the sensitivity of PCR-based HPV test starting from FFPE tissue samples (3). Despite small sample size has been analyzed, preliminary data obtained through EasyPap Direct HPV DNA test successfully showed that high sensitivity and specificity can be achieved without DNA extraction, with a remarkable decrease of assay cost and time. These results lead us to perform large scale EasyPap testing.

Breast Fine Needle Aspiration Cytology (FNAC) represents first choice diagnostic tool to determine mammographic lesions and clinically palpable breast nodules. FNAC is affected anyway by a certain degree of subjectiveness, that is more evident with the so- called “grey zone” cases (the ones not clearly benign or not clearly malignant) and when there are some doubts about material adequacy. These situations may create some misunderstandings between clinicians and pathologists with possible over or undertreatment for patients. The need for clear communication is extremely important, but it becomes a priority in senology because of the multidisciplinary aspects of the field. For this reasons a breast cytology reporting system has been proposed since 1993 within a U.K. mammographic breast screening programme. From our experience we think only a cytological exam with well demonstrated and well documented quality which satisfies those criteria suggested by guidelines will be able to play a key role in patients’ clinical management. It will allow reducing as much as possible diagnostic biopsies on benign lesions and frozen sections to confirm malignant lesions.We believe that a good quality cytology can provide an important and relevant contribution to define breast lesions’ nature without using other invasive procedures and to obtain in most cases good material for subsequent molecular investigations. Cytological approach to breast lesions was also validated in Breast Unit of Trieste during EUSOMA certification visit.

OBJECTIVE. Human Papillomavirus (HPV)-related squamous cell carcinoma of the oropharynx (HPV-OPSCC) usually originates from the reticulated epithelium lining the crypts of Waldeyer’s ring. The increasing epidemiologic evidence of this type of tumors and the peculiar positive prognosis, suggested to introduce a dedicated staging system (1). The 8th edition of the American Joint Committe on Cancer (AJCC) staging manual presented a new classification to discriminate HPV-related cancers based on p16 over-expression, since it demonstrated to be an important biomarker (2). The detection of HPV-DNA could not be used as a defining factor due to its cost and lack of universal applicability. The main purpose of this preliminary study was to evaluate the concordance between the results of immunohistochemistry for p16 overexpression and the HPV-DNA detection performed with a commercial kit as control. MATERIALS AND METHODS. We retrospectively collected 32 cases of OPSCC diagnosed in 2016 at the Academic Hospital of Cattinara in Trieste, whose samples had been tested with Real Time PCR (Pyro Mark System Q96 IDTM - CE-IVD Diatech Pharmacogentetics Iesi Italy). The study was performed using tumor samples fixed in buffered formalin, embedded in paraffin and stored at room temperature for 6 to 18 months. All samples were selected by a pathologist based on sections stained with hematoxylin and eosin to confirm the adequacy of the tissue. The samples were tested for p16 by immunohistochemistry (CINtech® p16 Histology E6H4 ROCHE) and the positivity was assessed when nuclear expression was ≥+2/+3 intensity and ≥75% distribution according to the 8th edition of the AJCC staging manual. In the context of oropharyngeal cancer, the optimal cut-off for p16 overexpression is ≥+2/+3 intensity (+/- cytoplasmic staining) with ≥75% distribution. RESULTS. Seven cases were excluded because of lack of histological stored material for p16 immunohistochemical analysis (only cytological specimen). Among the remaining 25 cases, there were 5 HPV-positive and 20 HPV-negative tumors. The sensibility and specificity of the p16 immunostaining was equal to 100% (95%, confidence interval 83.4%-100%) and 100% (95%, confidence interval 83.4%-97.8%), respectively; the positive predictive value was 100% (95%, confidence interval 83.4%-100%) and the negative predictive value 100% (95% confidence interval, 83.4%-100%). CONCLUSIONS. In agreement with the literature, p16 overexpression demonstrated to be a reliable parameter to identify the cases of OPSCC in which performing an in-depth analysis by means the detection of HPV-DNA would be indicated. Indeed, it is widely known that the association of p16 immunostaining and HPV-DNA detection by quantitative PCR increases the prognostic discriminator (3). We intend to carry on this study adopting a prospective design increasing the sample size and the test accuracy.