Cytological alterations of nasal mucosa after nasal packing.

PURPOSE: To assess the efficacy, tolerability and cytological alterations of nasal mucosa with various materials for nasal packing. MATERIALS AND METHODS: One hundred and five patients, submitted to a nasal surgery and post-operative packing with four different materials (Clauden®, Merocel®, two-fingered glove pack with gauzes inside and Lyofoam™) were prospectively enrolled in the study. All patients were submitted to a questionnaire of 12 questions which required the patients to refer to the tolerability of packing. Finally a nasal cytological test was performed after removal of packing and 10 days after, to evidence the alterations of ciliated cells and reduction of supranuclear stria. RESULTS: The questionnaire of tolerability of packing revealed a severe discomfort (grade III) in 13 patients (38.2%) with Clauden, 1 (2.4%) with Merocel, 7 (41.1%) with two-fingered glove packing and 6 (46.1%) with Lyofon. As regards cellular alterations, the two-fingered glove and the Merocel groups had the lowest percentage of cellular alterations which was particularly different from the Clauden group (p=.0014) The supranuclear stria was reduced in the Clauden group and the two-fingered glove packing group both after packing removal and after ten days with a statistical difference when compared to the other two groups (p<.001). CONCLUSIONS: Merocel packing appears to be well tolerated by the patients with the lowest percentage of cytological alteration, of ciliated cells. Also finger glove packing shows scarce damage of nasal mucosa with a lower percentage of nasal bleeding but its tolerability is not so good.

BRAF mutation and RASSF1A expression in thyroid carcinoma of southern Italy.

Aim of this work is to provide a detailed comparison of clinical-pathologic features between well-differentiated and poorly differentiated tumors according to their BRAF and RASSF1A status. We analyzed RASSF1A methylation by MSP and BRAF mutation by LCRT-PCR with LightMix® kit BRAF V600E in neoplastic thyroid tissues. Immunohistochemical evaluation of RASSF1A expression was also performed by standard automated LSAB-HRP technique. An overall higher degree of RASSF1A over-expression than normal thyroid parenchyma surrounding tumors (P < 0.05) has been found in all malignant well-differentiated lesions. Moreover, statistically significant higher levels of RASSF1A expression were observed in differentiated cancers associated to an inflammatory autoimmune background (P = 0.01). Amplifiable DNA for LC PCR with LightMix® kit BRAF V600E was obtained in nine PTCs, four FVPTCs, five ATCs, and one control. The V600E mutation was found in 13 of 18 (72%) tumors. BRAF was mutated in 6 of 9 (66%) classical PTC, in 2 of 4 (50%) follicular variant PTC and in all ACs (100%). The overall frequency of RASSF1A promoter methylation observed was 20.5% (9 cases out 44). Hypermethylation of RASSF1A in primary tumors was variable according to histotypes ranging from100% (5/5) in ACs to only 12.5% (4/32) in PTCs. We show a correlation between RASSF1A methylation status and RASSF1A protein expression. Finally, we conclude that BRAF V600E mutation and RASSF1A methylation were pathogenetic event restricted to a subgroup of PTC/FVPTCs in early stage and to clinically aggressive ATCs.

Cytofunctional changes in nasal ciliated cells in patients treated with hyaluronate after nasal surgery.

BACKGROUND: Hyaluronic acid (HA) plays a significant role in tissue repair of mucosal surfaces and, consequently, in surgical injury remodelling of nasal mucosa. OBJECTIVE: To assess the effect of high-molecular-weight HA administered by aerosol on the morphofunctional recovery of ciliated cells damaged by surgical trauma. METHODS: A single-blind, prospective, randomized trial was carried out with 94 patients who were randomly assigned, after endoscopic turbinoplasty, either to treatment with nasal saline solution irrigation (control group, n = 47) or to treatment with nasal douches based on high concentration (9 mg) and high-molecular-weight sodium hyaluronate (active treatment group, n = 47). All the patients were evaluated by using nasal fiberendoscopy, mucociliary transport time, nasal cytologic test, and a visual analog scale in terms of symptoms before and at 2 and 4 weeks after surgery. RESULTS: Visual analog scale values were significantly lower in the active treatment (AT) group at week 2 regarding each individual symptom. Mucociliary transport time was significantly reduced in patients in the AT group but only 1 month after surgery. Both the percentage of cellular impairments and the number of cells with hyperchromatic supranuclear stria showed significant improvements in the AT group in all postsurgery evaluations (p < 0.05). CONCLUSION: Intranasal use of sodium hyaluronate in patients who underwent functional nasal surgery improved both mucociliary clearance and nasal mucosa regeneration due to a faster recovery of the impaired ciliated cells.

Relationship between CK19 expression, deregulation of normal keratinocyte differentiation pattern and high risk-human papilloma virus infection in oral and oropharyngeal squamous cell carcinoma.

BACKGROUND: Simple epithelial keratins appear early during embryonic development and are expressed in non-stratified, ductal and pseudo-stratified epithelial tissues. CK19, the lowest molecular weight keratin, is also expressed in basal layer of squamous epithelia of mucosal surfaces. Previous studies have shown that High Risk-Human Papilloma Virus (HR-HPV) epithelial infection induces cell immortalization via E6 and E7 viral proteins and this, in turn, impairs cytokeratin expression in cancerous cells lines derived from uterine cervix. Here, we demonstrate the possible relationship between HR-HPV(+) oral/oropharyngeal cancer and the high levels of CK19 expression. METHODS: We analyzed 38 cases of Oral Squamous Cell Carcinomas/ Oro-Pharyngeal Squamous Cell Carcinomas (OSCCs/OPSCCs) by Immunohistochemistry (IHC) using specific antibody (Ab) detecting CK19, by In Situ Hybridization (ISH) and Polymerase Chain Reaction (PCR) based methods in order to define the HPV infectious status. We also evaluated the variation of CK19 expression in UPCI-SCC-131 (HPV(-)) and UPCI-SCC-154 (HPV(+)) cell lines by immunocytochemistry (ICC) and flow cytometry. RESULTS: CK19 OSCC/OPSCC score has been identified multiplying percentage of cancer expressing cells to staining intensity. CK19 expression score in OSCCs/OPSCCs was very different between HPV(+) (mean: 288.0 ± 24.3) and HPV(-) cancers (mean: 66.2 ± 96.9). This difference was statistically significant (p < 0.001) with a strong evidence of correlation (p < 0.001; Spearman's R: +0.72). ROC curve analysis was performed on CK19 expression index related to HPV positivity. Heterogeneous areas of immunoreactivity varying in percentage value, intensity and/or localization were observed in normal epithelium, both perilesional and distant from the tumor with important differences between HR-HPV(+) and HR-HPV(-) carcinomas. By ICC and flow cytometry, the two analyzed cell lines were both CK19 positive but showed a different level of expression, in particular it should be noted that the UPCI-SCC-154 (HPV(+)) cell line had a higher expression than UPCI-SCC-131 (HPV(-)). CONCLUSIONS: In this study we demonstrated, for the first time, strong association between CK19 up-regulation and HR-HPV(+) OSCCs/OPSCCs. This test has a good accuracy. We identified ROC curve with a cut-off > 195 for HR-HPV positive results (Sensitivity: 92.3 %; Specificity: 89.3 %). Furthermore, in OSCC/OPSCC, the CK19 test may be useful in identifying HR-HPV infection, the latter being related to HPV E7 potential to disrupt normal cytokeratin expression pattern.

Cell-mediated non-allergic rhinitis in children.

OBJECTIVE: Non-allergic rhinitis is a heterogeneous disease whose etiology is largely unknown. Nasal cytology only allows us to recognize different non-allergic rhinitis forms on the basis of the prevalent inflammatory cell infiltrate: non-allergic rhinitis with eosinophils, with neutrophils, with mast-cells and with both eosinophils and mast-cells. The aim of this study is to define the incidence, clinical features and comorbidity of the different types of cell-mediated non-allergic rhinitis in a pediatric age group. METHODS: One hundred and fourteen non-allergic children with chronic nasal obstruction and associated symptoms (rhinorrhea, sneezing and nasal itchiness) were retrospectively selected. All patients had been submitted to a clinical history, pediatric evaluation, anterior rhinoscopy and fiberendoscopy, rhinomanometry and nasal cytology. RESULTS: Non-allergic rhinitis with neutrophils was present in 46 (40.4%) children, non-allergic rhinitis with eosinophils in 53 (46.5%), non-allergic rhinitis with mast-cells in 12 (10.5%) and non-allergic rhinitis with both eosinophils and mast-cells in 3 (2.6%). Nasal obstruction was prevalent in non-allergic rhinitis with eosinophils and in non-allergic rhinitis with mast-cells patients (P<0.001) whereas rhinorrea and sneezing only in the form with eosinophils (P<0.0001). Nasal itching on the other hand was prevalent in the form with mast-cells (P<0.0003). Non-allergic rhinitis with eosinophils group showed a higher probability of asthma (P<0.02) and respiratory sleep disorders (P<0.04). CONCLUSIONS: In the pediatric age group the most frequent forms of non-allergic rhinitis are those with eosinophils or with neutrophils. A diagnosis of non-allergic rhinitis with eosinophils in children presumes more severe symptoms and a higher incidence of pulmonary disease and roncopathy.