Jacopo Galli

The Role of Acid and Alkaline Reflux in Laryngeal Squamous Cell Carcinoma

Hypothesis At present, main factors considered responsible for the onset of squamous cell carcinoma are tobacco smoking, alcohol abuse, and exposure to viral and toxic agents. In last years, great interest has been focused on gastroesophageal reflux as independent carcinogenic factor and co‐carcinogen in association with smoking and alcohol assumption.

Prognostic significance of cyclooxygenase-2 in laryngeal squamous cell carcinoma.

Epidermal growth factor receptor (EGFR) overexpression is an unfavorable prognostic marker in laryngeal squamous cell carcinoma (SCC). EGFR stimulates cyclooxygenase‐2 (COX‐2) expression in normal human keratinocytes and squamous carcinoma cells. Based on these observations a prognostic role of COX‐2 expression in laryngeal SCC can be hypothesized. Consequently, COX‐2 expression was studied in laryngeal SCC (median follow‐up = 47 months; range: 2–87 months) by quantitative immunohistochemistry (n = 61) and EGFR by binding assay (n = 51). Well‐differentiated regions of laryngeal SCC revealed strong COX‐2 immunostaining, whereas histologically normal areas neighboring tumor as well as poorly‐differentiated tumors were negative. Immunohistochemical results were confirmed by Western blot analyses. Coxs regression analysis showed that the combination of low levels of COX‐2 integrated density and high levels of EGFR covariates provided strong prediction, at 5‐year follow‐up, of both poor overall survival (χ2 = 12.905; p = 0.0016) and relapse‐free survival (χ2 = 9.209; p = 0.01). In vitro studies on CO‐K3 cell line, obtained from an EGFR positive, COX‐2 negative poorly‐differentiated laryngeal SCC, revealed that EGF stimulation failed to induce COX‐2 expression and PGE2 production suggesting a change in EGFR signaling pathway. These findings indicate that COX‐2 is overexpressed in less aggressive, low grade laryngeal SCC, whereas its expression is lost when tumors progress to a more malignant phenotype.

Postlaryngectomy Pharyngocutaneous Fistula: Incidence, Predisposing Factors, and Therapy

OBJECTIVE: The pharyngocutaneous fistula (PCF) is a serious complication after total laryngectomy, and its etiology is not well understood yet. The aim of our study was to evaluate predisposing factors, incidence, and management of this complication. STUDY DESIGN AND SETTING: This was a retrospective study of 268 patients who underwent total laryngectomy in our clinic (January 1990-December 2001). A number of factors potentially predisposing to PCF formation were evaluated. RESULTS: A PCF was observed in 16% of patients. Systemic diseases, previous radiotherapy, supraglottic origin of tumor, and concurrent radical neck dissection were significantly associated with PCF. Spontaneous closure was noted in 28 patients, whereas a surgical closure was necessary in 15 patients. CONCLUSIONS: In presence of a specific risk factor, PCF can be expected; nevertheless, its prevention remains very difficult. Moreover, given the high percentage of spontaneous closure, we suggest the “wait and see” approach for 28 days before proceeding with a surgical approach.

Epidermal growth factor receptor expression in primary laryngeal cancer: An independent prognostic factor of neck node relapse

Specimens of laryngeal squamous cell carcinoma (LSCC) were examined for epidermal growth factor receptor (EGFR) content using a radioreceptor method; 140 untreated consecutive patients with primary LSCC undergoing initial surgical resection were followed up for a median of 49 months (range 2–84 months) after surgery. Cox univariate regression analysis using EGFR as a continuous variable showed that EGFR levels were directly associated with the risk of lymph node metastasis. A significant relationship between EGFR status and cervical node metastasis was observed. The cutoff value of 20 fmol/mg protein was the best prognostic discriminator. The 5‐year metastasis‐free survival (MFS) was 66% for patients with EGFR− tumors compared with 15% for patients with EGFR+ tumors. By multivariate analysis, the EGFR status appeared to be a significant independent prognostic factor for MFS. Our results suggest that the assessment of EGFR status at the time of diagnosis may identify a subset of LSCC patients highly susceptible to neck node metastases thus defining therapy accordingly. Int. J. Cancer (Pred. Oncol.) 84:188–191, 1999.

Prognostic significance of the Ca2+ binding protein S100A2 in laryngeal squamous-cell carcinoma

We investigated by immunocytochemistry the expression of the Ca2+ binding protein S100A2 in 62 cases of laryngeal squamous‐cell carcinoma (SCC). S100A2 was detected in 18/19 (95%) low‐grade tumors and in 22/43 (51%) high‐grade tumors, which were partially keratinizing. The remaining 21/43 (49%) high‐grade tumors were non‐keratinizing, anaplastic tumors and clearly S100A2‐negative. In normal laryngeal squamous epithelium and in laryngeal SCC, S100A2 expression was strictly associated with that of cytokeratins 14 (P = 0.0002) and 17 (P = 0.0021), suggesting an association of S100A2 expression and cell commitment to squamous differentiation. A correlation was found between S100A2 tumor positivity and longer relapse‐free (P = 0.0005) and overall (P = 0.0095) survival. Int. J. Cancer 89:345–349, 2000.

HPV infection in squamous cell carcinomas arising from different mucosal sites of the head and neck region. Is p16 immunohistochemistry a reliable surrogate marker

Background:Human papillomavirus 16 infection has been proven to be associated with oropharyngeal squamous cell carcinomas (SCCs) and is probably the main reason of the reported increase in the incidence. The role of high-risk (HR) HPV for carcinogenesis of other sites in the head and neck awaits confirmation. With the aim to evaluate the prevalence of HPV infection and the reliability of different diagnostic tools in SCCs of different sites, 109 consecutive untreated head and neck SCCs were enroled, and fresh tumour samples collected.Methods:Human papillomavirus DNA was detected by Digene Hybrid Capture 2 (HC2). Human papillomavirus E6 and E7 mRNA were detected by NucliSENS EasyQ HPVv1. P16 expression was evaluated by immunohistochemistry.Results:In all, 12.84% of cases were infected by HR genotypes and 1.84% by low-risk genotypes. Human papillomavirus 16 accounted for 87% of HR infections. The overall agreement between DNA and RNA detection is 99.1%. Although p16 expression clearly correlates with HPV infection (P=0.0051), the inter-rater agreement is poor (k=0.27). The oropharynx showed the highest HR HPV infection rate (47.6%) and was also the only site in which p16 immunohistochemistry revealed to be a fair, but not excellent, diagnostic assay (κ=0.61).Conclusion:The prognostic role of HR HPV infection in oropharyngeal oncology, with its potential clinical applications, underscores the need for a consensus on the most appropriate detection methods. The present results suggest that viral mRNA detection could be the standard for fresh samples, whereas DNA detection could be routinely used in formalin-fixed, paraffin-embedded samples.

Magnetic resonance imaging findings in sudden sensorineural hearing loss.

OBJECTIVE To investigate the role of magnetic resonance imaging (MRI) in the diagnosis of sudden sensorineural hearing loss (SSNHL). METHODS Fifty-four consecutive patients affected by SSNHL were investigated using brain MRI. MRI was performed with an eight-channel phased-array head coil to study the entire audiovestibular pathway and the whole brain. The protocol study consisted of a high-resolution study of the temporal bone, internal auditory canal (IAC), cerebellopontine angle (CPA), and brainstem combining 2 mm thin-slice axial T(2)-weighted two-dimensional fast spin echo (FSE) and fluid-attenuated inversion recovery (FLAIR) sequences, pre- and postcontrast (gadolinium-diethylenetriamine pentaacetic acid) administration fat-suppressed axial T(1)-weighted two-dimensional FSE sequences, and a T(2)*-weighted three-dimensional Fourier transformation-constructive interference in steady state sequence (FT-CISS) , with 0.4 mm ultrathin partitions. The rest of the brain was studied with a 4 mm axial T(2)-weighted FLAIR sequence. RESULTS Thirty-one of 54 (57%) cases of SSNHL presented with MRI abnormalities. In 6 of 54 cases, the detected abnormality was directly correlated to the clinical picture (2 labyrinthine hemorrhage, 1 cochlear inflammation, 1 acoustic neuroma, 1 arachnoid cyst of the CPA, and 1 case of white matter lesions in the pons, compatible with demyelinating plaques along the central audiovestibular nervous pathway, as the first expression of multiple sclerosis). CONCLUSIONS An extensive MRI study of the audiovestibular nervous pathway and of the whole brain, pre- and postparamagnetic contrast administration, is recommended to rule out the wide spectrum of abnormalities that can cause SSNHL.

Human papillomavirus infection and cyclin D1 gene amplification in laryngeal squamous cell carcinoma: Biologic function and clinical significance†

Human papillomavirus (HPV) infection is suspected to be a risk factor for head and neck, and in particular for laryngeal, carcinogenesis. Cyclin D1 gene (CCND1) overexpression and amplification have been shown to play a role as prognostic factors in many human cancers, among which are head and neck cancers.

Damage to Ciliated Epithelium in Chronic Rhinosinusitis: What is the Role of Bacterial Biofilms?

Objectives: We assess the association between the presence of biofilms and cilial damage in patients with chronic rhinosinusitis (CRS), describe the microorganisms associated with samples that exhibited cilial loss and biofilms, and demonstrate the absence of ciliary injury and biofilms in similarly prepared “normal” controls. Methods: We examined samples of ethmoid mucosa obtained from 24 patients who underwent functional endoscopic sinus surgery for CRS. Samples from a control group (20 healthy subjects) were also examined. The specimens were divided into 2 fragments; the first was processed for bacterial cultures, and the second was subjected to scanning electron microscopy. Statistical analysis was performed. Results: All CRS samples had positive bacterial cultures. The scanning electron microscopy analysis showed bacterial biofilms in 10 of the 24 specimens. A marked destruction of the epithelium was observed in samples positive for biofilms (p < 0.001), and the presence of Haemophilus influenzae was associated with ciliary abnormalities (partial damage in 55.6% and absence of cilia in 50%; p = 0.041). Conclusions: The high percentage of biofilms in our specimens confirms the association between biofilms and CRS. Our data support the hypothesis that biofilm formation represents the latter phase of an inflammatory process that leads to complete epithelial destruction.

Low serum folate levels: A risk factor for sudden sensorineural hearing loss?

Objective Ischemic vascular damage of the inner ear is one of the known causes of sensorineural sudden hearing loss (SSHL). Folate is an emerging risk factor associated with an increased risk of vascular damage. The aim of this study was to investigate whether low serum folate levels are associated with SSHL. Material and Methods Serum folate levels were determined in 43 patients with SSHL and in 24 controls. Results Folate levels were found to be significantly lower in SSHL patients than in controls (mean difference −1.96 ng/ml; 95% CI −3.31, −0.59 ng/ml; p=0.006). No significant relationship between folate levels and either sex, age, cigarette smoking, alcohol consumption or hypertension was observed, while a significant relationship was found between low folate levels and high homocysteine (HCY) levels in all 43 patients (p<0.01). The potential influence of low folate levels on hearing impairment in SSHL patients can be explained by the effects on HCY metabolism and the diminution of folate antioxidant capacity. Conclusion Further studies are needed to elucidate whether low folate levels can be considered a risk factor for SSHL.