Arterial stiffness as an ultrasound biomarker of radiation-induced carotid artery disease.

Background: Radiation-induced carotid artery disease (RICAD) is an important issue in head and neck cancer (HNC) survivors after radiotherapy (RT). The risk of cerebrovascular disease in these patients is doubled. The aim of this study was to assess the effect of RT on carotid artery stiffness in HNC patients. Patients and methods: Conventional arterial stiffness parameters were measured in a total of 50 HNC survivors treated with RT for at least 5 years and compared to 50 unirradiated HNC patients. Elastic modulus (Ep) and Beta stiffness index (ß) were measured in proximal, mid and distal common carotid artery (CCA). Results: The mean age of the subjects was 68±9 years (range: 44-84) in the irradiated group and 67±10 years (range: 45-85) in the control group. The RT group was treated with a mean radiation exposure of 60.3±6.7 Gy (range: 44-72) in the neck. Carotid stiffness parameters showed significant group differences: Ep in the RT group was 2.329±1.222 vs 1.742±828 in the non-RT group (p=0.006) and ß index in the RT group was 23±11 vs 15±8 in the non-RT group (p<0.001). Radiation-induced carotid stiffness was quantified and cervical exposure to RT increased Ep in 575 kPa (p=0.014) and ß in 7 units (p<0.003). Conclusions: Ep and ß index could be suitable ultrasound biomarkers of radiation-induced atherosclerosis in HNC survivors. Further prospective studies are needed to feature RICD in this setting.

Prognostic significance of lymphangiogenesis in pharyngolaryngeal carcinoma patients.

BACKGROUND: Lymphatic vessel spread is considered a major route for head and neck squamous cell carcinoma metastasis. Formation of new lymphatic vessels could facilitate the process, raising the malignant potential of these tumours. Recent identification of lymphatic markers allows the study of the lymphangiogenesis phenomenon. We searched for molecular events involved in the lymphangiogenic process that could have prognostic value in laryngeal/pharyngeal carcinoma patients. METHODS: 104 paraffin-embedded pharyngeal/laryngeal tumour samples were studied. Immunohistochemical analysis of podoplanin and double immunofluorescence analysis of Ki-67 and D2-40 were performed. Lymph vessel density (inside the tumour mass, at its periphery or considered as a whole) and the presence of tumour emboli inside lymphatics were recorded. The proliferative state of endothelial lymphatic cells was evaluated. RESULTS: Lymphatic vessels were detected inside the tumour mass (75%) and in the surrounding tissue (80%); some of them in a proliferative state. Tumour emboli were detected in a high proportion of the cases (45%). Lymphatic vessel density was higher in the pharyngeal cases (p = 0.0029), in greater size (p = 0.039), more advanced stage primary tumours (p = 0.006) and in carcinomas of patients with affected nodes (p = 0.019). The presence of tumour emboli and a high global vessel density were indicators of poor prognosis (recorded as death from tumour) in the laryngeal group (p = 0.015 and p = 0.027, respectively), but notably not in the pharyngeal one. Interestingly, high global vessel density showed a negative prognostic value among pathologically staged N0 laryngeal carcinomas (p = 0.03). CONCLUSIONS: The lymphangiogenic process correlated with aggressive tumour features (pN category, tumour size, tumour stage), but might play different roles in tumours arising from different anatomic sites. Our results suggest that detection of tumour emboli and assessment of global vessel density using the D2-40 antibody, may be useful in the clinical practice, as predictors of reduced survival among pN0 laryngeal carcinoma patients.

Conservative treatment of retropharyngeal and parapharyngeal abscess in children.

Parapharyngeal and retropharyngeal infections, which can potentially cause life-threatening complications, could be treated conservatively with no need for surgical drainage. A retrospective analysis of all patients diagnosed with retropharyngeal and parapharyngeal infections was performed. Information regarding age, sex, presenting symptoms, physical examination, laboratory and imaging evaluations, management, duration of hospital stay, and complications were reviewed. Seven children were identified, 4 with retropharyngeal abscess and 3 with parapharyngeal abscess. All but 1 patient were under 7 years old, and all were treated with intravenous amoxicilin/clavulanic acid and corticosteroids. Torticollis and fever were present in all the patients. The mean length of hospital stay was 7 days. There were no complications associated. We demonstrate that retropharyngeal and parapharyngeal abscesses can be treated medically, reserving the surgical drainage for complicated cases. Treatment with intravenous antibiotics and corticosteroids is a safe option, reducing the durations of symptoms and the length of hospital stay.

Small cell and large cell neuroendocrine carcinoma of the larynx: A comparative analysis.

The poorly differentiated neuroendocrine carcinomas (NECs) of the larynx are rare tumors that comprise of a small cell-type (SCNEC) and a large cell-type (LCNEC). In order to consolidate the current knowledge about their characteristics and management a systematic review of the available literature was performed. The PubMed/Medline and Scopus databases search resulted in 141 articles published between 1972 and 2019, describing 273 patients: 230 cases were of SCNEC histology (84.2%) and 43 cases were LCNECs (15.8%). Comparing both histological entities, patients with LCNECs were older (63.2 vs. 58.7 years, p = 0.036) than SCNEC patients and had more often primary tumor in a supraglottic larynx (79.5 vs. 56.1%, p = 0.039), advanced-stage neck disease (N2-3, 56.8 vs. 40%, p = 0.061), surgery-based treatment (83.7 vs. 51.9%, p < 0.001) and had no radiotherapy (44.2 vs. 29%, p = 0.071). At presentation, systemic metastases were diagnosed in 12.1% of the patients, whereas disease relapse was experienced by two-thirds (65.3%) of those initially staged M0; systemic relapse, alone or in combination with local/regional recurrence, was the most frequent type of failure (in 71.9%). On multivariate analysis, more advanced disease stage and SCNEC histology adversely influenced disease-specific survival. Wide variations in the pattern of care, including radiotherapy doses and chemotherapy regimens, were found among long-term survivors without known disease at ≥ 24 months of follow-up (N = 36). We conclude that the most effective treatment for poorly differentiated NECs has yet to be determined.

Annexin A2 localizes to the basal epithelial layer and is down-regulated in dysplasia and head and neck squamous cell carcinoma.

Annexin A2 is a highly expressed gene with important roles in cell membrane physiology and is frequently dysregulated in cancer. The objective of this study was to determine the pattern of expression and prognostic significance of annexin A2 protein in head and neck squamous cell carcinoma. We assessed both quantitative changes and qualitative distribution of annexin A2 mRNA and protein expression in normal and diseased tissues by immunohistochemistry, immunofluorescence and in situ hybridization. Annexin A2 expression was confined to the basal and suprabasal cells of normal epithelium and the protein cellular location was consistently observed at the cell membrane. Expression levels correlated with histopathological grade, showing significant suppression in moderately and poorly differentiated tumours. We conclude that annexin A2 exhibits a characteristic pattern of expression, distinct from other annexins and suggestive of a cell-specific functional role. The marked reduction of annexin A2 in poorly differentiated tumours and dysplastic tissue is expected to result in a loss of function aimed at the coordination of membrane signalling enzyme complexes, actin polymerization and extracellular matrix proteolysis. The phenotypic consequences may become manifest in an alteration of epithelial tissue growth and remodelling with secondary influence on tumour development, progression and metastasis.

Epithelial cell nicotinic acetylcholine receptor expression in head and neck squamous cell carcinoma pathogenesis.

BACKGROUND: Molecular events following nicotinic acetylcholine receptor (nAChR) activation by nicotine are poorly understood. The phosphatidylinositol 3-kinase (PI3-K)/Akt/PTEN pathway has been suggested to play a role in the antiapoptotic responses to nicotine. MATERIALS AND METHODS: To elucidate the possible role of aalpha3, alpha5 and alpha7 nAChR subunit mediated PI3-K/Akt/PTEN pathway activation in squamous cell carcinoma of the head and neck (HNSCC) development, mRNA was isolated from 30 HNSCC tissues of known Akt activation state and were analyzed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: alpha3, alpha5 and alpha7 nAChR subunits were expressed in 1/30 (3.33%), 15/30 (50%) and 10/30 (33.33%), respectively. These results did not correlate with pAkt levels, previously assessed in our laboratory, or any of the clinicopathological parameters considered. CONCLUSION: This is the first report on nAChR subunit expression in human HNSCC surgical specimens of known pAkt levels. Our results suggest that nAChRs might exert their function through pathways different from PI3-K/Akt/PTEN and that alpha3, alpha5 or alpha7 nAChR subunit expression might not be useful prognostic markers in HNSCC.

Frequent genetic and biochemical alterations of the PI 3-K/AKT/PTEN pathway in head and neck squamous cell carcinoma.

We investigated the status of the PI 3-kinase/AKT/PTEN signaling pathway in a series of 117 head and neck squamous cell carcinomas (HNSCC) in a search for molecular alterations in genes/proteins with potential prognostic value. For this purpose, PIK3CA and AKT2 gene amplification was assessed by multiplex and Quantitative Real-Time PCR. Protein expression of AKT, p-AKT, p110alpha and PTEN was determined by Western blot. PTEN allelic loss was evaluated by microsatellite analysis. PTEN-exon 5 was screened for point mutations by PCR-SSCP. Homozygous deletions were determined by multiplex PCR. PIK3CA gene was amplified in 43/117 (37%) fresh tumor samples, a frequency that did not differ from that found in archival premalignant tissues: 15/38 (39%); 12/40 (30%) fresh tumors harbored AKT2 gene amplification. AKT was found activated in 6/36 (17%) fresh tumor samples, when compared to their normal tissue counterparts. Of these 6 cases, 1 showed p110alpha overexpression and 5 displayed PTEN protein downregulation. Neither allelic loss (found in 11/77 informative cases) nor point mutations or homozygous deletions accounted for the reduced PTEN protein expression observed in our tumor series. The histologically normal mucosa of 4 patients displayed some of the molecular alterations analyzed. Dysregulation of the PI 3-K/AKT/PTEN pathway might contribute to early HNSCC tumorigenesis and might constitute a potential clinical target. Overall, 17/36 (47%) cases showed at least 1 of the molecular alterations studied here, which makes the PI 3-kinase-initiated signaling pathway one of the most frequently altered in HNSCC.