Petr Čelakovský

High-risk human papillomavirus infection and p16INK4a protein expression in laryngeal lesions.

A total of 88 samples of laryngeal lesions (23 vocal cord nodules (VCNs), 23 papillomas (PAs), 18 dysplasias (DYs), and 24 carcinomas (CAs)) were analyzed for p16INK4a protein (p16) expression by immunohistochemistry and for high-risk human papillomavirus (HR-HPV) infection using chromogene in situ hybridization (CISH) and polymerase chain reaction (PCR). The series comprised 62 males and 26 females, aged 1-87 years (median 55 years). p16 expression was detected in 2 of 23 (9%) VCNs, 18 of 23 (78%) PAs, 9 of 18 (50%) DYs, and 14 of 24 (58%) CAs. Using CISH, HR-HPV DNA was detected in 3 of 23 (13%) VCNs, in 19 of 23 (83%) PAs, in 12 of 18 (67%) DYs, and in 14 of 24 (58%) CAs. HR-HPV DNA was found in six of nine (67%) PAs by PCR. A statistically significant difference in p16 expression and HR-HPV DNA presence detected by CISH was observed between VCNs and PAs (p<0.000001). The sensitivity and specificity of p16 expression for HR-HPV DNA presence detected by CISH was 0.612 and 0.773, respectively. Our study confirms a potential role of HR-HPV infection not only in the pathogenesis of malignant, but also in benign laryngeal lesions.

Histomorphologic study of nasal turbinates after surgical treatment: a comparison of laser surgery and radiofrequency-induced thermotherapy effects in animals.

Conclusions. From the clinical point of view, the healing process following radiofrequency-induced thermotherapy (RFITT) is faster and less painful for the patient. This corresponds with the histopathological findings, as in a case of diode laser treatment, the damage to the tissue was greater, and the regenerative and reparative processes were less prominent. Objective. Comparison of histopathologic changes and the healing process in the turbinate tissue regarding the kind of treatment and intensity of energy used for surgery. Materials and methods. Pigs were chosen as laboratory animals for this study. The animals were divided into two groups. Two different techniques were used: treatment with diode laser fiber applied submucosally in contact mode and RFITT. The first tissue samples were taken on the third postoperative day; further sampling was done on the eight postoperative day. Results. Histological features of the specimens taken from turbinates 8 days after the surgery included necrosis and ulceration, fibrin deposition in the mucosa, necrotizing sialometaplasia, as well as proliferation of the granulation tissue. Generally, the changes found in the turbinates treated by diode laser were more severe, with more intensive tissue damage and less prominent regenerative and reparative changes.

Risk factors for recurrent laryngeal nerve palsy after thyroidectomy

This study investigates the incidence of temporary and permanent recurrent laryngeal nerve palsy (RLNP) and possible risk factors for patients with different types of thyroid gland diseases. 1224 consecutive patients who underwent thyroidectomy for treatment of various thyroid diseases between the years 2001–2005. The rates of RLNP were evaluated. The surgeon and type of thyroid gland disorder were recognised as possible risk factors for RLNP. The incidence of temporary/permanent RLNP for the whole group was 4.5/0.8%. The rates of temporary RLNP for groups, classified as multinodular goitre, Graves’ disease, thyroid cancer or Hashimoto’s disease were 4.3%, 4.3%, 5.2% and 5.7%, respectively. The rates of permanent RLNP for the same groups were 0.4%, 0.9%, 1.6% and 1.9%, respectively. The frequency of temporary RLNP for individual surgeons ranges from 2.8 to 7.0% and the rates of permanent RLNP is between 0–3.1%. There was no relationship between the surgeon’s experience (the number of procedures performed) and RLNP rates. Total thyroidectomy is a safe procedure associated with a low incidence of RLNP not only for benign multinodular goitre, but also for Graves’disease, thyroid cancer and Hashimoto’s disease. The rates of RLNP among individual surgeons are acceptable with small inter-individual differences.

Unusual Manifestation of Anaplastic Thyroid Cancer

INTRODUCTION The authors present a rare case of a patient with symptoms consistent with retropharyngeal abscess. The diagnosis of anaplastic thyroid cancer was made after surgery and subsequent histological examination. CASE REPORT An 80-year-old woman was referred to Dpt. of Otolaryngology, Head and Neck Surgery, Charles University Medicine Faculty, Teaching Hospital in Hradec Králové with odynophagia and pain in the left side of the neck. The patient had pronounced swelling of the left side of her neck. We could also see swelling of the posterior pharyngeal wall, more pronounced on the left side. Inflammation markers were markedly elevated. Administration of antibiotics intravenously (amoxicillin combined with clavulan acid and gentamicin) was started. A computer tomography investigation (CT) was performed and a retropharyngeal abscess was found. The existence of a tumour was considered as well. An acute endoscopic examination and a puncture of the retropharyngeal space at the site of the swelling were performed, but no pus or any other liquid was found. On the sixth day of hospitalization a second CT scan was performed. As the retropharyngeal mass was still present along with continually elevated inflammatory markers, surgical revision of the retropharyngeal space from an external approach was performed. No abscess formation was found. During the surgery, retropharyngeal lymph nodes were removed for histological examination. The histological examination of the lymph nodes identified metastasis of anaplastic thyroid cancer. CONCLUSIONS The differential diagnosis of diseases affecting deep neck structures can be very difficult. Symptoms of inflammation dominating in the clinical picture do not exclude the possibility of malignancy. The most relevant imaging examination seems to be contrast enhanced computer tomography or magnetic resonance imaging.

SMARCB1/INI1-deficient sinonasal carcinoma shows methylation of RASSF1 gene: A clinicopathological, immunohistochemical and molecular genetic study of a recently described entity.

The aim of the study was detailed clinicopathological investigation of SMARCB1/INI1-deficient sinonasal carcinomas, including molecular genetic analysis of mutational status and DNA methylation of selected protooncogenes and tumor suppressor genes by means of next generation sequencing (NGS) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA). A total of 4/56 (7%) cases of SMARCB1/INI1-deficient carcinomas were detected among 56 sinonasal carcinomas diagnosed over a 19year period using immunohistochemical screening. The series comprised 3 males and 1 female, aged 27-76 years (median 64 years). All tumors arose in the nasal cavity. Three neoplasms were diagnosed in advanced stage pT4. During the follow-up period (range 14-111 months (median 72 months)), three tumors recurred locally, but none of the patients developed regional or distant metastases. Ultimately, two patients died due to the tumor. Microscopically, all tumors consisted of infiltrating nests of polygonal basaloid cells with a variable component of rhabdoid cells with eosinophilic cytoplasm. Immunohistochemically, there was almost diffuse expression of cytokeratins (CK), p16, p40 and p63 in all cases, while expression of CK5/6, CK7 and vimentin was only focal or absent. The detection of NUT gave negative results. In three cases, the absence of SMARCB1/INI1 expression was due to deletion of SMARCB1/INI1 gene. Methylation of SMARCB1/INI1 gene was not found. One tumor harbored HPV18 E6/E7 mRNA. All 12 genes (BRAF, BRCA1, BRCA2, KIT, EGFR, KRAS, NRAS, PDGFRA, PIK3CA, PTEN, RET, and ROS1) tested for mutations using NGS were wild-type. Regarding DNA methylation, all four SMARCB1/INI1-deficient tumors showed methylation of RASSF1 gene by means of MS-MLPA. There was a statistically significant difference in RASSF1 gene methylation between SMARCB1/INI1-deficient and SMARCB1/INI1-positive tumors (p=0.0095). All other examined genes (ATM, BRCA1, BRCA2, CADM1, CASP8, CD44, CDKN1B, CDKN2A, CDKN2B, CHFR, DAPK1, ESR1, FHIT, GSTP1, HIC1, KLLN, MLH1a, MLH1b, RARB, and VLH) were unmethylated. In summary, we described four cases of SMARCB1/INI1-deficient sinonasal carcinoma with detailed clinicopathological data indicating that these tumors can be regarded as a distinct entity with aggressive behaviour. For the first time, we performed analysis of DNA methylation in SMARCB1/INI1-deficient sinonasal carcinomas, reporting on significantly higher methylation of RASSF1 gene in this neoplasm.

Detection of Cervical Lymph Node Micrometastases in Patients with Squamous Cell Carcinoma of the Oral Cavity, Pharynx and Larynx.

BACKGROUND The goal of this prospective study was to determine the frequency of micrometastases in patients with squamous cell carcinoma (SCC) of the oral cavity, pharynx and larynx in whom elective neck dissection was indicated (cN0). PATIENTS AND METHODS A total of 12 patients (10 males and 2 females) were enrolled in the study. The age ranged 42-73 years (median 62 years). Elective neck dissection was performed in all patients (8 ipsilateral, 4 bilateral) and a total of 256 lymph nodes were removed and sent for microscopic examination. RESULTS The presence of tumor cells in cervical lymph nodes was found in 5/12 (42%) patients. Micrometastases of SCC were found in two patients and isolated tumor cells (ITC) in two other patients. In the remaining one patient with oropharyngeal SCC, a micrometastasis of papillary thyroid carcinoma (PTC) was detected. Positive lymph nodes were localized in level II in three patients with SCC of larynx, hypopharynx and tongue base, respectively, in level I in one patient with SCC of oral tongue and in level III in one patient with PTC. CONCLUSION Our results indicate that SCC of head and neck has a high potential for creating micrometastases which frequency is higher compared to clinically detected macrometastases. Therefore, elective neck dissection or radiotherapy of the neck should be considered in patients with high risk of occult metastases or micrometastases.

Oncocytic Schneiderian papilloma of frontal sinus.

Oncocytic Schneiderian papilloma (OSP) is one of the three morphologically distinct tumors that arise from Schneiderian membrane (the others include exophytic papilloma and inverted papilloma). OSP almost always occurs unilaterally in the paranasal sinuses, usually in the maxillary sinus, ethmoid cells or sphenoid sinus. We report a case of a 64-year-old woman with OSP arising from the left frontal sinus. In the report herein, we describe an OSP originating in the region of frontal sinus, which, to the best of our knowledge, represents the first documented example in English literature of OSP developing in this anatomical site.

Optic neuritis and paranasal sinus diseases

The relationship of sinusitis and paranasal sinus mucoceles to optic neuritis (ON) remains a challenge for both the otolaryngologist and the ophthalmologist. Currently, ON is assumed to be a rare complication of paranasal sinus diseases. Three new cases of ON associated with paranasal sinus diseases are described. We postulate different pathophysiologic mechanisms of ON in our three patients: compression of optic nerve due to sphenoid mucocele in the first patient; extension of bacterial infection to the optic nerve in the second patient; and invasion of aspergillosis to the orbital apex in the third patient. We assume that the complete recovery of the patient’s vision in the second case resulted from surgery, which had been performed immediately. A timely operation could possibly have prevented permanent blindness in cases 1 and 3 as well. Optic neuritis can occasionally be a complication of bacterial and mycotic sinusitis or paranasal sinus mucoceles, especially if located in the sphenoid sinus and posterior ethmoids. The evaluation of paranasal sinuses using imaging techniques is necessary to diagnose the problem, and immediate surgery can prevent permanent sequelae in indicated cases.