Towards effectiveness of cell free DNA based liquid biopsy in head and neck squamous cell carcinoma.

Liquid biopsy is a minimally invasive procedure, that uses body fluids sampling to detect and characterize cancer fingerprints. It is of great potential in oncology, however there are challenges associated with the proper handling of liquid biopsy samples that need to be addressed to implement such analysis in patients' care. Therefore, in this study we performed optimization of pre-analytical conditions and detailed characterization of cfDNA fraction (concentration, length, integrity score) in surgically treated HNSCC patients (n = 152) and healthy volunteers (n = 56). We observed significantly higher cfDNA concentration in patients compared to healthy controls (p < 0.0001) and a time dependent decrease of cfDNA concentration after tumor resection. Our results also revealed a significant increase of cfDNA concentration with age in both, healthy volunteers (p = 0.04) and HNSCC patients (p = 0.000002). Moreover, considering the multitude of HNSCC locations, we showed the lack of difference in cfDNA concentration depending on the anatomical location. Furthermore, we demonstrated a trend toward higher cfDNA length (range 35-10380 and 500-10380 bp) in the group of patients with recurrence during follow-up. In conclusion, our study provide a broad characterization of cfDNA fractions in HNSCC patients and healthy controls. These findings point to several aspects necessary to consider when implementing liquid biopsy in clinical practice including: (I) time required for epithelial regeneration to avoid falsely elevated levels of cfDNA not resulting from active cancer, (II) age-related accumulation of nucleic acids accompanied by less efficient elimination of cfDNA and (III) higher cfDNA length in patients with recurrence during follow-up, reflecting predominance of tumor necrosis.

Irreversible facial nerve palsy as a revelator ofparotid gland cancer.

<br><b>Introduction:</b> Bell's palsy is still the diagnosis of exclusion. In most patients it does not progress, it is unilateral, and selflimiting. Additionally, the majority of patients recover spontaneously within 3 weeks. It is well known that all patients with irreversible facial nerve paresis (FNP) need further examinations to exclude the organic, infectious, metabolic, and autoimmunological causes of the palsy. The goal of the study was to assess the frequency of malignancies hidden under the diagnosis of "Bell's palsy".</br> <br><b>Aim:</b> We aimed to create a diagnostic algorithm to avoid failures concerning patients whose only symptom of parotid gland cancer was irreversible FNP.</br> <br><b>Material and methods:</b> We analyzed 253 consecutive patients with FNP treated in our department in the last 5 years. The subject of the study was "Bell's palsy" cases. All patients with irreversible FNP were reassessed in 6-12 months. We underlined all shortcomings in the diagnostics of those in whom malignancies were found in MRI of the neck and presented the proposal for a diagnostic algorithm to avoid missing such an entity.</br> <br><b>Results:</b> Bell's palsy was observed in 157/253 patients (62.06%), in 36/157 (22.92%) it remained permanent. In 4/36 patients (11.11%) with irreversible FNP, which constituted 2.54% of all "Bell's palsy" cases, parotid gland deep lobe mass was found in MRI. In one patient, infiltration of the skull base was diagnosed. Adenoid cystic carcinoma was confirmed in final histopathology in all cases.</br> <br><b>Conclusions:</b> Our experience has shown that irreversible FNP can be a revelator of the malignant tumor located in the deep lobe of the parotid gland. Contrast-enhanced MRI covering intra- and extracranial segments of the facial nerve should be ordered in all cases of FNP without recovery after 4 months. Repeated imaging should be considered in undiagnosed cases. The main point of our study is to underline that the assessment of the deep lobe of the parotid gland with MRI should be included in the standard diagnostic protocol in all irreversible "Bell's palsy" cases.</br>.

The difficult "childhood era" of the liquidbiopsy as a diagnostic method in head and neckcancer patients.

<br><b>Aim:</b> Liquid biopsy (LB) is a method that detects circulating tumor cells or circulating tumor DNA or RNA in the body fluids of patients with cancer. Despite the developments in LB, it is still not used in clinical practice in head and neck cancers (HNC). The aim of our study was to analyze the epidemiological data of HNC patients and controls who were enrolled in an LB study based on circulating free DNA (cfDNA) detection.</br> <br><b>Material and methods:</b> A group of 152 patients diagnosed with HNC (128 men and 24 women) and 56 healthy volunteers (48 men and 8 women) were enrolled into the study. Peripheral blood samples were collected before treatment from HNC patients and controls. Plasma was isolated and cfDNA concentration was assessed in the range of 35-10,380 bp.</br> <br><b>Results:</b> The comparison of cfDNA concentration by gender between the HNC patients and the control group, and by comorbidities in the control group, showed no significant differences (p values: 0.13-0.69, 0.15-0.50 and 0.13-0.80, respectively).</br> <br><b>Conclusions:</b> Patients' gender and general status were found to have no effect on cfDNA concentration. Further analysis is necessary to define other correlations and the possible application of LB in HNC diagnosis, follow-up, and treatment.</br>.

The assesment of intraoperative use of mesna in canal wall-up cholesteatoma surgery - preliminary report.

<b>Introduction:</b> Surgery is still the method of choice in chronic otitis media with cholesteatoma. Except for some specific clinical situations, classic canal wall up technique (CWU), remains a gold standard as a primary treatment in most departments. Unfortunately, the risk of recurrence in such an approach is estimated at 9 to even 70%. This fact prompts researchers to look for ways to reduce those unfavourable statistics. One of the recognized methods supporting the removal of cholesteatoma is the intraoperative use of mesna (sodium 2-mercaptoethanesulfonate). This synthetic sulphur compound disrupts disulfide bridges in polypeptide chains, thanks to which it facilitates matrix preparation.</br></br> <b>Aim:</b> To evaluate the effect of intraoperative use of mesna on the treatment outcomes in patients with chronic otitis media with cholesteatoma operated on by means of the canal wall up technique (CWU).</br></br> <b>Material and methods:</b> 459 surgical reports of patients with middle ear cholesteatoma were analyzed. In total, 52 adult patients with no history of previous ear surgery operated on by means of the CWU technique by the same experienced otosurgeon with all follow-up data available were included in the study. Twenty-six were operated on with the use of mesna (mesna group) and 26 by means of the classic CWU technique (control / no-mesna group). There were 28 women and 24 men with a mean age of 41 years.</br></br> <b>Main Outcome Measure(s):</b> Postoperative hearing results and cholesteatoma recidivism rate.</br></br> <b>Results:</b> Overall recidivism rate was 21.15 %. It was higher in the no-mesna (26.9%) than in the mesna group (15.4%) - although the outcomes were better in the mesna group, the difference was not statistically significant (P = 0.49715). Hearing gain was better in the mesna than in the no-mesna group (10 dB vs 7 dB), but the difference was not statistically significant (P = 0.20089).</br></br> <b>Conclusions:</b> Our preliminary results show that mesna reduces recidivism rates in patients with cholesteatoma. Further study with the analysis of a larger group of patients is needed to prove it statistically.

Is Facial Nerve Decompression Justified in Malignant External Otitis? Literature Review and Own Experience.

OBJECTIVE: The aim of the study was to present the indications for facial nerve decompression in malignant external otitis, to analyze the results of such treatment, and to describe own experience in that field. METHODS: A search in the PubMed and Google Scholar databases for English language articles published between 1968 and May 2022 was performed. We focused on papers describing patients with malignant external otitis and facial nerve palsy treated by decompression. Moreover, retrospective analysis of 24 consecutive patients with malignant external otitis hospitalized in our department in the past 10 years was performed. RESULTS: In the literature, 48 cases with malignant external otitis and facial nerve paresis treated by decompression were identified. In total, 41 patients recovered (85.42%), 4 died (1 due to exacerbation of the disease, 2 for pneumonia, and 1 for heart failure), and in 3 cases the final outcome was not presented. In most cases (24; 50%), facial nerve function did not improve after decompression, in 8 patients (16.67%) partial recovery was observed, in 11 cases (22.92%) full improvement was observed, and data were not given for 5 patients. In only one case, the spread of infection and deterioration of local and general patient states were noted. In our material, we identified 13 patients with malignant external otitis and facial nerve palsy. Two of them were treated surgically, of which only one had facial nerve decompression. Partial improvement of facial nerve function was observed after 6 months of rehabilitation. CONCLUSION: The management of malignant external otitis is still difficult and not well defined. Facial nerve decompression seems justified in selected cases of malignant external otitis not responding to conservative treatment.

Challenges in the diagnosis and treatment of the malignant adnexal neoplasms of the head and neck.

PURPOSE OF REVIEW: The current review is to present the current knowledge regarding epidemiology, diagnostics, and management of malignant adnexal neoplasms (MANs). RECENT FINDINGS: Immunotherapy and gene-related therapies are still being developed as the methods of salvage treatment in advanced and disseminated cases: CACNA1S, ATP2A1, RYR1, and MYLK3, as well as p53 or the JAK/STAT pathways, may be therapeutic targets; the efficiency of talimogene laherparepvec and nivolumab is assessed. SUMMARY: MANs are rare tumors, but due to the aging of population their incidence is increasing. Their clinical presentation is unspecific, which makes the diagnosis challenging. Histopathological assessment is difficult even for experienced pathologists. Mohs micrographic surgery or wide local excision are recommended to treat primary lesions. Adjuvant radiotherapy may be beneficial in case of insufficient or positive surgical margins, in nodal metastases, in selected types of MANs like sebaceous, trichilemmal, and pilomatrix carcinomas, and as the induction treatment in large tumors located in medically fragile or cosmetically important regions. The role of chemotherapy is not well defined; however, it is recommended in distant metastases. Immunotherapy can improve the prognosis in advanced stage of the disease.

"Sandwich technique" enables preservation of hearing and antivertiginous effect in cholesteatomatous labyrinthine fistula.

PURPOSE: The aim of the study was to assess hearing, surgical and clinical results of the treatment in patients with cholesteatomatous labyrinthine fistula (LF) focusing on the different techniques and materials used in the management. METHODS: Study group included 465 patients. Cases with LFs discovered or confirmed during surgical procedure were thoroughly analyzed. RESULTS: LFs were noted in 11.4% of all cases. Thirty-eight patients, with all follow-up data available, were included into the further analysis. Most LFs were located in the lateral semicircular canal (87%). LFs were assessed as small in 2 cases, as medium in 24 patients while 12 were described as large. Based on Dornhoffer and Milewski classification, 50% of LFs were classified as IIa, 24% as IIb, 6 LFs were very deep (type III), while 4-superficial (type I). The size and type of LF did not influence postsurgical complaints (p = 0.1070, p = 0.3187, respectively). Vertigo was less frequent in LFs treated by "sandwich technique", especially those with opened endosteum. In 30 (79%) patients, hearing improved or did not change after surgery. Hearing outcomes were significantly better in the ears operated by means of CWU technique (p = 0.0339), in LFs with intact membranous labyrinth (p = 0.0139) and when "sandwich technique" was performed (p = 0.0159). Postsurgical bone conduction thresholds levels were significantly better in LFs covered by "sandwich method" (p = 0.0440). CONCLUSION: "Sandwich technique" (temporal fascia-bone pate-temporal fascia) enables preservation of hearing as well as antivertiginous effect in patients with cholesteatomatous labyrinthine fistula.

Facial nerve paresis in the course of masked mastoiditis as a revelator of GPA.

PURPOSE: The aim of this study was to present a series of 6 patients with facial nerve palsy and masked mastoiditis which constituted as revelators of localized granulomatosis with polyangiitis (GPA) and to evaluate the utility of the ACR/EULAR 2017 provisional classification criteria for GPA in such cases. METHODS: Study group included 58 patients with GPA. Cases with facial nerve palsy and masked mastoiditis were thoroughly analyzed. RESULTS: The mean age of patients was 37 years. All manifested unilateral facial nerve palsy and hearing loss, while only 2 reported aural complaints suggesting inflammatory cause of the disease. All cases were qualified for surgical intervention. Intraoperative findings were similar: granulation tissue in tympanic cavity and/or pneumatic spaces of the mastoid process. Only 50% of histopathological results suggested vasculitis. In all cases, elevated levels of antineutrophil cytoplasmic antibodies (ANCA) against peroxidase 3 (PR3-ANCA) were determined. Two patients presented rapid progression of the disease and died within 1 week and 2 months, respectively. Four other patients manifested gradual improvement of hearing and facial nerve function after treatment. CONCLUSION: GPA should be included into differential diagnosis in all cases of persistent facial nerve palsy especially when otological symptoms coexist. Even localized GPA could be very aggressive, revelating generalized form of the disease. Rapid systemic treatment of GPA can protect hearing and facial nerve from permanent severe dysfunction. The ACR/EULAR 2017 provisional classification criteria for GPA seem to be valuable tool in diagnosing ENT patients with localized otological form of the disease.

Efficacy of surgical treatment in patients with post-traumatic facial nerve palsy.

INTRODUCTION: The most common mechanism of post-traumatic facial nerve palsy are road accidents and falls. Treatment schemes as well as proper timing of surgery are still controversial. <br/><br/>Aim: The aim of the study was the evaluation of the effects of surgical treatment in patients with post-traumatic facial nerve palsy. Treatment results were correlated with epidemiological factors, mechanism of injury, level of nerve damage, time of surgery and its extent. <br/><br/>Material and methods: 9 patients with facial nerve palsy after head trauma were analyzed. In all patients complete paresis of the VII nerve occurred immediately after the injury. In 5 patients the nerve was damaged in the course of the longitudinal fracture of the temporal bone, in 3 as a result of its transverse fracture while in one woman there was no evident fracture line. In all cases, surgical treatment was performed between 4 days and 13 weeks after the trauma. In all cases transmastoid approach was used. Edema lesions of the nerve dominated in 6 patients, in two cases a bone fragment was noted along its course, in one person nerve was disrupted but primary reconstruction was not possible - the man was excluded from further analysis. The results of treatment were assessed by House-Brackmann (HB) scale 12 months after the procedure. <br/><br/>Results: Very good (HBI) or good (HBII) recovery of facial nerve function was achieved in 2 and 4 out of 8 patients respectively. Surgical timing, the extent of surgery, patient's age, mechanism of injury and level of nerve damage had no effect on the final outcome. <br/><br/>Conclusions: The management of post-traumatic facial nerve palsy should be individual. The commonly accepted recommendation on surgical treatment is to undertake it in patients with immediate-onset and complete paralysis. Patients who, due to their severe general condition, cannot undergo early facial nerve decompression may benefit from delayed treatment for up to 3 months after the injury.

Acinetobacter: An Enemy after Head and Neck Cancer Operations with Microvascular Free Flap Reconstruction?

Background: Patients after head and neck cancer reconstructive surgical procedures are predisposed to have post-operative surgical site infections (SSI) develop. They are very often caused by multi-drug resistant strains, including Acinetobacter baumannii as the most common one. Methods: The aim of the study was to determine important risk factors contributing to SSI of A. baumannii origin. The analysis included 134 head and neck cancer patients after salvage operations with microvascular free flap reconstruction. The A. baumannii was cultured in 27 of all 48 infected patients. Results: The following risk factors were significantly associated with A. baumannii infection: re-hospitalization before reconstructive operation (p = 0.00011), massive blood loss (p = 0.00277), and need of revision surgical procedure (p = 0.00419). Of patients with A. baumannii infection, 48% were hospitalized in a general intensive care unit (ICU) after operation that, together with prolonged intubation, constituted a strong risk factor of that infection (p = 0.01077). Mean time of hospital stay was significantly longer in the A. baumannii group (58 days vs. 35 days; p = 0.02697). Conclusions: Our analysis identified a subset of head and neck cancer patients after salvage operation with microvascular free flap reconstruction who are at high risk of A. baumannii infection developing. Previously hospitalized patients with extensive blood loss and need of surgical revision necessitate increased monitoring for the development of this complication. Mechanical ventilation and hospital stay in an ICU should be shortened maximally or avoided in that challenging group of patients. Early recognition of patients at high risk remains a key point to prevent or limit the spread of A. baumannii infections.

Copy number gains of the putative CRKL oncogene in laryngeal squamous cell carcinoma result in strong nuclear expression of the protein and influence cell proliferation and migration.

Laryngeal squamous cell carcinoma is a major medical problem worldwide. Although our understanding of genetic changes and their consequences in laryngeal cancer has opened new therapeutic pathways over the years, the diagnostic as well as treatment options still need to be improved. In our previous study, we identified CRKL (22q11) as a novel putative oncogene overexpressed and amplified in a subset of LSCC tumors and cell lines. Here we analyze to what extent CRKL DNA copy number gains correlate with the higher expression of CRKL protein by performing IHC staining of the respective protein in LSCC cell lines (n = 3) and primary tumors (n = 40). Moreover, the importance of CRKL gene in regard to proliferation and motility of LSCC cells was analyzed with the application of RNA interference (siRNA). Beside the physiological cytoplasmic expression, the analysis of LSCC tumor samples revealed also nuclear expression of CRKL protein in 10/40 (25%) cases, of which three (7.5%), presented moderate or strong nuclear expression. Similarly, we observed a shift towards aberrantly strong nuclear abundance of the CRKL protein in LSCC cell lines with gene copy number amplifications. Moreover, siRNA mediated silencing of CRKL gene in the cell lines showing its overexpression, significantly reduced proliferation (p < 0.01) as well as cell migration (p < 0.05) rates. Altogether, these results show that the aberrantly strong nuclear localization of CRKL is a seldom but recurrent phenomenon in LSCC resulting from the increased DNA copy number and overexpression of the gene. Moreover, functional analyses suggest that proliferation and migration of the tumor cells depend on CRKL expression.

Conductive hearing loss after surgical treatment of otosclerosis - long-term observations.

<b>Introduction:</b> For many years, surgical treatment of otosclerosis has been a widely accepted approach. Hearing improvement following stapes surgery is sometimes spectacular, and good treatment results are obtained in many centers in over 90% of patients. However, in the subsequent years after the treatment, some patients develop permanent or progressive conductive hearing loss. <br><b>Aim:</b> The aim of the study is to present a group of patients with conductive hearing loss after the first otosclerosis surgery and to analyze the causes of its occurrence. <br><b>Materials and Methods:</b> The retrospective review covered patients who underwent the initial surgery in the years 2000-2009. We analyzed the patients' medical records from before the end of 2019, which provided results of at least 10 years of postoperative follow-up. The group consisted of 1118 patients aged 14-82, including 802 women and 316 men.<br><b> Results:</b> Reoperations due to conductive hearing loss were performed on 93 patients, who accounted for 8.3% of the originally operated patients. They were much more common in patients after stapedectomies (19.7%) than in patients after stapedotomy (5.5%). Prosthesis dislocation was found to be the most frequent intraoperative observation (44.1%) and was often associated with erosion or necrosis of the long process of incus (28%). Less frequent reasons for hearing loss were: adhesions around the prosthesis (10.8%), too small hole in the stapes footplate (8.6%), too short prosthesis (8.6%), progression of otosclerosis (7.5%), too long prosthesis (6.4%), presence of a granuloma around the prosthesis (5.4 %), and displacement of incus (4.3%). <br>Conclusions:</b> Surgical treatment of otosclerosis is a widely accepted and good method. It allows to achieve an improvement in hearing in the vast majority of patients treated in this way. Unfortunately, over the years some patients develop recurrent conductive hearing loss. Reoperation creates an opportunity for finding the cause and improving hearing in the majority of cases.

Downregulation of CEACAM6 gene expression in laryngeal squamous cell carcinoma is an effect of DNA hypermethylation and correlates with disease progression.

We have turned our attention to CEACAM6 gene, already described as deregulated in various types of cancer. By using the expression microarrays performed on the set of 16 laryngeal squamous cell carcinoma (LSCC) samples: 11 cell lines and 5 primary tumors we have shown downregulation of CEACAM6 gene as compared to non cancer controls from head and neck region. CEACAM6 gene downregulation, further confirmed by quantitative PCR on 25 LSCC cell lines, was observed in cell lines derived from recurrent tumors in comparison to controls. A significant gene downregulation was observed in cell lines derived from advanced, high grade tumors in comparison to controls. Intrigued by the recurrent transcriptional loss of CEACAM6 we searched for the mechanism potentially responsible for its downregulation and hence we analyzed DNA copy number changes (a-CGH), promoter DNA methylation status and occurrence of gene mutations (in silico). Neither the analysis of gene copy number, nor the mutation screen has shown recurrent deletions or mutations, that could contribute to the observed downregulation of the gene. However, by using bisulfite pyrosequencing, we have shown DNA hypermethylation (mean DNA methylation > 78%) of CEACAM6 promoter region in 9/25 (36%) LSCC cell lines. Importantly, the 5-aza-2-deoxycytidine-induced inhibition of DNA methylation resulted in restoration of CEACAM6 expression in the two LSCC cell lines on mRNA level. In summary, we have shown that recurrent downregulation of CEACAM6 in LSCC is dependent on the gene's promoter DNA methylation and is observed predominantly in large, poorly differentiated tumors and recurrences.

Outcomes of CO2 laser-assisted posterior cordectomy in bilateral vocal cord paralysis in 132 cases.

The purpose of the study was to assess the role of laser-assisted posterior cordectomy in the management of patients with bilateral vocal cord paralysis. We aimed an analysis of 132 consecutive patients treated by CO2 laser posterior cordectomy, aged 38-91, 31% tracheotomized on admission. Cordectomy was performed under microlaryngoscopy using CO2 laser (Lumenis AcuPulse 40 CO2 laser, wavelength 10.6 µm, Lumenis Ltd., Yokneam, Israel). We looked at the number of laser glottic procedures necessary to achieve decannulation in tracheotomized patients and to achieve respiratory comfort in non-tracheotomized subjects and we evaluated the two groups for differences in patient characteristics. In tracheotomized patients, we also assessed factors affecting the success of decannulation and we evaluated the impact of tracheotomy on patients' lives. Decannulation was performed in 63% of tracheotomized patients. In terms of the number of procedures, 54% (14), 19% (5), and 27% (7) tracheotomized vs. 74% (61), 24% (20), and 2% (2) non-tracheotomized subjects underwent one, two, or three procedures, respectively. In the group of tracheotomized patients who were successfully decannulated, the number of multiple laser-assisted procedures was significantly higher than in the group of non-tracheotomized subjects with respiratory comfort after treatment (p = 0.04). Advanced age (> 66 years), comorbidities (diabetes, gastroesophageal reflux disease (GERD)), multiple thyroid surgeries, and tracheotomy below the cricoid cartilage were found to decrease the likelihood of successful decannulation. Posterior cordectomy is a simple method allowing for airway improvement and decannulation in patients with bilateral vocal cord paralysis. It is less effective in tracheotomized subjects with diabetes or GERD, older than 66 years old, after two or more thyroidectomies.

Simultaneous treatment of intracranial complications of paranasal sinusitis.

PURPOSE: The objective of this study was to analyse 51 patients with intracranial complications of sinusitis treated in the Department of Otolaryngology and Laryngeal Oncology at Poznan University of Medical Sciences from 1964 to 2016. MATERIALS AND METHODS: Males made up a significant portion of study participants at 70.5%. Treatment included simultaneous removal of inflammatory focal points in the paranasal sinuses and drainage of cerebral and epidural abscesses and subdural empyemas under the control of neuronavigation preceded by the implementation of broad-spectrum antibiotics continuously for 4 weeks. Seventy-three intracranial complications were found among 51 patients. Of the 51 patients, 25 had frontal lobe abscesses (including multiple abscesses). Other complications included the following: 16 epidural abscesses, 9 subdural empyemas, 15 meningitis cases, 3 intracerebral abscesses, 3 sinus thrombosis cases and 2 patients with cerebritis. Co-occurrence of these complications worsened the state of the patient and increased the duration of treatment. Patients with frontal lobe abscesses had a better prognosis and less pronounced neurological symptoms in recent years versus earlier treatment approaches. CONCLUSIONS: Simultaneous treatment of intracranial complications of sinusitis is an effective treatment method that has minimal burden for the patient. From 1964 to 1978, three deaths (17%) were reported among patients with these complications. Since 1978, no deaths were reported in the clinic.

Recurrent transcriptional loss of the PCDH17 tumor suppressor in laryngeal squamous cell carcinoma is partially mediated by aberrant promoter DNA methylation.

Protocadherins are cell-cell adhesion molecules encoded by a large family of genes. Recent reports demonstrate recurrent silencing of protocadherin genes in tumors and provide strong arguments for their tumor supresor functionality. Loss of protocadherins may contribute to cancer development not only by altering cell-cell adhesion, that is a hallmark of cancer, but also by enhancing proliferation and epithelial mesenchymal transition of cells via deregulation of the WNT signaling pathway. In this study we have further corroborated our previous findings on the involvement of PCDH17 in laryngeal squamous cell carcinoma (LSCC). We used bisulfite pyrosequencing to analyze a cohort of primary LSCC tumors for alterations in PCDH17 promoter DNA methylation as an alternative gene inactivation mechanism to the homozygous deletions reported earlier. Moreover, we analyzed primary LSCC samples by immunohistochemistry for PCDH17 protein loss. We identified recurrent elevation of PCDH17 promoter DNA methylation in 32/81 (40%) primary tumors (P < 0.001) and therein hypermethylation of 12 (15%) cases in contrast to no tumor controls (n = 24) that were all unmethylated. Importantly, DNA demethylation by decitabine has restored low level PCDH17 expression in LSCC cell lines. In conclusion, we provide a mechanistic explanation of recurrently observed PCDH17 silencing in LSCC by demonstrating the role of promoter methylation in this process. In light of these findings and recent reports showing that PCDH17 methylation is detectable in serum of cancer patients we suggest that testing PCDH17 DNA methylation might serve as a potential biomarker in LSCC.

Recurrent epigenetic silencing of the PTPRD tumor suppressor in laryngeal squamous cell carcinoma.

Cellular processes like differentiation, mitotic cycle, and cell growth are regulated by tyrosine kinases with known oncogenic potential and tyrosine phosphatases that downmodulate the first. Therefore, tyrosine phosphatases are recurrent targets of gene alterations in human carcinomas. We and others suggested recently a tumor suppressor function of the PTPRD tyrosine phosphatase and reported homozygous deletions of the PTPRD locus in laryngeal squamous cell carcinoma. In this study, we investigated other gene-inactivating mechanisms potentially targeting PTPRD, including loss-of-function mutations and also epigenetic alterations like promoter DNA hypermethylation. We sequenced the PTPRD gene in eight laryngeal squamous cell carcinoma cell lines but did not identify any inactivating mutations. In contrast, by bisulfite pyrosequencing of the gene promoter region, we identified significantly higher levels of methylation (p = 0.001 and p = 0.0002, respectively) in 9/14 (64%) laryngeal squamous cell carcinoma cell lines and 37/79 (47%) of primary laryngeal squamous cell carcinoma tumors as compared to normal epithelium of the upper aerodigestive tract. There was also a strong correlation (p = 0.0001) between methylation and transcriptional silencing for the PTPRD gene observed in a cohort of 497 head and neck tumors from The Cancer Genome Atlas dataset suggesting that DNA methylation is the main mechanism of PTPRD silencing in these tumors. In summary, our data provide further evidence of the high incidence of PTPRD inactivation in laryngeal squamous cell carcinoma. We suggest that deletions and loss-of-function mutations are responsible for PTPRD loss only in a fraction of cases, whereas DNA methylation is the dominating mechanism of PTPRD inactivation.

The role of brachytherapy in the treatment of squamous cell carcinoma of the head and neck.

Brachytherapy is a form of intensive local irradiation, allowing effective protection of surrounding structures with preservation of organ function and results in a favorable therapeutic ratio. It can be used alone, adjuvantly after surgery, and as a local boost in combination with external-beam radiation therapy. This paper is a literature review on the role of brachytherapy in the management of head and neck cancers with a special emphasis on papers published in the last 5 years. Technical details, effectiveness and potential toxicities of brachytherapy when used in different combinations with other therapeutic modalities and tumor sites are presented. Brachytherapy is an attractive treatment option in the management of primary malignancies and recurrent tumors in previously irradiated areas of the head and neck. It is effective and safe, and results in good functional and oncological outcomes.

The Knowledge of the Role of Papillomavirus-Related Head and Neck Pathologies among General Practitioners, Otolaryngologists and Trainees. A Survey-Based Study.

OBJECTIVES: The aim of the survey was to introduce knowledge of HPV's role in head and neck pathologies to general physicians (GPs), otorhinolaryngologists (ENTs) and newly graduated doctors, as well as to promote HPV-related diseases prevention. STUDY DESIGN: Cross-sectional study. METHODS: Self-designed questionnaire was sent to 2100 doctors. A total of 404 doctors, including 144 ENTs, 192 GPs and 68 trainees, responded. RESULTS: The majority of ENTs (86.8%) had contact with recurrent respiratory papillomatosis (RRP) and oropharyngeal cancers (OPCs) patients; in contrast, the majority of GPs (55.7%) did not (p = 0.00). The knowledge of HPV aetiology of cervical cancer versus OPCs and RRP was statistically higher. 7% of ENTs, 20% of GPs and 10% of trainees had not heard about HPV in oropharyngeal diseases. Women had greater knowledge than men. Both in the group of GPs and ENTs, 100% of respondents had heard about the impact of vaccination on the reduction of cervical cancer incidence. Only 39.11% of respondents had heard about the possibility of using vaccination against HPV in RRP-ENT doctors significantly more often than GPs and trainees (p = 0.00). Only 28.96% of physicians had heard about the potential value of HPV vaccination in preventing OPCs, including 44.44% of ENT doctors, 23.44% of GPs and 11.76% of trainees (p = 0.00). The doctors from district hospitals showed lower level of knowledge compared with clinicians (p = 0.04). CONCLUSIONS: The different levels of knowledge and awareness of HPV issues highlight the need for targeted awareness strategies in Poland with implementation of HPV testing and vaccination. The information should be accessible especially to those with lower education levels: ENTs from small, provincial wards, GPs from cities of < 200,000 inhabitants and older physicians. The incorporation of HPV issues into the studies curriculum would be fruitful in terms of improving the knowledge of trainees.

Intracranial complications from sinusitis.

INTRODUCTION: Despite increasingly better diagnostic and therapeutic methods intracranial sinogenic complications, invariably pose a direct threat to the lives of patients and a challenge for otolaryngologists. AIM: The aim of the study was to analyze patients with intracranial sinogenic complications treated at the Department of Otolaryngology and Otolaryngological Oncology of Poznan University of Medical Sciences in the years 2000-2013. MATERIAL AND METHODS: Analysis covered the period from January 2000 to December 2013. Twenty-one patients with intracranial sinogenic complications were treated at the Department of Otolaryngology in Poznan during that time. RESULTS: Material mainly included young men. Brain abscesses were the most common complications. Intracranial complications of sinusitis rarely occurred in isolation, often coexisting with other intracranial pathologies. A significant increase in the incidence was recorded in 2013. Treatment involved concurrently alleviating inflammation in the sinuses through implementation of broad-spectrum antibiotics for several weeks and decompressing the organized intracerebral abscesses, empyema, epidural and/or subdural abscesses under control of neuronavigation. There were no patient deaths recorded in the analyzed period. CONCLUSIONS: The risk of developing intracranial sinogenic complications is low but invariably present and should be included in the differential diagnosis. Since the incidence of intracranial complications may increase in the course of prevailing viral infection, it should raise diagnostic vigilance.