Cross validated serum small extracellular vesicle microRNAs for the detection of oropharyngeal squamous cell carcinoma.

BACKGROUND: Oropharyngeal squamous cell carcinoma (OPSCC) is often diagnosed at an advanced stage because the disease often causes minimal symptoms other than metastasis to neck lymph nodes. Better tools are required to assist with the early detection of OPSCC. MicroRNAs (miRNAs, miRs) are potential biomarkers for early head and neck squamous cell cancer diagnosis, prognosis, recurrence, and presence of metastatic disease. However, there is no widespread agreement on a panel of miRNAs with clinically meaningful utility for head and neck squamous cell cancers. This could be due to variations in the collection, storage, pre-processing, and isolation of RNA, but several reports have indicated that the selection and reproducibility of biomarkers has been widely affected by the methods used for data analysis. The primary analysis issues appear to be model overfitting and the incorrect application of statistical techniques. The purpose of this study was to develop a robust statistical approach to identify a miRNA signature that can distinguish controls and patients with inflammatory disease from patients with human papilloma virus positive (HPV +) OPSCC. METHODS: Small extracellular vesicles were harvested from the serum of 20 control patients, 20 patients with gastroesophageal reflux disease (GORD), and 40 patients with locally advanced HPV + OPSCC. MicroRNAs were purified, and expression profiled on OpenArray™. A novel cross validation method, using lasso regression, was developed to stabilise selection of miRNAs for inclusion in a prediction model. The method, named StaVarSel (for Stable Variable Selection), was used to derive a diagnostic biomarker signature. RESULTS: A standard cross validation approach was unable to produce a biomarker signature with good cross validated predictive capacity. In contrast, StaVarSel produced a regression model containing 11 miRNA ratios with potential clinical utility. Sample permutations indicated that the estimated cross validated prediction accuracy of the 11-miR-ratio model was not due to chance alone. CONCLUSIONS: We developed a novel method, StaVarSel, that was able to identify a panel of miRNAs, present in small extracellular vesicles derived from blood serum, that robustly cross validated as a biomarker for the detection of HPV + OPSCC. This approach could be used to derive diagnostic biomarkers of other head and neck cancers.

Does gene expression in laryngeal subsites differ between patients with laryngopharyngeal reflux and controls?

OBJECTIVE: To identify laryngeal mRNA gene changes in patients with laryngopharyngeal reflux (LPR). METHOD: Laryngeal biopsies from non-smoking LPR patients (n=10; Reflux Symptom Index (RSI) >12 and a Reflux Finding Score (RFS) >6) and controls (n=9; RSI <12 and RFS <6) were collected from four subsites (true vocal cord, false vocal cord, medial arytenoid and posterior commissure) of the larynx. qRT-PCR analyses were conducted on 20 reflux- and inflammation-related genes, including interleukins 6 and 8, cytokeratins 8 and 14, mucin genes MUC1, MUC2, MUC3B, MUC4, MUC5B, MUC6 and MUC7 and carbonic anhydrase III. Statistical analysis (Mann-Whitney U test) compared gene expression levels between LPR and control groups at each subsite. RESULTS: Site-specific differences in squamous metaplasia and gene expression were noted in LPR patients, with the majority present in the medial arytenoid region. Significant.differences were noted in genes related to mucosal defence and inflammation, including CRNN, CD1d, TGFß-1, MUC2, MUC5B and CDH1. CONCLUSION: Whilst the posterior commissure is commonly identified as the area demonstrating the most significant macroscopic change in LPR, the histological changes and genes assessed here showed more pronounced LPR associated differences in the medial arytenoid. We identified differences in expression of mucin genes, cytokeratin-14 and molecular markers of inflammation. Whilst some of these changes may be metaplasia-related, further evaluation of the mRNA expression of these genes may provide a useful biomarker panel for diagnosis and therapeutic monitoring of LPR.

Long-term quality-of-life outcomes following treatment for adult obstructive sleep apnoea: comparison of upper airway surgery, continuous positive airway pressure and mandibular advancement splints.

OBJECTIVE: Long-term quality-of-life (QOL) outcomes, complications and clinical effectiveness in patients undergoing treatment with upper airway surgery (UAS), continuous positive airway pressure (CPAP) and mandibular advancement splints (MAS) for adult obstructive sleep apnoea (OSA). DESIGN: Retrospective cohort study. SETTING: Multidisciplinary OSA clinic in University teaching hospital. PARTICIPANTS: Consecutive, simultaneously treated patients with OSA undergoing UAS (n = 83), CPAP (n = 83) and MAS (n = 79). MAIN OUTCOME MEASURES: Glasgow Benefit Inventory (GBI), Snoring Severity Scale (SSS), Epworth Sleepiness Score (ESS) and side-effects in all three groups were recorded at a mean of 34.5 months following start of treatment and compared via anova with Bonferroni's adjustment for pairwise comparisons. RESULTS: Upper airway surgery demonstrated a statistically significant QOL benefit over MAS. All three groups showed a significant improvement in SSS with CPAP significantly better than MAS, but equivalent to UAS. Uncomplicated UAS provided a greater QOL outcome than compliant MAS, non-compliant CPAP (P < 0.05) and comparable to compliant CPAP. Patients undergoing UAS with recorded complications still reported equivalent QOL outcomes to compliant CPAP and MAS, suggesting these surgical complications are relatively minor compared to the QOL benefit of OSA treatment. CONCLUSION: Upper airway surgery showed a significant improvement in QOL outcomes compared to non-compliant CPAP or MAS and equivalent benefit to compliant CPAP. This study strongly supports the role for contemporary UAS in OSA when CPAP is not or no longer an option.

Validation of virtual-reality-based simulations for endoscopic sinus surgery.

BACKGROUND: Virtual reality (VR) simulators provide an alternative to real patients for practicing surgical skills but require validation to ensure accuracy. Here, we validate the use of a virtual reality sinus surgery simulator with haptic feedback for training in Otorhinolaryngology - Head & Neck Surgery (OHNS). METHODS: Participants were recruited from final-year medical students, interns, resident medical officers (RMOs), OHNS registrars and consultants. All participants completed an online questionnaire after performing four separate simulation tasks. These were then used to assess face, content and construct validity. anova with post hoc correlation was used for statistical analysis. RESULTS: The following groups were compared: (i) medical students/interns, (ii) RMOs, (iii) registrars and (iv) consultants. Face validity results had a statistically significant (P < 0.05) difference between the consultant group and others, while there was no significant difference between medical student/intern and RMOs. Variability within groups was not significant. Content validity results based on consultant scoring and comments indicated that the simulations need further development in several areas to be effective for registrar-level teaching. However, students, interns and RMOs indicated that the simulations provide a useful tool for learning OHNS-related anatomy and as an introduction to ENT-specific procedures. CONCLUSIONS: The VR simulations have been validated for teaching sinus anatomy and nasendoscopy to medical students, interns and RMOs. However, they require further development before they can be regarded as a valid tool for more advanced surgical training.

Lysozyme expression is increased in the sinus mucosa of patients with chronic rhinosinusitis.

BACKGROUND: The presence of fungi and bacteria in the paranasal sinuses may contribute to ongoing inflammation. Lysozyme is an innate immune peptide with bactericidal and fungicidal activity. The expression of lysozyme in chronic rhinosinusitis (CRS) is poorly understood and deficiencies in lysozyme expression may contribute to the ongoing inflammation in CRS patients. OBJECTIVE: Determine lysozyme expression in sinus mucosa of normal and CRS patients with (CRSwNP) and without (CRSsNP) nasal polyps. METHODOLOGY: Sinus mucosa specimens (n = 82) were processed for standard histology, immunohistochemical localisation of lysozyme, immunofluorescent localisation of fungi, and qPCR analysis of lysozyme expression. RESULTS: CRS specimens displayed high-levels of lysozyme immunoreactivity in many of the abundant serous cells. Moderate levels were detected in some epithelial cells and inflammatory cells. Low levels were detected in some subepithelial glands of control specimens. No difference in immunoreactivity was detected between CRSwNP and CRSsNP specimens. Fungal elements were not visualised in any sinus specimen. qPCR analysis demonstrated variable lysozyme expression between individuals. CONCLUSIONS: Lysozyme protein expression is increased in patients with CRS, suggesting a defect in lysozyme expression is not responsible for the microbial colonisation often associated with CRS. The functional activity of lysozyme in CRS patients needs to be further investigated.

Reflux changes in adenoidal hyperplasia: a controlled prospective study to investigate its aetiology.

OBJECTIVES: To compare pepsin, carbonic anhydrase III (CAIII), cyclooxygenase-2 (COX-2) and mucin 5AC (MUC5AC) expression in children with adenoid hypertrophy and normal controls. DESIGN: A non-randomised, controlled prospective study. SETTING: Two paediatric hospitals in Adelaide, South Australia. PARTICIPANTS: Children aged 2-10 years, 21 undergoing adenoidectomy and 12 controls undergoing routine dental surgery. MAIN OUTCOME MEASURES: We measured expression of pepsin, CAIII, COX-2 and MUC5AC levels by real-time RT-PCR, immunohistochemistry, and Western blot to determine any difference between children with hyperplastic adenoids and controls. RESULTS: Pepsin was not detected in any study or control adenoid by immunohistochemistry or Western blot. Real-time RT-PCR analysis showed a statistically significant difference between groups with respect to COX-2 (P = 0.027) and MUC5AC (P = 0.02) but no difference in CAIII expression (P = 0.414). A significant correlation was also found between COX-2 and MUC5AC expression (Kendall Tau = 0.4, P = 0.005). CONCLUSION: Our results suggest that the biochemical changes seen in adenoid hypertrophy are different to those seen in reflux-affected tissues. The decreased COX-2 and MUC5AC expression may be due to squamous metaplasia and other inflammatory changes associated with adenoid hypertrophy. Our findings infer there is little evidence of reflux being a major contributory factor in the pathophysiology of adenoidal hypertrophy.

Microbiology of otitis media in Indigenous Australian children: review.

OBJECTIVES: To review research addressing the polymicrobial aetiology of otitis media in Indigenous Australian children in order to identify research gaps and inform best practice in effective prevention strategies and therapeutic interventions. METHODS: Literature review. RESULTS: Studies of aspirated middle-ear fluid represented a minor component of the literature reviewed. Most studies relied upon specimens from middle-ear discharge or the nasopharynx. Culture-based middle-ear discharge studies have found that non-typeable Haemophilus influenzae and Streptococcus pneumoniae predominate, with Moraxella catarrhalis, Staphylococcus aureus and Streptococcus pyogenes isolated in a lower proportion of samples. Alloiococcus otitidis was detected in a number of studies; however, its role in otitis media pathogenesis remains controversial. Nasopharyngeal colonisation is a risk factor for otitis media in Indigenous infants, and bacterial load of otopathogens in the nasopharynx can predict the ear state of Indigenous children. CONCLUSION: Most studies have used culture-based methods and specimens from middle-ear discharge or the nasopharynx. Findings from these studies are consistent with international literature, but reliance on culture may incorrectly characterise the microbiology of this condition. Advances in genomic technologies are now providing microbiologists with the ability to analyse the entire mixed bacterial communities ('microbiomes') of samples obtained from Indigenous children with otitis media.

Do tissue spears used to clear ear canal pus improve hearing? A case series study of hearing in remote Australian Aboriginal children with chronic suppurative otitis media before and after dry mopping with tissue spears.

OBJECTIVE: To determine whether the use of tissue spears to remove otorrhoea from Aboriginal children's ear canals improves hearing in the affected ear. DESIGN: Case series study with controls. METHODS: The study comprised 61 Aboriginal children from communities in the remote arid zone of South Australia who had otorrhoea obscuring the tympanic membrane in 1 or both ears. Eighty ears were treated with tissue spears, and hearing was assessed before and after treatment. RESULTS: Forty-two children had unilateral and 19 had bilateral active disease. An additional 13 ears without otorrhoea served as controls. Improvement by 5 dB HL or greater in a four-frequency pure tone average occurred in 41.3 per cent of treated ears. Subsequently, blinded audiologists made qualitative judgements that a functional improvement in hearing had occurred after tissue spear use in 28.4 per cent of ears. CONCLUSION: Tissue spears can improve hearing thresholds in a significant proportion of children with otorrhoea. However, the duration of the effect is unknown.

Bleeding following coblation tonsillectomy: a 10-year, single-surgeon audit and modified grading system.

OBJECTIVE: Coblation tonsillectomy can be controversial. This study assessed post-tonsillectomy haemorrhage outcomes for patients operated on by a single experienced coblation-trained ENT surgeon. STUDY DESIGN: A retrospective audit of coblation tonsillectomies was performed using the Flinders modification of Stammberger criteria for post-tonsillectomy haemorrhage. METHOD: Case note review, interview and database interrogation were utilised to obtain the dataset. Haemorrhage results were compared to reports in the current literature. RESULTS: Of those who underwent coblation tonsillectomy, 3.4 per cent were readmitted to hospital with haemorrhage and 1.3 per cent returned to the operating theatre (0.4 per cent primary haemorrhage and 0.9 per cent secondary haemorrhage). Younger children had a lower risk of returning to the operating theatre than older children or adults (0.3 per cent under the age of 12 years vs 2.0 per cent aged 12 years or older). CONCLUSION: Coblation can be a safe method for tonsillectomy with low complication rates when performed by an experienced ENT surgeon. The Flinders modification of the Stammberger criteria for post-tonsillectomy haemorrhage provides a simple system for data comparison.

Surgical management of airway stenosis by radiofrequency coblation.

BACKGROUND: Acquired airway stenosis can be challenging to manage endoscopically because of difficult field visualisation, instrument limitations and the risk of laser fire. At our institution, radiofrequency coblation has been successfully used for the resection of subglottic and tracheal stenosis in adults. This paper presents our experience with this technique. METHOD: A retrospective case note analysis of all cases of airway stenosis in adults from 2007 to 2012 was performed. RESULTS: Ten adult patients underwent coblation resection for airway stenosis. All lesions were classified as McCaffrey stage I (i.e. less than 1 cm long). Causes of stenosis included: idiopathic stenosis (40 per cent), previous tracheostomy (30 per cent) and endotracheal intubation (20 per cent). Six patients (60 per cent) required a single procedure and 4 (40 per cent) required multiple interventions. All patients reported significant improvement in their symptoms following treatment. All patients were alive at the time of writing and none have required open resection. CONCLUSION: Radiofrequency coblation is an attractive alternative technique for the treatment of idiopathic or acquired airway stenosis in adults.

Cochlear implantation in children with large vestibular aqueduct syndrome and a review of the syndrome.

OBJECTIVE: Children with Large Vestibular Aqueduct Syndrome (LVAS) frequently develop speech and language skills prior to deterioration of their hearing. Operations designed to halt the progression of hearing loss have largely failed so the question of Cochlear Implantation in these children has arisen. It had been suggested that there would be technical difficulties in implanting these patients and, therefore, there had been an initial reluctance to proceed to implantation. The aim of the present paper is to assess surgical and functional outcomes in implanted children with LVAS and review the related literature. MATERIAL AND METHODS: From the 170 children assessed by MRI in the Nottingham Paediatric Cochlear Implant Programme, seven (4%) were identified as having LVAS. Four of these children were implanted and had at least 12 months follow up. Two of the children are on the waiting list for implantation and one child was not implanted because of absence of the cochlear nerve. Operative findings, complications and outcome measures were recorded. The auditory skills of the children were assessed before implantation and 1 year following implantation. A literature search was done to identify other series with experience in implanting children with LVAS. RESULTS: Full insertion of the electrode array was achieved in all our cases. After cochleostomy two patients experienced a mild CSF leak that was easily controlled by the muscle graft. On the first day post-operation two patients were nauseous and one had an episode of vomiting, however, all were discharged within 24 h of surgery. Initial outcome measures at 12 months post-implantation were encouraging showing significant progress in children's auditory skills. CONCLUSIONS: The results of the present study and the review of the literature suggest that LVAS is not a contraindication to implantation as initial concerns about severe perilymph leaks and surgical complications have proved to be unfounded. The post-operative progress of these children in listening skills also suggest that these children are suitable for cochlear implantation

Mast cell distribution and morphology in human nasal turbinates following decalcification.

Whole inferior nasal turbinates were used to evaluate six different fixatives followed by decalcification. H and E staining was used to assess general fixation and toluidine blue and thionin to stain mast cells metachromatically. We show that neutral buffered formalin or paraformaldehyde give the best overall fixation. Very long times (six days) are needed in TB to show maximum mast cell numbers; these numbers are never as high, nor the cells as densely stained as those in undecalcified controls. The difference in number between the two supports the hypothesis that there is more than one subset of mast cells in nasal mucosa. The more superficial mast cells have a smaller size and may be dendritic, with relatively few granules which stain faintly. Those in deeper situations are often larger, ovoid, very granular and intensely stained.

The prevalence of allergic rhinosinusitis: a review.

A review of the literature indicates that there is an increase in the prevalence of self-reported symptoms suggestive of seasonal allergic rhinitis over the last 70 years. The reason for this remains unclear. Epidemiological studies which relate to the effect of pollution suggest that while pollution can exacerbate respiratory tract symptoms, there is no consistent evidence that high levels result in an increase in the prevalence of allergic rhinitis. The increased prevalence of house-dust mite, an alteration in immunity--possibly related to the number of childhood respiratory tract infections, and increased disease awareness, are all factors which may influence the reported prevalence of allergic rhinitis. At present the inconsistencies which exist in the literature mean that it is possible to 'pick' publications which support one view and ignore the rest, so that a variety of views may be held, each with apparently well referenced endorsement. The criteria defined by Bradford Hill (1965) to establish causation and not mere association between any of these factors and allergic rhinitis have, as yet, not been met (see Table V).

Atypically located submandibular gland diagnosed by Doppler ultrasound.

The anatomical location and relationships of the submandibular gland are well-known to most otolaryngologists and gross variations from the norm are rare. We report a case of an atypical submandibular gland located 2 cm below the mandible which presented as a painful neck swelling with non-diagnostic fine needle aspiration cytology. Using Doppler ultrasound the mass was noted to be closely located to the facial artery and vein and following the administration of oral lemon juice, the peak systolic velocity of a small artery within the mass rose from 8.5 cm/s to 16.4 cm/s, confirming the tissue's salivary nature. We review the literature on using Doppler ultrasound in this area of the neck and discuss realistic practical applications of the technique.

Intra-operative image guidance in otolaryngology--the use of the ISG viewing wand.

The ISG viewing wand is an intra-operative guidance system with a proprioceptive robotic-like jointed arm. It provides surgeons with almost instantaneously reconstructed computer-generated CT or MRI images in two or three dimensions and can correlate any point within the operative field to its corresponding locus on the reformatted scan images. In addition to having been used in over 400 neurosurgical patients in Bristol, 14 patients with skull-base, cerebello-pontine angle or temporal bone lesions have also undergone wand-guided resections. The wand has proved to be particularly useful in pre-operative planning, allowing minimally-invasive incisions, providing per-operative navigation, identifying the relationship and proximity of important anatomical structures and in assessing the extent of lesion resection. We illustrate the advantages of intra-operative image-guidance by discussing four cases. The potential applications of this form of imaging technology to other otolaryngological procedures are discussed.

Quality of life outcomes following dacryocystorhinostomy: external and endonasal laser techniques compared.

AIMS/BACKGROUND. Endonasal laser dacryocystorhinostomy (ELDCR) has recently become established as an alternative technique to conventional external DCR (EXDCR) for relief of epiphora. The aim of this study was to compare quality of life and symptomatic outcomes in patients treated by these two methods. METHODS. A postal questionnaire was sent to 204 ELDCR and 78 EXDCR patients who had undergone surgery under our care more than 6 months previously. The questionnaire included the validated 18-item Glasgow Benefit Inventory (GBI) and 5 further questions addressing ocular symptomatology. RESULTS. Fully completed questionnaires were received from 156 ELDCR and 50 EXDCR patients. Mean GBI scores of +16.8 (ELDCR) and +23.2 (EXDCR) were obtained (Mann-Whitney U test = NS). Ocular symptom scores were +33 (ELDCR) and +54 (EXDCR) (p=0.005). CONCLUSION. Patients undergoing either ELDCR or EXDCR experience significant benefit in their healthcare status as detected by the GBI. There is no significant difference between the scores obtained by the two groups of patients. There were improvements in ocular symptom scores in both groups, with patients who had EXDCR scoring significantly better than the ELDCR group.

Endonasal laser-assisted dacryocystorhinostomy.

Endonasal laser-assisted dacryocystorhinostomy allows relief of epiphora caused by nasolacrimal duct obstruction. It can be performed as a day-case procedure and has many advantages over conventional approaches. This article reviews conventional, endoscopic and laser-assisted dacryocystorhinostomy and describes the technical steps of an endonasal procedure.

Role of subtotal tonsillectomy ('tonsillotomy') in children with sleep disordered breathing.

INTRODUCTION: Sleep disordered breathing in children causes disturbance in behaviour and also in cardiorespiratory and neurocognitive function. Subtotal tonsillectomy ('tonsillotomy') has been performed to treat sleep disordered breathing, with outcomes comparable to established therapies such as total tonsillectomy or adenoidectomy. This review critically assesses the role of subtotal tonsillectomy in a paediatric setting. METHOD: The Medline database (1966 to October 2012) was electronically searched using key terms including subtotal or intracapsular tonsillectomy, tonsillotomy, tonsillectomy, paediatrics, and sleep disordered breathing. RESULTS: Eighteen papers were identified and reviewed. Subtotal tonsillectomy would appear to have an efficacy equal to that of total tonsillectomy for the treatment of sleep disordered breathing, and has significant benefits in reducing post-operative pain and analgesia use. Subtotal tonsillectomy patients appear to have less frequent post-operative haemorrhage compared with total tonsillectomy patients. CONCLUSION: In children, subtotal tonsillectomy is associated with fewer post-operative complications whilst having a comparable effect in improving sleep disordered breathing, compared with total tonsillectomy.