Differences in the transcriptome response in the gills of sea lamprey acutely exposed to 3-trifluoromethyl-4-nitrophenol (TFM), niclosamide or a TFM:niclosamide mixture.

Sea lamprey (Petromyzon marinus) control in the Laurentian Great Lakes of North America makes use of two pesticides: 3-trifluoromethyl-4-nitrophenol (TFM) and niclosamide, which are often co-applied. Sea lamprey appear to be vulnerable to these agents resulting from a lack of detoxification responses with evidence suggesting that lampricide mixtures produce a synergistic effect. However, there is a lack of information pertaining to the physiological responses of sea lamprey to niclosamide and TFM:niclosamide mixtures. Here, we characterized the transcriptomic responses of the sea lamprey to TFM, niclosamide, and a TFM:niclosamide (1.5 %) mixture in the gill. Along with a control, larval sea lamprey were exposed to each treatment for 6 h, after which gill tissues were extracted for measuring whole-transcriptome responses using RNA sequencing. Differential gene expression patterns were summarized, which included identifying the broad roles of genes and common expression patterns among the treatments. While niclosamide treatment resulted in no differentially expressed genes, TFM- and mixture-treated fish had several differentially expressed genes that were associated with the cell cycle, DNA damage, metabolism, immune function, and detoxification. However, there was no common differential expression among treatments. For the first time, we characterized the transcriptomic response of sea lamprey to niclosamide and a TFM:niclosamide mixture and identified that these agents impact mRNA transcript abundance of genes associated with the cell cycle and cellular death, and immune function, which are likely mediated through mitochondrial dysregulation. These results may help to inform the production of more targeted and effective lampricides in sea lamprey control efforts.

Transcriptomic impacts and potential routes of detoxification in a lampricide-tolerant teleost exposed to TFM and niclosamide.

Sea lamprey (Petromyzon marinus) control in the Laurentian Great Lakes of North America often relies on the application of 3-trifluoromethyl-4-nitrophenol (TFM) and niclosamide mixtures to kill larval sea lamprey. Selectivity of TFM against lampreys appears to be due to differential detoxification ability in these jawless fishes compared to bony fishes, particularly teleosts. However, the proximate mechanisms of tolerance to the TFM and niclosamide mixture and the mechanisms of niclosamide toxicity on its own are poorly understood, especially among non-target fishes. Here, we used RNA sequencing to identify specific mRNA transcripts and functional processes that responded to niclosamide or a TFM:niclosamide mixture in bluegill (Lepomis macrochirus). Bluegill were exposed to niclosamide or TFM:niclosamide mixture, along with a time-matched control group, and gill and liver tissues were sampled at 6, 12, and 24 h. We summarized the whole-transcriptome patterns through gene ontology (GO) term enrichment and through differential expression of detoxification genes. The niclosamide treatment resulted in an upregulation of several transcripts associated with detoxification (cyp, ugt, sult, gst), which may help explain the relatively high detoxification capacity in bluegill. Conversely, the TFM:niclosamide mixture resulted in an enrichment of processes related to arrested cell cycle and growth, and cell death alongside a diverse detoxification gene response. Detoxification of both lampricides likely involves the use of phase I and II biotransformation genes. Our findings strongly suggest that the unusually high tolerance of bluegill to lampricides is due to these animals having an inherently high capacity and flexible detoxification response to such compounds.

Variation in the Transcriptome Response and Detoxification Gene Diversity Drives Pesticide Tolerance in Fishes.

Pesticides are critical for invasive species management but often have negative effects on nontarget native biota. Tolerance to pesticides should have an evolutionary basis, but this is poorly understood. Invasive sea lamprey (Petromyzon marinus) populations in North America have been controlled with a pesticide lethal to them at lower concentrations than native fishes. We addressed how interspecific variation in gene expression and detoxification gene diversity confer differential pesticide sensitivity in two fish species. We exposed sea lamprey and bluegill (Lepomis macrochirus), a tolerant native species, to 3-trifluoromethyl-4-nitrophenol (TFM), a pesticide commonly used in sea lamprey control. We then used whole-transcriptome sequencing of gill and liver to characterize the cellular response in both species. Comparatively, bluegill exhibited a larger number of detoxification genes expressed and a larger number of responsive transcripts overall, which likely contributes to greater tolerance to TFM. Understanding the genetic and physiological basis for pesticide tolerance is crucial for managing invasive species.

Contrasting physiological responses between invasive sea lamprey and non-target bluegill in response to acute lampricide exposure.

Control of invasive sea lamprey (Petromyzon marinus) in the Laurentian Great Lakes of North America uses lampricides, which consist of 3-trifluoromethyl-4-nitrophenol (TFM) and niclosamide. Lampricides are thought to inhibit aerobic energy synthesis, with TFM having a relatively greater selective action against lampreys. While the toxicity and physiological effects of TFM are known, the impacts associated with exposure to niclosamide and TFM:niclosamide mixtures are poorly characterized in fishes. Therefore, focusing on energy metabolism, we quantified the physiological responses of larval sea lamprey and bluegill (Lepomis macrochirus), a non-target, native species. Exposures consisted of each lampricide alone (TFM at the species-specific 24 h LC10; niclosamide at 1.5% of the mixture's TFM concentration) or a mixture of the two (larval sea lamprey at TFM 24 h LC10 + 1.5% niclosamide; bluegill at sea lamprey's TFM 24 h LC99.9 + 1.5% niclosamide) for 24 h. Tissues (brain, skeletal muscle, and liver) were sampled at 6, 12, and 24 h of exposure and assayed for concentrations of ATP, phosphocreatine, glycogen, lactate, and glucose and tissue lampricide levels. In larval sea lamprey, TFM had little effect on brain and skeletal muscle, but niclosamide resulted in a depletion of high energy substrates in both tissues. Mixture-exposed lamprey showed depletion of high energy substrates, accumulation of lactate, and high mortality rates. Bluegill were largely unaffected by toxicant exposures. However, bluegill liver showed lower glycogen and lactate under all three toxicant exposures suggesting increased metabolic turnover. Bluegill also had lower concentrations of TFM and niclosamide in their tissues when compared to lamprey. Our results indicate that lampricide toxicity in sea lamprey larvae is mediated through a depletion of high energy substrates because of impaired aerobic ATP synthesis. We also confirmed that non-target bluegill showed high tolerance to lampricide exposure, an effect potentially mediated through a high detoxification capacity relative to lampreys.

A multi-centre analysis of a decade of endoscopic pharyngeal pouch surgery in Cheshire and Merseyside.

BACKGROUND: There are sparse data on the outcomes of endoscopic stapling of pharyngeal pouches. The Mersey ENT Trainee Collaborative compared regional practice against published benchmarks. METHODS: A 10-year retrospective analysis of endoscopic pharyngeal pouch surgery was conducted and practice was assessed against eight standards. Comparisons were made between results from the tertiary centre and other sites. RESULTS: A total of 225 procedures were performed (range of 1.2-9.2 cases per centre per year). All centres achieved 90 per cent resumption of oral intake within 2 days. All centres achieved less than 2-day hospital stays. Primary success (84 per cent (i.e. abandonment of endoscopic stapling in 16 per cent)), symptom resolution (83 per cent) and recurrence rates (13 per cent) failed to meet the standard across the non-tertiary centres. CONCLUSION: Endoscopic pharyngeal pouch stapling is a procedure with a low mortality and brief in-patient stay. There was significant variance in outcomes across the region. This raises the question of whether this service should become centralised and the preserve of either tertiary centres or sub-specialist practitioners.

Canal wall down mastoidectomy with obliteration versus canal wall up mastoidectomy in primary cholesteatoma surgery.

OBJECTIVE: This study sought to compare disease recidivism rates between canal wall up mastoidectomy and a canal wall down with obliteration technique. METHODS: Patients undergoing primary cholesteatoma surgery at our institution over a five-year period (2013-2017) using the aforementioned techniques were eligible for inclusion in the study. Rates of discharge and disease recidivism were analysed using chi-square statistics. RESULTS: A total of 104 ears (98 patients) were included. The mean follow-up period was 30 months (range, 12-52 months). A canal wall down with mastoid obliteration technique was performed in 55 cases and a canal wall up approach was performed in 49 cases. Disease recidivism rates were 7.3 per cent and 16.3 per cent in the canal wall down with mastoid obliteration and canal wall up groups respectively (p = 0.02), whilst discharge rates were similar (7.3 per cent and 10.2 per cent respectively). CONCLUSION: Our direct comparative data suggest that canal wall down mastoidectomy with obliteration is superior to a canal wall up technique in primary cholesteatoma surgery, providing a lower recidivism rate combined with a low post-operative ear discharge rate.

The BeSMART (Best Supportive Management for Adults Referred with Tonsillopharyngitis) multicentre observational study.

Introduction Tonsillopharyngitis is the most common ear, nose and throat emergency admission, with 80,000 episodes recorded in England in 2015-2016. Despite this, there is a paucity of evidence addressing the supportive management of tonsillopharyngitis in inpatients. The aim of this retrospective multicentre observational study was to consider the Best Supportive Management for Adults Referred with Tonsillopharyngitis (BeSMART) in the inpatient setting, and to establish any associations between practice and outcomes. Methods Seven hospitals in North West England and North East Scotland participated in the study. Overall, 236 adult patients admitted with tonsillopharyngitis were included. The main outcome measures were interval to return to soft diet, length of stay (LOS), pain scores and readmissions. Results Women were more likely to seek professional help before presenting to secondary care (p=0.04). Patients admitted at the weekend were more likely to have a shorter LOS (p=0.03). There was no relationship between day of admission and seniority or specialty of the doctor initially seen. Prescription of corticosteroid, analgesia and a higher initial intravenous fluid infusion rate were not related to a shorter LOS. Conclusions This study is the first to yield valuable insights into the inpatient management of tonsillopharyngitis. This work represents part of an ongoing project to establish the evidence for common medical interventions for sore throat. Patient and professional surveys as well as a prospective interventional study are planned for the future.

Pathology-based staging for HPV-positive squamous carcinoma of the oropharynx.

OBJECTIVE: The rapid worldwide rise in incidence of human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC) has generated studies confirming this disease as an entity distinct from traditional OPSCC. Based on pathology, surgical studies have revealed prognosticators specific to HPV-positive OPSCC. The current AJCC/UICC staging and pathologic nodal (pN)-classification do not differentiate for survival, demonstrating the need for new, HPV-specific OPSCC staging. The objective of this study was to define a pathologic staging system specific to HPV-positive OPSCC. METHODS: Data were assembled from a surgically-managed, p16-positive OPSCC cohort (any T, any N, M0) of 704 patients from five cancer centers. Analysis was performed for (a) the AJCC/UICC pathologic staging, (b) newly published clinical staging for non-surgically managed HPV-positive OPSCC, and (c) a novel, pathology-based, "HPVpath" staging system that combines features of the primary tumor and nodal metastases. RESULTS: A combination of AJCC/UICC pT-classification and pathology-confirmed metastatic node count (⩽4 versus ⩾5) yielded three groups: stages I (pT1-T2, ⩽4 nodes), II (pT1-T2, ⩾5 nodes; pT3-T4, ⩽4 nodes), and III (pT3-T4, ⩾5 nodes), with incrementally worse prognosis (Kaplan-Meier overall survival of 90%, 84% and 48% respectively). Existing AJCC/UICC pathologic staging lacked prognostic definition. Newly published HPV-specific clinical stagings from non-surgically managed patients, although prognostic, showed lower precision for this surgically managed cohort. CONCLUSIONS: Three loco-regional "HPVpath" stages are identifiable for HPV-positive OPSCC, based on a combination of AJCC/UICC primary tumor pT-classification and metastatic node count. A workable, pathologic staging system is feasible to establish prognosis and guide adjuvant therapy decisions in surgically-managed HPV-positive OPSCC.

Pharmacological Management of Secondary Hyperparathyroidism in Patients with Chronic Kidney Disease.

Secondary hyperparathyroidism (SHPT) is a common complication of chronic kidney disease (CKD) and is part of the CKD-mineral bone disorder (CKD-MBD). SHPT is associated with increased risk of fracture and mortality; thus, SHPT control is recommended as kidney function declines. Effective SHPT management becomes more difficult once skeletal and cardiovascular adverse effects associated with severe SHPT have become established. However, interventional studies to lower parathyroid hormone (PTH) have so far shown inconsistent results in improving patient-centred outcomes such as mortality, cardiovascular events and fracture. Pharmacological treatment effect on PTH level is also inconsistent between pre-dialysis CKD and dialysis patients, which adds to the complexity of SHPT management. This review aims to give an overview on the pathophysiology, pharmacological and non-pharmacological treatment for SHPT in CKD including some of the limitations of current therapeutic options.

Laser-assisted tympanomastoidectomy for active mucosal chronic otitis media.

Laser-assisted tympanomastoidectomy for active mucosal chronic otitis media. OBJECTIVES: Achieving disease control in the context of active granulating mucosal chronic otitis media (COM) is typically challenging. Adjuvant laser use in cholesteatomatous COM surgery enhances disease clearance, yet its role in mucosal COM remains unexplored. The purpose of this study is to present the outcomes and our experience of the latter, and to attempt to define the role of this approach in managing mucosal COM. METHODS: Patients that underwent primary laser-assisted tympanomastoidectomy (2007-2012) for active granulating mucosal COM with at least six months follow-up were studied. Disease control was defined by successful tympanic membrane repair and resolution of otorrhoea. Time-to-event analysis using the Kaplan-Meier method was used to gain a temporal estimate of disease control. RESULTS: Seventy ears (66 patients) were included with mean follow-up of 22 months (6-50 months). Disease control was achieved in 94.3% of cases at latest follow-up, while one- and two-year temporal estimates of control were 97.1% and 91.8% respectively. A mean post-operative four-frequency air-bone gap of 20 dB was achieved in 52 cases (74.3%). Surgical complications included transient vertigo (n = 2), wound infection (n = 2), and temporary facial nerve palsy (n = 1). CONCLUSIONS: We present disease control rates favourable to other published series of active mucosal COM, supporting the use of the laser as an efficacious adjunct to tympanomastoid surgery in this context. This initial report should foster further appraisal of this approach in larger prospective studies.

Injection of botulinum toxin for the treatment of post-laryngectomy pharyngoesophageal spasm-related disorders.

Introduction Pharyngoesophageal spasm (PES) can cause dysphagia, central valve leak (CVL), and dypshonia in post-laryngectomy patients. Botulinum toxin has been used effectively for the treatment of PES, but data regarding patient-reported outcomes and efficacy for CVL are limited. We evaluated the results of botox injection for PES spasm using subjective and objective measures. Methods Data were collected prospectively (February 2010 to August 2013) on 13 patients undergoing botox injection for PES as identified by video fluoroscopy. We collected digital voice recordings, air-pressure measurements (APMs) for speech, and quality of life (QoL) data before and after the procedure: University of Washington QoL questionnaire (UW-QoL), MD Anderson Swallowing Inventory (MDADI) and the Voice Handicap Index (VHI-30). Results APMs for a sustained vowel decreased by 18% after botox injection, whereas maximum phonatory times increased by 63% (mean increase, 8 to 13 seconds). Sustained vowel amplitude decreased (mean, 87db to 83db) with an associated reduction in sustained vowel frequency (117Hz to 77Hz). MDADI scores improved by 10.2% overall, and UW-QoL scores showed an improvement in score of 7.6%. Mean scores for VHI-30 deteriorated by 2% overall but, when considering only those patients experiencing dysphonia, an improvement of 9.4% was seen. There was an overall net reduction for the CVL cohort of 12 speech valves in the year after injection. Conclusions Our series confirm the safety and objective efficacy of botox injection for PES. QoL measurements were less convincing, and this disparity between subjective and objective measurements must be considered when treating such patients.

Prognostic implications of pathologically determined tumour volume in glottic carcinomas treated by transoral laser microsurgery.

OBJECTIVES: The TNM classification system for squamous cell carcinoma (SCC) of the head and neck neglects to incorporate volumetric analysis of the primary tumour. Tumour volume (TV) has been implicated prognostically in laryngeal SCC treated by primary radiotherapy (RT), but data for patients treated surgically are lacking. We evaluated such for glottic SCCs resected by transoral laser microsurgery (TLM). DESIGN: Retrospective cohort study utilising TVs calculated as the product of tumour resection dimensions and time-to-event analyses using the Kaplan-Meier method. The prognostic ability of variables was estimated using log-rank statistics, univariate Cox regression and receiver-operating characteristics analysis where appropriate. SETTING: Tertiary referral head and neck cancer centre. PARTICIPANTS: Patients undergoing primary TLM for glottic SCC with curative intent (2007-2011) with at least 12 months follow-up data. MAIN OUTCOME MEASURES: Prognostic impact of TV on local control (LC), overall survival (OS) and disease-specific survival (DSS). RESULTS: Eligible patients (n = 129) had a median follow-up of 40 months (range 14-79 months). Median TV for all cases was 300 mm(3) (range 2-19800 mm(3)). Three-year LC, OS and DSS were 92%, 92% and 98%, respectively. Tumour volume was not a significant predictor of any oncological outcome measure. Otherwise, a significant influence of pT stage on DSS was observed and of age on OS. CONCLUSIONS: In contrast to laryngeal SCC treated by RT, TV does not appear to portend oncological outcome in glottic SCC managed specifically by TLM and consequently does not warrant incorporation into current prognostic models for such patients.

The rising rate of admissions for tonsillitis and neck space abscesses in England, 1991-2011.

INTRODUCTION: Sore throats and tonsillitis represent a considerable health burden as well as a significant source of expenditure for the National Health Service (NHS). As part of the recent NHS savings drive, the introduction of 'procedures of low clinical effectiveness' (PoLCE) lists has reinforced a large reduction in the number of tonsillectomies performed. We carried out a cross-sectional study of trends in emergency sore throat admissions in the context of the number of tonsillectomies performed. METHODS: Hospital Episode Statistics (HES) data were extracted. Office for National Statistics data were also used. RESULTS: Between 1991 and 2011, the overall tonsillectomy rate fell by 44%. In the same time, the admission rate for tonsillitis rose by 310% (Pearson's r=-0.67, p=0.01). The peritonsillar abscess admission rate rose by 31% (r=-0.79, p<0.01). Between 1996 and 2011, the overall tonsillectomy rate fell by 41% and the retro and parapharyngeal abscess admission rate rose by 39% (r=-0.55, p=0.026). There was a 14% overall increase in tonsillectomy and sore throat associated bed days. This was despite the large fall in tonsillectomy numbers and the reduction in length of hospital stay. CONCLUSIONS: Efforts to reduce the tonsillectomy rate are correlated with a significant rise in emergency admissions. The rise in the retro and parapharyngeal abscess rate is perhaps most alarming given the very high mortality of these conditions. Bed day data suggest that no net saving has been made despite the new measures.

Characterisation of patients with endoscopy-negative, computer tomography-negative midfacial segment pain using the sino-nasal outcome test.

BACKGROUND: The purpose of this study was to qualitatively characterise patients with midfacial segment pain (MSP) using the Sino-Nasal Outcome Test (SNOT). The data will provide a detailed overview of the physical and psychological impact on patients'well-being, and how it compares with the normal, healthy population. METHODS: Suitable patients were prospectively identified from the Multi-disciplinary Facial Pain Clinic at the Royal Liverpool University Hospital, based on the diagnostic criteria for MSP. The pre-treatment SNOT-22 of these patients were also compared to patients with chronic rhinosinusitis and normal healthy volunteers. RESULTS: Twenty-nine consecutive patients with a diagnosis of MSP were identified, and compared with 30 CRS patients and 34 healthy volunteers. The average SNOT-22 scores of MSP and CRS patients were higher than normal healthy volunteers. Patients with CRS had the highest rhinological subscale SNOT scores compared to normal healthy volunteers and MSP. Conversely, the reported ear and facial symptoms of MSP patients were most unfavourable. A similar trend was observed in reported sleep function where MSP patients recorded higher subscale scores than the other two cohorts. The subscale mean score for psychological function of MSP patients was not significant when compared to the mean score of patients diagnosed with CRS. CONCLUSION: MSP has an adverse impact on both physical and psychological well-being. The subtle differences in the SNOT subscores between MSP and CRS have provided greater insight into the character and disease impact of MSP. We propose that the SNOT may be suitably utilised in MSP to document disease severity and measure response to treatment.

Infections of cerebrospinal fluid diversion devices in adults: the role of intraventricular antimicrobial therapy.

OBJECTIVES: The precise role for intraventricular (IVT) antimicrobials in combination with systemic antibiotics in management of cerebrospinal fluid (CSF) diversion device-associated infections is uncertain. We evaluated our current practice, comparing dual therapy against systemic antimicrobials alone. METHODS: All adult patients with at least two consecutive CSF isolates who were treated for CSF diversion device-related infection over a 5-year period (2005-2010) were identified retrospectively. Clinical and laboratory parameters, microbiology, surgical and antimicrobial management, and treatment outcomes were analysed. RESULTS: Forty-eight patients were identified - 25 received IVT and systemic antibiotics (group A), and 23 systemic antibiotics alone (group B). Clinical features were similar between groups, as were causative organisms. CSF leucocyte counts differed slightly (A > B, p = 0.067) but no laboratory parameters differed significantly. Infected devices were generally revised (A = 92%, B = 91%). Mean times to CSF sterilisation and normalisation of CSF microscopy were significantly shorter for group A (p < 0.05 and p < 0.005 respectively), as was duration of hospital stay (p < 0.002) and required length of systemic antimicrobial therapy (p < 0.001). CONCLUSIONS: Our findings indicate that IVT antibiotics enhance clinical and microbiological recovery and should therefore be considered for patients with CSF infection associated with a CSF diversion device. We recommend further evaluation of this approach in a prospective, randomised, controlled trial.

Influence of natural organic matter (NOM) quality on Cu-gill binding in the rainbow trout (Oncorhynchus mykiss).

Natural organic matter (NOM) in aquatic environments reduces metal toxicity to fish by forming metal-NOM complexes, which reduce metal bioavailability, metal-gill binding and toxicity. However, differences in the chemical composition of different types of NOM (quality) could also affect metal-NOM binding and toxicity. We predicted that Cu-gill binding would vary in trout exposed to Cu in the presence of NOM of different qualities. NOM was collected from three sources: Luther Marsh (terrigenous approximately allochthonous), Bannister Lake (nominally autochthonous), and from a local sewage treatment plant (designated Preston effluent). Excitation-emission matrix spectroscopy (EEMS) revealed that terrigenous Luther Marsh NOM was primarily humic acid-like material (74%), whereas Bannister Lake and Preston sewage effluent NOM had lower humic acid-like material but greater fulvic acid-like material (30% and 50%, respectively). The specific absorption coefficient (SAC) of Luther Marsh NOM was also much higher (SAC=37.8), consistent with its darker color, compared to more autochthonous, lightly coloured Bannister Lake (SAC=12.4) NOM, and Preston effluent NOM (SAC=9.2). At low-moderate waterborne Cu (0-2,000 nmol L(-1)), all NOM isolates reduced Cu-gill accumulation by 70-90%. Surprisingly, there were no measurable differences in Cu-gill binding amongst the three NOM treatments when fish were exposed to Cu in the low-moderate range. Only at higher Cu (>2,000 nmol L(-1)) were differences observed, where terrigenous Luther Marsh and Preston effluent NOM reduced Cu-gill binding by 40-50% more than the more autochthonous Bannister Lake NOM. Although Cu-gill binding estimates using the HydroQual BLM showed similar trends, the BLM consistently underestimated Cu-gill binding. We conclude that differences in Cu toxicity at lower-moderate Cu concentrations in the presence of different types of NOM are not necessarily related to measurable differences in Cu-gill accumulation. Rather, we suggest that differences in Cu toxicity reported in the presence of different types of NOM might be explained by direct actions of NOM on the gills, which are quality dependent.