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The honeybee, Apis mellifera is economically important for its products (honey, wax, and propolis) and for its role in pollination. This insect is threated due to high population losses in both agriculture and beekeeping. Within causes involved in the loss of honeybees is the increased pesticide use on agriculture. Although current testing for the regularization of insecticide use considers its acute toxic effects on pollinators, little is known about the effects of chronic exposure to sublethal concentrations that may persist in the environment. This study investigated the effect of chronic exposure to sublethal concentrations of lambda-cyhalothrin on the midgut, hypopharyngeal glands, and brain of A. mellifera. Honey bees were fed for eight days with LC 50/100 insecticide. Subsequently, the midgut, hypopharyngeal glands, and brain were analyzed in light and transmission electron microscopies. The midgut was not affected after exposure, except in the posterior region with cell fragments in the lumen and changes in the mitochondria. The hypopharyngeal glands were severely affected by the insecticide with changes in the rough endoplasmic reticulum and cell death. The brain has extensive gaps in the neuropil as well as in the cellular bodies, especially in the corpora pedunculata. These resembled cellular alterations similar to those seen in death processes. The results of this study indicate that lambda-cyhalothrin is toxic to bees at sublethal concentrations and ingested chronically, causing damage to the midgut, hypopharyngeal glands, and brain, and may affect physiological and behavioral aspects of these insects., Competing Interests: Declaration of competing interest All authors declare that they have no competing interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

As a eusocial species, Apis mellifera , the European honey bee, is effectively a superorganism -a group of genetically related individuals functioning as a collective unit. Because the unit of selection is the colony and not the individual, standard methods for assessing toxicologic pathology can miss colony-level responses to stress. For over a decade, US populations of honeybees have experienced severe annual losses attributed to a variety of environmental stressors varying temporally and geographically; differentiating among those stressors is accordingly a high priority. Social interactions among individuals in this social species, however, mean that the "footprint" of stressors such as pesticides, phytochemicals, pathogens, and parasites may be most discernible in individuals that did not themselves directly encounter the stressor. For example, neurotoxic effects of pesticides on nurse bees may impair their behavioral responses to queen-destined larvae, which may then emerge as adults with altered anatomy or physiology. Similarly, pesticide-induced size alterations in nurse hypopharyngeal glands, which produce royal jelly, the exclusive food of larval and adult queens, may disproportionately affect the queen's (and thus colony) health. Thus, evaluating toxicologic pathology in the honeybee requires a new perspective and development of assays that preserve the social context that ultimately determines colony health.

Biliary reflux has been considered a potential risk factor in upper aerodigestive tract malignancies. It is not yet clearly known how pH affects the bile-induced activation of NF-κB and its related oncogenic pathway previously linked to hypopharyngeal carcinogenesis. In this study, repetitive applications of conjugated primary bile acids with unconjugated secondary bile acid, deoxycholic acid (DCA), on human hypopharyngeal primary cells reveal that strongly acidic pH (4.0) optimally enhances the tumorigenic effect of bile, by inducing activation of NF-κB, STAT3 nuclear translocation, bcl-2 overexpression and significant overexpression of the oncogenic mRNA phenotype, compared to weakly acidic pH (5.5) or neutral pH (7.0). As the pH becomes less acidic the partially activated primary bile acids and activated DCA begin to exert their influence; however, with significantly less intensity compared to bile acids at strongly acidic pH. Our findings suggest that biliary tumorigenic effect is strongly pH dependent. Controlling pH during reflux events may be therapeutically effective in reducing the potential risk of bile-induced hypopharyngeal cancer., (© 2019 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

The presence of bile is not an uncommon finding in acidic oesophageal and extra-oesophageal refluxate, possibly affecting the hypopharyngeal mucosa and leading to neoplastic events. We recently demonstrated that acidic bile (pH ≤ 4.0) can induce NF-κB activation and oncogenic mRNA phenotype in normal hypopharyngeal cells and generate premalignant changes in treated hypopharyngeal mucosa. We hypothesize that curcumin, a dietary inhibitor of NF-κB, may effectively inhibit the acidic bile-induced cancer-related mRNA phenotype, in treated human hypopharyngeal primary cells (HHPC), supporting its potential preventive use in vivo. Luciferase assay, immunofluorescence, Western blot, qPCR and PCR microarray analysis were used to explore the effect of curcumin in HHPC exposed to bile (400 μmol/L) at acidic and neutral pH. Curcumin successfully inhibited the acidic bile-induced NF-κB signalling pathway (25% of analysed genes), and overexpression of NF-κB transcriptional factors, c-REL, RELA(p65), anti-apoptotic bcl-2, oncogenic TNF-α, EGFR, STAT3, WNT5A, ΔNp63 and cancer-related IL-6. Curcumin effectively reduced bile-induced bcl-2 overexpression at both acidic and neutral pH. Our novel findings suggest that, similar to pharmacologic NF-κB inhibitor, BAY 11-7082, curcumin can suppress acidic bile-induced oncogenic mRNA phenotype in hypopharyngeal cells, encouraging its future in vivo pre-clinical and clinical explorations in prevention of bile reflux-related pre-neoplastic events mediated by NF-κB., (© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

Purpose: Bile-containing gastroesophageal reflux may promote cancer at extraesophageal sites. Acidic bile can accelerate NF-κB activation and molecular events, linked to premalignant changes in murine hypopharyngeal mucosa (HM). We hypothesize that short-term in vivo topical application of NF-κB inhibitor BAY 11-7082 can prevent acidic bile-induced early preneoplastic molecular events, suggesting its potential role in disease prevention., Experimental Design: We topically exposed HM (C57Bl/6j wild-type) to a mixture of bile acids at pH 3.0 with and without BAY 11-7082 3 times/day for 7 days. We used immunofluorescence, Western blotting, immunohistochemistry, quantitative polymerase chain reaction, and polymerase chain reaction microarrays to identify NF-κB activation and its associated oncogenic mRNA and miRNA phenotypes, in murine hypopharyngeal cells in vitro and in murine HM in vivo., Results: Short-term exposure of HM to acidic bile is a potent stimulus accelerating the expression of NF-κB signaling (70 out of 84 genes) and oncogenic molecules. Topical application of BAY 11-7082 sufficiently blocks the effect of acidic bile. BAY 11-7082 eliminates NF-κB activation in regenerating basal cells of acidic bile-treated HM and prevents overexpression of molecules central to head and neck cancer, including bcl-2, STAT3, EGFR, TNF-α, and WNT5A. NF-κB inhibitor reverses the upregulated "oncomirs" miR-155 and miR-192 and the downregulated "tumor suppressors" miR-451a and miR-375 phenotypes in HM affected by acidic bile., Conclusion: There is novel evidence that acidic bile-induced NF-κB-related oncogenic mRNA and miRNA phenotypes are generated after short-term 7-day mucosal exposure and that topical mucosal application of BAY 11-7082 can prevent the acidic bile-induced molecular alterations associated with unregulated cell growth and proliferation of hypopharyngeal cells., (Copyright © 2018. Published by Elsevier Inc.)

Because of its ecological and economic importance, the honey bee Apis mellifera is commonly used to assess the environmental risk of insect-resistant, genetically modified plants. In the present study, feeding-exposure experiments were used to determine whether pollen from transgenic rice harms A. mellifera worker bees. In 1 experiment, the survival and mean acinus diameter of hypopharyngeal glands of adult bees were similar when bees were fed on pollen from Bt rice lines or from a non-Bt rice line, but bee survival was significantly reduced when they received pollen that was mixed with potassium arsenate as a positive control. In a second experiment, bee survival and hypopharyngeal gland development were not reduced when adult bees were fed on non-Bt pollen and a sucrose solution supplemented with Cry2A at 400 µg/g, Cry1C at 50 µg/g, or bovine serum albumin (BSA) at 400 µg/g, but bee survival and hypopharyngeal gland development were reduced when the diet was supplemented with soybean trypsin inhibitor as a positive control. In both experiments, the uptake of Cry proteins by adult bees was confirmed. Overall, the results indicate that the planting of Bt rice lines expressing Cry2A or Cry1C protein poses a negligible risk to A. mellifera worker bees. Environ Toxicol Chem 2017;36:1243-1248. © 2016 SETAC., (© 2016 SETAC.)

Myeloid cell leukemia-1 (Mcl-1) plays an important role in survival, chemo- and radioresistance of head and neck squamous cell carcinoma (HNSCC). Cyclin-dependent kinase 9/cyclin T (CDK9) promotes excessive production of multiple pro-survival proteins including Mcl-1, leading to impaired apoptosis of cancer cells. As such, CDK9 is an emerging therapeutic target in cancer therapy. We herein report the first study of targeting CDK9 as a treatment strategy for hypopharyngeal squamous cell carcinoma (HSCC), an aggressive malignancy associated with one of the worst prognoses within HNSCC. We showed that mRNA levels of Mcl-1 were significantly higher in HSCC tumor tissues than in the adjacent non-tumor mucosae. In addition, the levels of Mcl-1 mRNA correlated with the tumor size and clinical stage of HSCC patients. CDKI-73, a potent CDK9 inhibitor, was capable of downregulating the expression of Mcl-1 in the HSCC cells by suppression of the CDK9 mediated phosphorylation of RNA polymerase II. CDKI-73 effectively induced apoptosis as a single agent and synergized anti-tumor activity of cisplatin in HSCC cells. Taken together, our study presents compelling evidence for developing CDK9 inhibitors, such as CDKI-73, as new therapeutic strategy for HSCC., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Angioedema (AE) of the upper airways is a severe and potentially life-threatening condition. The incidence has been increasing in the past two decades, primarily due to increased use of medications inhibiting the degradation of vasoactive peptides. Acquired angioedema related to angiotensin-converting enzyme inhibitors (ACEI-AAE) is well known, but other pharmaceutical agents also affect the degradation of bradykinin and substance P. We present a middle-aged man with recurrent episodes of severe AE of the oral cavity, hypopharynx and larynx due to pharmacological inhibition of dipeptidyl peptidase IV., Competing Interests: ERR has received payment for lectures from MSD Norway and Shire. She has received research grants for HAE research from Shire and genetic studies regarding angioedema from CSL Behring and ViroPharma. AB has been involved in clinical research or educational events involving CSL Behring, Jerini, AG/Shire, Sobi, and ViroPharma. TH has participated in a conference on HAE organised and paid by Shire., (2017 BMJ Publishing Group Ltd.)

The worldwide use of neonicotinoid pesticides has caused concern on account of their involvement in the decline of bee populations, which are key pollinators in most ecosystems. Here we describe a role of non-neuronal acetylcholine (ACh) for breeding of Apis mellifera carnica and a so far unknown effect of neonicotinoids on non-target insects. Royal jelly or larval food are produced by the hypopharyngeal gland of nursing bees and contain unusually high ACh concentrations (4-8 mM). ACh is extremely well conserved in royal jelly or brood food because of the acidic pH of 4.0. This condition protects ACh from degradation thus ensuring delivery of intact ACh to larvae. Raising the pH to ≥5.5 and applying cholinesterase reduced the content of ACh substantially (by 75-90%) in larval food. When this manipulated brood was tested in artificial larval breeding experiments, the survival rate was higher with food supplemented by 100% with ACh (6 mM) than with food not supplemented with ACh. ACh release from the hypopharyngeal gland and its content in brood food declined by 80%, when honeybee colonies were exposed for 4 weeks to high concentrations of the neonicotinoids clothianidin (100 parts per billion [ppb]) or thiacloprid (8,800 ppb). Under these conditions the secretory cells of the gland were markedly damaged and brood development was severely compromised. Even field-relevant low concentrations of thiacloprid (200 ppb) or clothianidin (1 and 10 ppb) reduced ACh level in the brood food and showed initial adverse effects on brood development. Our findings indicate a hitherto unknown target of neonicotinoids to induce adverse effects on non-neuronal ACh which should be considered when re-assessing the environmental risks of these compounds. To our knowledge this is a new biological mechanism, and we suggest that, in addition to their well documented neurotoxic effects, neonicotinoids may contribute to honeybee colony losses consecutive to a reduction of the ACh content in the brood food.

The hypopharyngeal glands (HPGs) of worker honeybees undergo physiological changes along with the age-dependent role change from nursing to foraging: nurse bee HPGs secrete mainly major royal jelly proteins, whereas forager HPGs secrete mainly α-glucosidase III, which converts the sucrose in the nectar into glucose and fructose. We previously identified two other genes, Apis mellifera buffy (Ambuffy) and Apis mellifera matrix metalloproteinase 1 (AmMMP1), with enriched expression in nurse bee and forager HPGs, respectively. In the present study, to clarify the molecular mechanisms that coordinate HPG physiology with worker behavior, we first analyzed whether Ambuffy, AmMMP1, mrjp2 (a gene encoding one of major royal jelly protein isoforms), and Hbg3 (a gene encoding α-glucosidase III) expression, is associated with worker behavior in 'single-cohort colonies' where workers of almost the same age perform different tasks. Expression of these genes correlated with the worker's role, while controlling for age, indicating their regulation associated with the worker's behavior. Associated gene expression suggested the possible involvement of some hormonal factors in its regulation. We therefore examined the relationship between ecdysone- and juvenile hormone (JH)-signaling, and the expression profiles of these 'indicator' genes (nurse bee HPG-selective genes: mrjp2 and Ambuffy, and forager HPG-selective genes: Hbg3 and AmMMP1). Expression of both ecdysone-regulated genes (ecdysone receptor, mushroom body large type Kenyon cell specific protein-1, and E74) and JH-regulated genes (Methoprene tolerant and Krüppel homolog 1) was higher in the forager HPGs than in the nurse bee HPGs, suggesting the possible roles of ecdysone- and JH-regulated genes in worker HPGs. Furthermore, 20-hydroxyecdysone-treatment repressed both nurse bee- and forager-selective gene expression, whereas methoprene-treatment enhanced the expression of forager-selective genes and repressed nurse bee-selective genes in the HPGs. Our findings suggest that both ecdysone- and JH-signaling cooperatively regulate the physiological state of the HPGs in association with the worker's behavior.

Biodegradable crosslinked polyurethane (cPU) was synthesized using polyethylene glycol (PEG), L-lactide (L-LA), and hexamethylene diisocyanate (HDI), with iron acetylacetonate (Fe(acac)3) as the catalyst and PEG as the extender. Chemical components of the obtained polymers were characterized by FTIR spectroscopy, (1)H NMR spectra, and Gel Permeation Chromatography (GPC). The thermodynamic properties, mechanical behaviors, surface hydrophilicity, degradability, and cytotoxicity were tested via differential scanning calorimetry (DSC), tensile tests, contact angle measurements, and cell culture. The results show that the synthesized cPU possessed good flexibility with quite low glass transition temperature (T g , -22°C) and good wettability. Water uptake measured as high as 229.7 ± 18.7%. These properties make cPU a good candidate material for engineering soft tissues such as the hypopharynx. In vitro and in vivo tests showed that cPU has the ability to support the growth of human hypopharyngeal fibroblasts and angiogenesis was observed around cPU after it was implanted subcutaneously in SD rats.

Background: Coordination between breathing and swallowing helps prevent aspiration of foreign material into the respiratory tract. The authors examined the effects of anesthesia and hypercapnia on swallowing-breathing coordination., Methods: In a randomized controlled crossover study, general anesthesia with propofol or sevoflurane was titrated using an up-down method to identify the threshold for suppression of the motor response to electrical stimulation of the forearm. Additional measurements included bispectral index, genioglossus electromyogram, ventilation (pneumotachometer), and hypopharyngeal pressure. During wakefulness and at each level of anesthesia, carbon dioxide was added to increase the end-tidal pressure by 4 and 8 mmHg. A swallow was defined as increased genioglossus activity with deglutition apnea and an increase in hypopharyngeal pressure. Spontaneous swallows were categorized as physiological (during expiration or followed by expiration) or pathological (during inspiration or followed by an inspiration)., Results: A total of 224 swallows were analyzed. Anesthesia increased the proportion of pathological swallows (25.9% vs. 4.9%) and decreased the number of swallows per hour (1.7±3.3 vs. 28.0±22.3) compared to wakefulness. During anesthesia, hypercapnia decreased hypopharyngeal pressure during inspiration (-14.1±3.7 vs. -8.7±2 mmHg) and increased minute ventilation, the proportion of pathological swallows (19.1% vs. 12.3%), and the number of swallows per hour (5.5±17.0. vs. 1.3±5.5)., Conclusions: Anesthesia impaired the coordination between swallowing and respiration. Mild hypercapnia increased the frequency of swallowing during anesthesia and the likelihood of pathological swallowing. During anesthesia, the risk for aspiration may be further increased when ventilatory drive is stimulated.

Background: To evaluate the efficacy and adverse events of cisplatin, tegafur, and leucovorin concomitantly with radiotherapy for patients with advanced, non-metastatic squamous cell carcinoma (SCC) of the oropharynx and hypopharynx., Methods: The PTL regimen consisted of cisplatin (P) 50 mg/m 2 on day 1, oral tegafur (T) 800 mg/day plus leucovorin (LV) 60 mg/day on days 1 through 14. It was repeated every 2 weeks through the radiotherapy course. Conventional radiotherapy with 1.8-2.0 Gy/day, 5 days per week, was delivered in a total dose of between 70 and 72 Gy., Results: Sixty-five patients with stage III or IV of SCC of the head and neck were consecutively treated between May 2002 and November 2005. Forty-six (70.7%) patients had complete response after concomitant chemoradiotherapy (CCRT). With a median follow-up of 54.0 months (range 1-103 months), the 5-year locoregional control, progression-free survival, and overall survival rates were 50.6%, 40.7%, and 59.7%, respectively. Three (4.6%) patients had toxic death during treatment. Fifty-one (80.0%) patients experienced grade 3-4 mucositis which occurred in about 35% of the CCRT duration. The functional preservation rate among post-CCRT complete responders was 93.5% (43/46). The median cisplatin accumulated dosage was 150 mg, and the rate of hearing impairment among the survivors was 7.8%., Conclusion: CCRT with outpatient-based PTL for advanced SCC of oropharynx and hypopharynx is feasible and has comparative efficacy and acceptable adverse events.

Outcome Objectives: (1) Investigate the role of reflux, specifically pepsin, in laryngopharyngeal carcinogenesis. (2) Evaluate effects of chronic pepsin exposure on cell migration, apoptosis, and colony-forming ability in hypopharyngeal cells., Study Design: Translation research., Setting: Academic research laboratory., Methods: Human hypopharyngeal squamous carcinoma FaDu cells were chronically exposed to nonacidic pepsin (exposed for 24 hours, 4 times over 2 weeks at the following concentrations: 0.01 mg/mL, 0.1 mg/mL, or 1 mg/mL). Precise wounds were created in confluent cell plates, and rates of cell migration into wounds were quantified. Separately, cell viability of chronic pepsin-exposed FaDu cells acutely treated with paclitaxel was measured. Finally, a clonogenic assay was performed on these cells to measure effects of chronic pepsin exposure on colony-forming ability., Results: An increased rate of relative wound density was observed in chronic pepsin-treated (0.01 mg/mL, 0.1 mg/mL) cells compared with control (P < .001), suggesting greater rates of cell migration. Pepsin-treated (0.1 mg/mL) cells demonstrated on average greater cell viability compared with control after exposure to paclitaxel, suggesting possible apoptotic resistance; however, this was not statistically significant. Chronic pepsin exposure (0.1 mg/mL, 1 mg/mL) was associated with a dose-dependent increase in colony-forming ability relative to control (P < .001)., Conclusion: Hypopharyngeal squamous cell line chronically exposed to pepsin demonstrated increased cell migration and colony-forming ability relative to control cells. These experiments indicate that chronic pepsin exposure acts as a promoter of tumorigenesis and metastasis of airway epithelium, suggesting a role for pepsin in laryngopharyngeal carcinogenesis attributed to gastric reflux.

Background: Cough hypersensitivity may be related to the pathogenesis of upper airway cough syndrome (UACS). The purpose of the study was to investigate the role of capsaicin-sensitive cough receptors on the laryngopharynx and lower airway in the cough hypersensitivity of patients with UACS., Material and Methods: 59 patients with UACS, 33 patients with rhinitis/sinusitis without cough, and 39 healthy volunteers were recruited for the study. Cough threshold C5, defined as the lowest concentration of capsaicin required for the induction of ≥ 5 coughs upon exposure to capsaicin, were determined at baseline and after laryngopharngeal anesthesia with lidocaine in all the subjects. After induced sputum cytology, the concentrations of histamine, prostaglandin E2 (PGE2), and calcitonin-gene-related peptide (CGPR) in the induced sputum were measured by ELISA. In 15 patients with UACS, sputum cytology and measurement of the above mediators were repeated after successful therapy., Results: C5 response to capsaicin was significantly lower in the UACS group than in the rhinitis/sinusitis group and healthy control groups [3.9 (0.98, 7.8) µmol/L vs. 7.8 (3.9, 93.75) µmol/L vs. 31.2 (15.6, 62.5) µmol/L, H=40.12, P=0.000]. Laryngopharngeal anesthesia with lidocaine dramatically increased C5 to capsaicin in the subjects of all 3 groups by a similar degree, but the increase in the UACS group was still the lowest, with an increased level of histamine, PGE2, and CGRP in the induced sputum. When cough resolved with the treatment of cetirizine alone or in combination with erythromycin, the levels of CGRP and histamine in the induced sputum decreased significantly in 15 patients with UACS, with no obvious change in cell differential or concentration of PGE2 in the induced sputum., Conclusions: Laryngeal TRPV1 plays an important role in cough sensitivity, but sensitization of capsaicin-sensitive cough receptors in the lower airway may be more responsible for the cough hypersensitivity in patients with UACS.

Objective/hypothesis: Laryngopharyngeal reflux (LPR) is thought to be a significant risk factor for laryngeal squamous cell carcinoma (SCC), but causality has never been proven. It is accepted that chronic reflux into the esophagus can induce metaplastic changes in esophageal mucosa with subsequent increased risk of esophageal adenocarcinoma, but no similar associations have been established for LPR and laryngopharyngeal SCC. The objective of this study was to test the hypothesis that reflux of pepsin into the laryngopharynx can promote carcinogenesis., Study Design: Translational research study., Methods: Normal human laryngeal primary epithelial cell cultures and hypopharyngeal FaDu SCC cells were exposed to human pepsin and analyzed by Human Cancer PathwayFinder and miRNA Superarrays, flow cytometry, and Western blot to determine the effect of pepsin on carcinogenesis. Laryngeal biopsy specimens taken from cancer patients and normal control subjects were analyzed for the presence of pepsin by Western blot., Results: Microarray analysis demonstrated that pepsin significantly altered the expression of 27 genes implicated in carcinogenesis and also affected the expression of 22 microRNAs known to be altered in human head and neck cancers. Pepsin increased proliferation in both FaDu SCC cells and cultured normal laryngeal epithelial primary cells by increasing S phase distribution on flow cytometry analysis in a time- and dose-dependent manner. Furthermore, pepsin was detected in 60% (3/5) human laryngeal cancer biopsies, absent in all (0/5) normal control specimens., Conclusions: These data support a role for refluxed pepsin in the promotion of epithelial proliferation and carcinogenesis of the larynx and pharynx., (Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.)

The objective of the present study was to estimate the efficacy of laripront intended for the treatment of inflammatory diseases of the laryngopharynx in the children. Available for the observation were 50 patients aged between 4 and 14 years suffering from the following ENT pathologies: adenoiditis, lacunar tonsillitis, acute laryngitis, chronic tonsillitis, oropharyngeal candidiasis, chronic hypertrophic pharyngitis, atrophic pharyngolaryngitis after the chemical burn of the mouse cavity and laryngopharynx or in the case of gastroesophageal reflux disease. All the patients enjoyed the positive outcome of the treatment that was especially efficacions in the patients with acute pathologies. No adverse effects of the treatment were documented.

Iron pill-induced mucosal injury of the airways with massive necrosis and stricture of the lower airways is known to occur, but symptomatic injury of the hypopharyngeal mucosa secondary to therapeutic oral iron ingestion has not been described. We report iron-sulfate-induced mucosal injury of the hypopharynx in a patient receiving therapeutic oral iron supplementation. The patient presented with dysphagia when swallowing pills, but not when eating or drinking. Imaging studies and clinical examination revealed an ulcerated and protuberant hypopharyngeal mass. Histology showed mucosal ulceration with deposits of extracellular crystalline iron particles. The histologic changes were identical to those seen in patients with "iron pill" gastritis.

Stimulation of water receptors in the laryngopharynx (LP) with water facilitates voluntary swallowing in humans. Based on measures of swallowing intervals (SIs) in repetitive swallowing, we investigated characteristics of laryngopharyngeal water receptors in humans. Healthy adult volunteers were instructed to perform repetitive swallowing as quickly as possible during infusion of a solution into the LP. Infusion of water shortened SI, suggesting that water excites water receptors. Infusion of 0.3 M NaCl solution prolonged SI, suggesting that the NaCl solution inhibits activity of water receptors. SI increased with increasing concentration of NaCl. Anion or cation substitutions indicated that excitation of water receptors is due to absence or reduced concentration of Cl(-). With diminution of peripheral inputs, cortical inputs would play a dominant role in voluntary swallowing. With infusion of a nonstimulating solution (0.3 M NaCl at 0.2 mL/min), SI varied greatly from subject to subject, suggesting that the ability of central regulation of swallowing to initiate repetitive voluntary swallowing varies among subjects. Facilitation of swallowing by chemosensory inputs from water receptors appeared strongly in subjects with longer SI with infusion of the nonstimulating solution. It appears that chemosensory activation compensates for the difficulty in initiating swallowing via the central neural mechanism.

In this study, we assessed the effects of halofuginone and methylprednisolone on hypopharyngeal and esophageal stricture that can develop following radiation to the head and neck of rats. Rats were divided into four groups randomly and 18 Gy radiation was given to the head and neck regions of all rats except the control group. Group 1 (Control Group): No radiation or drugs were administered. Group 2 (Radiation Group): only radiation was applied without any drugs. Group 3 (Halofuginone Group): halofuginone 100 microg/kg per day was given intraperitoneally. Group 4 (Methylprednisolone Group): methylprednisolone 1 mg/kg per day was administered intramuscularly. In all groups, 90 days after application of radiation, sections of the proximal esophagus and hypopharynx were examined for fibrosis, fibroblast proliferation, vascularization, epithelial atypia, necrosis, polymorphonuclear leukocytes, mononuclear cells, and stenosis index by light microscope and the hydroxyproline levels were assessed biochemically. Fibrosis, epithelial atypia and hydroxyproline levels were found to be significantly higher in the radiation group compared to the control group (P < 0.05). We did not observe fibrosis in either the halofuginone or the control groups. Fibrosis was also significantly lower in the methylprednisolone group than the radiation group (P < 0.05). The differences of the stenosis index scores between the groups were not statistically significant (P < 0.05). Vascularization was similar in all groups. We think that especially halofuginone is a drug that can be used safely to prevent fibrosis due to radiotherapy, but further studies are needed.

Objective: To investigate the contribution of pepsin to inflammation attributed to nonacidic gastric reflux via analysis of inflammatory cytokine and cytokine receptor gene expression in pepsin-treated human hypopharyngeal epithelial cells in vitro., Study Design: Translational research., Setting: This study was performed in an academic research laboratory., Subjects and Methods: Human hypopharyngeal epithelial cells were incubated with or without pepsin (0.1 mg/mL) at pH 7.4, 37 degrees C, overnight. Expression of 84 inflammatory cytokines and cytokine receptors was analyzed via RT(2) qPCR array., Results: Expression of a number of inflammatory cytokines and receptors was altered in human hypopharyngeal epithelial cells following overnight treatment with pepsin at neutral pH. Greater than 1.5-fold change in gene expression was detected for CCL20, CCL26, IL8, IL1F10, IL1A, IL5, BCL6, CCR6, and CXCL14 (P < 0.05)., Conclusion: Exposure of hypopharyngeal cells to pepsin in a nonacidic environment induces the expression of several pro-inflammatory cytokines and receptors, including those known to be involved in inflammation of esophageal epithelium in response to reflux and which contribute to the pathophysiology of reflux esophagitis. These data indicate that refluxed pepsin may contribute to laryngeal inflammation associated with nonacidic gastric reflux, including that experienced by patients despite maximal acid suppression therapy.

The aim of this clinical study was to evaluate the effect of laryngopharyngeal reflux (LPR) on the healing process of surgical laryngeal trauma. A total of 112 consecutive patients, who suffered from LPR and were scheduled for operation of Reinke edema or laryngeal polyps/nodules (40 and 72 patients, respectively) during a period of 5 years, were included. Diagnosis of LPR was made on the basis of both history and dual pH probe recording during 24 h in the inferior esophagus and the hypopharynx. The reflux finding score (RFS) and the reflux symptom index (RSI) were used to estimate the clinical severity of LPR. In patients with LPR, proton pump inhibitors (PPI) were initiated in half of them, randomly chosen. Fifty LPR-free subjects operated for Reinke edema or laryngeal polyps during the same time period (19 and 31 patients, respectively) were used as controls. In six patients who had been administered PPI, resolution of the disease was observed and no surgical treatment was undertaken. The remaining patients were operated on under general anesthesia by a single surgeon. All patients had 1-year postoperative follow-up. Epithelization was complete in all vocal cords of both the control group and the group of patients who had been administered PPI. Within the group of patients who had not taken PPI, six patients presented granulation tissue or recurrence of the polyps and in two of them revision surgery was needed. RFS and RSI scores showed significant improvement postoperatively, across all the three groups of patients, with major differences observed in the group treated by PPI. Comparison of the postoperative RFS and RSI scores between the two groups of patients with LPR showed statistically significant differences in both, indicating better treatment outcome in those patients who had received PPI. It may be thus concluded that LPR influences epithelization and recurrence of laryngeal polyps or Reinke edema in vocal cords, after partial or total decortication. Surgical outcome is superior in patients with LPR with preoperative and postoperative anti-reflux treatment.

Laryngopharyngeal reflux (LPR) refers to the backflow of stomach contents into the laryngopharynx. Increasing evidence has demonstrated that LPR is a contributing factor in some cases of hoarseness, vocal fatigue, voice breaks, cough and globus and chronic throat clearing. However, several randomised placebo-controlled trials of proton pump inhibitors in the treatment of LPR have been reported with the majority showing no significant benefit in patient symptom scores over placebo. The aim of this pilot clinical study was to investigate whether any improvement in LPR-related symptoms, using the Reflux Symptom Index (RSI), and clinical findings, using the Reflux Finding Score (RFS), could be achieved with treatment with a liquid alginate suspension compared to control (no treatment). Patients presenting with the symptoms of LPR to the Otorhinolaryngology Outpatient Department at the Queen's Medical Centre, Nottingham, UK were considered eligible if they had an RSI of greater than 10 and an RFS greater than 5 based on a fibreoptic examination of the larynx. A total of 49 patients were randomised into the open, parallel group study; 24 patients were randomised to receive 10 ml liquid alginate suspension (Gaviscon Advance) four times daily after meals and at bedtime, and 25 patients into the control group (no treatment). Patients were assessed pre-treatment and at 2, 4 and 6 months post treatment. Mean (SD) RSI and RFS pre-treatment scores were 23.9 (7.0) and 10.4 (3.6) for the treatment group and 24.6 (7.4) and 10.3 (3.3) for the control group, respectively. Significant differences between treatment and control were observed for RSI at the 2-month (11.2 (7.0) vs. 16.8 (6.4), P=0.005) and 6-month (11.2 (8.1) vs. 18.3 (9.4), P=0.008) assessments and for RFS at the 6-month (7.1 (2.8) vs. 9.5 (3.4), P=0.005) assessment. Significant improvement in symptom scores and clinical findings were achieved with liquid alginate suspension (Gaviscon Advance) compared to control and further evaluation for the management of patients presenting with LPR is warranted.

Objective: To determine the efficacy of proton pump inhibitor (PPI) therapy with esomeprazole on symptoms and signs associated with laryngopharyngeal reflux (LPR)., Study Design and Methods: Prospective, double-blind, randomized, placebo-controlled study. Sixty-two patients with a reflux finding score (RFS)>7 and a reflux symptom index (RSI)>13 were enrolled and received either esomeprazole 20 mg twice daily or placebo for three months. RSI and RFS were assessed at baseline, after six weeks, and after three months., Results: Reductions of total RSI and RFS as well as of several subscores were significantly higher in the treatment group compared to placebo after three months (P<0.05 each). The difference between study groups was most pronounced for posterior commissure hypertrophy (P<0.01)., Conclusion: In the treatment of LPR-related symptoms a high placebo effect can be observed. However, compared to control, twice-daily PPI treatment for three months demonstrated a significantly greater improvement in laryngeal appearance and LPR symptoms.

Objective: The objective of this study was to define the outcome of empiric treatment of suspected laryngopharyngeal reflux (LPR) symptoms with proton pump inhibitors (PPIs)., Design: The authors conducted a systematic review of the English and foreign literature. Studies that used PPIs as an empiric treatment modality for suspected LPR, whether alone or in combination with other acid suppressants and/or placebo, were included. Studies that did not include PPIs as a treatment option were excluded., Main Outcome Measures: A lack of common outcome measures was evident in the uncontrolled studies. In the randomized, controlled trials, outcome measures included symptom questionnaires and videolaryngoscopy. Only one study used computerized voice analysis., Results: Fourteen uncontrolled studies together with one unblinded, nonrandomized study with a control group of healthy volunteers and six double-blind, placebo-controlled randomized trials were identified from 1994 to 2004. Selection bias, blinding of the results, and lack of common outcome measures were some of the problems preventing a formal metaanalysis. Although uncontrolled series reported positive results, randomized, controlled trials demonstrated no statistically significant differences for changes in severity or frequency of symptoms associated with suspected reflux between PPIs and placebo., Conclusions: Recommendations for empiric treatment of suspected LPR with PPIs, by far the most common ear, nose and throat practice in the United Kingdom, are based on poor levels of evidence from uncontrolled studies. The few randomized, controlled trials have failed to demonstrate superiority of PPIs over placebo for treatment of suspected LPR.

Purpose: To identify the anatomic structures whose damage or malfunction cause late dysphagia and aspiration after intensive chemotherapy and radiotherapy (RT) for head-and-neck cancer, and to explore whether they can be spared by intensity-modulated RT (IMRT) without compromising target RT., Methods and Materials: A total of 26 patients receiving RT concurrent with gemcitabine, a regimen associated with a high rate of late dysphagia and aspiration, underwent prospective evaluation of swallowing with videofluoroscopy (VF), direct endoscopy, and CT. To assess whether the VF abnormalities were regimen specific, they were compared with the VF findings of 6 patients presenting with dysphagia after RT concurrent with high-dose intra-arterial cisplatin. The anatomic structures whose malfunction was likely to cause each of the VF abnormalities common to both regimens were determined by literature review. Pre- and posttherapy CT scans were reviewed for evidence of posttherapy damage to each of these structures, and those demonstrating posttherapy changes were deemed dysphagia/aspiration-related structures (DARS). Standard three-dimensional (3D) RT, standard IMRT (stIMRT), and dysphagia-optimized IMRT (doIMRT) plans in which sparing of the DARS was included in the optimization cost function, were produced for each of 20 consecutive patients with advanced head-and-neck cancer., Results: The posttherapy VF abnormalities common to both regimens included weakness of the posterior motion of the base of tongue, prolonged pharyngeal transit time, lack of coordination between the swallowing phases, reduced elevation of the larynx, and reduced laryngeal closure and epiglottic inversion, contributing to a high rate of aspiration. The anatomic structures whose malfunction was the likely cause of each of these abnormalities, and that also demonstrated anatomic changes after RT concurrent with gemcitabine doses associated with dysphagia and aspiration, were the pharyngeal constrictor muscles (median thickness near midline 2.5 mm before therapy vs. 7 mm after therapy; p = 0.001), the supraglottic larynx (median thickness, 2 mm before therapy vs. 4 mm after therapy; p < 0.001), and, similarly, the glottic larynx. The constrictors and the glottic and supraglottic larynx were, therefore, deemed the DARS. The lowest maximal dose delivered to a stricture volume was 50 Gy. Reducing the volumes of the DARS receiving > or =50 Gy (V(50)) was, therefore, a planning and evaluation goal. Compared with the 3D plans, stIMRT reduced the V(50) of the pharyngeal constrictors by 10% on average (range, 0-36%, p < 0.001), and doIMRT reduced these volumes further, by an additional 10% on average (range, 0-38%; p <0.001). The V(50) of the larynx (glottic + supraglottic) was reduced marginally by stIMRT compared with 3D (by 7% on average, range, 0-56%; p = 0.054), and doIMRT reduced these volumes by an additional 11%, on average (range, 0-41%; p = 0.002). doIMRT reduced laryngeal V(50) compared with 3D, by 18% on average (range 0-61%; p = 0.001). Certain target delineation rules facilitated sparing of the DARS by IMRT. The maximal DARS doses were not reduced by IMRT because of their partial overlap with the targets. stIMRT and doIMRT did not differ in target doses, parotid gland mean dose, spinal cord, or nonspecified tissue maximal dose., Conclusions: The structures whose damage may cause dysphagia and aspiration after intensive chemotherapy and RT are the pharyngeal constrictors and the glottic and supraglottic larynx. Compared with 3D-RT, moderate sparing of these structures was achieved by stIMRT, and an additional benefit, whose extent varied among the patients, was gained by doIMRT, without compromising target doses. Clinical validation is required to determine whether the dosimetric gains are translated into clinical ones.

Purpose of Review: Laryngopharyngeal reflux is of great interest to otolaryngologists, speech and language therapists, and gastroenterologists. This is a brief review of recent publications in the diagnosis of laryngopharyngeal reflux., Recent Findings: Otolaryngologic signs and symptoms can be found in 4 to 10% of patients with gastroesophageal reflux and those presenting for ear, nose, and throat evaluations. Laryngeal signs are not pathognomonic for laryngopharyngeal reflux because many of these signs can be found in healthy volunteers. A combination of signs and symptoms should be sought before suspecting this diagnosis. Most investigators consider pH monitoring the best currently available instrument to diagnose gastroesophageal reflux, even though it is not considered to be 100% sensitive and specific. Studies in normal volunteers indicate that a minimal number of reflux episodes reach the hypopharynx. The correlation between laryngeal signs and symptoms and pH-documented reflux is less than perfect, whereas the combination of pH testing and signs and symptoms is better in detecting patients with a favorable response to acid-suppressing therapy. Using an empiric trial of high-dose proton pump inhibitors over a prolonged period of time to diagnose laryngopharyngeal reflux is supported mainly by uncontrolled studies. To date, double-blind, placebo-controlled studies suggest that empiric trials of proton pump inhibitors may not have high accuracy for the diagnosis of laryngopharyngeal reflux., Summary: Multidisciplinary trials are needed to establish the optimal combination of sign and symptom scores, reflux monitoring results, and empiric treatment trials for the most accurate diagnosis of laryngopharyngeal reflux.

Background: The cricopharyngeal muscle is of the skeletal type and, in this way, unable to sustain continuous contraction for long periods. Despite of this it has been considered as the responsible by the high pressure area, registered by manometry into the pharyngoesophageal transition. For this reason, it has been the object of therapeutics that promote the rupture of its integrity., Aims: To give the anatomical bases to define the limits of participation of the cricopharyngeal muscle in the pharyngoesophageal transition function. To consider a morphological and functional alternative to explain the high pressure area on pharyngoesophageal transition and the implications of the myotomy, use of the botulinum toxin and balloon dilatation on pharyngoesophageal transition function., Material and Method: Study of the laryngopharyngeal region in their morphologic characteristics and relationships on 24 pieces obtained from adults' corpses of both sexes fixed in 10% formaldehyde solution., Results: The cricopharyngeal muscle presenting its anterior-lateral insertion, with a C-shaped outline, on the posterior-lateral edge of the cricoid cartilage. This kind of morphology blocks the possibility to generate a predominant anterior and posterior high pressure during its contraction like that we find at the pharyngoesophageal transition. The observation of this kind of pressure has its explanation in a tweezers-like relationship exerted on one side by the vertebral body and on the other side by the posterior contour of the cricoid cartilage., Conclusions: The muscular organization of the laryngopharyngeal segment allowed us to sustain that a large myotomy of the pharyngoesophageal transition, that takes more than just the cricopharyngeal transversal fasciculus, hinders the ejection function in a region where the dimension do not need any parietal sectioning. Myotomy that encompasses only the transversal fasciculus can contribute to improve the pharyngoesophageal flux by a decrease of the local resistance. The efficiency of this myotomy depends mostly on some residual pharyngeal ejection force and also on a slight hyolaryngeal displacement. The transversal fasciculus of the cricopharyngeal muscle is a narrow strip of muscular mass to be injected by percutaneous way with solution of botulinum toxin; maybe endoscopically. For this reason, dose, dilution and injection sites have an important meaning in the cricopharyngeal therapeutics using botulinum toxin. The efficiency of this procedure, like myotomy, depends on some residual pharyngeal ejection force and on, at least, some hyolaryngeal displacement. The dilation of the pharyngoesophageal transition with pneumatic balloon does not seem to be an adequate procedure for a region that does not present a narrow lumen determined by fibrosis. For anatomical characteristics of the TFE region, mean pressure as registered by the manometric method does not evaluate either the effectiveness or inadequacy of surgical myotomy, denervation or dilation using pneumatic balloon.

We present a case report of a 2-year-old child with a hypopharyngeal stricture secondary to caustic soda ingestion. We discuss the initial and long-term management of caustic soda ingestion. In this case the hypopharynx was successfully reconstructed using a pedicled pectoralis major flap. We demonstrate the versatility of this flap, which is traditionally known for its use in adult head and neck surgery.

Laryngeal evoked brainstem responses (LBRs) were recorded in normal human subjects in an attempt to develop a central laryngeal function test and enhance our understanding of neurolaryngologic disorders. The results showed that the human LBR consists of five positive peaks and five negative peaks reproducible within 10 ms after a vibratory stimulation to the superior laryngeal nerve (SLN). The waveform reproducibility was verified by blocking the SLN and topically anesthetizing the hypopharyngeal cavity. The morphology and latency of peak 5 were similar to results obtained in animal LBR experiments. It was concluded that a vibratory stimulation to the SLN was a noninvasive method to elicit far-field potentials from the central laryngeal pathway. These findings encourage further effort to establish normative data and explore clinical correlations.

In a honeybee colony, brood stimulates development of hypopharyngeal glands of nurse bees. A chemical signal, a blend of 10 fatty acid esters, has been identified on larval cuticle. We demonstrate that the blend of 10 esters, ethyl oleate, and methyl palmitate stimulates the protein synthesis of hypopharyngeal glands of nurses. Thus, in Apis mellifera the chemical signal from the brood acts as a primer pheromone in addition to its previously shown role as a releaser pheromone.

The presence of metacyclical promastigotes of Leishmania pifanoi in the hypopharyngeal duct of Lutzomyia youngi is reported. The insects were experimentally infected by engorgement on the tarsal lesions of hamsters. The metacyclics, whose morphology is illustrated, appeared in the hypopharynx 5 to 9 days after engorgement; they were more frequently found in the insects fed on unrefined commercial sugar. They role of amino derivates of glucose and galactose, as well as of amino acids in the development and migration of metacyclics, is investigated.

Background: Previous studies suggest that upper airway neuromuscular activity can be affected by changes in blood pressure via a baroreceptor-mediated mechanism. It was hypothesized that increases in blood pressure would increase upper airway collapsibility predisposing to airway obstruction at a flow-limiting site in the hypopharynx., Methods: To examine the effect of blood pressure on upper airway function, maximal inspiratory air flow was determined through the isolated feline upper airway before, during, and after intravenous infusion of phenylephrine (10-20 micrograms.kg-1.min) in six decerebrate, tracheotomized cats. Inspiratory flow, hypopharyngeal pressure, and pressure at the site of pharyngeal collapse were recorded as hypopharyngeal pressure was rapidly decreased to achieve inspiratory flow limitation in the isolated upper airway. Pressure-flow relationships were used to determine maximal inspiratory air flow and its mechanical determinants, the upper airway critical pressure (a measure of pharyngeal collapsibility), and the nasal resistance upstream to the site of flow limitation., Results: An increased mean arterial blood pressure of 71 +/- 16 mmHg (mean +/- SD) was associated with significant decrease in maximal inspiratory air flow from 147 +/- 38 ml/s to 115 +/- 27 ml.sec-1 (P < 0.01). The decrease in maximal inspiratory air flow was associated with an increase in upper airway critical pressure from -8.1 +/- 3.8 to -5.7 +/- 3.7 cm H2O (p < 0.02), with no significant change in nasal resistance. When blood pressure was decreased to baseline by discontinuing the phenylephrine infusion, maximal inspiratory air flow and upper airway critical pressure returned to their baseline values., Conclusions: Increased blood pressure increased the severity of upper airway air flow obstruction by increasing pharyngeal collapsibility. Previous studies relating baroreceptor activity to neuromuscular regulation of upper airway tone, are consistent with this effect being mediated by afferent activity from baroreceptors. These findings warrant further study because they suggest the possibility that upper airway obstruction in postoperative patients could either be caused or exacerbated by an increase in blood pressure.

Thorium dioxide, widely used as a contrast material, is a producer of alpha particle radiation. This is well demonstrated by autoradiography. The case described illustrates that life-threatening, thorium dioxide-induced pharyngeal haemorrhage may occur even with an occluded carotid artery. The radiation exposure caused an intense foreign body reaction with a marked cell-deficient fibrosis. The alpha particles are well demonstrated by autoradiography. In addition, we were able to show a defect in the wall of the carotid artery due to the Thorotrast injection, which was closed by cell-depleted connective tissue. As the radioactivity of the nuclides of thorium dioxide peaks 30-40 years after its first application, the morbidity will increase and the disease has to be taken into careful consideration in head and neck tumour lesions.

We report two cases of cervical thorotrastoma. Thorotrast is no longer used as a radiographic contrast agent, so that the rare thorotrastoma is often forgotten in the differential diagnosis of hypopharyngeal tumours. Surgical treatment of a thorotrastoma is not recommenced.