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The human paranasal sinuses are the major source of intrinsic nitric oxide (NO) production in the human airway. NO plays several roles in the maintenance of physiological homeostasis and the regulation of airway inflammation through the expression of three NO synthase (NOS) isoforms. Measuring NO levels can contribute to the diagnosis and assessment of allergic rhinitis (AR) and chronic rhinosinusitis (CRS). In symptomatic AR patients, pro-inflammatory cytokines upregulate the expression of inducible NOS (iNOS) in the inferior turbinate. Excessive amounts of NO cause oxidative damage to cellular components, leading to the deposition of cytotoxic substances. CRS phenotype and endotype classifications have provided insights into modern treatment strategies. Analyses of the production of sinus NO and its metabolites revealed pathobiological diversity that can be exploited for useful biomarkers. Measuring nasal NO based on different NOS activities is a potent tool for specific interventions targeting molecular pathways underlying CRS endotype-specific inflammation. We provide a comprehensive review of the functional diversity of NOS isoforms in the human sinonasal system in relation to these two major nasal disorders' pathologies. The regulatory mechanisms of NOS expression associated with the substrate bioavailability indicate the involvement of both type 1 and type 2 immune responses.

Objective: It remains controversial whether nasal nitric oxide (NO) serves as a reliable parameter to evaluate treatment efficacy in patients with allergic rhinitis (AR). The measurement of local nasal NO levels has been shown to be a sensitive marker for the diagnosis of symptomatic AR patients. Here we assessed the applicability of nasal NO to evaluations of the efficacy of intranasal steroids (INS) in a prospective design., Methods: We enrolled 25 patients with perennial AR and 10 age-matched healthy participants. The AR patients received fluticasone furoate (FF) once daily for 2 months. Fractional exhaled NO and nasal NO measurements were carried out using an electrochemical analyzer at pretreatment and at 2 weeks and 2 months after treatment. Nasal NO levels were directly measured at two different areas of the nasal cavity: the surface of the inferior turbinate (IT area) and the front of the middle meatus (MM area). Subjective nasal symptoms were also recorded at each visit., Results: The mean total nasal symptom score in the AR patients decreased significantly after FF treatment (p<0.0001). The mean nasal NO levels in the IT area in the AR patients were significantly higher at pretreatment than those of the healthy participants (109 vs. 62.5 ppb, respectively; p<0.001). After FF administration, the nasal NO levels in the IT area of the AR group showed a significant reduction at both 2 weeks and 2 months (79.1 and 71.9 ppb, respectively; p<0.05 and p<0.01). There was no significant difference in nasal NO levels in the MM area between the controls and the AR group at any visit timepoint. When the ratio of the MM area to the IT area (MM/IT ratio) was plotted for each subject, the untreated AR patients showed a marked decrease in the ratio, whereas after the FF treatment, the AR patients' mean MM/IT ratios showed a significant increase. No significant difference compared to the control group existed at 2 months., Conclusion: Nasal NO measurement around the inferior turbinate is an objective measure to evaluate allergic conditions and is useful to monitor therapeutic effects of INS., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: Patients with allergic rhinitis (AR) show augmented activity of nitric oxide (NO) metabolism, similar to those in bronchial asthma (BA). We hypothesized that measurements of nasal fractional exhaled NO (FeNO) could be used as an objective marker to detect the presence of AR. Our objective was to clarify the influence of nasal airflow resistance (NAR) on nasal FeNO levels through an exhalation maneuver in symptomatic AR patients. We also examined the diagnostic test validity of the mean nasal FeNO level for disease discrimination by means of a receiver operating characteristic (ROC) curve analysis., Methods: Fifty-nine untreated perennial AR patients without BA and 60 healthy controls were enrolled in this retrospective cross-sectional study. The subjective symptoms were recorded and the disease severity was classified according to the Japanese guideline for AR. The oral and nasal FeNO measurements were carried out using a handheld electrochemical analyzer according to the ATS/ERS guidelines. NAR was measured using a rhinomanometer by the anterior method., Results: The patients in the moderate-to-most severe AR group showed significantly higher levels of oral FeNO compared to the controls. The AR patients in both the mild (n=25) and the moderate-to-most severe (n=34) groups showed significantly higher levels of nasal FeNO compared to the controls (44.1ppb, 54.5ppb, and 26.5ppb, respectively). There was no significant difference in total NAR between the AR patients and the controls. The results of our comparison of nasal FeNO and NAR values of the ipsilateral nasal cavity for each individual indicated no significant correlation between the two-paired parameters. The optimal cut-off point of the mean nasal FeNO level was calculated as 38.5ppb (with 71% sensitivity and 86% specificity) to discriminate the presence of AR., Conclusion: Nasal FeNO measurements can be an objective parameter for the diagnosis and classification of perennial AR in Japanese individuals. Nasal FeNO and NAR appear to be two independent measures that can be used to objectively evaluate nasal functions., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Objective: Although human paranasal sinuses are critical organs for nitric oxide production, little information is available regarding the role of arginase in alterations of arginine metabolism and nasal nitric oxide levels that may be informative for classifying chronic rhinosinusitis subtypes., Methods: The expression and localisation of arginase and nitric oxide synthase isoforms in paranasal sinus mucosa were examined, and the fractional exhaled nitric oxide was measured in chronic rhinosinusitis without nasal polyps (n=18) and chronic rhinosinusitis with nasal polyps (n = 27) patients., Results: Increased arginase-2 activities in chronic rhinosinusitis without nasal polyps patients were associated with significantly lower levels of nasal fractional exhaled nitric oxide. Chronic rhinosinusitis with nasal polyps patients showed significant NOS2 messenger RNA upregulation with concomitant higher levels of oral and nasal fractional exhaled nitric oxide., Conclusion: These results indicate that fractional exhaled nitric oxide is a valid marker for differentiating chronic rhinosinusitis phenotypes based on a delicate balance between arginase and nitric oxide synthase activities in nitric oxide production.

Background: Because of the anatomical complexity and the high output of the human nose, it has been unclear whether nasal nitric oxide (NO) serves as a reliable marker of allergic rhinitis (AR). We examined whether nasal NO levels in the inferior turbinate (IT) surface and the middle meatus (MM) differ in symptomatic AR patients., Methods: We measured fractional exhaled NO (FeNO) and nasal NO in normal subjects (n = 50) and AR patients with mild symptoms (n = 16) or moderate or severe symptoms (n = 27). Nasal NO measurements were obtained using an electrochemical analyzer connected to a catheter and an air-suction pump (flow rate 50mL/sec)., Results: Compared to the normal subjects, the AR patients showed significantly higher nasal FeNO and nasal NO levels in the IT area. No significant difference in the MM area was observed among the three groups. The MM area showed higher NO levels than the IT area in all three groups. The ratio of nasal NO levels of the MM area to the IT area (MM/IT ratio) was significantly lower in the AR groups. The moderate/severe AR patients showed significantly higher nasal NO in the IT area (104.4 vs. 66.2ppb) and lower MM/IT ratios than those in the mild AR patients. The analysis of nasal brushing cells revealed significantly higher eosinophil cationic protein and nitrotyrosine levels in the AR groups., Conclusions: Nasal NO assessment in the IT area directly reflects persistent eosinophilic inflammation and may be a valid marker to estimate the severity of AR.

Objective: Monitoring of fractional concentrations of exhaled nitric oxide (FeNO) has become a reliable marker of inflammation in human nose and paranasal sinuses. However, it is still unknown to what extent nasal NO levels contribute to the pathology of chronic rhinosinusitis (CRS). In the present study, we aimed to examine FeNO levels and the underlying mechanism of NO production and metabolism in patients with eosinophilic chronic rhinosinusitis (ECRS) and non-ECRS., Methods: Thirty-three untreated ECRS patients, 16 non-ECRS patients, and 38 normal subjects were enrolled in this cross-sectional study of FeNO levels. Oral and nasal FeNO levels were measured before treatment using an electrochemical NO analyzer (NObreath(®)) with a nose adaptor. The mRNA expression of three nitric oxide synthase (NOS) isoforms, interleukin-5 (IL-5), and transforming growth factor-beta (TGF-β) in the ethmoid sinus mucosa and nasal polyps were analyzed by real-time PCR. Immunohistological localization of inducible NOS (iNOS) and nitrotyrosine (NT), a marker for oxidized NO metabolites, was also examined., Results: ECRS patients showed significantly higher oral FeNO levels compared to non-ECRS patients and normal subjects (mean values, 47.6, 13.5, and 15.3ppb, respectively). Nasal FeNO levels of the non-ECRS patients (30.5ppb) were significantly lower than those of the ECRS patients (53.9ppb) and normal subjects (45.5ppb). Positive correlations existed between the blood eosinophil percentage and FeNO levels in ECRS patients. Histologically, ECRS patients showed higher eosinophil accumulation in the ethmoid mucosa than non-ECRS patients (103.1 vs. 16.3cells/HPF). Real-time PCR analysis showed significant upregulation of iNOS and IL-5 mRNA expression in the ethmoid mucosa of the ECRS patients compared to those of non-ECRS patients. Positive iNOS immunoreactivity was observed in ciliated epithelial cells, submucosal glands and associated inflammatory cells in both groups. NT immunoreactivity was detected in the epithelium and around inflammatory cells. Intense NT staining was co-localized with eosinophil accumulation and ECRS patients showed significantly higher rates of NT-positive cells than non-ECRS patients., Conclusion: A combination of oral and nasal FeNO measurement is a valid marker for the classification and definition of different CRS subtypes in Japan. Higher levels of oral and nasal FeNO in ECRS patients may reflect the persistence of eosinophilic inflammation in sinus mucosa with concomitant iNOS upregulation and accompanying deposition of oxidized NO metabolites., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Background: We aimed to examine the effect of different therapeutic modalities on levels of fractional concentrations of exhaled nitric oxide (FeNO) in patients with eosinophilic chronic rhinosinusitis (ECRS)., Methods: Thirty-six ECRS patients with nasal polyps were treated either medically or surgically. Oral and nasal FeNO levels were measured using an electrochemical NO analyzer initially and at 1 and 6 months. The mRNA expression and localization of nitric oxide synthase (NOS) isoforms in sinus mucosa and nasal polyps were analyzed by real-time polymerase chain reaction (PCR) and immunohistochemistry., Results: The mean oral FeNO levels in the surgical group had decreased significantly from 50.9 to 36.8 ppb 6 months after endoscopic sinus surgery. All patients in this group showed significantly higher nasal FeNO levels after treatment. The mean nasal FeNO levels were 62.3 ppb at 1 month and 93.6 ppb at 6 months. Mean oral and nasal FeNO levels in the medical group after treatment remained unchanged when compared with the baseline levels. Positive immunoreactivity of inducible NOS (iNOS) was observed in both epithelial cells and submucosal inflammatory cells. Real-time PCR analysis showed significant up-regulation of iNOS and IL-5 mRNA expression., Conclusion: A combination of oral and nasal FeNO measurements is useful to monitor the extent of inflammation in CRS patients. The increase in nasal FeNO in the surgical group indicates prompt recovery of NO release from healed sinus mucosa through the opened sinus ostia. Reduction of oral FeNO levels may reflect a cessation of the underlying lower airway inflammation that is characteristic of ECRS.

Background: There has been an increasing interest in monitoring the fractional concentrations of exhaled NO (FeNO) levels in allergic rhinitis (AR) patients. In the present study, we examined whether the nasal FeNO measurement might reflect the degree of local allergic inflammation as well as subjective symptoms., Methods: The FeNO measurement was performed using a handheld electrochemical analyzer (NObreath®) with a nose adaptor. In the cross-sectional study, 56 patients with perennial AR patients, 18 AR patients with bronchial asthma (BA), 12 patients with vasomotor rhinitis, and 30 normal subjects were enrolled. For the follow-up study, 12 seasonal allergic rhinitis (SAR) patients against Japanese cedar and 10 perennial AR patients who underwent laser surgery were examined., Results: The AR patients and vasomotor rhinitis patients showed significantly higher oral FeNO levels as compared with the normal subjects. The nasal FeNO levels were significantly higher in the perennial AR patients with or without BA than in the normal subjects and vasomotor rhinitis patients. There were positive correlations between the nasal symptom scores and FeNO levels. The SAR patients showed a significant decrease in the nasal FeNO level after the pollen dispersion season. In addition, the therapeutic effects of laser surgery in the AR patients accompanied a significant reduction in the nasal FeNO levels one month after treatment., Conclusions: The nasal FeNO measurement by NObreath® is easy to perform and suitable for monitoring AR patients in various treatment modalities. Furthermore, it may have potential usefulness as a tool to improve daily clinical care.

Background: Nitric oxide (NO) is believed to participate in the regulation of airway clearance and non-specific cellular immunity. Recent studies have suggested that airway epithelial cells of allergic and non-allergic individuals may differ in their ability to produce this molecule., Objective: The aim of this study was to detect the difference in NO production in human nasal epithelial cells between normal subjects and patients with perennial allergic rhinitis (AR), and to assess the relationship between the expression of nitric oxide synthase (NOS) isoforms and the severity of the disease., Methods: Nasal epithelial cells were obtained from the inferior turbinate. The expression of mRNAs encoding constitutive endothelial NOS (eNOS) and inducible NOS (iNOS) was studied by reverse transcription-polymerase chain reaction (RT-PCR). Direct NO production in living cells was visualized and quantified by a fluorescent indicator, DAF-2 DA., Results: RT-PCR analysis demonstrated that AR patients with a RAST score of 5 or 6 showed significant increases in the levels of iNOS mRNA and slight reductions in those of eNOS mRNA. Patients with a RAST score of 2-4 also revealed the same tendency however, the difference was not significant. DAF-2 DA imaging demonstrated that epithelial cells, especially the ciliated cells, produced a larger amount of NO than non-epithelial inflammatory cells. Preincubation with L-NAME resulted in an approximate 40% decrease in both groups., Conclusion: These results directly indicate that nasal epithelial cells of AR patients overall produce higher levels of NO through the concomitant expression of different NOS isoforms. Continuous NO production by the epithelial cells in normal subjects further support the hypothesis that NO derived from epithelium may play dual roles in the regulation of nasal airway clearance and in the host defense. In addition, the use of DAF-2 DA provides a reliable method to visualize and quantify the direct NO production of living cells.

Nitric oxide (NO) has been found to have various actions in the body. Recently, considerable attention has been focused on the intimate relationship between the intracellular production of NO and morphological or functional changes in ciliated cells. The aim of this study was to clarify the functional significance of NO in the nasal mucosa. Healthy, adult, pigmented guinea pigs were randomly divided into one control and three experimental groups. The animals were instilled with either lipopolysaccharide (LPS) only or LPS plus dexamethasone or NG-nitro L-arginine methyl ester (L-NAME). The effect of NO on the nasal epithelium was analyzed morphologically by scanning electron microscopy and physiologically by ciliary beat frequency (CBF) measurement. The origin of NO was also investigated using a fluorescent indicator for NO, namely 4,5-diaminofluorescein diacetate. LPS induced damage of cilia 3 days after the first instillation, while dexamethasone or L-NAME seemed to attenuate the effect of LPS. NO production was localized in ciliated cells and the main source of NO in ciliated cells is suggested to be inducible NO synthase. The greater number of ciliated cells of LPS-treated animals produced a larger amount of NO compared with normal animals. LPS also induced a decrease in CBF, which was inhibited by dexamethasone or L-NAME. It is suggested that NO may play an important role in pathological changes in the nasal mucosa.

The primary function of well-differentiated ciliated epithelium in the paranasal sinus is to eliminate harmful agents through the beating action of cilia. Respiratory epithelium also contributes to local inflammatory processes through the release of various proinflammatory cytokines. Recently, considerable attention has been focused on the intimate relationship between the cytokine-dependent regulation of the ciliary beat frequency (CBF) and intra-cellular production of nitric oxide (NO) in ciliated epithelial cells. The aims of this study are to examine the effect of tumor necrosis factor-alpha (TNF-alpha), one of the major proinflammatory cytokines, on the ciliary activity of human sinus epithelial cells and to assess the hypothesis that NO is involved in this regulatory mechanism. Human maxillary or ethmoidal sinus mucosa (n = 23) were cultured by the explant-outgrowth method. CBF of the outgrowth ciliated cells was measured by the photoelectrical method before and after being treated with TNF-alpha (0.1, 1 and 10 ng/ml) or dexamethasone (10(-6) M and 10(-7) M). We also investigated the expression of nitric oxide synthase (NOS) isoforms, enzymes responsible for NO synthesis, by fluorescent immunohistochemistry. TNF-alpha increased CBF at relatively low concentrations (0.1 and 1 ng/ml) and decreased CBF at a high concentration (10 ng/ml). Dexamethasone decreased CBF at a concentration of 10(-6) M. Fluorescent immunohistochemistry demonstrated that the expression of inducible NOS was augmented by TNF-alpha and attenuated by dexamethasone, whereas that of endothelial NOS remained unchanged. We conclude that human sinus epithelial cells potentially contribute to the inflammatory process by regulating their ciliary motility through an NO-dependent pathway. Proinflammatory cytokines and steroids are able to modulate this mechanism by the induction or inhibition of expression of different NOS isoforms.

Nitric oxide (NO) plays an important role in the regulation of upper respiratory function. In the nasal cavity, the concentration of NO in the air in patients with untreated allergic rhinitis is higher than that in normal individuals. NO is produced by the action of NO synthase (NOS) using L-arginine as a substrate. To investigate the expression of NOS in human nasal mucosa, histochemical staining for NADPH diaphorase and immunohistochemical staining for NOS isoforms were carried out in nasal inferior turbinate mucosa from patients with nasal allergy. Those without nasal allergy served as controls. NADPH diaphorase histochemical study revealed that NOS was expressed in the nasal epithelium, submucosal glands, nerve fibres and the endothelium in specimens of both allergic and control groups. Immunoreactivity to endothelial NOS (eNOS) was localized to epithelial and endothelial cells in both allergic and control groups. In some specimens in both groups, nerve fibres around submucosal glands stained positively for eNOS. Immunoreactivity to eNOS, however, was slightly stronger in the epithelia of the allergic group than in those of the controls. Immunoreactivity to inducible NOS (iNOS) was localized to epithelial cells, endothelial cells, nasal glands and inflammatory cells. The staining of epithelial cells and inflammatory cells was more marked in the allergic group than the controls. These findings may suggest that the greater amounts of NO in the nasal air of patients with allergic rhinitis are mainly induced by iNOS activity.