Spectrofluorometric determination of calcium in the human nasal secretions investigating the association with olfactory function.

Ionic microenvironment of the nasal secretions especially calcium ions play essential role in the olfactory transmission. However, there is a critical need to determine the free calcium levels in healthy people's nasal secretions in contrast to those of patients with olfactory impairment. A selective spectrofluorometric method was created to quantify nasal calcium levels utilizing its quenching ability to the fluorescence of the functionalized carbon quantum dots. The surface of carbon quantum dots was functionalized with calcium ionophore A23187 and ion association complex, calcium phosphotungstate, to improve the selectively to quantify calcium ions. The functionalized carbon quantum dots exhibited a concentration-dependent fluorescence quenching upon interaction with calcium ions. Different factors influencing the quenching process were done to provide efficient analytical process. The new method, demonstrated accurate calcium determination over the concentration range of 200-4000 ng/mL. The suggested technique was used to measure the calcium in the nasal secretions of both healthy people and patients with olfactory impairment. The findings revealed significantly higher calcium levels in the patient with olfactory dysfunction (healthy vs. patient; 735 ± 20 ng/mL vs. 2987 ± 37 ng/mL, p < 0.05).

Secretory IgA as Biomarker for Gastrointestinal Nematodes Natural Infection in Different Breed Sheep.

Specific IgA antibody has been shown to play an important role in resistance to gastrointestinal nematode (GIN) infections in sheep, particularly in Teladorsagia circumcincta parasitosis. In some breeds, negative associations have been shown between IgA levels and worm burden in experimentally infected sheep. In the present study, we have studied the relationship between IgA levels in naturally infected sheep (582 ewes in total; 193 younger than one year old and 389 older than one year old) and fecal egg count (FEC) in the Assaf, Castellana, and Churra breeds. ELISA assays were performed to measure IgA levels against the somatic antigen of T. circumcincta third larval stage (L3) and a 203-amino-acid fragment of the protein disulfide isomerase from the same GIN species. A multilevel random intercept model was developed to predict the infection risk according to age or breed. Spearman's correlation rank was used for statistical analysis. The prediction model showed that breed was not an influential factor in this study, although the Assaf breed could be considered slightly more susceptible than the others. In addition, age affected the infection risk, with the young ewes more susceptible to infection than the adult groups, except for the Castellana breed, whose risk of infection was similar at all ages. The most significant positive association was found between FEC and IgA measured in the nasal secretions of young ewes using both antigens (Rho = 0.5; p = 0.00); the correlation of FEC with IgA in serum was moderately significant (Rho = 0.306; p = 0.00). Comparing both antigens, the protein disulfide isomerase antigen was less reactive than the somatic antigen from L3. In conclusion, under natural conditions, specific IgA against GIN was positively associated with FEC in sheep, with nasal secretions from young animals being the sample where this association is stronger, which, therefore, could be used as a marker of infection in further studies.

Development of a method of nasal secretions sampling for local nasal inflammation studies.

BACKGROUND: Analysis of immune markers in nasal secretions has become crucial in the study of nasal diseases. We proposed the cotton piece method, a modified method, for the collection and processing of nasal secretions. METHODS: The nasal secretions of 31 healthy control participants and 32 patients with nasal diseases were collected by the traditional sponge method and the cotton piece method, respectively. The concentrations of 14 cytokines and chemokines related to nasal diseases were detected. RESULTS: The properties of nasal secretions collected by the cotton piece method were more uniform than the sponge method. The concentration of IL-6 in the disease group collected by the cotton piece method was significantly higher than that in the control group (P = 0.002), and the cotton piece method could distinguish the positive detection rates of IL-1ß (P = 0.031) and TNF-α (P = 0.001) between the control and disease groups. The levels of inflammatory mediators in nasal secretions could preliminarily distinguish different nasal diseases. CONCLUSIONS: The cotton piece method is a noninvasive and reliable method for collecting nasal secretions, which is beneficial for detecting local inflammatory and immune responses of the nasal mucosa.

Challenges in Local Allergic Rhinitis Diagnosis, Management, and Research: Current Concepts and Future Perspectives.

Local allergic rhinitis (LAR) is diagnosed based on the presence of clinical symptoms such as rhinorrhea, sneezing, and nasal itching using negative skin prick testing and serum IgE assessment. Several novel studies have shown that it is possible to use the assessment of nasal sIgE (specific immunoglobulin E) secretion as an additional diagnostic criterion for local allergic rhinitis. Additionally, allergen immunotherapy is a promising-albeit still not fully assessed and evaluated-future method of managing patients with LAR. In this review, the historical background, epidemiology, and main pathophysiological mechanisms of LAR shall be presented. Additionally, we address the current state of knowledge based on selected articles regarding the assessment of the local mucosal IgE presence in response to exposure to such allergens as mites, pollen, molds, and others. The impact of LAR on quality of life as well as the possible options of management (including allergen immunotherapy (AIT), which showed promising results) will then be presented.

Predictive Value of Eosinophil Cationic Protein in Nasal Secretions in Eosinophilic Chronic Rhinosinusitis.

OBJECTIVES: To investigate the value of secretions Eosinophilic cationic protein (ECP) detection in the diagnosis of endotypes of Chronic rhinosinusitis (CRS) and its correlation with clinical symptoms, so as to provide guidance for the clinical application of EOS and ECP detection in secretions. METHODS: Patients' nasal secretions and polyps (or middle turbinate for control) were collected and their EOS% and ECP levels were measured. Correlation analysis was performed for EOS% and ECP levels in secretions and tissues, respectively. The correlation between secretions EOS% and ECP and clinical symptom scores (symptomatic visual analog scale (VAS) scores, Lanza-kennedy scores from nasal endoscopy and Lund-Mackay scores from sinus CT) was further analyzed. Receiver operating characteristic curves were used to assess the predictive potential of EOS% and ECP in nasal secretions. RESULTS: Eosinophilic chronic rhinosinusitis (ECRS) patients had higher concentrations of ECP in nasal secretions than healthy subjects and NECRS (non-eosinophilic CRS) (p < 0.0001;0.0001); EOS% in nasal secretions was higher in ECRS than healthy subjects (p = 0.0055), but the differences between ECRS and NECRS were not statistically significant (p = 0.0999). Correlation analysis showed that tissue EOS% was correlated with ECP concentration and EOS% in nasal secretions (R = 0.5943;0.2815). There was a correlation between EOS% in secretions with a total LM score (R = 0.3131); ECP concentration in secretions with a total LK score (R = 0.3792). To diagnose ECRS, the highest area under the curve (0.8230) was determined for ECP in secretions; the highest area under the curve (0.6635) was determined for EOS% in secretions. CONCLUSION: Measurement of ECP in nasal secretions is useful for non-invasive diagnosis of ECRS. LEVEL OF EVIDENCE: 3 Laryngoscope, 133:3304-3312, 2023.

Diagnostic value of myeloperoxidase and eosinophil cationic protein in nasal secretions for endotypes of chronic rhinosinusitis.

OBJECTIVES: To explore associations between inflammatory endotypes and clinical presentations in CRS. To investigate the value of secretions myeloperoxidase (MPO) and eosinophilic cationic protein (ECP) detections in the diagnosis of endotypes of chronic rhinosinusitis (CRS), so as to provide guidance for the clinical application of MPO and ECP detection in secretions. METHODS: We collected clinical symptom scores from patients with CRS and examined the differences between endotypes in clinical features. Patients' nasal secretions and polyps (or middle turbinate for control) were collected and their NEU number, EOS%, MPO and ECP levels were measured. Correlation analysis was performed for these biomarkers in secretions and tissues, respectively. Receiver operating characteristic curves were used to assess the predictive potential of the biomarkers mentioned above in nasal secretions. RESULTS: Patients with Eos+Neu+ and Eos+Neu-CRS scored highest in most clinical symptom scores, while Eos-Neu+ and Eos-Neu-CRS scored lowest. Correlation analysis showed that tissues NEU number was correlated with NEU number and MPO level in nasal secretions (R = 0.4088; 0.6613); tissues EOS % was correlated with EOS% and ECP level in nasal secretions (R = 0.2344; 0.5774). To diagnose Neu+CRS, the highest area under the curve (AUC) (0.8961) was determined for MPO in secretions; the highest AUC (0.7400) was determined for NEU number in secretions. To diagnose Eos+Neu-CRS from Eos-Neu-CRS in Neu-CRS, the highest AUC (0.8801) was determined for ECP in secretions. CONCLUSIONS: Clinical presentations are directly associated with CRS endotypes. Measurement of MPO and ECP in nasal secretions is useful for the endotypes diagnosis of CRS.

Voluntary Biosurveillance of Streptococcus equi Subsp. equi in Nasal Secretions of 9409 Equids with Upper Airway Infection in the USA.

This study aimed to describe selected epidemiological aspects of horses with acute onset of fever and respiratory signs testing qPCR-positive for S. equi and to determine the effect of vaccination against S. equi on qPCR status. Horses with acute onset of fever and respiratory signs from all regions of the United States were included in a voluntary biosurveillance program from 2008 to 2020 and nasal secretions were tested via qPCR for S. equi and common respiratory viruses. A total of 715/9409 equids (7.6%) tested qPCR-positive for S. equi, with 226 horses showing coinfections with EIV, EHV-1, EHV-4, and ERBV. The median age for the S. equi qPCR-positive horses was 8 ± 4 years and there was significant difference when compared to the median age of the S. equi qPCR-negative horses (6 ± 2 years; p = 0.004). Quarter Horse, Warmblood, and Thoroughbred were the more frequent breed in this horse population, and these breeds were more likely to test qPCR-positive for S. equi compared to other breeds. There was not statistical difference for sex between S. equi qPCR-positive and qPCR-negative horses. Horses used for competition and ranch/farm use were more likely to test qPCR-positive for S. equi (p = 0.006). Horses that tested S. equi qPCR-positive were more likely to display nasal discharge, fever, lethargy, anorexia, and ocular discharge compared to horses that tested S. equi qPCR-negative (p = 0.001). Vaccination against S. equi was associated with a lower frequency of S. equi qPCR-positive status.

[Expression and predictive value of type â ¡ inflammatory cytokines in nasal secretion in eosinophilic chronic rhinosinusitis with nasal polyps].

Objective:To analyze the expression of type Ⅱ inflammatory-related cytokines in nasal secretions of patients with eosinophilic chronic rhinosinusitis with nasal polyps（ECRSwNP）, and to preliminarily explore the role of type Ⅱ inflammatory cytokines in nasal secretions in predicting ECRSwNP. Methods:A prospective analysis was made of 91 patients with CRSwNP who underwent endoscopic sinus surgery in Peking University Third Hospital from November 2020 to June 2021. All the selected patients had their SNOT-22 score, Lund-Mackay score and blood eosinophilia collected before surgery. Percentage and absolute value; the nasal secretions of patients were collected before operation, and enzyme-linked immunosorbent assay was used to detect the typeⅡinflammatory cytokines（IL-4, IL-5, IL-13, IL-25, IL-33, Eotaxin-3, periostin）, intraoperative nasal polyp tissue was collected for eosinophil count. According to the proportion of eosinophils in the tissue≥10%, they were divided into ECRSwNP group and nECRSwNP group. The clinical baseline data and type Ⅱ inflammatory cytokines were compared between the two groups, and the related factors of ECRSwNP were evaluated by univariate logistic regression analysis. The receiver operating characteristic（ROC） curve was used to evaluate the predictive potential of each clinical index. Results:The SNOT-22 score, Lund-Mackay score, blood eosinophil percentage and absolute value in the ECRSwNP group were higher than those in the nECRSwNP group（P<0.05）. In the nECRSwNP group（P<0.01）. Logistic regression analysis found that IL-5, Eotaxin-3 and blood eosinophil percentage were risk factors for ECRSwNP（P<0.05）. ROC analysis found that IL-5, Eotaxin-3 and blood eosinophil percentage had predictive diagnostic value（P<0.01）, among which blood eosinophil percentage had the greatest predictive value（AUC=0.756）. The prediction model composed of Eotaxin-3, SNOT-22 score, sinus CT Lund-Mackay score, blood eosinophil percentage and blood eosinophil absolute value had better prediction effect on ECRSwNP（AUC=0.873）. Conclusion:Type Ⅱ inflammatory cytokines IL-5 and Eotaxin-3 in nasal secretions may be involved as biomarkers for early diagnosis of ECRSwNP.

Role of microRNAs in type 2 diseases and allergen-specific immunotherapy.

MicroRNAs (miRs) have gained scientific attention due to their importance in the pathophysiology of allergic diseases as well as their potential as biomarkers in allergen-specific treatment options. Their function as post-transcriptional regulators, controlling various cellular processes, is of high importance since any single miR can target multiple mRNAs, often within the same signalling pathway. MiRs can alter dysregulated expression of certain cellular responses and contribute to or cause, but in some cases prevent or repress, the development of various diseases. In this review article, we describe current research on the role of specific miRs in regulating immune responses in epithelial cells and specialized immune cells in response to various stimuli, in allergic diseases, and regulation in the therapeutic approach of allergen-specific immunotherapy (AIT). Despite the fact that AIT has been used successfully as a causative treatment option since more than a century, very little is known about the mechanisms of regulation and its connections with microRNAs. In order to fill this gap, this review aims to provide an overview of the current knowledge.

Levels of eicosanoids in nasal secretions associated with nasal polyp severity in chronic rhinosinusitis.

Severe nasal polyposis and mucosal inflammation, in patients with chronic rhinosinusitis (CRS) may include a dysregulated eicosanoid profile, but a clinical role for eicosanoids in CRS with nasal polyps (NP; CRSwNP) remains to be elucidated. This study focused on assessing levels and clinical implications of inflammatory mediators in nasal secretions and urine from patients with different NP severity or Aspirin Exacerbated Respiratory Disease (AERD). Levels of leukotrienes E4 and B4, prostaglandins D2 and E2 as well as 15(S)-hydroxyeicosatetraenoic acid were measured with enzyme immunoassays and cytokines with magnetic bead immunoassays. Patients with CRSwNP were subdivided based on NP score; CRSwNP-low (NP score ≤ 4, n = 11) or CRSwNP-high (NP score ≥ 5, n = 32) and compared to CRS without polyps (CRSsNP, n = 12), CRSwNP-AERD (n = 11) and individuals without CRS (n = 25). Smell test score, fractional exhaled nitric oxide (FeNO), blood eosinophils and Sinonasal outcome test-22 were assessed as clinical markers. Leukotriene E4, prostaglandin D2 and 15(S)-hydroxyeicosatetraenoic acid in nasal secretions correlated with NP score. Nasal leukotriene E4 also correlated with FeNO and smell test score, with highest levels found in CRSwNP-AERD. Levels of prostaglandin D2 in nasal secretion as well as urinary levels of the prostaglandin D2 metabolite 11ß-prostaglandin F2α differed between CRSNP-high and CRSwNP-low. Urinary 11ß-prostaglandin F2α was associated with asthma comorbidity whereas a similar association with prostaglandin D2 in nasal secretions was not observed. In conclusion, subdividing patients based on NP severity in combination with analysis of eicosanoids in non-invasively collected nasal secretions, may have clinical implications when assessing CRS disease severity.

Investigation of the Role of Healthy and Sick Equids in the COVID-19 Pandemic through Serological and Molecular Testing.

More and more studies are reporting on the natural transmission of SARS-CoV-2 between humans with COVID-19 and their companion animals (dogs and cats). While horses are apparently susceptible to SARS-CoV-2 infection based on the homology between the human and the equine ACE-2 receptor, no clinical or subclinical infection has yet been reported in the equine species. To investigate the possible clinical role of SARS-CoV-2 in equids, nasal secretions from 667 horses with acute onset of fever and respiratory signs were tested for the presence of SARS-CoV-2 by qPCR. The samples were collected from January to December of 2020 and submitted to a commercial molecular diagnostic laboratory for the detection of common respiratory pathogens (equine influenza virus, equine herpesvirus-1/-4, equine rhinitis A and B virus, Streptococcus equi subspecies equi). An additional 633 serum samples were tested for antibodies to SARS-CoV-2 using an ELISA targeting the receptor-binding domain of the spike protein. The serum samples were collected from a cohort of 587 healthy racing Thoroughbreds in California after track personnel tested qPCR-positive for SARS-CoV-2. While 241/667 (36%) equids with fever and respiratory signs tested qPCR-positive for at least one of the common respiratory pathogens, not a single horse tested qPCR-positive for SARS-CoV-2. Amongst the racing Thoroughbreds, 35/587 (5.9%) horses had detectable antibodies to SARS-CoV-2. Similar to dogs and cats, horses do not seem to develop clinical SARS-CoV-2 infection. However, horses can act as incidental hosts and experience silent infection following spillover from humans with COVID-19. SARS-CoV-2-infected humans should avoid close contact with equids during the time of their illness.

Invaplex functions as an intranasal adjuvant for subunit and DNA vaccines co-delivered in the nasal cavity of nonhuman primates.

Development of intranasal vaccines for HIV-1 and other mucosal pathogens has been hampered by the lack of adjuvants that can be given safely to humans. We have found that an intranasal Shigella vaccine (Invaplex) which is well tolerated in humans can also function as an adjuvant for intranasal protein and DNA vaccines in mice. To determine whether Invaplex could potentially adjuvant similar vaccines in humans, we simultaneously administered a simian immunodeficiency virus (SIV) envelope (Env) protein and DNA encoding simian-human immunodeficiency virus (SHIV) with or without Invaplex in the nasal cavity of female rhesus macaques. Animals were intranasally boosted with adenoviral vectors expressing SIV env or gag,pol to evaluate memory responses. Anti-SIV antibodies in sera and nasal, genital tract and rectal secretions were quantitated by ELISA. Intracellular cytokine staining was used to measure Th1-type T cells in blood. Macaques given DNA/protein immunizations with 0.5 mg Invaplex developed greater serum IgG, nasal IgA and cervicovaginal IgA responses to SIV Env and SHIV Gag,Pol proteins when compared to non-adjuvanted controls. Rectal IgA responses to Env were only briefly elevated and not observed to Gag,Pol. Invaplex increased frequencies of IFNγ-producing CD4 and CD8 T cells to the Env protein, but not T cell responses induced by the DNA. Ad-SIV boosting increased Env-specific polyfunctional T cells and Env- and Gag,Pol-specific antibodies in serum and all secretions. The data suggest that Invaplex could be highly effective as an adjuvant for intranasal protein vaccines in humans, especially those intended to prevent infections in the genital or respiratory tract.

Portable Electronic Nose for Analyzing the Smell of Nasal Secretions in Calves: Toward Noninvasive Diagnosis of Infectious Bronchopneumonia.

The paper demonstrates a new approach to identify healthy calves ("healthy") and naturally occurring infectious bronchopneumonia ("sick") calves by analysis of the gaseous phase over nasal secretions using 16 piezoelectric sensors in two portable devices. Samples of nasal secretions were obtained from 50 red-motley Holstein calves aged 14-42 days. Calves were subjected to rectal temperature measurements, clinical score according to the Wisconsin respiratory scoring chart, thoracic auscultation, and radiography (Carestream DR, New York, USA). Of the 50 calves, we included samples from 40 (20 "healthy" and 20 "sick") in the training sample. The remaining ten calves (five "healthy" and five "sick") were included in the test sample. It was possible to divide calves into "healthy" and "sick" groups according to the output data of the sensor arrays (maximum sensor signals and calculated parameters Ai/j) using the principal component linear discriminant analysis (PCA-LDA) with an accuracy of 100%. The adequacy of the PCA-LDA model was verified on a test sample. It was found that data of sensors with films of carbon nanotubes, zirconium nitrate, hydroxyapatite, methyl orange, bromocresol green, and Triton X-100 had the most significance for dividing samples into groups. The differences in the composition of the gaseous phase over the samples of nasal secretions for such a classification could be explained by the appearance or change in the concentrations of ketones, alcohols, organic carboxylic acids, aldehydes, amines, including cyclic amines or those with a branched hydrocarbon chain.

What have we learned from 7 years of equine rhinitis B virus qPCR testing in nasal secretions from horses with respiratory signs.

BACKGROUND: Equine rhinitis B virus (ERBV) has been given little attention by practitioners compared to other respiratory viruses, mainly because of the lack of diagnostic modalities and association with clinical disease. The objective of the study was to determine the frequency of detection of ERBV in nasal secretions from 6568 horses with acute onset of respiratory signs. METHODS: ERBV-positive qPCR results from nasal secretions submitted to a molecular diagnostic laboratory from 2013 to 2019 were reviewed. RESULTS: A total of 333 ERBV qPCR-positive samples (5.1%) were detected with increasing yearly frequency since the introduction of the assay in 2013. In comparison, only three of 356 (0.8%) healthy horses tested qPCR-positive for ERBV. Median age for ERBV qPCR-positive horses was 3 years of age, and fever, coughing and nasal discharge were the most common signs reported. Further, co-infections with other respiratory pathogens were reported in 73 (21.9%) of ERBV qPCR-positive samples. CONCLUSION: ERBV is a commonly detected respiratory virus from nasal secretions of young horses presenting with fever, nasal discharge and coughing.

Charcot-Leyden Crystal Protein in Nasal Secretions of Patients with Nonallergic Rhinitis with Eosinophilia Syndrome.

INTRODUCTION: Charcot-Leyden crystal (CLC) protein has been regarded as a hallmark of eosinophilic inflammation. OBJECTIVE: The purpose of this study was to investigate the role and levels of CLC protein in patients with nonallergic rhinitis with eosinophilia syndrome (NARES). METHODS: Overall, 39 NARES patients and 19 controls were recruited. The severity of nasal symptoms was measured by visual analogue scale and serum and local specific immunoglobulin E were determined in all patients. Nasal eosinophilia was assessed by semiquantitative analysis of eosinophils in nasal scrapings. Nasal secretion CLC protein concentrations were evaluated by ELISA. RESULTS: CLC protein concentrations were significantly higher in NARES patients than in controls (p < 0.0001). Nasal secretion CLC protein levels were significantly correlated with the degree of eosinophilia in nasal scrapings (rs = 0.331; p = 0.04) in NARES patients. Patients with high CLC protein concentrations displayed more severe nasal symptoms than patients with low CLC protein concentrations (p = 0.0080), particularly, nasal itching (p = 0.0029). Pilot study in 8 NARES patients demonstrated that treatment for 1 month with intranasal fluticasone propionate significantly decreased the nasal secretion CLC protein concentrations from baseline levels (p = 0.0335) and markedly attenuated the degree of swelling of inferior turbinate. CONCLUSIONS: CLC protein levels are significantly higher in nasal secretions of NARES patients and associated with the degree of nasal eosinophilia and the severity of nasal symptoms. Significantly, nasal secretion CLC protein levels obviously decreased after treatment with intranasal corticosteroids, suggesting its possible role in evaluating the medical treatment.

IgE antibody repertoire in nasal secretions of children and adults with seasonal allergic rhinitis: A molecular analysis.

BACKGROUND: There is growing interest both in testing IgE in nasal secretions (NS) and in molecular diagnosis of seasonal allergic rhinitis (SAR). Yet, the reliability of nasal IgE detection with the newest molecular assays has never been assessed in a large cohort of pollen allergic patients. OBJECTIVE: To investigate with microarray technology and compare the repertoires of specific IgE (sIgE) antibodies in NS and sera of a large population of children and adults with SAR. METHODS: Nasal secretions were collected with an absorbent device (Merocel 2000® , Medtronic) and a minimal dilution procedure from 90 children and 71 adults with SAR. Total IgE (tIgE) (ImmunoCAP, Thermo Fisher Scientific (TFS)) and sIgE antibodies against 112 allergen molecules (ISAC-112, TFS) were measured in NS and serum. RESULTS: Nasal sIgE was detectable in 68.3% of the patients. The detected nasal sIgE antibodies recognized airborne (88%), vegetable (10%), and animal food or other (<1%) allergen molecules. The prevalence and average levels of sIgE in NS and serum were highly interrelated at population level. A positive nasal sIgE antibody to a given molecule predicted the detection of the same antibody in the patient's serum with a specificity of 99.7% and a sensitivity of 40%. CONCLUSIONS: The concentration of sIgE is much lower in nasal secretions than in the serum. sIgE assays with very high analytical sensitivity and sampling methods with minimal dilution will be therefore needed to validate nasal secretions as alternative to serum in testing the sIgE repertoire.

Development of an effective sample transfer device for biomarker detection in nasal secretions.

Nasal secretions (NS) reflect inflammatory activity of the nasal mucosa and thus can be utilized for disease diagnosis and determining treatment effects in Allergic rhinitis (AR). However, non-standardized collection of samples can affect the measured concentration of inflammatory biomarker in NS. In this study, we aimed to develop and evaluate new devices capable of standardizing the collection, storage, and preprocessing methods of NS samples. First, we chose the best swab as polyester (PE) and selected a stimulation method, twirling for 10 s at 1 Hz, to efficiently release AR biomarkers from a PE swab. Storage of sample solutions at -20 °C was optimal for the stability of biomarkers for the detection of AR. The new swab sample transfer device showed excellent concentration recovery efficiency (90-100%) for tryptase (Trp) and eosinophil cationic protein (ECP) without crosstalk between the two biomarkers. Finally, we compared the concentration of Trp in human NS samples of AR patients (n = 6) pre-processed by the new device with that by centrifuge as a standard method. As a result, the concentrations of Trp in NS were very similar in both groups. Therefore, this device can be utilized as an effective sample transfer and pre-processing device for point-of-care testing of AR.

An ultrasensitive electrochemical detection of tryptase using 3D macroporous reduced graphene oxide nanocomposites by one-pot electrochemical synthesis.

In this study, we introduce an ultrasensitive and stable sensing platform based on gold nanoparticle decorated 3D reduced graphene oxide - carbon nanotube nanocomposites (GNP/CNT/rGO) for detection of tryptase (Tryp), which is a potential biomarker within nasal secretions for allergic rhinitis (AR). The GNP/CNT/rGO on the glassy carbon electrode (GCE) was fabricated through single-step one-pot electrochemical co-reduction and deposition. The GNP/CNT/rGO exhibited excellent electrocatalytic activity as the working electrode compared with bare GCE, GNP/rGO and CNT/rGO on GCEs. The enhanced performance of GNP/CNT/rGO may be attributed to 3D macroporous structures, which may provide increased surface area, faster charge transfer and lower mass transport resistance, as well as high electrocatalytic activity and good biocompatibility due to GNP. A GNP/CNT/rGO-based sandwich-type electrochemical immunosensor was fabricated for detection of Tryp, which requires high sensitivity and specificity. The GNP/CNT/rGO-based immunosensor exhibited a detection range from 100 pg/mL to 100 ng/mL, a detection limit of 50 pg/mL and a sensitivity of 1.64 µA/(ng/mL). This immunosensor possessed high selectivity, excellent reproducibility (RSD 2.1%) and high stability over approximately 1 month. Therefore, we expect that the GNP/CNT/rGO and the modified immunosensor will be useful tools for detecting Tryp in nasal secretions for clinical diagnosis of AR.

Dupilumab reduces local type 2 pro-inflammatory biomarkers in chronic rhinosinusitis with nasal polyposis.

BACKGROUND: Chronic rhinosinusitis with nasal polyposis (CRSwNP) is a type 2-mediated inflammatory disease associated with significant clinical, social, and economic burdens and high unmet therapeutic need. Dupilumab, a fully human monoclonal antibody targeting the interleukin-4 receptor α (IL-4Rα) subunit, demonstrated efficacy and acceptable safety in CRSwNP and other type 2 diseases (eg, atopic dermatitis and asthma). We now report the local effects of dupilumab on type 2 inflammatory biomarkers in nasal secretions and nasal polyp tissues of patients with CRSwNP in a randomized, placebo-controlled, phase 2 trial (NCT01920893). METHODS: Cytokines, chemokines, and total immunoglobulin E (IgE) levels were measured using immunoassay techniques in nasal secretions and nasal polyp tissue homogenates of CRSwNP patients receiving dupilumab 300 mg or placebo weekly for 16 weeks. RESULTS: With dupilumab, type 2 biomarker concentrations decreased in nasal secretions (least squares mean area under the curve from 0 to 16 weeks for the change from baseline) vs placebo for eotaxin-3 (-30.06 vs -0.86 pg/mL; P = 0.0008) and total IgE (-7.90 vs -1.86 IU/mL; P = 0.022). Dupilumab treatment also decreased type 2 biomarker levels in nasal polyp tissues at Week 16 vs baseline for eosinophilic cationic protein (P = 0.008), eotaxin-2 (P = 0.008), eotaxin-3 (P = 0.031), pulmonary and activation-regulated chemokine (P = 0.016), IgE (P = 0.023), and IL-13 (P = 0.031). CONCLUSIONS: Dupilumab treatment reduced multiple biomarkers of type 2 inflammation in nasal secretions and polyp tissues of patients with CRSwNP, demonstrating that antagonism of IL-4Rα signaling suppresses IL-4-/IL-13-dependent processes, such as mucosal IgE formation, as well as the expression of chemokines attracting inflammatory cells to the nasal mucosa.

Diagnostic value of Eos in nasal secretion of allergic rhinitis patients to predict serum slgE level / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the relationship between the distribution of eosinophils(Eos) of nasal secretion and serum specific IgE(sIgE) titer in patients with allergic rhinitis, and to evaluate the diagnostic value of Eos in nasal secretion for allergic rhinitis. METHODS A total of 60 allergic rhinitis patients from Department of Otolaryngology, Beijing Shunyi Hospital of Traditional Chinese Medicine during Jan.2016 to June 2017 were selected as treatment group, and another 60 cases of non-allergic rhinitis and 30 normal persons in the corresponding time period were chosen as control group. Eos in nasal secretions and serum sIgE were measured in all the subjects and the relationship between Eos distribution and serum sIgE level was analyzed. RESULTS The Eos distribution and serum level of sIgE was consistent(κ=0.264, P=0.000) and statistical significance was found. The area of ROC of the Eos in nasal secretion was 0.881, the standard deviation was 0.025 and 95％ confidence interval was 0.841 to 0.924. The Udden index of ROC reached the maximum when the Eos in nasal secretion smear was graded as level 3, the sensitivity was 88.5％ and specificity was 99.4％. CONCLUSION Eos cytological examination of nasal secretions has some auxiliary value in the diagnosis of allergic rhinitis. It is a cheap, simple and rapid method, which can be used as an effective reference index for diagnosing allergic rhinitis in primary hospital.