An Analysis of Neuropeptides at Nasal Contact Points of Patients With Secondary Headache.

OBJECTIVES: This prospective research study was designed to analyze the surgical outcomes and the intensity of substance P (SP), neurokinin A (NA), and calcitonin gene-related peptide (CGRP) in contact and noncontact nasal mucosa of patients with headache. METHODS: Twenty adults with secondary headache and correctible nasal obstruction were included in this study. The patients had nasal contact points between the nasal septum and the middle or inferior turbinates on nasal endoscopy and computed tomography scan. During surgical procedures, sample tissues were obtained from the nasal contact point and the noncontact area of the lateral nasal wall of these patients. Fluorescein staining intensity for antibodies against SP, NA, and CGRP was analyzed using image J software. Headaches were evaluated using a visual analog scale preoperatively and postoperatively. RESULTS: The differences between the preoperative and the postoperative 3rd month (P < 0.001) and 12th month (P < 0.001) visual analog scale scores were statistically significant. However, fluorescein staining intensity for SP (P = 0.631), NA (P = 0.546), and CGRP (P = 0.683) did not show statistically significant differences between the contact mucosa and the noncontact mucosa groups. CONCLUSIONS: Although in selected patients significant relief of headache can be obtained by surgery, there is no evidence from this study that SP, NA, and CGRP are responsible for the initiation of headache.

Nasal air-conditioning after partial turbinectomy: myths versus facts.

BACKGROUND: Turbinectomy, although a common procedure, is often accused of having a negative impact in all nasal functions. This study is the first in vivo study that evaluates objectively the effect of partial turbinectomy on nasal air-conditioning capacity. METHODS: In total, 57 patients with prior partial inferior turbinectomy and 28 healthy controls were examined. Intranasal temperature and humidity values were measured at the level of the head of inferior and middle turbinate. Nasal patency was evaluated by means of acoustic rhinometry. The clinical assessment was completed with nasal endoscopy and the Nasal Obstruction Symptom Evaluation questionnaire for subjective evaluation of nasal patency. RESULTS: Significant changes of temperature were found in both detection sites with 13% reduced heating capacity of the air at the level of the inferior and 19% at the level of the middle turbinate, respectively. No similar results were found for humidity measurements. No correlations were found between air-conditioning values and acoustic rhinometry results for both study groups. Nasal endoscopy revealed normal healing in all patients. No major complications were reported by the patients. Their subjective ratings of nasal obstruction were similar to healthy controls. CONCLUSION: Partial turbinectomy seems to have a negative impact on intranasal air heating but not to humidification. This effect has no impact on clinical condition and subjective perception of surgical outcome.

A novel device for delivery of intranasal particulate medication: a pilot study.

BACKGROUND: Intranasal medication delivery for allergic rhinitis (AR) is considered a mainstay of therapy but is hampered by poor compliance. Among reasons given are unpleasant sensations associated with spray penetration into the pharynx. Our objective was to study a novel method of particle delivery to the nose that would abrogate these issues. METHODS: This was a double-blind, randomized study. Subjects who met study criteria underwent intranasal particle delivery using a novel device (Trivair Nasal Deposition System; Trimel Pharmaceuticals, Toronto, Canada) that delivered anhydrous lactose particles into the nose via a transoral air puff (thus elevating soft palate and blocking the nasopharynx). Subjects had nostrils randomized into 4 groups (particle sizes 5 µm and 50 µm × doses 12.5 mg and 25 mg). Particle deposition was assessed at 1 minute, 10 minutes, and 30 minutes on the inferior turbinate, middle turbinate, and nasopharynx, respectively, using high-definition endoscopic photography. Each image was compared using an expert blinded 2-person panel for percentage particles remaining. Nonparametric data was assessed using the Wilcoxon signed-rank test via Strata software. RESULTS: Twelve nostrils in total met study criteria. The results showed no difference in effectiveness of nasal particle retention between the groups based on particle size or dose. No particles entered the nasopharynx or oropharynx. CONCLUSION: This study provides proof-of-principle data that the Trivair Nasal Deposition System is effective at retaining medication in the nose without pharyngeal penetration. Larger studies on this device are warranted.

Nasal gel and olfactory cleft.

OBJECTIVE: To evaluate whether a nasal gel, administrated using a radial-hole inhaler, reaches the olfactory cleft and if a different administration method influences distribution. MATERIAL AND METHOD: Sixteen healthy volunteers underwent a nasal endoscopy at 1 and 7minutes after the administration of a intranasal gel, with a different method in each fossa. RESULTS: No dye deposition was identified at the olfactory cleft, middle turbinate or middle meatus. In all cases the gel was identified at the nasal vestibule. On the right side, the second most frequent dye identification area was the inferior turbinate, with a rate of 87% at the first minute and 75% at 7 minutes. It was followed by the septum (75 and 62%) and the inferior meatus (6.2 and 12.5%). On the left side, the second most frequent stained area was the septum (18.7 and 13.5%), followed by the inferior meatus (6.5 and 65%). No inferior turbinate staining was found in the left side. There was a significant difference in the deposition rate at the septum (P<.01) and inferior turbinate (P<.001), when both administration methods were compared. CONCLUSIONS: No nasal gel, administrated using a radial-hole inhaler, was found at the olfactory cleft, middle turbinate or middle meatus. Gel distribution was located at the anterior and inferior portion of the nose, independent of the administration method used. Significantly different gel distribution rates were found at the septum and inferior turbinate when the 2 administration methods were compared.

Inmunohistochemical evaluation of estrogen receptors alpha and beta in normal inferior turbinate mucosa / Evaluación inmunohistoquímica de los receptores de estrógenos alfa y beta en la mucosa normal de concha nasal inferior

It has been postulated that the nasal mucosa, like other human tissues, is affected by a complex interactive network of neuropeptides, cytokines, allergic and inflammatory mediators and hormones such as estrogen, in which associations between symptoms (e.g. nasal stuffiness and coryza) and hormonal variations deriving from pregnancy, use of hormonal contraceptives and menstrual cycle phases are observed. The objective is evaluating the presence of specific estrogen receptors (types alpha and beta) in inferior turbinate mucosa in healthy subjects without nasal symptoms. Samples of nasal inferior turbinate were removed from patients undergoing aesthetic nasal surgery, and analyzed using hematoxylin-eosin staining, followed by immunohistochemical preparations on paraffin-embedded sections from the material sample, to detect estrogen receptors alpha and beta. Positive immunohistochemical reactions for both beta and alpha receptors were found in various regions of the inferior nasal turbinate. In conclusion both alpha and beta receptors were found, though the expression of beta was greater and more intense in the anterior portion of the inferior turbinate. No difference was found between male and female patients regarding the intensity of expression of receptors in the inferior turbinate.

Se ha postulado que la mucosa nasal, al igual que otros tejidos humanos, se ve afectada por una compleja red interactiva de neuropéptidos, citoquinas, mediadores alérgicos e inflamatorios, y hormonas como el estrógeno, en el que las asociaciones entre los síntomas (por ejemplo, congestión nasal y catarro) y hormonales las variaciones derivadas del embarazo, se observó el uso de anticonceptivos hormonales y las fases del ciclo menstrual. El objetivo es evaluar la presencia de receptores de estrógenos específicos (tipos de alfa y beta) en la mucosa de la concha nasal inferior en sujetos sanos sin síntomas nasales. Las muestras de la concha nasal inferior fueron retirados de los pacientes sometidos a cirugía nasal estética y analizados mediante hematoxilina-eosina, seguidos de cortes de preparados de inmunohistoquímica incluídos en parafina de la muestra de material, para detectar los receptores de estrógenos alfa y beta. Las reacciones de inmunohistoquímica fueron positiva para ambos receptores alfa y beta, éstas se encuentran en diversas regiones del cornete nasal inferior. En conclusión, tanto los receptores alfa y beta se encuentran, aunque la expresión de la beta fue mayor y más intensa en la porción anterior de la concha nasal inferior. No se encontraron diferencias entre pacientes hombres y mujeres en relación con la intensidad de la expresión de los receptores en el concha nasal inferior.

The presence of ovarian hormone receptors in the nasal mucosa and their relationship to nasal symptoms.

BACKGROUND: There is evidence in the literature showing a link between ovarian hormones and changes to nasal physiology. OBJECTIVES: The aim of this pilot study was to identify and quantify female hormone receptor positive cells in the nasal mucosa and to establish if there is a correlation with rhinitic symptoms. METHODS: Twenty-five adult patients attending a university hospital for routine, elective nonrhinological ENT procedures under general anaesthetic (mainly tonsillectomy) were recruited pre-operatively. Background information about each participant was recorded. Biopsies were taken from the inferior turbinates. These were analysed using immunohistochemistry techniques to assess for the presence of Progesterone, Oestrogen-alpha (ERalpha) and Oestrogen-beta (ERbeta) receptors. The mean number of cells positive for the receptors in each biopsy was deduced using a stratified random sampling technique. RESULTS: All nasal biopsies were negative for progesterone and ERalpha receptors. ERbeta receptors were present in the mucosal glands in 24 out of the 25 biopsies. Using unpaired t-tests to compare the sexes, smoking status and atopic history no statistical difference was shown between any of these groups (p > 0.05). However, the rhinitis quality of life questionnaire score and the mean number of ERbeta receptor positive cells per biopsy showed a positive correlation (Pearson correlation of 0.4, p < 0.05). CONCLUSIONS: The number of oestrogen receptor positive cells appears unaffected by sex, smoking history, hormone status, age or atopy. However, there is a significant positive relationship between the mean number of ERbeta positive cells and nasal symptoms. Pharmacological downregulation of ERbeta positive cells may reduce rhinitic symptoms and is the subject of further research.

Spontaneous leiomyosarcoma arising from the ethmoid turbinate of a rat.

Spontaneous leiomyosarcoma arising from the left ethmoid turbinate was observed microscopically in an 83-week-old male F344 rat. The tumor cells showed smooth-muscle differentiation with prominent nuclear pleomorphism and a small number of mitotic figures. The tumor cells were also immunohistochemically positive for smooth-muscle actin. The tumor protruded slightly into the nasal cavity and invaded the surrounding tissues. The present article is the first case of spontaneous leiomyosarcoma in the rat nasal cavity.

Analysis of particle deposition in the turbinate and olfactory regions using a human nasal computational fluid dynamics model.

The human nasal passages effectively filter particles from inhaled air. This prevents harmful pollutants from reaching susceptible pulmonary airways, but may leave the nasal mucosa vulnerable to potentially injurious effects from inhaled toxicants. This filtering property may also be strategically used for aerosolized nasal drug delivery. The nasal route has recently been considered as a means of delivering systemically acting drugs due to the large absorptive surface area available in close proximity to the nostrils. In this study, a computational fluid dynamics (CFD) model of nasal airflow was used with a particle transport and deposition code to predict localized deposition of inhaled particles in human nasal passages. The model geometry was formed from MRI scan tracings of the nasal passages of a healthy adult male. Spherical particles ranging in size from 5 to 50 microm were released from the nostrils. Particle trajectories and deposition sites were calculated in the presence of steady-state inspiratory airflow at volumetric flow rates of 7.5, 15, and 30 L/min. The nasal valve, turbinates, and olfactory region were defined in the CFD model so that particles depositing in these regions could be identified and correlated with their release positions on the nostril surfaces. When plotted against impaction parameter, deposition efficiencies in these regions exhibited maximum values of 53%, 20%, and 3%, respectively. Analysis of preferential deposition patterns and nostril release positions under natural breathing scenarios can be used to determine optimal particle size and flow rate combinations to selectively target drug particles to specific regions of the nose.

Expression and distribution of ion transport mRNAs in human nasal mucosa and nasal polyps.

CONCLUSIONS: Our results indicate that the electrogenic kidney Na(+)/HCO(3)(-) cotransporter (kNBC), KCl cotransporter (KCC1 and -4) and Ca(2+)-activated Cl(-) channel (CaCC1, -2, -3) mRNAs are expressed in normal nasal mucosa and nasal polyp, suggesting that altered expression of all CaCC mRNAs in nasal polyp may cause impaired electrolyte and water transport across the epithelial cells. OBJECTIVE: Electrolyte transport by nasal epithelia has been suggested to be important for controlling the quantity and composition of the nasal fluid and may play an important role in the development of nasal polyps. Transepithelial transport of ions and water in various fluid-transporting epithelia is strictly dependent on the localization of specific membrane proteins in the polarized epithelial cells. In this study we investigated the expression and distribution of mRNA transcripts for kNBC, pancreatic NBC, KCC1, -2, -3, -4 and CaCC1, -2, -3 gene families in human nasal mucosa and nasal polyp. MATERIAL AND METHODS: The expression and localization of these gene families were investigated in inferior turbinate tissues and nasal polyp using reverse transcriptase polymerase chain reaction (RT-PCR), semiquantitative RT-PCR and in situ hybridization. RESULTS: mRNAs for kNBC, KCC1 and -4 and all the CaCC families (CaCC1, -2 and -3) are expressed in human turbinate mucosa and nasal polyp. The expression levels of kNBC and KCC1 and -4 mRNAs did not differ between nasal mucosa and nasal polyp. However, the expression levels of all the CaCC genes were significantly decreased in nasal polyp. In situ hybridization revealed that the expression of these genes was mainly localized in the epithelial layer and submucosal glands of inferior turbinate mucosa and in the epithelial layer of nasal polyp.

Neuropeptide immunofluorescence in human nasal mucosa: assessment of the technique for vasoactive intestinal peptide (VIP).

UNLABELLED: Neuropeptides are important neurotransmitters in nasal physiology and the increasing knowledge of their role in nasal diseases brings new therapeutic perspectives. The investigation of human nasal mucosa neuropeptides is based mostly on immunocytochemistry, a complex approach whose resulting factors may be variable. Aiming to make this kind of research available, an immunofluorescence approach for vasoactive intestinal peptide (VIP) in human nasal mucosa is proposed and evaluated. STUDY DESIGN: Transversal cohort. MATERIAL AND METHOD: Human inferior turbinate samples were obtained at time of nasal surgery from eight patients. The samples were fixed in Zamboni solution (4% phosphate-buffered paraformaldehyde and 0.4% picric acid), snap-frozen and stored at -70 degrees C. 14 microm sections were then obtained. Immunofluorescence staining for VIP (Peninsula Laboratories) was performed and its images documented by conventional photography. The method's specificity, sensitivity and reproducibility of execution were evaluated. Additionally, the reproducibility of interpretation of results was evaluated through the comparison of staining scores (0 to 4) attributed to the images by six observers. RESULTS: The results showed the approach to be very specific and sensible, besides being reproducible in its execution. The interpretation of results may depend on the observer's accuracy in judging immunofluorescence images, but it showed uniformity. CONCLUSION: The proposed method was highly useful for research purposes in neuropeptides in human nasal mucosa.

Component analysis and growth process of nasopharyngeal calculus as revealed by Fourier transform infrared (FT-IR) spectroscopy.

A quite rare case of nasopharyngeal calculus in a woman in her twenties associated with the nasal discharge of pseudomonas infection was reported. As the substance was irregularly large in size, we extracted it partially by piecemeal resection using forceps and also by cracking technique using the holmium yttrium-aluminum-garnet (YAG) laser, under saline irrigation and stereotactic microscopic navigator (SMN) system under endoscopic observation. The substance was firmly fixed to the pharyngeal tonsil bed. The final extract was a small piece of singly folded bandage, which is probably the focal background for calculus formation. In a cross section of calculus specimen removed during surgery, Fourier transform infrared (FT-IR) analysis revealed that a) signal ratio of methylene group (organic substance) to amide I (protein) was 21.6% at the nasal cavity side, gradually decreased toward nasal mucous membrane showing approximate 50%, b) signal ratio of amide I to P04(3-) (inorganic substance) ranged between 17.7% and 26.7% at the different sites and inside the calculus, the protein content was approximate 1/5 of the inorganic substance, and c) signal ratio of the methylene group to amide I at the nasal cavity site showed that their contents were almost equal. The quantity of the organic substance was estimated at approximate 1/2 quantity of the protein at both the central part and the part contacted with the mucous membrane. From these results, it seems that throughout the course of calculus growth, both inorganic substance and protein remain almost constant inside the calculus, while organic substance is released from the internal part of the calculus being probably formed at an early stage.

Concentration, distribution and expression of interleukin-5 in human nasal polyp tissues.

OBJECTIVES: To study the concentration, distribution and expression of IL-5 in nasal polyp tissues and explore its significance in micro-environment differentiation of eosinophil accumulation. METHODS: The concentration and expression of IL-5 in nasal polyp tissues of 40 patients were determined by ELISA and immunohistochemistry and inferior turbinate mucosa from patients with nasal polyps and healthy volunteers were used as control. RESULTS: IL-5 concentration in polyp tissues was significantly higher than that in turbinate mucosa (P < 0.05). There was no significant difference in the turbinate mucosae between patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentrations in polyp tissues were markedly higher in patients with allergic rhinitis compared with those without (P < 0.05). IL-5 concentrations had no correlation with age and sex (P > 0.05). 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of lymphocytes and neutrophils were positive for IL-5; IL-5 was not detectable in epithelial cells. IL-5 expression in eosinophils of polyp tissues was remarkably stronger than that of the turbinate mucosa (P < 0.05); there was no significant difference in the the turbinate mucosae between patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissue was significantly stronger in patients with allergic rhinitis compared with those without (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and turbinate nasal mucosa (both P > 0.05). CONCLUSION: IL-5 is the key cytokine in eosinophilic pathologic mechanisms in nasal polyp tissues.

Levels of intracellular protein and messenger RNA of mucin and lysozyme in normal human nasal and polyp epithelium.

OBJECTIVES: Mucus hypersecretion is a characteristic feature in chronic sinusitis with nasal polyps. The objective of this study is to examine whether the polyp epithelium itself contributes to a certain extent to the increased mucous secretions in chronic sinusitis with nasal polyps, and if it does, to determine which mucin genes are responsible for the increased mucin secretion. METHODS: Three pooled samples of normal nasal epithelial cells from each subject were obtained by scrapings from the inferior turbinates of 30 healthy adult volunteers and nasal polyps from 6 patients who underwent intranasal ethmoidectomy and polypectomy. Isolated epithelial cells were used for total RNA isolation for reverse transcriptase polymerase chain reaction and cell lysates for immunoblotting. RESULTS: The intracellular level of mucin from polyp epithelium was 2.9 times higher than that of normal nasal epithelium (P < .05). Interestingly, MUC2 and MUC8 messenger RNA (mRNA) levels were clearly upregulated in polyp epithelium compared with those of normal turbinate epithelium. CONCLUSIONS: Polyp epithelium can be considered to contribute in part to increased secretion in chronic sinusitis with polyps, and increased mucous secretion might be related to the increased mRNA level of MUC2 or MUC8 or both.

[Significance and detection of interleukin-8 in human nasal polyps].

OBJECTIVE: To investigate the distribution and content of immunoreaction interleukin-8(IL-8) antigen in the human nasal polyps. METHODS: Twenty-two cases of nasal polyp and 10 cases of normal inferior turbinate were studied with immunohistochemical SABC method and ELISA method. RESULTS: Immunohistochemical results demonstrated that IL-8 antigen staining occurred predominantly within inflammatory and epithelium cell plasm. The level of IL-8 in nasal polyp tissue was higher than in normal control specimens. The value in nasal polyp tissue was in 322.06-2091.41 pg/ml, but in normal with a range of 29.31-332.19 pg/ml. The results demonstrated a remarkable difference between the two groups (P < 0.001). CONCLUSION: These data demonstrated the presence of IL-8 antigen in nasal polyps. Thus IL-8 likely plays a significant role in the pathogenesis of this disease process. There is an important significance of finding a potential treatment method.

Increased expression of IL-4, IL-5, IFN-gamma, IL-6, IL-8, and TGF-beta mRNAs in maxillary mucosa of patients with chronic sinusitis.

This study aimed to investigate expression of various cytokine mRNAs, including IL-6, IL-8, TGF-beta, IL-4, IL-5, and IFN-gamma in maxillary sinus mucosa of patients with chronic sinusitis. Maxillary sinus mucosae of six patients with chronic sinusitis and turbinate mucosae of six healthy subjects were obtained. We performed RT-PCR and Southern blot to examine gene expression of the cytokines IL-6, IL-8, TGF-beta, IL-4, IL-5, and IFN-gamma in maxillary sinus mucosa and compared the results with cytokine gene expressions in normal turbinate mucosa. IL-6, IL-8, TGF-beta, IL-4, IL-5, and IFN-gamma mRNAs were expressed more frequently in maxillary sinus mucosa from patients with chronic sinusitis than in normal turbinate mucosa. All the maxillary sinus mucosa specimens revealed relatively higher mean density ratio for each cytokine investigated than did normal turbinate mucosa. IL-6, IL-8, TGF-beta, IL-4, IL-5, and IFN-gamma mRNAs were expressed simultaneously in maxillary sinus mucosa of chronic sinusitis. These cytokines may be responsible for recruitment of inflammatory cells and for mucosal thickening in chronic sinusitis, and thus chronicity of the disease.

Transforming growth factor-alpha and rhinitis.

OBJECTIVES: Transforming growth factor-alpha (TGF-alpha) has been implicated in diverse physiologic and pathophysiologic functions including immunological, inflammatory, and neoplastic processes. TGF-alpha has been localized in the hyperproliferative, inflammatory environment of chronic otitis media, cholesteatoma, and asthmatic airways. TGF-beta1, which must be present with TGF-alpha to transform fibroblasts, has been found in rhinitic mucosa and in asthma in prior studies. The authors sought to identify whether TGF-alpha also played a role in the inflammatory cascade and fibrosis of rhinitis. STUDY DESIGN: A nonrandomized, prospective study was carried out in which samples of inferior turbinate and nasal polyps from rhinitic and nonrhinitic patients were subjected to immunohistochemistry and Western blotting to determine the presence of TGF-alpha. METHODS: Twenty-seven subjects undergoing surgery for rhinitis, obstructive sleep apnea, nasal fracture, and rhinoplasty were recruited for this study, the latter three groups acting as controls. Immunohistochemical and Western blotting techniques were employed to identify the presence of TGF-alpha in inferior-turbinate and nasal-polyp samples of rhinitic subjects. RESULTS: Immunohistochemistry demonstrated the selective staining of TGF-alpha in the basement membrane and extracellular matrix, including lymphatic, vascular, and glandular structures, in most turbinate samples and the absence of staining in corresponding controls. Further, TGF-alpha was isolated to a discrete 30-kD band in both inferior turbinate and polyp tissues by Western blotting without staining in the corresponding controls. CONCLUSIONS: These results suggest that TGF-alpha may play a role in the inflammatory derangement of rhinitis.

Developmental analysis of the peripheral olfactory organ of the opossum Monodelphis domestica.

The gray, short-tailed opossum, Monodelphis domestica, is born in a very immature state after a brief (14-day) gestation. As a result, the species provides a unique opportunity to examine very early periods of mammalian development. The present study provides the first detailed morphometric analysis of the development of the olfactory mucosa and the nasal cavity in Monodelphis. The extent of the sensory mucosa increases dramatically across development, covering a growing nasal cavity and increasingly elaborate turbinates. Both nasal cavity convolution (a measure of turbinate complexity) and mucosal surface area show extensive growth between birth and adulthood. These measurements are greatest in the central portion of the mucosa (in the caudal portion of the nose) at all ages examined. A developmental BrdU study reveals a robust decrease in cellular proliferation with age; proliferation decreases to near adult-like patterns by postnatal day (P) 40. Results from these studies show that there is dramatic structural and cellular postnatal growth in the opossum peripheral olfactory organ.

Olfactory neuron-specific expression of NeuroD in mouse and human nasal mucosa.

Human olfactory neuroepithelium (OE) is situated within the olfactory cleft of the nasal cavity and has the characteristic property of continually regenerating neurons during the lifetime of the individual. This regenerative ability of OE provides a unique model for neuronal differentiation, but little is known about the structure and biology of human olfactory mucosa. Thus, to better understand neurogenesis in human OE, we studied the expression of olfactory marker protein (OMP), TrkB and NeuroD in human nasal biopsies and autopsy specimens and compared these data with those obtained from normal and regenerating mouse OE. We show that NeuroD and TrkB are coordinately expressed in human OE. Thus, by using these markers we have been able to extend the known boundaries of the human OE to include the inferior middle turbinate. In normal mouse OE, TrkB and OMP expression overlap in cells closest to the superficial layer, but TrkB is expressed more strongly in the lower region of this layer. In contrast, NeuroD expression is more basally restricted in a region just above the globose basal cells. These characteristic expression patterns of OMP, TrkB and NeuroD were also observed in the regenerating mouse OE induced by axotomy. These results support a role of NeuroD and brain-derived neurotrophic actor (BDNF), the preferred ligand for TrkB, in the maintenance of the olfactory neuroepithelium in humans and mice.

Detection of anionic antimicrobial peptides in ovine bronchoalveolar lavage fluid and respiratory epithelium.

Three small antimicrobial anionic peptides (AP) were originally isolated from an ovine pulmonary surfactant. However, their presence in bronchoalveolar lavage (BAL) fluid and tissues of the respiratory tract is unknown. In this study, we made affinity-purified rabbit polyclonal and mouse monoclonal antibodies to synthetic H-DDDDDDD-OH. Antibody specificity was assessed by a competitive enzyme-linked immunosorbent assay (ELISA), and the exact epitope binding sites were determined with analog peptides synthesized on derivatized cellulose. These antibodies were used to detect AP in BAL fluid by ELISA and in respiratory tissues by Western blot analysis and immunocytochemistry. BAL fluid from 25 sheep contained 0.83 +/- 0.33 mM AP (mean +/- standard deviation; range, 0.10 to 1.59 mM) and was antimicrobial. The presence of AP in BAL fluid was confirmed by reverse-phase high-pressure liquid chromatography fractionation followed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry on those fractions which were positive by competitive ELISA and demonstrated antimicrobial activity. In Western blots, polyclonal antibody PAB96-1 and monoclonal antibody 1G9-1C2 (5.0 micrograms/ml) detected four bands in solubilized turbinate and tracheal epithelial cells (53.7, 31.2, 28.0, and 25.7 kDa) and five bands in lung homogenates (53.5, 37.1, 31.2, 28.0, and 25.7 kDa). Only a single band was seen in solubilized liver and small-intestine homogenates, and no bands were seen in blots containing BAL fluid, albumin, or kidney or spleen homogenates. In pulmonary-tissue sections, both antibodies PAB96-1 and 1G9-1C2 identified accumulated protein in the apical cytoplasm of the bronchial and bronchiolar epithelia, in the cytoplasm of pulmonary endothelial cells, and in an occasional alveolar macrophage. As a first step in identifying a candidate AP precursor gene(s), degenerate oligonucleotides representing all possible coding combinations for H-GADDDDD-OH and H-DDDDDDD-OH were synthesized and used to probe Southern blots of sheep genomic DNA. Following low-stringency washes and a 2-day exposure, strongly hybridizing bands could be identified. One degenerate oligonucleotide, SH87, was used as a hybridization probe to screen a sheep phage genomic library. Two independent phage contained the H-GADDDDD-OH coding sequence as part of a larger predicted protein. AP may originate as part of an intracellular precursor protein, with multistep processing leading to the release of the heptapeptide into mucosal secretions. There it may interact with other innate pulmonary defenses to prevent microbial infection.

Immunohistochemical localisation of tear lipocalin in human nasal mucosa.

Lipocalins are low molecular weight soluble proteins, a sub-class of this family are the odorant binding proteins (OBPs) which are postulated as having an important role in the perireceptor events of olfactory transduction, though their specific physiological function has yet to be defined. From the nasal mucus of normal subjects we recently isolated a 19kDa protein, the amino-acid sequence of which, limited to the first 20 residues, is identical to that of tear lipocalin. In this study we performed an immuno-histochemical investigation on the nasal localisation of this protein, using tissue specimens taken from the inferior (27 samples), middle (7 samples) and superior turbinates (6 samples) in 27 patients undergoing septoplastic surgery. The protein was detected in the sub-epithelial tubulo-acinar glands of the nasal mucosa, particularly in the mucoserous glands, in 74% of the specimens taken from the inferior turbinate, in 71.4% of those from the middle turbinate and in 66.6% of the samples of superior turbinate tissue. The homogeneous distribution of this protein in the nasal mucus could imply that it functions as a general protection agent rather than as an odour carrier or transducer.