Characterization and Treatment of SARS-CoV-2 in Nasal and Bronchial Human Airway Epithelia.

In the current COVID-19 pandemic context, proposing and validating effective treatments represents a major challenge. However, the scarcity of biologically relevant pre-clinical models of SARS-CoV-2 infection imposes a significant barrier for scientific and medical progress, including the rapid transition of potentially effective treatments to the clinical setting. We use reconstituted human airway epithelia to isolate and then characterize the viral infection kinetics, tissue-level remodeling of the cellular ultrastructure, and transcriptional early immune signatures induced by SARS-CoV-2 in a physiologically relevant model. Our results emphasize distinctive transcriptional immune signatures between nasal and bronchial HAE, both in terms of kinetics and intensity, hence suggesting putative intrinsic differences in the early response to SARS-CoV-2 infection. Most important, we provide evidence in human-derived tissues on the antiviral efficacy of remdesivir monotherapy and explore the potential of the remdesivir-diltiazem combination as an option worthy of further investigation to respond to the still-unmet COVID-19 medical need.

The Nasal Gland in Turkeys (Meleagris gallopavo): Anatomy, Histology, and Ultrastructure.

This study provides a detailed description of the major morphoanatomic and ultrastructural features of the nasal gland in turkeys. In this avian species, nasal or salt glands are bilateral, pale pink, elongated to spindle-shaped, serous, tubuloalveolar structures, with a mean length ranging from 0.64 ± 0.15 cm in poults of 4 days of age to 2.15 ± 0.17 cm at 22 weeks. Instead of having a supraorbital location as commonly seen in waterfowl and other avian species, these glands run underneath the lacrimal, frontal, and nasal bones in turkeys. The reference point for sample collection for histologic examination is just before the rostral edge of the eyelid. Each gland adheres to the surrounding bone through a thick capsule of dense connective tissue merging with the skull periosteum. Histologically, the salt gland consists of secretory tubuloalveolar structures, lined by cuboidal epithelial cells with a central canaliculus and ducts. There are small and large ducts lined by a bilayered epithelium consisting of large apical columnar secretory cells occasionally admixed with rare cuboidal cells. These cells are periodic acid Schiff negative and slightly Alcian blue positive. Both alveolar and secretory ductal cells contain slightly electrondense granular vesicles, highly folded lateral surfaces, and large numbers of mitochondria, characteristic of ion-transporting epithelia. This study provides valuable information for the accurate identification and localization of the nasal gland during necropsy, as well as its correct histologic interpretation, ultimately improving our understanding of the role of this gland in the pathophysiology of specific diseases in turkeys.

La glándula nasal en pavos (Meleagris gallopavo): Anatomía, histología y ultraestructura Este estudio proporciona una descripción detallada de las principales características morfo-anatómicas y ultraestructurales de la glándula nasal en pavos. En esta especie aviar, las glándulas nasales o glándulas salinas son estructuras bilaterales, tubuloalveolares de color rosa pálido, alargadas y serosas, con una longitud media que varía de 0.64 ± 0.15 centímetros en los pavipollos de 4 días de edad hasta 2.15 ± 0.17 centímetros en aves a las 22 semanas. En lugar de tener una ubicación supraorbital como se ve comúnmente en las aves acuáticas y otras especies de aves, estas glándulas corren por debajo de los huesos lagrimales, frontales y nasales en los pavos. El punto de referencia para la recolección de muestras para el examen histológico es justo antes del borde rostral del párpado. Cada glándula se adhiere al hueso circundante a través de una gruesa cápsula de tejido conectivo denso que se fusiona con el periostio del cráneo. Histológicamente, la glándula salina consiste en estructuras tubulo-alveolares secretoras, revestidas por células epiteliales cuboidales con un canalículo central y conductos. Hay conductos pequeños y grandes revestidos por un epitelio de dos capas que consiste en grandes células secretoras columnares apicales ocasionalmente mezcladas con escasas células cuboidales. Estas células son ácido periódico de Schiff negativas y ligeramente positivas para el azul de alcián. Las células ductales alveolares y secretoras contienen vesículas granulares ligeramente electrondensas, superficies laterales altamente plegadas y grandes cantidades de mitocondrias, características de los epitelios transportadores de iones. Este estudio proporciona información valiosa para la identificación y localización exacta de la glándula nasal durante la necropsia, así como su correcta interpretación histológica, mejorando en última instancia nuestra comprensión del papel de esta glándula en la fisiopatología de enfermedades específicas en pavos.

Microvascular anatomy of olfactory and accessory olfactory (vomeronasal) organs in adult Xenopus laevis: scanning electron microscopy of vascular corrosion casts / Anatomía microvascular de órganos olfatorios y accesorios (vomeronasal) en adultos Xenopus laevis: microscopía electrónica de barrido de moldes de corrosión vascular

Microvascular anatomy and histomorphology of olfactory and vomeronasal organs in adult Xenopus laevis Daudin were studied by scanning electron microscopy of vascular corrosion casts and paraplast embedded stained serial tissue sections. Results show that the arterial supply is bilaterally by terminal arterioles of the medial branch of the nasal artery and by the palatal artery. Arterioles give rise to a capillary meshwork characteristic for respiratory surfaces in principal chambers and in dorsal and caudal areas of middle chambers. Anterior and inferior areas of the middle chambers own a distinctly different capillary network with conspicuous short capillary loops. Loops have a dilated tip and extend in acute angles towards the chamber lumen. The vomeronasal organ (VNO) locates beneath the olfactory organ. It has a medial to lateral extension and attaches with its caudal circumference to the medial nasal glands. Its capillary bed displays rectangular meshes which preferentially orientate along the long axis of the VNO. Locally, capillaries form short hairpin-like or strongly twisted loops with dilated tips which point towards the lumen of the VNO. These capillaries slow-down blood velocity and may lead to an increased exchange of oxygen, nutrients and water-borne odorants in the middle chambers and of pheromones in the VNO. In the latter vascular structures are present which might serve as a vascular pump.

Se estudiaron la anatomía microvascular e histomorfología de los órganos olfatorios y vomeronasales de Xenopus laevis Daudin adultos, mediante microscopía electrónica de barrido de moldes de corrosión vascular y secciones de tejido seriadas, teñidas e incluídas en paraplast. Los resultados muestran que el suministro arterial es bilateral por arteriolas terminales de la rama medial de la arteria nasal y por la arteria palatina. Las arteriolas dan lugar a un lecho capilar característico de las superficies respiratorias en las cámaras principales y en las áreas dorsal y caudal de las cámaras intermedias. Las áreas anterior e inferior de las cámaras centrales poseen una red capilar significativamente diferente con llamativos bucles capilares cortos. Los bucles tienen una punta dilatada y se extienden en ángulos agudos hacia la luz de la cámara. El órgano vomeronasal (VNO) se ubica debajo del órgano olfatorio. Se extiende de medial a lateral y se une con su circunferencia caudal a las glándulas nasales mediales. El lecho capilar muestra mallas rectangulares que se orientan preferentemente a lo largo del eje longitudinal del VNO. Localmente, los capilares forman bucles cortos en forma de horquilla o fuertemente retorcidos con puntas dilatadas que apuntan hacia la luz del VNO. Estos capilares ralentizan la velocidad de la sangre y pueden conducir a un mayor intercambio de oxígeno, nutrientes y odorizantes, a base de agua en las cámaras intermedias y de feromonas, en el VNO. En este último, están presentes estructuras vasculares que podrían servir como una bomba vascular.

Epithelial crypts: A complex and enigmatic olfactory organ in African and South American lungfish (Lepidosireniformes, Dipnoi).

African lungfish (Protopterus) seem unique among osteognathostomes in possessing a potential vomeronasal organ homolog in form of accessory epithelial crypts within their nasal cavity. Many details regarding structural and functional properties of these crypts are still unexplored. In this study, we reinvestigate the issue and also present the first data on epithelial crypts in the South American lungfish Lepidosiren paradoxa. The nasal cavities of L. paradoxa and Protopterus annectens were studied using histology, scanning electron microscopy, and alcian blue and PAS staining. In both species, the epithelial crypts consist of a pseudostratified sensory epithelium and a monolayer of elongated glandular cells, in accordance with previously published data on Protopterus. In addition, we found a new second and anatomically distinct type of mucous cell within the duct leading into the crypt. These glandular duct cells are PAS positive, whereas the elongated glandular cells are stainable with alcian blue, suggesting distinct functions of their respective secretions. Furthermore, the two lungfish species show differently structured crypt sensory epithelia and external crypt morphology, with conspicuous bilaterally symmetrical stripes of ciliated cells in L. paradoxa. Taken together, our data suggest that stimulus transport into the crypts involves both ciliary movement and odorant binding mucus.

Ultrastructural characterization of normal Merkel cells in the dog.

BACKGROUND: Involvement of Merkel cells (MKs) in different cutaneous diseases as well as in the growth, differentiation and homeostasis of the skin has been previously documented. HYPOTHESIS/OBJECTIVES: The aim was to assess the ultrastructural features of MKs in canine skin, including morphometrics, highlighting their similarities with and differences from those described for other mammals. ANIMALS: Hard palate, nasal planum, lower lip and whisker pad samples were taken from two healthy young dogs destined for academic purposes. METHODS: Ultrathin sections of samples fixed in osmium tetroxide and embedded in Epon 812 resin were stained with uranyl acetate and lead citrate and examined using a JEOL JEM 2010 transmission electron microscope. RESULTS: Ultrastructural characteristics included the following: (i) arrangement in clusters in the basal layer of the epidermis, oral mucosa and external follicular root sheath; (ii) inconstant link with nerve terminal; (iii) oval (10.27 ± 1.64 µm major axis) cell shape with large lobulated nuclei (5.98 ± 1.16 µm major axis); (iv) spine-like and thick cytoplasmic processes interdigitating with surrounding keratinocytes; (v) presence of desmosomes in the cell body or at the base of spine-like processes attaching to neighbouring keratinocytes; and (vi) cytoplasm containing loosely arranged intermediate filaments (10.04 ± 1.17 nm) and numerous dense-core granules (100.1 ± 17.12 nm) arranged in the basal portion of the cytoplasm. CONCLUSIONS AND CLINICAL IMPORTANCE: This study provides the first complete description of the ultrastructural characteristics of MKs in the dog, enhancing our knowledge of the skin structure in this species and providing a basis for future physiological and pathological studies of the role of these cells in normal and damaged canine tissues.

The sense of touch in the star-nosed mole: from mechanoreceptors to the brain.

Star-nosed moles are somatosensory specialists that explore their environment with 22 appendages that ring their nostrils. The appendages are covered with sensory domes called Eimer's organs. Each organ is associated with a Merkel cell-neurite complex, a lamellated corpuscle, and a series of 5-10 free nerve endings that form a circle of terminal swellings. Anatomy and electrophysiological recordings suggest that Eimer's organs detect small shapes and textures. There are parallels between the organization of the mole's somatosensory system and visual systems of other mammals. The centre of the star is a tactile fovea used for detailed exploration of objects and prey items. The tactile fovea is over-represented in the neocortex, and this is evident in the modular, anatomically visible representation of the star. Multiple maps of the star are visible in flattened cortical preparations processed for cytochrome oxidase or NADPH-diaphorase. Star-nosed moles are the fastest known foragers among mammals, able to identify and consume a small prey item in 120 ms. Together these behavioural and nervous system specializations have made star-nosed moles an intriguing model system for examining general and specialized aspects of mammalian touch.

Morphometric and ultrastructural comparison of the olfactory system in elasmobranchs: the significance of structure-function relationships based on phylogeny and ecology.

This study investigated the relationship between olfactory morphology, habitat occupancy, and lifestyle in 21 elasmobranch species in a phylogenetic context. Four measures of olfactory capability, that is, the number of olfactory lamellae, the surface area of the olfactory epithelium, the mass of the olfactory bulb, and the mass of the olfactory rosette were compared between individual species and groups, comprised of species with similar habitat and/or lifestyle. Statistical analyses using generalized least squares phylogenetic regression revealed that bentho-pelagic sharks and rays possess significantly more olfactory lamellae and larger sensory epithelial surface areas than benthic species. There was no significant correlation between either olfactory bulb or rosette mass and habitat type. There was also no significant difference between the number of lamellae or the size of the sensory surface area in groups comprised of species with similar diets, that is, groups preying predominantly on crustaceans, cephalopods, echinoderms, polychaetes, molluscs, or teleosts. However, some groups had significantly larger olfactory bulb or rosette masses than others. There was little evidence to support a correlation between phylogeny and morphology, indicating that differences in olfactory capabilities are the result of functional rather than phylogenetic adaptations. All olfactory epithelia exhibited microvilli and cilia, with microvilli in both nonsensory and sensory areas, and cilia only in sensory areas. Cilia over the sensory epithelia originated from supporting cells. In contrast to teleosts, which possess ciliated and microvillous olfactory receptor types, no ciliated olfactory receptor cells were observed. This is the first comprehensive study comparing olfactory morphology to several aspects of elasmobranch ecology in a phylogenetic context.

Three-dimensional fine structural reconstruction of the nose sensory structures of Acrobeles complexus compared to Caenorhabditis elegans (Nematoda: Rhabditida).

Nematode sensory structures can be divided into two classes; cuticular sensillae, with dendrites ending outside the epidermis, and internal receptors, that typically are single dendrites terminating within the body cavity. Fine structure of the former has been described completely in more than a dozen nematode taxa, while the latter were previously only well understood in the microbial feeder Caenorhabditis elegans. The distantly related nematode Acrobeles complexus has a similar ecology and together the two span a clade representing a large proportion of nematode biodiversity. The cuticular sensillae and internal receptors of A. complexus are here shown to be remarkably similar in number, arrangement, and morphology to those of C. elegans. Several key differences are reported that likely relate to function, and suggest that this nematode has a cuticular sensillum morphology that is closer to that of the common ancestor of the two taxa. Internal sensory receptors have more elaborate termini than those of C. elegans. The existence of a novel form of mechanoreceptor in A. complexus and spatial relationships between sensillum dendrites suggest differences between two classes of sensillae in how a touch-response behavior may be mediated.

Rinometria acústica: correlacão anatômica dos dois primeiros entalhes encontrados no rinograma / Acoustic rhinometry: anatomic correlation of the first two notches found in the nasal echogram

O gráfico obtido pela rinometria acústica em indivíduos adultos, caucasianos, sem alteracões nasais, mostra, com clareza, dois entalhes no início do rinograma; porém, na literatura existe controvérsia sobre sua correlacão anatômica. OBJETIVO: Realizamos este estudo com o objetivo de obtermos dados que contribuam para a correlacão anatômica destes dois entalhes. FORMA DE ESTUDO: Clínico prospectivo. CASUíSTICA E MÉTODO: Foram analisados os rinogramas de 35 indivíduos em condicão basal, após o uso de vasoconstritor tópico nasal e após obliteracão da válvula nasal com algodão embebido em vaselina. RESULTADOS: Identificou-se diminuicão e aumento, estatisticamente significante, da área de seccão transversal apenas do segundo entalhe após obliteracão da válvula nasal e após o uso de vasoconstritor tópico nasal, respectivamente. CONCLUSAO: A análise dos resultados sugeriu que o primeiro entalhe do rinograma se refere à narina e o segundo à válvula nasal como um todo.

[Study of mucociliary transport and nasal ciliary ultrastructure in patients with Kartagener's syndrome]. / Estudio del transporte mucociliar y de la ultraestructura ciliar nasales en pacientes con síndrome de Kartagener.

OBJECTIVE: Kartagener's syndrome (KS) is a clinical variant of primary ciliary dyskinesia involving situs inversus associated with chronic airway infections. The ciliary defect associated with this syndrome is the absence of dynein arms. The aim of this study was to evaluate mucociliary transport and ciliary ultrastructure in 14 patients with KS. PATIENTS AND METHODS: We studied nasal mucociliary transport using a radioisotopic technique and ciliary ultrastructure in 14 patients with KS. RESULTS: Thirteen patients had mucociliary stasis and 1 had severely slowed transport (1.3 mm/min). Four patients (29%) had cilia with normal dynein arms, 2 patients (14%) had short inner dynein arms, and 8 patients (57.1%) had total absence of inner and outer dynein arms. CONCLUSIONS: We conclude that the typical clinical presentation, together with altered mucociliary transport as identified by an isotopic technique, is diagnostic of KS, even when the ciliary ultrastructure is normal. KS is clinically homogenous and morphologically heterogenous.

Estudio del transporte mucociliar y de la ultraestructura ciliar nasales en pacientes con síndrome de Kartagener / Study of mucociliary transport and nasal ciliary ultrastructure in patients with Kartagener's syndrome

OBJETIVO: El síndrome de Kartagener (SK) es una variante clínica de la discinesia ciliar primaria que asocia a las infecciones crónicas de las vías respiratorias un situs inversus. La ausencia de brazos de dineína ha sido el defecto ciliar asociado a este síndrome. El objeto de este trabajo es el estudio del transporte mucociliar y de la ultraestructura ciliar en 14 pacientes con SK. PACIENTES Y MÉTODOS: Hemos estudiado el transporte mucociliar nasal, mediante una técnica radioisotópica, y la ultraestructura ciliar en 14 pacientes con SK. RESULTADOS: En 13 pacientes había estasis mucociliar y en uno, un transporte muy enlentecido (1,3 mm/min). Mostraban cilios con brazos de dineína normales 4 pacientes (29%); brazos internos de dineína cortos, 2 pacientes (14%), y ausencia completa de brazos internos y externos de dineína, 8 casos (57,1%). CONCLUSIONES: Concluimos que la presentación clínica típica junto con un transporte mucociliar alterado, objetivado con una técnica isotópica, es diagnóstica del SK, aunque la ultraestructura ciliar sea normal. El SK es clínicamente homogéneo y morfológicamente heterogéneo

OBJECTIVE: Kartagener’s syndrome (KS) is a clinical variant of primary ciliary dyskinesia involving situs inversus associated with chronic airway infections. The ciliary defect associated with this syndrome is the absence of dynein arms. The aim of this study was to evaluate mucociliary transport and ciliary ultrastructure in 14 patients with KS. PATIENTS AND METHODS: We studied nasal mucociliary transport using a radioisotopic technique and ciliary ultrastructure in 14 patients with KS. RESULTS: Thirteen patients had mucociliary stasis and 1 had severely slowed transport (1.3 mm/min). Four patients (29%) had cilia with normal dynein arms, 2 patients (14%) had short inner dynein arms, and 8 patients (57.1%) had total absence of inner and outer dynein arms. CONCLUSIONS: We conclude that the typical clinical presentation, together with altered mucociliary transport as identified by an isotopic technique, is diagnostic of KS, even when the ciliary ultrastructure is normal. KS is clinically homogenous and morphologically heterogenous

Development of the basement membrane and formation of collagen fibrils below the placodes in the head of anuran larvae.

The development of the basement membrane and collagen fibrils below placodes, including the corneal region of the ectoderm, lens epithelium, nasal plate, and auditory vesicle in anuran larvae was observed by transmission electron microscopy and compared with that in nonplacodal regions such as the epidermis, neural tube, and optic vesicle. In the corneal region the lamina densa becomes thick concomitantly with the development of the connecting apparatuses such as hemidesmosomes and anchoring fibrils. The collagen fibrils increase in number and form a multilayered structure, showing similar morphology to the connective tissues below the epidermis. These two areas, i.e., the corneal region and epidermis, possess much collagenous connective tissue below them. On the other hand, the neural tube and ophthalmic vesicle that originated from the neural tube each have a thin lamina densa and a small number of underlying collagen fibrils. The lamina densa does not thicken and the number of collagen fibrils do not significantly increase during development. These two areas possess little extracellular matrix. The nasal plate and auditory vesicle show intermediate characteristics between the epidermis-type and the neural tube-type areas. In these areas, the lamina densa becomes thick and hemidesmosomes and anchoring fibrils develop. The number of collagen fibrils increases during development, but does not show an orderly arrangement; rather, they are randomly distributed. It is thought that the difference in the arrangement of collagen fibrils in different tissues is due to differences in the extracellular matrix around the collagen fibrils. Placodal epithelia have the same origin as epidermis, but during development their morphological characteristics differ and they are not associated with the pattern of extracellular matrix with characteristics of epidermal and corneal multilayered collagen fibril areas.

The embryonic development of sensory organs and the skull in the trisomy 16 mouse, an animal model for Down's syndrome.

The trisomy 16 mouse is a widely accepted animal model for the study of the embryonic development of human trisomy 21. While the development of the brain and heart has been thoroughly studied, there are hardly any data on the development of sensory organs like the eye, nose and ear. By studying scanning electron microscopic pictures and semithin sections from the tenth to the 15th day of development, we found delayed development of the nose, and, in particular, of the vomer. Sensory structures of the otic vesicle also showed a marked developmental delay. Pigmentation of the outer layer of the otic cup starts later in trisomic animals. Cleared specimens on day 16 showed retarded development of ossification centres in all areas of the skull. These findings correspond with the abnormal facial morphology found in Down's syndrome and may also provide new insights into the hearing impairment commonly found. The observations in the eye and skull bones indicate that neural crest tissue maldevelopment is not the sole cause of malformations.

A comparative study of perichondrial tissue in mammalian cartilages.

The general organization, cellular and extracellular components, and structural variation of perichondrium have been studied in different mammalian cartilages by polarized light and transmission electron microscopy. The overall structure is that of a dense connective tissue composed of variable numbers of thin, stratified, closely-packed lamellae, themselves composed of closely-matted collagen fibres running in the plane of the cartilage surface, but oriented at various angles to each other. Variations mainly concern the arrangement of the fibre bundles in the transition zones between perichondrial and cartilage matrices, and between perichondrium and surrounding tissues. Perichondrial cells have the characteristics of fibrocytes. A cambial layer of undifferentiated stem cells was never observed. A layer of 'perichondrial lining cells' with distinctive ultrastructural characteristics was observed in some cartilage units, which separates the perichondrium from the surrounding loose connective tissue. The ultrastructural results demonstrate that the cartilage and perichondrial extracellular matrices are distinct, and what have been designated perichondrial 'transition' and 'proliferative' zones are in fact parts of the most superficial cartilage layer. Variations in perichondrial structure appear to correlate with diversity of cartilage function and we conclude that each cartilage unit plus perichondrium forms a tightly-integrated entity, best regarded as a unitary organ within the skeletal system.

Comparative morphology and cytology of the olfactory organs in Moray eels with remarks on their foraging behavior.

BACKGROUND: This study compares the morphology and cytology of olfactory organs in moray eels (Muraenidae), particularly Siderea grisea and some species of the genera Echidna, Gymnothorax, and Lycodontis, fishes that are top predators in shallow-water marine habitats. Some of the species search visually for food while others search by olfaction. METHODS: The morays were collected in the Red Sea; the nasal olfactory organs were dissected and fixed in Bouin's solution for light-microscopy, and 3.5% glutaraldehyde for electron-microscopy studies. RESULTS: In each studied species the olfactory rosettes are elongated structures situated in closed olfactory chambers between anterior tubular inlet nares and slit-form posterior outlet openings. The double row of lamellae constituting these rosettes are round in Siderea and Echidna and elongated in the other species. They are attached at their base to a median raphe and range in number from 20 in the youngest observed Siderea to 168 in Gymnothorax of 1,500 mm total length. As in other teleosts, the lamellae are covered by a ciliated epithelium composed of three types of sensory cells: two of these, ciliated sensory neurons and ciliated supporting cells, differ in detail, length, and thickness of their cilia and intracellular rootlet system; the third type of sensory cells bears microvillae as well as cilia. Proximal, axonal extensions of the ciliated cells cross the basal lamina in bundles and combine to form fila olfactoria from which the two olfactory nerves extend towards the olfactory bulbs. Lateral extensions at the basal parts of these ciliated cells, the so-called spines, cross the membranes of neighboring cells as dendrites, possibly changing part of all of the ciliated epithelium into an olfactory field. The density and number of sensory cells on the lamellae, as well as observed differences in their foraging behavior in nature and captivity, enable the morays to be divided into two groups: one group, in which the lamellae are richly covered with stereocilia, includes species of the genera Siderea and Echidna, that search for food by olfaction, and the second group, which has a great deal less cells with stereocilia and includes the studies species of Gymnothorax and Lycodontis, locates its food visually.

Evaluation of the human cell line RPMI 2650 as an in vitro nasal model.

We have investigated a human nasal cell line, RPMI 2650 (ATCC), as an in vitro model to assess the absorption and tolerability of nasally administrated peptides. This cell line was cultured on different filters (i.e. Nunc, Costar, Falcon, Millipore, and Becton) which were uncoated or coated with collagen types I or IV, laminin, fibronectin and extracellular matrix. Cell line morphology, capability of forming a cell layer and phenotype were analyzed by light-, fluorescence- and electron microscopy. The morphological analysis showed that RPMI 2650 cells were forming cell clusters on one group of filters (i.e., Nunc, Costar, Falcon and Becton filters coated with collagen types I or IV, laminin and extracellular matrix). On the second group of filters (i.e., Becton and Millipore filters, coated with fibronectin) the cells had the tendency to spread in a cell layer. In both groups of filters, the cells never showed cell polarization, nor microvilli and tight junctions. Phenotyping of this cell line was performed by indirect immunofluorescence using monoclonal antibodies against human cytokeratins 10, 17 and 18 (markers of epithelial cells), desmin (markers of mesothelial cells) and vimentin (marker of mesothelial cells). Vimentin was strongly expressed, cytokeratins 10, 17 and 18 were weakly expressed, and desmin was not expressed. The human nasal cell line RPMI 2650 was not able to form a tight cell layer under these cell culture conditions. The limits of this cell line as an in vitro nasal model for drug absorption is discussed.

Differential expression of vomeromodulin and odorant-binding protein, putative pheromone and odorant transporters, in the developing rat nasal chemosensory mucosae.

Expression of the putative pheromone and odorant transporter, vomeromodulin, was characterized in developing rat nasal mucosae using in situ hybridization and immunocytochemistry. Initial expression of vomeromodulin mRNA and protein was detected at embryonic day (E)16 in the maxillary sinus component of the lateral nasal glands. The abundance of mRNA and protein in the lateral nasal glands increased with age and reached a peak at postnatal day (P)27. Also at P27, vomeromodulin mRNA and protein expression was initiated in vomeronasal glands and posterior glands of the nasal septum. Comparison of the developmental expression of odorant-binding protein, another carrier protein synthesized in the lateral nasal glands, with that of vomeromodulin demonstrated major differences. In contrast to vomeromodulin, odorant-binding protein was not detected until postnatal day 2 in the ventral component of the lateral nasal glands and anterior glands of the nasal septum. These results suggest that the expression of vomeromodulin and odorant-binding protein is developmentally and differentially regulated and confirms the suggestion that vomeromodulin may function in olfactory and vomeronasal perireceptor processes as a transporter for pheromones and odorants. In addition, the embryonic expression of vomeromodulin suggests its involvement in olfactory perireceptor processes in utero.

[The structural-functional changes in the epithelium of the respiratory area of the nose and intrapulmonary bronchi in human ontogeny]. / Strukturno-funktsional'nye izmeneniia épiteliia dykhatel'noi oblasti nosa i vnutrilegochnykh bronkhov v ontogeneze cheloveka.

The epithelia of nasal respiratory area and intrapulmonary bronchi were studied in human ontogenesis by means of light and electron microscopy. The results indicate the stageness of morphofunctional organization of epithelia of the organs studied. The succession of the respiratory tract epithelium cytodifferentiation, dynamics of correlation between the epithelial layer cell elements in ontogenesis and peculiarities of ultrastructural organization of the nasal mucosa epithelial cell were established.