Clinical and Histopathologic Features of a Feline SARS-CoV-2 Infection Model Are Analogous to Acute COVID-19 in Humans.

The emergence and ensuing dominance of COVID-19 on the world stage has emphasized the urgency of efficient animal models for the development of therapeutics for and assessment of immune responses to SARS-CoV-2 infection. Shortcomings of current animal models for SARS-CoV-2 include limited lower respiratory disease, divergence from clinical COVID-19 disease, and requirements for host genetic modifications to permit infection. In this study, n = 12 specific-pathogen-free domestic cats were infected intratracheally with SARS-CoV-2 to evaluate clinical disease, histopathologic lesions, and viral infection kinetics at 4 and 8 days post-inoculation; n = 6 sham-inoculated cats served as controls. Intratracheal inoculation of SARS-CoV-2 produced a significant degree of clinical disease (lethargy, fever, dyspnea, and dry cough) consistent with that observed in the early exudative phase of COVID-19. Pulmonary lesions such as diffuse alveolar damage, hyaline membrane formation, fibrin deposition, and proteinaceous exudates were also observed with SARS-CoV-2 infection, replicating lesions identified in people hospitalized with ARDS from COVID-19. A significant correlation was observed between the degree of clinical disease identified in infected cats and pulmonary lesions. Viral loads and ACE2 expression were also quantified in nasal turbinates, distal trachea, lungs, and other organs. Results of this study validate a feline model for SARS-CoV-2 infection that results in clinical disease and histopathologic lesions consistent with acute COVID-19 in humans, thus encouraging its use for future translational studies.

The activity of N-acetyl-ß-d-hexosaminidase A and B and ß-glucuronidase in nasal polyps and hypertrophic nasal concha.

UNLABELLED: Nasal polyps and hypertrophic lower nasal conchae are common disorders of nasal cavity. The majority of etiopathogenetic theories indicate inflammatory background of polyps and hypertrophic concha. N-acetyl-ß-D-hexosaminidase and ß-glucuronidase are lysosomal exoglycosidases revealing accelerated activity in inflammatory processes. AIM: The aim of the study was to evaluate the catabolism of glycoconjugates in nasal polyps and hypertrophic nasal concha basing on the activity of N-acetyl-ß-D-hexosaminidase (HEX) and ß-glucuronidase (GLU). MATERIAL AND METHODS: Material consisted of nasal polyps taken from 40 patients during polypectomy in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and hypertrophic lower nasal conchae taken from 20 patients during mucotomy. The activity of HEX, HEX A, HEX B and GLU in supernatant of homogenates of nasal polyps and hypertrophic lower nasal concha tissues has been estimated using colorimetric method. RESULTS: Statistically significant decrease has been observed in concentration of the activity (per 1mg of tissue) of HEX (p<0.05), HEX B (p<0.001) and specific activity (per 1mg of protein) of HEX B (p<0.001) in nasal polyps tissue in comparison to hypertrophic lower nasal conchae tissue. CONCLUSIONS: Decrease in the activity and specific activity concentration of the majority of examined lysosomal exoglycosidases (increasing in inflammations) in comparison to hypertrophic lower nasal conchae suggests electrolytes disorders and questions the inflammatory background of nasal polyps.

Expression of nitric oxide synthases in leukocytes in nasal polyps.

BACKGROUND: Nitric oxide (NO) has various roles in airway physiology and pathophysiology. Monitoring exhaled NO levels is increasingly common to measure airways inflammation and inhaled NO studied for its therapeutic value in premature infants and adult respiratory distress syndrome. NO is produced by 3 isoforms of NO synthase (NOS1, 2, 3), and each can play distinct and perhaps overlapping roles in the airways. However, the distribution, regulation, and functions of NOS in various cells in the upper airways, particularly in leukocytes, are incompletely understood. OBJECTIVE: To characterize the expression of NOS isoforms in leukocytes in normal middle turbinate tissues (MT) and in inflammatory nasal tissue (nasal polyps, NP). METHODS: Normal MT tissue was collected from surgical specimens that were to be discarded. The NP samples were from surgical tissue archives of 15 patients with chronic rhinosinusitis. Isoforms of NOS in cells were identified by double immunostaining using NOS isoform-specific and leukocyte-specific (mast cell, eosinophil, macrophage, neutrophil, or T cell) antibodies. RESULTS: The proportion of total cells below the epithelium that were positive for each isoform of NOS was higher in NP than in MT. Each isoform of NOS was found in all leukocyte populations studied, and there were significant differences in the percentage of leukocytes expressing NOS isoforms between MT and NP. CONCLUSION: All isoforms of NOS are expressed in leukocytes in MT and NP, and their expression varies among leukocyte types. Our data provide a basis to investigate the regulation, cell distribution, and distinct functions of NOS isoforms in normal and inflamed nasal tissues.

Chitinolytic activity in nasal polyps.

BACKGROUND: Chitin is a recognition element for tissue infiltration by innate cells implicated in allergy and immunity. This process can be negatively regulated by vertebrate chitinases. Both acidic mammalian chitinase (AMCase) and chitotriosidase (ChT) have chitinolytic activity. This study aimed to determine the activities of AMCase and ChT in nasal polyps (NPs), as well as their in situ localization in NP tissue. METHODS: AMCase and ChT activities in NPs were compared with those in inferior turbinate tissue samples. Tissue samples were measured for AMCase and ChT activities at a range of pHs using the fluorogenic substrate 4-methylumbelliferyl-beta-d-N,N',N''-triacetyl-chitotriose. Double immunofluorescent staining for the localization of both AMCase and ChT was performed using NP cryosections. RESULTS: Both AMCase and ChT displayed markedly increased chitinolytic activity in all NPs, compared with inferior turbinate tissues. Double immunofluorescent staining revealed that CD68 highlighted monocytes in the submucosa of NP and these cells disclosed coexpression of AMCase and ChT. CD31 detected capillary endothelial cells, but did not express any AMCase and ChT. CONCLUSION: The increased chitinolytic activities of AMCase and ChT in NPs may be important in NP pathogenesis, suggesting that inhibition of chitinolytic activity may be a novel therapeutic strategy for the treatment of NPs.

Metalloproteinases in juvenile angiofibroma--a collagen rich tumor.

Matrix metalloproteinases (MMPs) act in diverse physiological and pathological conditions such as tumor growth and angiogenesis by cleaving extracellular matrix and nonmatrix substrates. MMPs with gelatinase/collagenase activity have not yet been studied in juvenile angiofibroma, a unique fibrovascular tumor with prominent collagen expression. Quantitative real-time polymerase chain reaction studies, Western blot analysis, immunofluorescence studies, gel zymography, and in situ zymography were used to analyze MMP-1, MMP-2, MMP-9, MMP-13, MMP-14, TIMP-1, and TIMP-2 in 9 juvenile angiofibromas and 2 inferior nasal turbinate specimens. Quantitative real-time polymerase chain reaction found significantly elevated expression of MMP-2, MMP-9, and MMP-14 (P < .05) in tumor tissue compared with the inferior nasal turbinate specimens. Western blot analysis detected more prominent MMP-1, MMP-2, and MMP-9 protein levels in juvenile angiofibromas compared with inferior nasal turbinates, but not MMP-13, MMP-14, TIMP-1, and TIMP-2. Immunofluorescent staining proved a mainly stromal localization of the analyzed MMPs. Only MMP-9 and MMP-14 were also detected in vessel walls. MMP-1, MMP-2, and MMP-13 also stained mast cells. Gel zymography indicated increased MMP-2 and MMP-9 gelatinase activity in juvenile angiofibromas compared with inferior nasal turbinates. Finally, in situ zymography detected very high stromal gelatinase/collagenase activity. This study indicates significant expression of MMPs with gelatinase/collagenase activity in juvenile angiofibromas with evidence of a disturbed balance of MMPs to TIMPs toward enhanced MMP activity. These MMPs are assumed to be involved in tumor pathology with an influence on tumor growth and angiogenesis.

Telomerase in nasal polyps.

BACKGROUND: The aim of this study was to determine the presence and location of telomerase activities and the possible influence of elevated human telomerase reverse transcriptase (hTERT) mRNA levels on the outcome after surgical treatment in nasal polyposis. METHODS: Telomerase activity in nasal polyps of 21 patients was quantified by measuring the hTERT mRNA contents with one-step real-time polymerase chain reaction. Inferior turbinates of 12 patients served as controls. Immunohistochemistry with specific antibodies was performed against hTERT. The number of hTERT marked cells was determined in 15 randomly selected fields. All patients were followed up after surgery for 60 months. RESULTS: Elevated hTERT mRNA expression and number of hTERT+ cells was detected in nasal polyps in comparison with inferior turbinates (p < 0.001). hTERT+ cells were detected in the basal layer of the epithelia, the endothelia, and in some seromucous glands. During follow-up, it was discovered that tissue samples of five patients with recurrent polyposis did not have higher amounts of hTERT when compared with patients without relapse. CONCLUSION: Telomerase activity is elevated in nasal polyps. Elevated hTERT expression does not predict the recurrence of nasal polyposis after surgical treatment.

Malondialdehyde level and adenosine deaminase activity in nasal polyps.

OBJECTIVE: Although there are many reports on adenosine deaminase (ADA) activities in different tissues, no information is available about the enzyme activity in nasal mucosa and polyp tissues. Whereas ADA is related to the production of free radicals by neutrophils, malondialdehyde (MDA) is an indicator of lipid peroxidation that is a general mechanism of tissue damage by free radicals. This study is aimed at determining and comparing the ADA activity and MDA level in nasal polyps and normal mucosa. STUDY DESIGN AND SETTING: Twenty-three patients with nasal polyps and a control group consisting of 14 patients with septal deviation and lower turbinate hypertrophy were included in the study. Tissue MDA level was measured by the method of Okawa with modification and tissue ADA activity by the method of Giusti. RESULTS: In patients with nasal polyp, mean tissue MDA level and ADA activity were 2.43 +/- 0.38 nmol/mg protein (Pr) and 0.235 +/- 0.055 U/mg Pr, respectively, which were significantly higher than those of control nasal mucosa (1.03 +/- 0.41 nmol/mg protein and 0.056 +/- 0.011 U/mg Pr, respectively) (P < 0.05). In addition, tissue MDA level was positively correlated to ADA activity in nasal polyps (r = 0.701, P < 0.001). CONCLUSIONS: The present study showed the presence of detectable ADA activity in nasal mucosa, and also significant increases in both tissue MDA level and ADA activity in NP tissue when compared to normal turbinate tissue. EBM RATING: B-2b.

Expression and distribution of thioredoxin and thioredoxin reductase in human nasal mucosa and nasal polyp.

CONCLUSIONS: The results of this study indicate that thioredoxin (Trx) and thioredoxin reductase (TrxR) may play a role in the defense of normal human nasal mucosa against external noxious stimuli. Based on the fact that normal nasal mucosa is continuously exposed to inhaled toxicants and contains a considerable number of inflammatory cells, Trx and TrxR may be upregulated even in normal nasal mucosa and perhaps the difference in their expression levels between normal nasal mucosa and nasal polyp, if it exists at all, is small and therefore difficult to detect. Further studies will be needed to clarify the roles of Trx and TrxR in the pathogenesis of nasal polyp. OBJECTIVES: The cellular antioxidant defense system includes thiol-containing proteins such as Trx and TrxR, which have recently attracted much attention due to their strong antioxidant radical quenching capabilities and other important biological functions related to the regulation of the cellular redox state. This study was undertaken to investigate the expression and distribution of Trx and TrxR in normal human nasal mucosa and nasal polyp, and to improve understanding of the significance of the Trx system in these conditions. MATERIAL AND METHODS: The expression and distribution of Trx and TrxR in normal human inferior turbinate mucosa and nasal polyp were investigated using reverse transcriptase polymerase chain reaction (RT-PCR), semiquantitative RT-PCR, Western blotting and immunohistochemistry. RESULTS: mRNAs and protein for both Trx and TrxR were detected in normal human inferior turbinate mucosa and nasal polyp. Semiquantitative RT-PCR and Western blotting revealed that there was no significant difference in the expression levels of Trx and TrxR between inferior turbinate mucosa and nasal polyp. Immunoreactivity for both Trx and TrxR was seen in nasal epithelial cells, glands and vascular endothelium of inferior turbinate mucosa and nasal polyp. Trx and TrxR immunoreactivity was also found in inflammatory infiltrating cells in inferior turbinate mucosa and nasal polyp.

[Expression of the inducible isoform of nitric oxide synthase mRNA and the role in nasal polyps].

OBJECTIVE: To explore the pathogenesis of nasal polyps. The aim of this study is to detect the expression of inducible isoform of nitric oxide synthase (iNOS) in nasal polyp tissue, and to compare these findings with that in normal nasal turbinate. METHODS: We examined the expression of iNOS in human nasal polyps from patients undergoing nasal polypectomy (n = 8) and nasal turbinectomy(n = 6). The iNOS mRNA expression was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The iNOS mRNA expression was significantly stronger in the nasal polyps (NP) than that in turbinate tissues (P < 0.01). CONCLUSIONS: iNOS expression is upregulated in NP, which indicate that iNOS and NO may play a potential role in the formation and growth of NP.

[Study of the enzyme peptidyl peptidase IV in nasal mucosa]. / Etude de l'enzyme dipeptidyl peptidase IV dans la muqueuse nasale.

INTRODUCTION: The endothelial serine protease dipeptidyl peptidase IV (DPP IV) cleaves the tyrosin-prolin dipeptide of several inflammatory mediators and neuropeptides, including neuropeptide Y (NPY), yielding the endogenous Y2-receptor agonist NPY (3-36) which modulates sensory and parasympathetic nerve activity. The aims of the study were to investigate the localisation of DPP IV in human nasal mucosa and to measure in vitro activity of DPP IV in nasal mucosa biopsies from patients suffering from chronic rhinosinusitis. PATIENTS AND METHODS: Using immunohistochemistry we have studied the localisation of DPP IV in human nasal biopsies. The activity of DPP IV was measured in vitro in nasal mucosa samples obtained from 45 patients suffering from chronic rhinosinusitis and compared with the density of inflammatory cell infiltration. RESULTS: Positive immunoreactivity for DPP IV was observed in the human nasal mucosa. Low activity of DPP IV was associated with high density of inflammatory cells in the mucosa of patients suffering from chronic rhinosinusitis. The regressive correlation was statistically significant (p < 0.001). CONCLUSION: Low level DPP IV activity is associated with inflammation of the nasal mucosa. This enzyme may be involved in the pathophysiological mechanism of nasal hyperreactivity and chronic rhinosinusitis.

Metabolism of chloroform by cytochrome P450 2E1 is required for induction of toxicity in the liver, kidney, and nose of male mice.

Chloroform is a nongenotoxic-cytotoxic liver and kidney carcinogen and nasal toxicant in some strains and sexes of rodents. Substantial evidence indicates that tumor induction is secondary to events associated with cytolethality and regenerative cell proliferation. Therefore, pathways leading to toxicity, such as metabolic activation, become critical information in mechanism-based risk assessments. The purpose of this study was to determine the degree to which chloroform-induced cytotoxicity is dependent on the cytochromes P450 in general and P450 2E1 in particular. Male B6C3F(1), Sv/129 wild-type (Cyp2e1+/+), and Sv/129 CYP2E1 knockout (Cyp2e1-/- or Cyp2e1-null) mice were exposed 6 h/day for 4 consecutive days to 90 ppm chloroform by inhalation. Parallel control and treated groups, excluding Cyp2e1-null mice, also received an i.p. injection (150 mg/kg) of the irreversible cytochrome P450 inhibitor 1-aminobenzotriazole (ABT) twice on the day before exposures began and 1 h before every exposure. Cells in S-phase were labeled by infusion of BrdU via an implanted osmotic pump for 3.5 days prior to necropsy, and the labeling index was quantified immunohistochemically. B6C3F(1) and Sv/129 wild-type mice exposed to chloroform alone had extensive hepatic and renal necrosis with significant regenerative cell proliferation. These animals had minimal toxicity in the nasal turbinates with focal periosteal cell proliferation. Administration of ABT completely protected against the hepatic, renal, and nasal toxic effects of chloroform. Induced pathological changes and regenerative cell proliferation were absent in these target sites in Cyp2e1-/- mice exposed to 90 ppm chloroform. These findings indicate that metabolism is obligatory for the development of chloroform-induced hepatic, renal, and nasal toxicity and that cytochrome P450 2E1 appears to be the only enzyme responsible for this cytotoxic-related metabolic conversion under these exposure conditions.

Laser surgery for allergic rhinitis: the effect on seromucinous glands.

Ten patients with perennial allergic rhinitis were subjected to CO2 laser turbinectomy. Tiny (1 mm3) biopsy specimens were taken at the time of surgery and 1 month thereafter. The biopsy specimens were processed for transmission electron microscopy. Also, the activity of succinic dehydrogenase and cholinesterase enzymes was measured. The study showed that laser turbinectomy was followed by reduction in the number and activity of the glandular acini in the laser-treated areas. This reduction is ascribed to the local destructive effect of laser energy on the glandular acini and on the surrounding cholinergic nerve fibers. The enzymatic activity of the cholinergic nerve fibers themselves, however, did not diminish, indicating that laser surgery has no inhibitory effect on the local allergic reaction.

In vitro expression of inducible nitric oxide synthase in the nasal mucosa of guinea pigs after incubation with lipopolysaccharides or cytokines.

In order to demonstrate the involvement of nitric oxide synthases (NOS)--in particular the inducible isoform (iNOS)--in inflammatory processes within the nasal airways, we used organ-bath incubation to study isolated inferior turbinates and mucosa of the maxillary sinus of guinea pigs. The pattern of the expression in various substructures of the nasal mucosa was of special interest. Mucosa was incubated for 6 h with lipopolysaccharides (LPS) produced by E. coli, interleukin II (IL-2) or tumor necrosis factor-alpha (TNF-alpha). Saline was used as the control solution. Following incubation the specimens were fixed in buffered 4% formaldehyde solution over a period of 4 h. Tissues were next exposed to nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase-reaction and immunostained with specific antibodies to iNOS. Results then showed a clearly increased or initiated expression of iNOS in epithelium, glands, leucocytes and blood vessels of treated tissues in comparison to the control specimens. The inflammatory mediator LPS and the cytokines Il-2 or TNF-alpha alone were found to be capable of increasing the expression of iNOS, although the effects of LPS clearly exceeded those of the cytokines. This finding implicates iNOS-generated nitric oxide as a key factor for causing nasal swelling, secretion and obstruction during nasal infections and allergic episodes.

Aminopeptidase activity in human nasal mucosa.

BACKGROUND: Aminopeptidases activate bradykinin and degrade many inflammatory peptides. OBJECTIVE: The objective of this study was to identify the types of aminopeptidase activities in human nasal mucosa. METHODS: Human nasal mucosa was homogenized (n = 12), and cytoplasmic (S2) and membrane-rich (P2) fractions were obtained. Several aminopeptidase (Ap) activities were defined by (1) substrate specificity with leucine-enkephalin (leu-Ap) and alanine-nitroanilide (ala-Ap), (2) inhibitor studies with puromycin and bestatin, (3) enzyme activity histochemistry (zymography), (4) immunohistochemistry, and (5) gel electrophoresis. Human volunteers had methacholine, histamine, and allergen nasal provocations to determine the mechanisms controlling nasal aminopeptidase secretion in vivo. RESULTS: P2 was the largest reservoir of puromycin-resistant aminopeptidase activity (630 pmol leu-enk/min/mg protein). S2 contained 32 pmol leu-enk/min/mg activity, with 80% representing puromycin-resistant activity and 20% puromycin-sensitive aminopeptidase (PS-Ap). Ala-Ap was detected in both P2 and S2 fractions and was localized by zymography to epithelial and gland cells. Anti-rat brain-soluble PS-Ap IgG detected immunoreactive material in epithelium, glands, and endothelium. In nasal provocation studies, leu-AP correlated with glandular exocytosis but not vascular leak. CONCLUSIONS: The predominant aminopeptidase in human nasal epithelial and submucosal gland cells was membrane-bound puromycin-resistant aminopeptidase. A novel soluble puromycin-resistant aminopeptidase and lower amounts of soluble PS-Ap were also detected.

Neutral endopeptidase activity and concentration of sensory neuropeptide in the human nasal mucosa.

Human nasal mucosa biopsy samples were studied by biochemical and histological methods to determine whether the concentration of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) as a marker of sensory nerves was dependent on the activity of neutral endopeptidase-like enzyme (NEP-LE). Mucosal samples from the middle turbinate were obtained from 32 patients undergoing functional endoscopic nasal surgery for non-allergic chronic rhinosinusitis. The degree of symptoms related to nasal obstruction, rhinorrhea and headaches was recorded. The number of inflammatory cells in each biopsy sample was evaluated by conventional histopathological examination. CGRP-LI was measured by radioimmunoassay. The activity of NEP-LE was evaluated in vitro using [3H] Leu5-enkephalin as substrate. A good correlation was observed between increased concentrations of CGRP, abundant inflammatory cells and the intensity of symptoms (R2 = 0.80). A low activity of NEP-LE was associated with a high concentration of both inflammatory cells and CGRP, suggesting that NEP-LE activity was reduced during inflammation. These observations further support the hypothesis that reduced degradation of sensory neuropeptides could be involved in the pathophysiological mechanisms of non-specific chronic rhinosinusitis.

An in vitro model for measuring tryptase release from nasal mucosa in response to allergen challenge.

A simple in vitro nasal mucosal culture model has been developed to measure release of the mast cell specific enzyme tryptase in response to allergen challenge. Patients who were undergoing inferior turbinectomy were skin-tested for commonly inhaled allergens. The mucosa from the inferior turbinates was kept viable using Minimal Essential Medium. Tryptase release into the medium was measured using the Pharmacia Riact Assay. There was a significant increase in tryptase release in response to allergen challenge from the mucosa harvested from skin-test positive patients. Mucosa from skin-test negative patients failed to demonstrate an increase in tryptase release. This could prove to be a useful research model for the study of nasal type I hypersensitivity and drugs that affect it.

Nasal secretion of the ozone scavenger uric acid.

Uric acid, an important scavenger of ozone, has been identified as the major low molecular weight antioxidant in baseline and cholinergically induced nasal secretions. The purpose of this study was to determine the specific tissue source of uric acid in airway secretions. The secretion of uric acid is increased by cholinergic stimulation and correlates closely with the secretion of lactoferrin (a nasal glandular protein), suggesting that submucosal glands are involved. Indeed, nasal turbinate tissue was found to contain uric acid. However, careful analysis of nasal turbinate tissue failed to reveal the presence of xanthine oxidase, the enzyme responsible for uric acid synthesis. These data suggest that uric acid might be taken up secondarily by glands from plasma. This possibility was strengthened by the observation that lowering the plasma urate level with probenecid concomitantly lowered urate secretion. These findings are consistent with the hypotheses that the principal source of uric acid in nasal secretions is plasma and that uric acid is taken up, concentrated, and secreted by nasal glands.

Purified Pasteurella multocida protein toxin reduces acid phosphatase-positive osteoclasts in the ventral nasal concha of gnotobiotic pigs.

To study the in vivo response of conchal (turbinate) osteoclasts to Pasteurella multocida toxin, four gnotobiotic pigs (7 days of age) were inoculated subcutaneously with 0.2 microgram/kg of purified toxin. One toxin-treated pig along with one control pig were necropsied at 2, 5, 9, and 14 days postinoculation. The entire length of nasal concha from the nasal planum toi ethmoid region was removed, blocked by transverse cuts into five areas, decalcified, sectioned, and then stained with tartrate-resistant acid phosphatase (TRAP) to identify osteoclasts. In each section, total area of concha, total osteoclast cytoplasmic area, and number of osteoclasts were determined using an image analysis morphometric unit. Also collected from pigs were blood and serum for complete blood counts, electrolyte levels, liver enzymes, and TRAP levels. Conchal atrophy increased in severity with time after 2 days postinoculation. In general, the ventral conchae from toxin-treated pigs at 9 and 14 days postinoculation had decreased surface area, osteoclast cytoplasmic area, and numbers of osteoclasts. Serum levels of TRAP were mildly elevated when compared with age-matched controls. No other significant alterations in blood cells or chemistries occurred and no lesions were present histologically in tissues (liver, kidney, lung, heart, and spleen) other than concha. This study shows that the P. multocida toxin induces rapid bone resorption and increases serum levels of acid phosphatase but leads to diminished acid phosphatase expression and presumably, numbers of osteoclasts.

Effects of several selected odorants on the sodium- and potassium-dependent adenosine triphosphatase activities of two different chicken olfactory tuberinals.

The NaK-adenosine triphosphatase (ATPase)-rich nerve-ending particle preparations (B fractions) of the epithelial tissue of chicken olfactory tubercle and the olfactory main concha were isolated by differential centrifugation. These tissues were exposed to the following odorants: 2-nonanone, 1-nonanol, 1-octanol, (+)2-octanol, and (-)2-octanol. There was a significant stimulation of NaK-ATPase by 2-nonanone (1 x 10(-3) M) in the B fraction of the olfactory tubercle and olfactory main concha. The NaK-ATPase increased significantly in the B fraction response of the olfactory main concha with 1 x 10(-3) M of 1-octanol but NaK-ATPase declined significantly in the presence of 1 x 10(-3) M of 1-nonanol. Both optical and structural isomers of odorants were shown to elicit different responses in NaK-ATPase activity. It is proposed that interactions between the odorant molecules and the membrane bound NaK-ATPase complex lead to recognition of odor by chickens.

Xenobiotic-metabolizing enzymes in human respiratory nasal mucosa.

Study of oxidative and non-oxidative xenobiotic-metabolizing enzymes was undertaken in microsomal and cytosolic fractions of two human livers, 10 individual and several pooled samples of human respiratory nasal mucosa obtained by surgical operation of male and female patients affected by hypertrophy of the inferior turbinates. The purity of nasal microsomes was checked by electron microscopy and marker enzyme assay. The pooled samples of respiratory nasal epithelium contained, relative to liver, a low amount of cytochrome P450 (about 25 pmol/mg protein) and associated biotransformation activities, and a low level of other components of the mixed-function oxidase system such as cytochrome b5, NADH and NADPH-cytochrome c reductase however the NADH-cytochrome b5 reductase activity was comparable to that of liver. The P450-dependent monooxygenase activities such as ethoxycoumarin O-deethylase, ethoxyresorufin O-deethylase and the dimethylnitrosamine N-demethylase were found in nearly all nasal microsomal specimens. The aniline hydroxylase and the aminopyrine or hexamethylphosphoramide N-demethylases were detected only in the pooled nasal samples. With regard to the non-oxidative enzymes, the activities of glutathione S-transferase, DT-diaphorase, epoxide hydrolase, UDP-glucuronyl-transferase, carbonyl reductase, benzaldehyde and propionaldehyde dehydrogenases, were investigated both in the individual and pooled nasal tissues and livers. These activities were similar in nasal and liver tissue, except for UDP-glucuronyltransferase which was not detected in nasal mucosa. The present findings demonstrate that the respiratory section of human nose contains a wide array of oxidative and non-oxidative enzymes, which could play a crucial role in the bioactivation or detoxication in situ of inhaled xenobiotics.