Human Nasal Tissue Research Articles

Differential Expression of Surfactant Protein A in the Nasal Mucosa of Patients With Allergy Symptoms

To characterize surfactant protein A (SP-A) expression in human nasal tissue and correlate differential expression of SP-A with symptoms suggestive of allergic rhinitis. Allergic rhinitis symptom data were prospectively collected in the form of the Rhinitis Symptom Utility Index, the Rhinoconjunctivitis Quality of Life Questionnaire, and a Visual Analog Scale. Immunohistochemical staining for SP-A was performed on resected nasal tissue. Quantitative polymerase chain reaction amplification of the SP-A gene referenced to beta-actin was performed on complementary DNA samples synthesized from total RNA isolates. Academic tertiary referral center, department of otolaryngology laboratories. Twenty-five consecutive patients undergoing nasal surgery. Immunohistochemical staining of SP-A in human nasal mucosa and submucosa, polymerase chain reaction amplification of SP-A messenger RNA, and rhinitis symptom scores. Immunostaining localized SP-A to the mucosa and submucosal glands in specimens. Quantitative polymerase chain reaction demonstrated correlation between SP-A messenger RNA concentration and the total Rhinitis Symptom Utility Index score (0.51, P = .009) as well as "sneezing over the previous week" (0.40, P = .049), "runny nose over the previous week" (0.55, P = .005), and "sneezing today" (0.47, P = .02). To our knowledge, this is the first report of SP-A expression in human nasal tissue. Furthermore, the degree of expression correlated with severity of disease as measured by the Rhinitis Symptom Utility Index in patients with allergic rhinitis symptoms.

Heme Oxygenase (HO)‐1 Is Upregulated in the Nasal Mucosa With Allergic Rhinitis

Heme oxygenase (HO) is considered to be an antioxidant enzyme that catabolizes heme to produce carbon monoxide (CO) and biliverdin. Three isoforms of HO have been discovered. Recently, HO-1 has been found to be upregulated after allergic inflammations of the lower airway. The objective of this study was to address the expression of HO isoenzymes 1 and 2 in the nasal mucosa of patients with allergic rhinitis as well as normal control subjects. Nasal mucosa from 30 patients with persistent allergic rhinitis as well as from 10 normal volunteers was used in this study. We used immunofluorescent technique, Western blotting, and real-time quantitative polymerase chain reaction to localize and quantify the expression of these isoenzymes in normal and allergic human nasal tissues. We found that HO-1 is expressed in the epithelial cells of seromucinous glands and macrophages with significant upregulation of its glandular expression in allergic rhinitis but with no difference in its macrophage expression between the study groups in contrast to HO-2 that is expressed in the vascular endothelial lining cells as well as macrophages with no marked difference between the study groups. We demonstrated that expression of HO-1, but not HO-2, was upregulated within the nasal tissues in allergic rhinitis inflammation, and understanding the induction of HO-1 expression may provide for better management of allergic rhinitis that involves oxidative stress.

Human Nasal Polyp Microenvironment Maintained in Viable and Functional States as Xenografts in SCID Mice

We undertook to maintain human nasal polyp tissue in a viable and functional state in SCID (severe combined immunodeficiency) mice. Small, nondisrupted pieces of human nasal polyp tissues were subcutaneously implanted into SCID mice depleted of natural killer cells. The resulting xenografts were examined histologically, and the sera were evaluated for the presence of human protein. The original histologic architecture of the polyp was maintained in the xenografts. The tissues, including pseudostratified columnar epithelial-lined polyps and subepithelial stroma, remained viable, and goblet cells continued to produce mucin for up to 26 weeks after engraftment. Human inflammatory leukocytes, including CD3+ T cells, CD20+ B cells, CD138+ plasma cells, and CD68+ monocytes and/or macrophages, were present. Identification of human immunoglobulin and human interferon-gamma in the sera of xenograft-bearing mice indicated that the B cells or plasma cells and T cells within the xenografts remained functional for 2 weeks after engraftment. The ability to engraft and maintain nasal polyps provides an in vivo human/mouse chimeric model with which to investigate the role of inflammatory leukocytes and stromal cells in the maintenance and progression of polyposis and to determine how exogenous cytokines may alter the interaction of inflammatory cells, stromal cells, and epithelial cells in the polyp.

The Expression of Human Antimicrobial Peptide LL-37 in the Human Nasal Mucosa

LL-37, an antimicrobial peptide, has been discovered to be produced by a number of epithelial cells. It is identified as a key element in the innate host defense mechanism. Because little is known about the expression of LL-37 in human sinonasal tract, we conducted this study to investigate the expression of LL-37 in human nasal mucosa. We investigated the expression of LL-37 gene by the reverse transcription polymerase chain reaction (RT-PCR) and localization of LL-37 peptide by immunohistochemistry in the inferior turbinate mucosa of 7 normal subjects and the nasal polyps of 12 patients with chronic paranasal sinusitis. The transcripts of the LL-37 gene were detected in all human nasal tissues analyzed by RT-PCR. There is a significant increase of LL-37 mRNA expression in nasal polyps as compared with the normal nasal mucosa. Using immunohistochemistry, LL-37 peptide was localized in surface epithelial cells and submucosal glands. Our findings suggest that LL-37 is expressed by nasal mucosa and is upregulated during inflammation.

Gene expression profiles in human nasal polyp tissues studied by means of DNA microarray

Nasal polyposis (NP) is a chronic inflammatory disease of the sinuses. Its pathogenesis is unknown. DNA microarray analysis allows simultaneous measurement of expression of thousands of genes in the same tissue sample and might help to identify gene alterations in various disorders. We sought to screen for disease-related genes in NP by using DNA microarrays and to validate the altered expression of selected genes at the mRNA and protein level. Expression microarrays containing approximately 10,500 genes were used to compare individual gene profiles of NP samples (n=10) and normal mucosal samples obtained from sphenoid sinuses in patients undergoing pituitary surgery (n=4). Four of the 5 most upregulated, and the single most downregulated, genes were retested by means of quantitative RT-PCR and immunohistochemistry in a different set of NP and normal mucosal samples obtained from the ethmoid and sphenoid sinuses. Compared with normal sinus tissue, 192 genes were upregulated at least 2-fold, and 156 genes were downregulated by at least 50% in NP samples (approximately 3% of genes evaluated). Four of the top 5 overexpressed genes (statherin, 48.0-fold; prolactin-induced protein [PIP] , 24.9-fold; lactoferrin, 26.6-fold; and deleted in malignant brain tumor 1 [DMBT1] , 30.3-fold) and the most underexpressed gene (Clara cell 10-kd protein [CC10] , -20.1-fold) were selected and retested by means of quantitative RT-PCR and immunohistochemical staining. Quantitative RT-PCR and immunohistochemical staining confirmed the differential expression of all except statherin in NP tissue. DNA microarrays can provide new insight into the possible pathophysiologic processes involved in NP.

Gene expression profiles in human nasal polyp tissues studied by DNA microarray

Rationale The pathogenesis of nasal polyposis (NP) is unknown, but it may be the result of the activation of a unique set of genes. DNA microarray analysis allows simultaneous measurement of expression of thousands of genes in the same tissue sample, and may help to identify gene alterations in various disorders. Methods HG-U95Av2 GeneChips were used to compare individual gene profiles of NP samples and normal mucosal samples obtained from sphenoid sinuses in patients undergoing pituitary surgery. The four most up-regulated and the one most down-regulated genes were retested for confirmation by quantitative RT-PCR and immunohistochemistry in a different set of NP and normal mucosal samples obtained from ethmoid and sphenoid sinuses. Results Compared with normal sinus tissue, 358 genes were up regulated at least twofold, and 63 genes were down regulated by at least 50% in NP samples (=4% of genes evaluated). The top four over-expressed genes were statherin (123.4 fold), prolactin-induced protein (100.8 fold), lactoferrin (87.4 fold), and Deleted in malignant brain tumor-1 (85.9 fold). The most under expressed gene was Clara cell 10-kDa protein (11.5 fold). Quantitative RT-PCR and immunohistochemical staining confirmed the differential expression of all except statherin in NP. Conclusion DNA microarrays can provide new insight into the pathophysiological processes involved in NP.

Nasal-associated lymphoid tissue (NALT): frequency and localization in young children.

In mucosal immunology nasal-associated lymphoid tissue (NALT) is taken as a constitutive structure of the nasal immune system and as a target tissue in strategies of local defence and an induction site for vaccination. These concepts are based on findings in rodents, but it has not been investigated systematically whether NALT also is present in humans and if so in which amount and localization. In a postmortem study the presence of NALT in humans is documented as a morphologically distinct structure additional to the lymphoid structures of the Waldeyer's ring. Human nasal tissue blocks of 150 children who had died in the first two years of life either of sudden infant death (n = 109) without signs of respiratory tract infections or of different traumatic (n = 22) and natural causes of death (n = 19) were obtained using a specific autopsy-technique and were investigated systematically using histology. Clearly in contrast to rodents human NALT was found disseminated in the nasal mucosa with typical morphological features in 38% of all children, mainly in the middle concha, with similar morphology and frequency in the examined groups. No correlation was found between the presence of NALT and the cause of death and especially the grade of inflammation in general. Therefore, NALT might be the morphological basis for inhalative vaccination strategies in young children and play a role in mucosal host defence.

Olfactory function in workers exposed to styrene in the reinforced-plastics industry.

Impairment of olfactory function in humans has been associated with occupational exposure to volatile chemicals. To investigate whether exposure to styrene was associated with olfactory impairment, olfactory function was examined in workers with a minimum of 4 years exposure to styrene in the reinforced-plastics industry (current mean exposure: 26 ppm, range: 10-60 ppm; historic mean dose: 156 ppm-years, range: 13.8-328 ppm-years) and in a group of age- and gender-matched, unexposed controls. Olfactory function was assessed using a standardized battery that included tests of threshold sensitivity for phenylethyl alcohol (PEA), odor identification ability, and retronasal odor perception. Odor detection thresholds for styrene were also obtained as a measure of specific adaptation to the work environment. No differences were observed between exposed workers and controls on tests of olfactory function. Elevation of styrene odor detection thresholds among exposed workers indicated exposure-induced adaptation. The present study found no evidence among a cross-section of reinforced-plastics industry workers that current or historical exposure to styrene was associated with impairment of olfactory function. Taken together with anatomical differences between rodent and human airways and the lack of evidence for styrene metabolism in human nasal tissue, the results strongly suggest that at these concentrations, styrene is not an olfactory toxicant in humans.

Molecular Cloning and Characterization of spurt, a Human Novel Gene That Is Retinoic Acid-inducible and Encodes a Secretory Protein Specific in Upper Respiratory Tracts

Retinoids, such as all-trans-retinoic acid, play an essential role in the regulation of airway epithelial cell growth, differentiation, and gene expression. Using cDNA microarray, we identified a clone, DD4, that contains the cDNA of a novel gene, spurt (secretory protein in upper respiratory tracts) that was significantly induced by all-trans-retinoic acid in primary cultured human tracheobroncheal epithelia. Two alternatively spliced spurt transcripts of 1090 and 1035 base pairs exist that contain the same open reading frame expressing a 256-amino acid peptide. The full-length spurt cDNA sequence spans a genomic DNA fragment of 7,313 bp, and the gene is located on chromosome 20q11.21. spurt mRNA is notably expressed at high levels in human nasal, tracheal, and lung tissues. In situ hybridization demonstrated that spurt message is often present in secretory cell types. The human spurt gene product is a secretory protein that contains a distinct signal peptide sequence in its first 19 amino acids. Mono-specific antibodies were generated to characterize spurt expression. Our data demonstrate that spurt is secreted onto the apical side of primary human airway epithelial cultures and is present in clinical sputum samples. spurt gene expression is higher in sputum and tissue samples obtained from patients with chronic obstructive lung disease. Our results provide the cloning and characterization of this tissue-specific novel gene and its possible relationship with airway diseases.

Functional expression of the C-X-C chemokine receptor CXCR4 by human bronchial epithelial cells: regulation by proinflammatory mediators.

CXCR4 and its ligand stromal cell-derived factor 1alpha (SDF-1alpha) have recently been implicated in the development of airway inflammation in a mouse model of allergic airway disease. Here we report, for the first time, the expression of a functional CXCR4 in primary human normal bronchial epithelial cells and the regulation of CXCR4 gene expression by proinflammatory mediators. Both bradykinin (BK) and IL-1beta induced an accumulation of CXCR4 mRNA in normal bronchial epithelial cells in a time-dependent manner, with peak levels of CXCR4 mRNA reached between 4 and 24 h after stimulation. Ligand activation of CXCR4 in airway epithelial cells resulted in the activation of the extracellular signal-regulated kinase and stress-activated protein kinase/c-Jun amino-terminal kinase signaling pathways and calcium mobilization. Pretreatment of airway epithelial cells with BK or IL-1beta enhanced SDF-1alpha induced phospho-extracellular signal-regulated kinase and calcium mobilization, in addition to increasing the level of CXCR4 protein. Finally, we describe the expression of CXCR4 mRNA and its regulation by BK in vivo in human nasal tissue. CXCR4 mRNA levels are significantly higher in the nasal tissue of symptomatic allergic rhinitis subjects compared with normal subjects. Moreover, BK challenge significantly increased CXCR4 mRNA levels in nasal tissue of mild allergic rhinitis subjects in vivo, but not normal controls. In conclusion, this study demonstrates that human airway epithelial cells respond to proinflammatory mediators by up-regulating the chemokine receptor CXCR4, thus enabling the cells to respond more effectively to constitutively expressed SDF-1alpha. This may lead to enhanced activation of intracellular signaling pathways resulting in the release of mediators involved in inflammatory allergic airway disease.

Physiologically Based Pharmacokinetic (PBPK) Models for Nasal Tissue Dosimetry of Organic Esters: Assessing the State-of-Knowledge and Risk Assessment Applications with Methyl Methacrylate and Vinyl Acetate

Mathematical models have been developed to describe nasal epithelial tissue dosimetry with two compounds, vinyl acetate (VA) and methyl methacrylate (MMA), that cause toxicity in these tissues These models couple computational fluid dynamics (CFD) calculations that map airflow patterns within the nose with physiologically based pharmacokinetic (PBPK) models that integrate diffusion, metabolism, and tissue interactions of these compounds. Dose metrics estimated in these models for MMA and VA, respectively, were rates of MMA metabolism per volume of tissue and alterations in pH in target tissues associated with VA hydrolysis and metabolism. In this article, four scientists who have contributed significantly to development of these models describe the many similarities and relatively few differences between the MMA and VA models. Some differences arise naturally because of differences in target tissues, in the calculated measures of tissue dose, and in the modes of action for highly extracted vapors (VA) compared with poorly extracted vapors (MMA). A difference in the approach used to estimate metabolic parameters from human tissues provides insights into interindividual extrapolation and identifies opportunities for studies with human nasal tissues to enhance current risk assessments. In general, the differences in model structure for these two esters were essential for describing the biology of the observed responses and in accounting for the different measures of target tissue dose. This article is intended to serve as a guide for understanding issues of optimum model structure and optimal data sources for these nasal tissue dosimetry models. We also hope that it leads to greater international acceptance of these hybrid CFD/PBPK modeling approaches for improving risk assessment for many nasal toxicants. In general, these models predict either equivalent (VA) or lower (MMA) nasal tissue doses in humans compared with tissue doses at equivalent exposure concentrations in rats.

Styrene Respiratory Tract Toxicity and Mouse Lung Tumors Are Mediated by CYP2F-Generated Metabolites

Mice are particularly sensitive to respiratory tract toxicity following styrene exposure. Inhalation of styrene by mice results in cytotoxicity in terminal bronchioles, followed by increased incidence of bronchioloalveolar tumors, as well as degeneration and atrophy of nasal olfactory epithelium. In rats, no effects on terminal bronchioles are seen, but effects in the nasal olfactory epithelium do occur, although to a lesser degree and from higher exposure concentrations. In addition, cytotoxicity and tumor formation are not related to blood levels of styrene or styrene oxide (SO) as measured in chronic studies. Whole-body metabolism studies have indicated major differences in styrene metabolism between rats and mice. The major differences are 4- to 10-fold more ring-oxidation and phenylacetaldehyde pathways in mice compared to rats. The data indicate that local metabolism of styrene is responsible for cytotoxicity in the respiratory tract. Cytotoxicity is seen in tissues that are high in CYP2F P450 isoforms. These tissues have been demonstrated to produce a high ratio of R-SO compared to S-SO (at least 2.4 : 1). In other rat tissues the ratio is less than 1, while in mouse liver the ratio is about 1.1. Inhibition of CYP2F with 5-phenyl-1-pentyne prevents the styrene-induced cytotoxicity in mouse terminal bronchioles and nasal olfactory epithelium. R-SO has been shown to be more toxic to mouse terminal bronchioles than S-SO. In addition, 4-vinylphenol (ring oxidation of styrene) has been shown to be highly toxic to mouse terminal bronchioles and is also metabolized by CYP2F. In human nasal and lung tissues, styrene metabolism to SO is below the limit of detection in nearly all samples, and the most active sample of lung was approximately 100-fold less active than mouse lung tissue. We conclude that styrene respiratory tract toxicity in mice and rats, including mouse lung tumors, are mediated by CYP2F-generated metabolites. The PBPK model predicts that humans do not generate sufficient levels of these metabolites in the terminal bronchioles to reach a toxic level. Therefore, the postulated mode of action for these effects indicates that respiratory tract effects in rodents are not relevant for human risk assessment.

Immunocytochemical and biochemical identification of angiotensin II in human nasal tissue.

Angiotensin II (ANG II) was identified immunocytochemically and biochemically in biopsy samples of human nasal tissue. Staining for ANG II was predominantly found in structures similar to a string of pearls with consecutive short varicose areas, which is characteristic for neuronal tissue. The localization of ANG II in neurons was confirmed by positive staining of adjacent tissue sections with a specific antibody to neurofilament or doublestaining with both antibodies in one section. Likewise, ANG II-like material was also determined radioimmunologically in nasal tissue extracts. The concentrations of ANG II varied form 1.28 to 332.78 fmol/g wet tissue weight with an average concentration of 79.61+/-44.09 fmol ANG II/g wet tissue weight (mean+/-SEM, n=7). The ANG II-immunoreactive material was further characterized biochemically by HPLC on a reversed phase C(18) column in an acetonitrile and methanol gradient as Ile(5)-ANG II and ANG II metabolites such as Ile(4)-ANG III, Ile(3)-ANG II(3-8)hexapeptide and Ile(2)-ANG II(4-8)pentapeptide.

Benzalkonium chloride as a preservative in nasal solutions: re-examining the data

Recent studies have suggested that benzalkonium chloride (BKC), an antimicrobial agent used as a preservative in nasal sprays, lacks deleterious effects on the nasal ciliated epithelium. Other data, including recent in vivo findings, suggest that BKC may, in fact, produce adverse clinical effects on human nasal tissue, including the aggravation of rhinitis medicamentosa. Toxic effects have also been reported. In light of the discrepancy between negative results and studies suggesting no safety concerns, we consider the possibility of problems in the design and methodology of some of the studies and in the interpretation of results. Clearly, further research is warranted to clarify the significance of conflicting findings. In the meantime, without conclusive data regarding BKC and the possibility of harmful effects, the use of nasal formulations without BKC might be a reasonable alternative.

The toxicity of styrene to the nasal epithelium of mice and rats: studies on the mode of action and relevance to humans

Inhaled styrene is known to be toxic to the nasal olfactory epithelium of both mice and rats, although mice are markedly more sensitive. In this study, the nasal tissues of mice exposed to 40 and 160 ppm styrene 6 h/day for 3 days had a number of degenerative changes including atrophy of the olfactory mucosa and loss of normal cellular organisation. Pretreatment of mice with 5-phenyl-1-pentyne, an inhibitor of both CYP2F2 and CYP2E1 completely prevented the development of a nasal lesion on exposure to styrene establishing that a metabolite of styrene, probably styrene oxide, is responsible for the observed nasal toxicity. Comparisons of the cytochrome P-450 mediated metabolism of styrene to its oxide, and subsequent metabolism of the oxide by epoxide hydrolases and glutathione S-transferases in nasal tissues in vitro, have provided an explanation for the increased sensitivity of the mouse to styrene. Whereas cytochrome P-450 metabolism of styrene is similar in rats and mice, the rat is able to metabolise styrene oxide at higher rates than the mouse thus rapidly detoxifying this electrophilic metabolite. Metabolism of styrene to its oxide could not be detected in human nasal tissues in vitro, but the same tissues did have epoxide hydrolase and glutathione S-transferase activities, and were able to metabolise styrene oxide efficiently, indicating that styrene is unlikely to be toxic to the human nasal epithelium.

Presence of fluticasone propionate on human nasal mucosal surface and in human nasal tissue over a period of 24 h after intranasal application.

Once-daily use of nasally applied glucocorticoids was demonstrated to be effective in the treatment of allergic rhinitis. The aim of the study was to measure concentrations of fluticasone propionate (FP) in nasal secretion and nasal tissue over a period of 1 day after a single application of 100 microg FP. Twenty-six patients applied nasal FP spray at different time intervals before surgery. Cotton swabs, used to clean the mucosal surface, and resected nasal tissue were extracted. FP concentrations were determined by RIA. FP was found in nasal secretions in concentrations from 15 to 1 microg/g over a period of 20 h, and in nasal tissue in concentrations from 200 to 13 ng/g up to 24 h after the single application. The long-persisting high concentrations of FP provide the pharmacokinetic basis for once-daily treatment.

Expression of the Voltage-Dependent Chloride Channel ClC-3 in Human Nasal Tissue

In the present study, ClC-3, one of the voltage-dependent chloride channels, was identified in human nasal tissue. In situ hybridization and immunohistochemical investigations demonstrated the localization of ClC-3 in the serous acini and ductal portions of submucosal nasal glands, which are the primary source of nasal secretion. Our data suggest that this channel contributes to nasal secretion via chloride transport. Its dysfunction might lead to abnormal nasal secretion in such pathological states as sinusitis.

Methyl methacrylate toxicity in rat nasal epithelium: studies of the mechanism of action and comparisons between species

Female F344 rats exposed to 200 ppm methyl methacrylate for 6 h developed a lesion in the nasal olfactory epithelium which was characterised by degeneration and atrophy. The severity of the lesion was markedly reduced by pre-treatment of the rats with an intraperitoneal dose of 100 mg/kg bis-( p-nitrophenyl)phosphate, an inhibitor of carboxylesterase enzymes, thus demonstrating that the lesion is caused by the carboxylesterase mediated metabolism of methyl methacrylate to methacrylic acid, an irritant and corrosive metabolite. The distribution of the carboxylesterases in nasal tissues has been investigated and the metabolism of methyl methacrylate to methacrylic acid has been compared in rat, hamster and human nasal tissue fractions in vitro. Histocytochemistry showed that the carboxylesterases are heavily localised in the sustentacular cells and Bowman's glands of the rat olfactory region, but are more generally distributed in human olfactory epithelium. Consistent with this, the enzyme activity in all three species was higher in fractions prepared from olfactory tissue than from respiratory tissue, 3-fold in rat and human and 12-fold in the hamster. The maximum rates ( V max) of metabolism in rat and hamster olfactory tissue fractions were comparable, whereas those in human olfactory tissue fractions were at least 13-fold lower. The rate of metabolism in rat olfactory tissue was also comparable to that in rat liver whereas in humans, the rate in olfactory tissue was 500-fold lower than that in the liver. In respiratory tissues, the rate in humans was at least 6-fold lower than that in the rat. These results suggest that humans are significantly less sensitive than rodents to the nasal toxicity of methyl methacrylate.

Interaction of Bordetella pertussis with human respiratory mucosa in vitro

The human respiratory tract pathogen Bordetella pertussis is the major cause of whooping cough in infants and young children, and also causes chronic cough in adults. B. pertussis infection damages ciliated epithelium in the respiratory tract. However, the interaction of the bacterium with the respiratory mucosa is poorly understood, and previous studies have either utilized animal tissue which may not be appropriate, or isolated cell systems which lack the complexity of the respiratory mucosa.We have studied the interaction of B. pertussis strain BP536 with human nasal turbinate tissue in an air-interface organ culture over 5 days. We have also compared infection by BP536 with two other strains, Tohama I and CN2992, to determine whether the interactions observed with BP536 are consistent, and, in both nasal turbinate and adenoid organ cultures at 24 h, to determine whether there were differences between tissue from different parts of the respiratory tract.BP536 adhered to cilia, most commonly at their base, and disorganized their spatial arrangement; they also adhered to damaged tissue and muscus, but very rarely to unciliated cells. Within the first 24 h there was a five-fold increase in bacterial density on ciliated cells, and the total number of adherent bacteria increased up to 96 h. Infection caused increased mucus at 24 h and an increase in damaged epithelium from 72 h which involved both ciliated and unciliated cells. The number of residual ciliated cells did not decrease after 72 h. The three different strains of B. pertussis exhibited similar interactions with the mucosa, and there was no tissue specificity for adenoid or turbinate tissue. We conclude that B. pertussis adhered to multiple sites on the mucosa and caused hypersecretion and epithelial damage which are the pathological changes described in vivo.

Acetochlor-Induced Rat Nasal Tumors: Further Studies on the Mode of Action and Relevance to Humans

The herbicide acetochlor, and its analogue alachlor, have similar toxicological properties, the most significant being the induction of nasal adenomas in rats in 2-year feeding studies. Previous investigations have proposed a mode of action involving metabolism to a quinone-imine, the formation of protein adducts, cell death, and compensatory hyperplasia leading to the observed adenomas. Comparisons between rats and humans of the metabolic cascade leading to the quinone-imine indicate that these chemicals do not pose a threat to humans. Further investigations with acetochlor, presented here, have revealed an additional activation pathway in which a sulfoxide metabolite of acetochlor plays a key role. The sulfoxide was found to be the major plasma metabolite in rats dosed with acetochlor. Whole-body autoradiography studies established that this metabolite selectively accumulates and persists in the olfactory epithelium of rats. Radiolabeling of the sulfoxide molecule in the phenyl ring and in the sulfoxide side-chain demonstrated that the metabolite accumulating in nasal tissues retains the sulfoxide side-chain. The formation of a quinone-imine from the sulfoxide was facilitated by hydroxylation of the phenyl ring by a cytochrome P450 isoenzyme which was specific to the nasal epithelium in the rat. This metabolic conversion could not be detected in 33 fresh human nasal tissue samples, supporting the earlier view that the acetochlor-induced rat nasal tumors do not represent a hazard for humans.