Gene-expression profiles in human nasal polyp tissues and identification of genetic susceptibility in aspirin-intolerant asthma.

BACKGROUND: Aspirin-intolerant asthma (AIA) is a subtype of asthma induced by non-steroidal anti-inflammatory drugs and characterized by an aggressive mucosal inflammation of the lower airway (asthma) and the upper airways (rhinitis and nasal polyp). The lower airway lesion and the nasal polyp in AIA are postulated to have common pathogenic features involving aspirin sensitivity that would be reflected in the gene expression profile of AIA polyps. OBJECTIVE: This study was conducted to clarify the pathogenesis of AIA using gene expression analysis in nasal polyps, and identify genetic susceptibilities underlying AIA in a case-control association study. METHODS: Global gene expression of nasal polyps from nine AIA patients was examined using microarray technology in comparison with nasal polyps from five eosinophilic sinusitis (ES) patients, a related disease lacking aspirin sensitivity. Based on the AIA-specific gene expression profile of nasal polyp, candidate genes for AIA susceptibility were selected and screened by a case-control design of 219 AIA patients, 374 non-asthmatic control (CTR), and 282 aspirin-tolerant asthmatic (ATA) subjects. RESULTS: One hundred and forty-three elevated and three decreased genes were identified as AIA-specific genes that were enriched in immune response according to Gene Ontology analysis. In addition, a k-means-based algorithm was applied to cluster the genes, and a subclass characteristic of AIA comprising 18 genes that were also enriched in immune response was identified. By examining the allelic associations of single nucleotide polymorphisms (SNPs) of AIA candidate genes relevant to an immune response with AIA, two SNPs, one each of INDO and IL1R2, showed significant associations with AIA (P=0.011 and 0.026 after Bonferroni's correction, respectively, in AIA vs. CTR). In AIA-ATA association analysis, modest associations of the two SNPs with AIA were observed. CONCLUSION: These results indicate that INDO and IL1R2, which were identified from gene expression analyses of nasal polyps in AIA, represent susceptibility genes for AIA.

[A case of pediatric recurrent acute mastoiditis caused by penicillin-resistant Streptococcus pneumonia complicated by primary immunodeficiency].

Penicillin-resistant Streptococcus pneumoniae (PRSP) is a frequently detected pathogen of intractable acute otitis media and is associated with prolonged or recurrent infection. The use of antibiotics has made the incidence of secondary acute mastoiditis following acute otitis media relatively rare, but when it does occur, its severe complications may be life-threatening. We report a case of pediatric recurrent acute mastoiditis caused by PRSP in a 6-year-old boy suffering from PRSP acute mastoiditis on 4 occasions, twice undergoing simple mastoidectomy. Although we initially suspected PRSP to be the chief factor in iterative infection, immunological analysis demonstrated significantly decreased IgG and IgA antibodies in serum and the patient was diagnosed as having common variable immunodeficiency (CVID). As the first middle ear infection occurred at the age of 6 and there was no history of upper respiratory tract infection, CVID may be the main pathological factor of recurrent mastoiditis, although infection occurred, only in the ear and did not involve other organs. This suggests that recurrent mastoiditis in the present case involved the coexistence of PRSP and CVID.

Lymphoscintigraphy for interpretation of changes of cervical lymph node function in patients with oral malignant tumors: comparison of Tc-99m-Re and Tc-99m-HSA-D.

OBJECTIVE: The purpose of this study is to compare the usefulness of technetium-99m-rhenium colloid (Tc-99m-Re) and technetium-99m-human serum albumin diethylene-triamine-pentaacetic acid (Tc-99m-HSA-D) as lymphoscintigraphic agents and to discuss the significance of lymphoscintigraphy in comparison with computed tomography (CT), magnetic resonance imaging (MRI), and ultrasonography (US). STUDY DESIGN: Dynamic and static lymphoscintigraphies were performed with Tc-99m-Re and Tc-99m-HSA-D. The usefulness of the 2 agents was evaluated statistically in comparison with pathologic findings and palpation. The significance of lymphoscintigraphy is discussed in comparison with CT, MRI, and US (by the literature of CT, MRI, and US). RESULTS: Lymphoscintigraphy was superior to palpation, and Tc-99m-Re was superior to Tc-99m-HSA-D in accuracy, specificity, and the incidence of true-positive and false-positive. Statistical significance was shown between the static lymphoscintigraphy with Tc-99m-Re and palpation. The reliability of lymphoscintigraphy seemed to be slightly inferior to CT and MRI in specificity and accuracy. However, lymphoscintigraphy had some advantages that CT and MRI lacked; for example, it showed very high sensitivity (100%) and no false-negative (0%). It also showed changes of lymph node function, showed all levels of neck nodes at one time, and showed a possibility of detecting small lymph node metastases. CONCLUSION: Lymphoscintigraphy was more useful than palpation in detecting lymph node metastases, and Tc-99m-Re was superior to Tc-99m-HSA-D as an agent. Lymphoscintigraphy is significant when it is performed as a preliminary examination before CT or MRI.

The role of thymidine phosphorylase in the pathogenesis of allergic rhinitis.

The activity and distribution of thymidine phosphorylase (TP) in the nasal mucosa of patients with nasal allergy was examined and compared with those in healthy subjects. TP activity was analyzed by spectrophotometry and expression was examined by immunoblotting and immunohistochemical staining using a monoclonal antibody specific to TP. The expression level of TP detected by immunoblotting showed a correlation with the activity detected by spectrophotometry. In nasal mucosa obtained from patients with nasal allergy, the level of TP was significantly higher than that from normal subjects. Eosinophils, basal cells in mucosal epithelium and fibroblasts in nasal mucosa obtained from patients with nasal allergy were stained with anti-TP monoclonal antibody. Strong staining of eosinophils present in nasal discharge was observed. The present results indicate that an increased number of TP-expressing cells, especially eosinophils in nasal mucosa, might be associated with the pathogenesis of nasal allergy.

Application of YAMIK sinus catheter for patients with paranasal sinusitis with and without nasal allergy.

PURPOSE: YAMIK sinus catheter (YAMIK) has already been reported to be a useful therapeutic device for sinusitis cases. The purpose of this study was to compare the effectiveness of YAMIK in sinusitis cases with and without nasal allergy in order to contribute toward establishing its indication. METHODS: YAMIK was tried in 10 chronic sinusitis cases complicated with nasal allergy and 20 cases without nasal allergy. Clinical symptoms, endoscopic findings and pathological opacification in X-ray photographs were compared in these cases with and without nasal allergy. RESULTS: No significant differences were seen in the effectiveness of YAMIK between cases with and without nasal allergy. CONCLUSION: These findings suggest that YANIK is useful and applicable even in sinusitis cases complicated with nasal allergy.

Uptake of BrdU in olfactory and respiratory epithelium of rabbits with experimental sinusitis.

Sinusitis was induced in six rabbits in order to evaluate its influence on the proliferation of cells in the olfactory epithelium compared with the respiratory epithelium during conservative antibiotic therapy. Then 1% ofloxacin was injected into the paranasal sinuses. Three normal rabbits were not administered any treatment and served as controls. The rabbits were sacrificed under intravenous anesthesia and the olfactory and respiratory mucosa was excised 24 hours after intravenous administration with the labeling reagent 5-bromo-2'-deoxyuridine (BrdU). The extent of cell proliferation in these tissues was estimated by immunohistochemical staining with BrdU-specific antibody. The uptake of BrdU was significantly increased (p = 0.0099) in the respiratory mucosa, but not in the olfactory mucosa. Furthermore, in olfactory epithelium, 79.2% and 16.7% of all BrdU-positive cells were olfactory and basal, respectively. Thus, turnover of epithelial cells due to sinusitis was not as accelerated in the olfactory mucosa as in the respiratory mucosa during antibiotic treatment.

Clinical evaluation of lymphoscintigraphy with a new technetium compound for metastatic cervical lymphadenopathy.

OBJECTIVES: To evaluate the potential usefulness of lymphoscintigraphy for the detection of metastatic cervical lymphadenopathy. METHODS: Dynamic and static lymphoscintigraphy with 99Tcm-HSA-D was performed in 23 patients with malignant oropharyngeal tumors. The internal jugular nodes were excised and examined pathologically. The results of the lymphoscintigraphy were compared with the pathological examination and clinical palpation for accuracy. RESULTS: All cases showed abnormal findings on lymphoscintigraphy. The accuracy on dynamic scintigraphy was 69% for asymmetric drainage, 69% for delayed drainage and 100% for inverse drainage. The overall accuracy for each side was 71%. With static scintigraphy, the accuracy was 69% for asymmetry, 86% for fitting defect, 67% for mottled-patchy appearance, 70% in swelling and 100% for collateral pathway. The overall accuracy for each side was 74%. The agreement between lymphoscintigraphy and palpation was 44%. CONCLUSIONS: Lymphoscintigraphy with 99Tcm-HSA-D was superior to palpation. The method is useful as a supplementary modality for the evaluation of metastatic cervical lymphadenopathy because of its relatively high accuracy and a low false-negative rate.

Effects of clarithromycin on cultured human nasal epithelial cells and fibroblasts.

OBJECTIVE/METHODS: Long-term administration of clarithromycin has been reported to be effective in the treatment of chronic sinusitis. To investigate the mechanism underlying the anti-inflammatory activity of clarithromycin, the authors evaluated the effect of clarithromycin on the gene expression of proinflammatory cytokine and the DNA-binding activity of nuclear factor (NF)-kappa B in cultured human nasal epithelial cells and fibroblasts. Cells were incubated with endotoxin purified from nontypable Haemophilus influenzae or interleukin (IL)-1 beta in the presence of clarithromycin. RESULTS: Northern blot analysis revealed that clarithromycin suppressed IL-1 beta gene expression in human nasal epithelial cells stimulated by H. influenzae endotoxin (HIE). Intercellular adhesion molecule-1 gene expression in nasal fibroblasts stimulated by IL-1 beta was also suppressed by clarithromycin. Furthermore, electrophoretic mobility shift assay demonstrated that clarithromycin reduced DNA-binding activity of NF-kappa B in both human nasal epithelial cells and fibroblasts stimulated by HIE or IL-1 beta, respectively. CONCLUSION: The present results suggest that clarithromycin may reduce gene expression of proinflammatory cytokines and adhesion molecules from nasal mucosa at the transcriptional factor level and exert an anti-inflammatory effect on nasal mucosa in chronic sinusitis.

Interpretation of scintigraphy of papillary cystadenoma lymphomatosum (Warthin's tumor) on the basis of histopathologic findings.

OBJECTIVES: The accumulation of 99mTc-pertechnetate in Warthin's tumor was estimated scintigraphically and histopathologically to determine the role of the epithelial component in scintigraphy. STUDY DESIGN: Six cases underwent histopathologic examination and scintigraphic evaluation with 99mTc-pertechnetate. Histopathologically the tumors were classified into types according to the epithelial component and cystic space and compared with scintigraphic results evaluated by dynamic radioactive index and wash-out image. RESULTS: Histopathologic and scintigraphic observations showed an adequate correlation. Cases with a large epithelial component and poor cystic space showed a large radioactive index of dynamic scintigraphy and hot accumulation of wash-out image. CONCLUSION: The scintigraphic results were chiefly due to the epithelial component, but the influence of the cystic space could not be disregarded.

Mucosa-associated lymphoid tissue in middle ear and Eustachian tube in children.

The presence of mucosal lymphoid follicles with germinal centers, so called mucosa-associated lymphoid tissue (MALT), in the Eustachian tube (ET) and middle ear (ME) was investigated in 23 human temporal bones containing the entire ET obtained from 23 children, 3 months to 10 years old at death. Greater numbers of MALTs were found in specimens from children with otitis media (OM) than from those without OM. MALT showed a wedge-shaped distribution through the ME and ET: in all 3 specimens that had MALTs in the ME, these tissues were seen throughout the ET; in all 4 specimens with MALTs in the bony portion of the ET, these tissues were present both in the tympanic and pharyngeal portions of the cartilagines ET; all 7 specimens that had MALTs in the tympanic half of the cartilagines ET had MALTs in the pharyngeal portion of the cartilagines ET as well. Furthermore, MALTs were noted in the pharyngeal portion of the ET in more than half of the specimens without OM but in none of the MEs of specimens without OM. These results support the hypothesis that persistent and recurrent inflammation in the nasopharynx spreads through the ET to play a role in the pathogenesis of chronic OM in children.

Histological changes in elastic components of soft palate scars after CO2 and contact Nd:YAG laser incisions in the dog as an experimental model.

Soft palate stiffening operations--often with CO2 or contact Nd:YAG lasers--have been used recently to treat patients with pronounced snoring. Differences in soft palate scars formed after CO2 and contact Nd:YAG laser incisions were studied in an animal model to determine which of these lasers produces a more rigid scar and consequently might be more effective for a soft palate stiffening operation. Six mongrel dogs were placed in each laser group, after which either the CO2 or Nd:YAG laser was used to make a 1.5-cm-long soft palate incision. The scars produced were then excised and studied histologically 4 weeks postoperatively. Special attention was paid to collagen density and elastin fibers in the scar tissue. After contact Nd:YAG laser incisions scar tissue was found to contain densely aligned collagen fibers and practically no elastin fibers. CO2 laser incisions produced significantly different scars: irregularly arranged collagen fibers with some elastin fibers also inside the scar tissue. Differences in the shapes of the scars and wound contractions were also found that were only seen after contact laser incisions. These findings suggest that the contact Nd:YAG laser might be more effective for soft palate stiffening operations.

[Immunohistochemical study on temporal bone of autopsy cases with Mycobacterium leprae infection].

Three temporal bones were obtained en bloc from autopsy cases with lepromatous leprosy from the middle cranial fossa side after removing the brain. After fixation with 10% formalin followed by sufficient decalcification, the specimens were embedded in paraffin en bloc and cut serially to stain every 10th section with hematoxylin and eosin (H&E) for anatomical orientation. An immunohistochemical study with anti-neurofilament, anti-MBP (myelin basic protein), anti-BCG (Bacillus Calmette-Gue'rin) and anti-PGL (Mycobacterium leprae-specific antiphenolic glycolipid-I) antibodies were performed to vestibular, cochlear and facial nerves, respectively, on the basis of anatomical orientation of the adjacent H&E sections. In one of three cases, positive staining by anti-PGL antibodies was recognized only in the facial nerve both in its internal auditory meatal and tympanic portions. However, even in this case, no neural damage was observed either by anti-neurofilament nor anti-MBP stainings. This finding supports the possibility of central neural infection by Mycobacterium leprae.

Effect of exogenous ATP and physical stimulation on ciliary function impaired by bacterial endotoxin.

The effects of exogenous ATP and physical stimulation on ciliary beat depressed by lipopolysaccharide from Pseudomonas aeruginosa were studied in cell culture. Both the ciliary beat frequency (CBF) and the amplitude (CBA) of human respiratory cells in monolayer cell cultures were studied by using a differential interference microscope equipped with a high speed video. Both the ATP and the physiol stimulation stimulated temporarily the depressed CBF and CBA. In both groups the CBF and CBA increased in 1 min to the initial level and then gradually decreased to the level before the stimulation. The duration of the ATP stimulation on the CBF (10.1 +/- 1.8 min) and CBA (10.1 +/- 2.3 min) were significantly better than the duration of physical stimulation on CBF (8.0 +/- 1.8 min) and CBA (6.8 +/- 1.6 min). However, the areas under the CBF/CBA curves (from the beginning of stimulation until the initial level of CBF or CBA was reached again) did not differ significantly. After the removal of the bacterial toxin the CBF was restored to the initial level. In the present study the adenosine receptor antagonist could not prevent the ciliostimulative effect of exogenous ATP combined with the physical stimulation. Both exogenous ATP and physical stimulation have a clear but temporary stimulative effect on the ciliary beat depressed by bacterial toxin, ATP being slightly more effective. It seems that the effects of ATP are not mediated by adenosine receptors.

Localization of function in the eustachian tube: a hypothesis.

This communication uses our recent quantitative morphometric studies to develop a hypothesis for localization of ventilatory, clearance, and protective functions in different portions of the eustachian tube (ET). We hypothesize that the ET roof is involved mainly with ventilation and that the ET floor is involved mainly with clearance, whereas both portions are involved with protection of the middle ear. The hypothesis states that 1) the cross-sectional shape of the ET lumen and the attachment of the tensor veli palatini muscle to the tip of the lateral lamina are important for ventilation, 2) the ciliated cells in the ET floor subserve clearance, 3) the richly distributed elastin in the hinge portion of the ET cartilage, Ostmann's fatty tissue, and rich mucosa-associated lymphoid tissue are protective, 4) the well-developed ET cartilage lateral lamina is important for both ventilation and protection, and 5) the rich mucosal folding, goblet cells, and glands subserve both clearance and protection. Finally, the utility of the hypothesis for elucidating anatomic factors in otitis media is discussed.

Effect of exogenous ATP on ciliary beat of human ciliated cells studied with differential interference microscope equipped with high speed video.

To study the detailed effects of exogenous adenosine triphosphate (ATP) on ciliary function, we used the differential interference microscope equipped with high speed video and evaluated ciliated cells from the human sinus mucosa in monolayer culture. With this system it was possible to evaluate all parts of ciliary motility with a minimum of interference from the mucous membrane, secretory cells and the autonomic nervous system. The best direct ciliostimulative effect of exogenous ATP on ciliary beat frequency (CBF) and ciliary beat amplitude (CBA) was observed at concentrations of ATP ranging from 10(-5) M to 10(-3) M. Exogenous ATP appeared to normalize the slightly damaged ciliary motility in groups with an initial CBF of 14 Hz or higher. When the CBF was less than 14 Hz, ATP produced an increase in CBF greater than 40%, an increase in CBA greater than 30%, but these did not reach the normal level in 5 min. The biggest increases: 59.9% in CBF and 40.7% in CBA were seen in the group with an initial CBF less than 9 Hz. In the cells with a low initial CBF and unsynchronized motility exogenous ATP increased the synchrony of ciliary movement together with an increase in CBF and CBA.

Degeneration of human respiratory cell ciliary beat in monolayer cell cultures.

The degeneration of ciliary beat of human respiratory cells was studied in monolayer cell cultures by using a differential interference microscope equipped with a high speed video system. This method for studying ciliary beat in cell cultures on collagen-coated cover glasses is quite advantageous, because it allows for detailed study of all parts of ciliary function and not just ciliary beat frequency (CBF). In the present study both CBF and ciliary beat amplitude (CBA) were found to decrease continuously from the 1st day after plating but the wave form of ciliary beat did not change. Cultures with high cell density provided better preservation of normal ciliary beat for a longer period. In contrast, ciliary beat degenerated quickly in cultures with low cell density. CBF and CBA in cell cultures less than 5 days after plating were always high, supporting use of these cultures for studies of normal ciliary motility.

Mucosal folds in human eustachian tube: a hypothesis regarding functional localization in the tube.

Differences in the extent of mucosal folding in the superior versus the inferior half of the human eustachian tube (ET) lumen were studied by measuring, as it appeared in histologic cross section, the length of the mucosal margin in the pharyngeal portion, the midcartilaginous portion, the isthmus portion, and the osseous portion of each half of the ET in 16 normal ET temporal bone specimens from 16 individuals 12 to 70 years of age. The mucosal margin of the ET was significantly longer in the inferior half than in the superior half in all four portions (paired t test: pharyngeal portion t[15] = 3.59, p < .01; midcartilaginous portion t[15] = 3.52, p < .01; isthmus portion t[15] = 4.15, p < .01; osseous portion t[15] = 2.24, p < .05). These results indicate that the inferior half of the ET mucosa has more folds than the superior half, and that the surface area of the inferior half of the ET is larger than that of the superior half. This, together with the finding previously reported that secretory components of the ET such as glands and goblet cells are more richly distributed in the inferior (floor) portion than in the superior (roof) portion of the ET, may be related to the fact that the floor of the ET has an active role in clearance function. A hypothesis regarding localization of the ventilatory and clearance functions within the ET is proposed.

Distributions of eustachian tube goblet cells and glands in children with and without otitis media.

The distributions of the goblet cells and submucosal glands in eustachian tubes (ETs) from children with or without otitis media (OM) were studied. From 12 temporal bone-ET specimens obtained from children 1 to 7 years old at death, representative vertically cut histology sections were selected, stained with periodic acid-Schiff-Alcian blue and hematoxylin, and studied by light microscopy. In children without OM, the density of the goblet cells was significantly lower in the roof than in the floor of the ET and was highest in the midcartilaginous portion; glands were most richly distributed near the pharyngeal orifice, and became less densely distributed toward the tympanic portion. In children with OM, the density of the goblet cells was significantly higher both in the roof and floor portions, and the glands were more richly distributed than in the ETs of children without OM. The possibility that the observed increase in ET secretory components in children with OM impairs the passage of air through these tubes is discussed.

Ciliary beat of cultured human respiratory cells studied with differential interference microscope and high speed video system.

The ciliary beating of upper respiratory tract cells cultured on cover glasses was studied by using differential interference microscope equipped with high speed video. By culturing the cells on collagen coated cover glasses, objectives with higher magnifications could be used. With this system we could evaluate not only ciliary beat frequency, but also amplitude, wave form, orientation and synchrony of ciliary beating. Also some structural anomalies such as compound cilia and tide cilia bundles could be recognized. Ciliary beat frequency measured from 1,026 ciliated cells was 20.6 +/- 4.7 Hz (mean +/- SD). The orientation of ciliary beat directions was random and the mean standard deviation for measured angles was 73.0 degrees +/- 28.9 degrees (mean SD +/- SD). When the ciliary beat frequency was 20 Hz, the time used for effective phase was 0.022 +/- 0.002 s (mean +/- SD), and 0.028 +/- 0.004 s (mean +/- SD) for the recovery phase of beat. This system is advantageous for studying ciliary function because all parts can be studied simultaneously with higher magnification, and the effects of chemical physical mediators can be studied without disturbing effects of the autonomic nervous system or secretory cells. Also, the same cells could be observed before and after challenge with test medication and thus evaluated more accurately.

Elastin at the hinge portion of the eustachian tube cartilage in specimens from normal subjects and those with cleft palate.

The density of elastin in the intermediate portion between the lateral lamina and the medial lamina of human eustachian tube (ET) cartilage was examined in six normal adults, seven normal children, and six children with cleft palate (CP) in order to obtain information about how the physical properties of the ET cartilage differ as a function of age and presence of CP. Cross sections of the midcartilaginous portions of the ETs that had been stained with Weigert's elastin stain were photographed at uniform magnification, the area for study was projected, and the meshlike are of the ET cartilage that stained elastin-positive was represented on paper by lines. A digitizer was used to measure the total length of all the lines representing elastin in each photomicrograph, and the mean was determined for each of the three groups. The mean density of elastin was significantly greater in normal adults than in normal children (Student's t test, t = 2.781; p less than .02). It was also significantly greater in normal children than in CP children (Wilcoxon t = 24.0; p less than .05). These results appear to indicate that CP children have poorer elasticity in this area of the ET cartilage, which might cause functional obstruction (floppiness) of the ET in those children.