The pH Value as a Factor Modifying Bacterial Colonization of Sinonasal Mucosa in Healthy Persons.

OBJECTIVE: The aim of the present study is to determine and compare the range of pH value in nasal and sinus cavities in vivo regarding the presence of bacteria colonizing sinonasal mucosa among healthy subjects. METHODS: The nasal pH value measurement using a portable pH meter (Dx-pH System, Restech) and the microbiological culture swab were taken from beneath the middle turbinate and in the sinus cavity in 39 healthy subjects during maxillary bone corrective osteotomy with the Le Fort I technique. RESULTS: The mean pH value (independently of sex, P = .441) in the healthy sinus cavity was statistically higher than in the nasal middle meatus: 7.96 (SD ± 0.29) versus 7.83 (SD ± 0.30) (P = .032). Forty-eight strains of bacteria were cultured from sinus maxillaries cavities-aerobic 36.8%, aerobic and anaerobic 52.6%, anaerobic only 10.5%-and 23 strains from the nasal meatus-aerobic 25%, aerobic and anaerobic 75%. A statistically significant correlation was found between the type and location of 8 microorganisms, especially Propionibacterium acnes, identified only in the sinus cavities. CONCLUSIONS: Differences in the pH value between the middle nasal meatus and the maxillary sinus are characteristic of healthy subjects and could be associated with the diverse bacterial flora. The role of bacteria Propionibacterium acnes seems to be crucial for the pH range and sinus flora in healthy subjects.

Use of pharmacodynamic endpoints for the evaluation of levofloxacin for the treatment of acute maxillary sinusitis.

Sinusitis remains 1 of the most common reasons for antimicrobial prescriptions in the United States, with health care costs approaching $4 billion annually. We utilized the serial sinus aspirate sampling (SSAS) technique to obtain daily specimens to evaluate the time course of drug effect in patients with acute maxillary sinusitis. Eighteen patients with a radiologically confirmed acute maxillary sinusitis were enrolled into a study evaluating the relationship between levofloxacin exposure and the time course of antimicrobial effect using SSAS. SSAS was performed daily during therapy for bacteriologic evaluation. Six steady-state levofloxacin concentrations were obtained. Levofloxacin plasma and sinus aspirate concentrations were modeled using Monte Carlo Parametric Expectation Maximization algorithm implemented in S-ADAPT 1.53. Endpoints evaluated included time to resolution of signs and symptoms and time to sinus sterilization. Among the 18 enrolled patients, 15 were clinically evaluable. From these, 1 Streptococcus pneumoniae, 3 Haemophilus influenzae, 1 Moraxella catarrhalis, 1 Corynebacterium spp., and 1 coagulase-negative Staphylococcus organisms were isolated, with the latter 2 organisms being likely contaminants. For the pathogens, levofloxacin MIC values ranged from 0.03 to 2 mg/L. All pathogens were eradicated by the 4th day of therapy. The median and mean time to sinus sterilization (pathogens only) was 1 and 1.4 days, respectively. The median time to resolution of each sign and symptom ranged from 1.5 to 12-19 days, with the 83% of total signs and symptoms resolved by the end of therapy (day 5). The mean plasma area under the concentration-time curve (AUC) (mg x h/L) was 100.1 (n = 14, %CV = 27). Plasma AUC/MIC ratios ranged from 33.9 to 1696 for isolated pathogens. In this pilot SSAS study, levofloxacin rapidly eradicated isolated pathogens from the maxillary sinus.

The influence of the size of the maxillary sinus ostium on the nasal and sinus nitric oxide levels.

BACKGROUND: Nitric oxide (NO) is produced in significant quantities in the nasal sinuses and is thought to have a beneficial effect on the mucociliary transport of the sinuses and nose and to have significant antibacterial properties that contribute to the health of the sinuses. Recently, the concept of "mini-functional endoscopic sinus surgery" has been introduced where the uncinate is removed without enlargement of the maxillary ostium. Although no scientific evidence has been published, enlargement of the ostium is thought to possibly disrupt the mucociliary pathway and decrease the concentration of NO in the nose and sinuses. The aim of this study was to establish the effect of enlargement of the maxillary ostium on sinus and nasal NO. METHODS: Twenty-nine patients who were post-endoscopic sinus surgery were included with 52 who were maxillary sinus ostia cannulated. There were 22 large maxillary sinus ostia and 30 small ostia. Smoking, allergy status, and topical steroid use were recorded. NO levels were measured in the nose and maxillary sinus after decongestion with patients mouth breathing and breath holding. RESULTS: This study shows that enlargement of the maxillary sinus ostium above its normal size (20 mm2) produces a significant decrease in both the maxillary sinus and the nasal cavity NO levels. In addition, the size of the ostium showed a significant correlation to the sinus NO level. Use of topical nasal steroid sprays and topical decongestants were shown to effect NO levels in the sinuses and nasal cavity. The lowered levels of NO were found irrespective of the technique of measurement of the NO. CONCLUSIONS: The effect of this lowered NO level on the susceptibility of the maxillary sinuses to recurrent infection is yet to be determined.

The paranasal sinuses as reservoirs for nitric oxide.

OBJECTIVE: Nitric oxide (NO) is an important mediator and inflammatory marker in human upper airways. Enzymes responsible for NO production have been demonstrated both in the nose and in the paranasal sinuses, but NO levels in the sinuses are reported to be several times higher than those in the nose. It has been postulated that the paranasal sinuses may be the primary sites for NO production in the upper respiratory tract. The present study was designed to compare the NO levels sampled from the nose with those found in the paranasal sinuses. MATERIAL AND METHODS: NO levels in the maxillary sinus and nose were determined using a continuous chemiluminescence measuring technique in seven healthy volunteers. RESULTS: When NO was sampled, via a drainage tube inserted into the maxillary sinus, a transient peak in NO level was recorded. The maximal NO level (5,761 +/- 1,513 ppb; n = 7) was reached within 10 s and was followed by the establishment of a lower steady-state level (304 +/- 51 ppb). When NO was continuously sampled from the nose a steady-state level, similar to that found in the sinus, was immediately established (313 +/- 52 ppb). CONCLUSION: The data presented confirm previous findings of extremely high NO levels in the paranasal sinuses and suggest that these cavities may also function as reservoirs for NO.

Production of beta-defensin antimicrobial peptides by maxillary sinus mucosa.

beta-Defensins are endogenous cationic peptides with broad-spectrum antimicrobial activity that are thought to play a role in the innate immune response. Two human beta-defensins, beta-defensin-1 (HBD-1) and beta-defensin-2 (HBD-2), have been identified. These peptides have recently been characterized in several human tissues. The presence of these peptides in the paranasal sinuses has not been investigated. We examined maxillaary sinus secretions from six patients with sinusitis and 10 patients without signs, symptoms, or radiologic evidence of sinus disease for the presence of beta-defensins. Cationic peptides were extracted from antral lavage specimens and examined for the presence of HBD-1 and HBD-2 by Western blot. Normal maxillary sinus epithelium was obtained from two patients and analyzed by RT-PCR for the presence of HBD-1 and HBD-2 mRNA. Tissue immunostaining for the two peptides was also used. Western blot analysis identified HBD-1 in two of 10 patients in the control group and in three of six patients in the sinusitis group. HBD-2 was identified in one of 10 patients in the control group and in four of six patients in the sinusitis group. RT-PCR revealed HBD-1 mRNA in one of two normal controls tested. Immunostaining localized HBD-1 and HBD-2 to the epithelial cell cytoplasm. This is the first demonstration of HBD-1 and HBD-2 production in the paranasal sinuses. In the present study, HBD-1 and HBD-2 were detected more frequently in the maxillary sinus fluid of patients with inflamed sinuses than in normal controls.

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

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

Surfactant protein A in rabbit sinus and middle ear mucosa.

In the present study, pulmonary surfactant protein A (SP-A) messenger RNA (mRNA) and protein were characterized in adult rabbit middle ear and maxillary sinus. Fifteen adult rabbits were used for the study: 6 with evidence of acute middle ear infections and maxillary sinusitis, 6 with infections that were successfully treated with tetracycline, and 3 that were pathogen-free. We detected SP-A mRNA in maxillary sinus and middle ear tissues by Northern blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR). The RT-PCR also revealed the presence of SP-B and SP-C mRNA in middle ear and sinus tissues. We detected SP-A protein, of molecular weight approximately 29 and 70 kd, in middle ear and sinus tissues by immunoblot analysis. Unlike the SP-A protein present in the lung, the molecular weight of the SP-A protein present in the middle ear and paranasal sinus was not altered by digestion with an enzyme that cleaves N-linked carbohydrates. Immunostaining and in situ hybridization showed that SP-A protein and mRNA, respectively, were present in surface epithelial cells of the middle ear and in epithelial cells of submucosal glands in sinus tissues. These data provide the first evidence of the presence of pulmonary surfactant proteins in the paranasal sinuses and confirm previous reports of SP-A in the middle ear epithelium.

Urokinase-type plasminogen activator and plasminogen activator inhibitor antigen in tissue extracts of paranasal sinus mucous membranes affected by chronic sinusitis and antrochoanal polyps.

We examined the effect of pH on the extraction of urokinase-type plasminogen activator (u-PA) and plasminogen activator inhibitor-1 (PAI-1) from paranasal sinus mucous membrane associated with chronic sinusitis and antrochoanal polyps. The specific activity of u-PA extracted with buffer at pH 7.4 was stronger than that extracted with buffer at pH 4.2. The antigen level of u-PA extracted with the acidic buffer was significantly higher than that extracted with the neutral buffer. In contrast, the difference in antigen levels of PAI-1 extracted with the acidic buffer and neutral buffer was not significant. Based on these results, we inferred that the u-PA-PAI-1 complex was extracted by the acidic buffer and the activity of u-PA was therefore decreased.

ICAM-1 expression in upper respiratory mucosa is differentially related to eosinophil and neutrophil inflammation according to the allergic status.

BACKGROUND: Chronic non-infectious sinusitis is an inflammatory disease sharing numerous features with both allergic and non-allergic rhinitis and asthma. METHODS: In an attempt to analyse the underlying mechanisms of sinus and nasal inflammation, we assessed ICAM-1 immunoreactivity on maxillary sinus, nasal polyp and turbinate biopsies obtained surgically from 21 patients with sinusitis and chronic allergic rhinitis (AR), 23 patients with sinusitis and chronic non-allergic rhinitis (NAR) and 17 non-atopic control subjects. We compared ICAM-1 expression with both eosinophil (EG1) and neutrophil (NP57 elastase) infiltrates. RESULTS: In nasal turbinates and polyps, ICAM-1 was broadly distributed within the epithelium, on ciliated and basal epithelial cells and in the submucosa on endothelial and inflammatory cells. ICAM-1 was rarely expressed in the mucosa of the sinus, compared with polyps (P < 0.01) and turbinates (P < 0.0001). Submucosal and epithelial eosinophils were increased in both sinus and nasal biopsies from AR by comparison with NAR (P=0.003 and P=0.02) and non-atopic control subjects (P<0.0001 and P=0.014). Submucosal and epithelial neutrophil numbers were increased in the two chronic rhinitis groups and correlated with ICAM-1 submucosal expression in NAR only (P<0.01). On the other hand, epithelial eosinophil numbers correlated with ICAM-1 epithelial expression in AR only (P = 0.002). CONCLUSION: This study further characterizes sinus and nasal granulocyte inflammation in chronic non-infectious sinusitis and rhinitis and suggests a differential role for ICAM-1 in neutrophil and eosinophil recruitment according to the allergic status.

Allergic fungal sinusitis in the southeastern USA: involvement of a new agent Epicoccum nigrum Ehrenb. ex Schlecht. 1824.

Alternaria alternata, Aspergillus spp., Bipolaris spicifera, Curvularia lunata, Epicoccum nigrum and Fusarium solani were isolated repeatedly from groups of patients among 96 diagnosed with allergic fungal sinusitis (AFS). Epicoccum nigrum was obtained consistently from four patients, one of whom yielded mycelial masses consistent in morphology with E. nigrum. Fifteen of the predominant fungi recovered from air samples from selected patients' residences included the same species isolated from the mucin of its inhabitants. Air samples from other buildings, whose occupants (non-AFS individuals) complained of poor indoor air quality or of symptoms of the sick building syndrome (SBS), yielded some of the same species involved in AFS. An association of SBS with AFS was not established. Eight of the species implicated in AFS were found to colonize the surfaces of indoor construction and finishing materials at sites other than the residence of the patient. To our knowledge, this is the first report that E. nigrum can colonize nasal sinuses and cause AFS.

Gaseous dynamics of the rabbit maxillary sinus.

To better understand the mechanisms underlying maxillary sinus function, the gas composition of the sinus antrum in spontaneously breathing and tracheotomized rabbits (n = 17) was compared. The gas composition of samples (n = 117) obtained from rabbit maxillary sinuses was determined by gas chromatography. Results demonstrated significant differences (P < .005) in sinus gas composition between nasal breathing and tracheotomized animals for oxygen (02) and carbon dioxide (CO2). In tracheotomized animals O2 levels decreased while CO2 levels increased markedly to suprasystemic levels. This unexpected finding may be due to reduced sinus blood flow and the effects of nasal versus tracheal respiration. We conclude that the gaseous dynamics and perhaps the function of the maxillary sinus vary under different respiratory conditions. A better understanding of these processes may lead to earlier diagnosis and the development of improved treatments for sinus disease.

Calcium phosphate in aspergillosis of the maxillary sinus.

The unique appearance of apatite in fungus balls of aspergillosis in the maxillary sinus was investigated using scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), X-ray microdiffraction and Fourier transform infrared spectroscopy (FT-IR). Rod-shaped fragments with tubular structures, and globular vesicles covered with conidia were observed in the fungus balls. Massive fragments of a solid substance were demonstrated inside the fungus balls. Calcium and phosphate were detected in necrotic areas of the fungus balls by EDS. X-ray microdiffraction and FT-IR showed the presence of an apatite-like substance, but failed to demonstrate the presence of calcium oxalate crystals usually found in such fungus balls.

Dexamethasone treatment reduces sensory neuropeptides and nerve sprouting reactions in injured teeth.

Dental injuries have been shown to generate extensive structural and cytochemical changes in sensory fibers that contain neuropeptides such as calcitonin gene-related peptide (CGRP) or substance P (SP). The present study was designed to test whether the anti-inflammatory drug dexamethasone (DEX) can alter neural responses to dental injuries. DEX (20 micrograms/100 g body weight) was given to adult rats (n = 10) prior to dental surgery and daily thereafter for 4 days. Control animals received sterile saline vehicle (n = 6) or no injection (n = 1). Each rat was then anesthetized for dental surgery and a cavity was drilled partway through dentin on the anterior side of the right maxillary first molar. Pulp exposure injuries were also made on two right mandibular molars in 14 of 17 rats. After 4 days of daily drug treatment, the rats were anesthetized and fixed by perfusion with formaldehyde-picric acid, and their jaws were prepared for immunocytochemistry. Neural CGRP immunoreactivity near the maxillary cavity injury site of DEX-treated rats was reduced more than 50% compared to controls, as determined both qualitatively and by digital analysis. The SP immunoreactive (IR) fibers in molar pulp also had extensive inhibition of neural reactions to cavity injury. DEX also reduced the immunoreactivity for CGRP and SP in normal contralateral rat molars of all treated rats, and it caused a postoperative loss of weight. Pretreatment for 1-5 days prior to the 4 day injury gave the same results as pretreatment for 1 h. The mandibular pulp exposure injuries induced a chronic abscess and advancing pulpal necrosis but did not show differences in nerve reactions between DEX-treated rats and the controls. In conclusion, the synthetic steroid dexamethasone suppressed the CGRP and SP neuropeptide immunoreactivity in normal dental nerves and it reduced nerve-sprouting responses to dentin cavity injuries; however, sensory nerve reactions to pulpal exposure injuries were not affected by DEX in these experiments.

Arachidonic acid metabolites in human nasal polyps.

Arachidonic acid metabolites (AAMs) are known to be involved in inflammation. It is suggested that AAMs play an important role in the pathogenesis of nasal polyp. We have measured the levels of prostaglandin E2, 6-keto prostaglandin F1 alpha, thromboxane B2, leukotriene B4 and a mixture of leukotriene C4, D4 and E4 in both nasal polyp and maxillary sinus mucosa by radioimmunoassay. Our results showed that arachidonic acid metabolism in nasal polyps from allergic patients was more active than that from non-allergic patients. The arachidonic acid metabolism in nasal polyp was more active than in maxillary sinus mucosa among allergic patients. On the other hand, arachidonic acid metabolism in maxillary sinus mucosa was more active than that in nasal polyps among non-allergic patients. On the basis of these results, we hypothesized the causal mechanisms of nasal polyps as follows: The nasal polyp in allergic patients is caused by primary inflammation of the nasal mucosa, and sinusitis occurs secondarily. In non-allergic patients, the primary side of inflammation is located in the maxillary sinus mucosa, leading to the secondary formation of nasal polyp.

Presence of urokinase-type plasminogen activator (u-PA) in tissue extracts of antrochoanal polyp.

Using a biochemical technique, the authors characterized and identified a plasminogen activator (PA) derived from tissue extracts of antrochoanal polyp (AP) and paranasal mucous membrane (PMM) with chronic sinusitis. The results of fibrin zymography indicated that the tissue extracts of AP revealed two lytic zones and that those of PMM revealed a single lytic zone on fibrin-agarose plates. One of the AP zones exhibited the same relative mobility as the PMM zone (molecular weight: 65 kd), while the other AP zone had a smaller molecular weight (about 54 kd). Goat immunoglobulin G (IgG) fraction of antihuman uterine tissue-type plasminogen activator (t-PA) inhibited the 65-kd lytic zones of AP and PMM. Antihuman low-molecular-weight urokinase inhibited only the 54-kd lytic zone of AP, and nonspecific goat IgG failed to inhibit any of the lytic zones. On the other hand, 10(-2) mol trans 4-(aminomethyl)cyclohexane-carboxylic acid (t-AMCHA) inhibited all of the lytic zones. No lytic zones could be observed on plasminogen-free fibrin-agarose plates. These findings confirmed that the tissue extracts of PMM contained t-PA, and that those of AP contained both t-PA and urokinase-type plasminogen activator (u-PA). In addition, it appeared that u-PA in inflammatory tissue was related to proliferative changes of the mucous membrane.

Neuropeptide Y in the rabbit maxillary sinus modulates cholinergic acceleration of mucociliary activity.

The distribution of neuropeptide Y (NPY)-immunoreactivity was investigated in the rabbit maxillary sinus and adjacent ganglia. A moderate supply of NPY-containing nerve fibers occurred around seromucous glands and a denser supply around small blood vessels. Only a few immunoreactive nerve fibers were seen beneath the epithelium. Double immunostaining showed that vasoactive intestinal peptide (VIP) coexisted with NPY in the nerve fibers surrounding blood vessels and seromucous glands. NPY-containing nerve cell bodies were numerous in the superior cervical ganglion, and moderately numerous in the sphenopalatine ganglion. The finding of NPY-containing neurons in the latter parasympathetic ganglion suggests that NPY may influence the cholinergic regulation of mucociliary activity. The effect of NPY on the mucociliary activity of the maxillary sinus in connection with cholinergic stimulation has therefore been investigated in vivo using a photoelectric technique. At dosages of 2.5 and 5.0 micrograms/kg, the ganglionic stimulant nicotine bitartrate, which increases mucociliary activity by a cholinergic pathway, accelerated mucociliary activity by 28.0 +/- 7.5% and 36.8 +/- 6.2%, respectively. In the same experiment repeated during infusion of NPY (0.1 microgram/kg/min), the increase in mucociliary activity was reduced to 10.8 +/- 2.3% and 28.9 +/- 7.1%, respectively. Infusion of NPY did not affect the stimulating effect on mucociliary activity by bolus injections (0.1 and 0.5 microgram/kg) of the cholinergic agonist, methacholine. It is concluded that NPY-like immunoreactivity is present in nerve fibers in the rabbit maxillary sinus and in neurons in the sympathetic and parasympathetic ganglia that supply the nose and paranasal sinuses. NPY attenuates the effect of nicotine on mucociliary activity, probably via a prejunctional mechanism, and may act as a modulator of cholinergic regulation of the mucociliary system.

Immunohistochemical distribution of fibronectin in the mucosa with chronic sinusitis.

The distribution of fibronectin (FN) in operative specimens of maxillary sinus mucosa from 35 patients with chronic sinusitis was examined immunohistochemically. Mouse monoclonal antibody to human FN was used for the peroxidase-antiperoxidase staining technique. FN was found in most of the serous cells and in some (about 10%) of the mucous cells in the nasal glands, and also in the nasal fluid in the lumina. It was not found elsewhere, be it in the epithelial cells, the basement membrane, or the interstitial tissue, except in two patients. These results suggest that FN is produced in the nasal glands and secreted into the nasal fluid in the maxillary sinus in chronic sinusitis.