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Biomedical subjects

W K Dolen

Publications and source records attributed to W K Dolen.

At least 19 recordsLinked to original sources

Local and systemic eosinophil activation in allergic fungal sinusitis.

BACKGROUND: In allergic fungal sinusitis diagnostic and monitoring criteria are not firmly established, and the role of eosinophils in pathogenesis is not clear. OBJECTIVE: To determine whether assessment of eosinophil activation by measurement of eosinophil cationic protein in serum or allergic mucin would be useful in distinguishing patients with allergic fungal sinusitis from patients with chronic sinusitis of other etiologies. METHODS: Thirteen patients referred for possible allergic fungal sinusitis were evaluated and given a definite allergic fungal sinusitis diagnosis if they met five of the following six criteria: (1) history and physical not suggesting another etiology, (2) sinus computed tomography consistent with allergic fungal sinusitis, (3) typical allergic mucin, (4) fungus isolated from allergic mucin, (5) presence of fungal-specific IgE, and (6) elevated total IgE. Eosinophil cationic protein, a marker of eosinophil activation, was measured in serum and allergic mucin. RESULTS: Nine patients met criteria for allergic fungal sinusitis. All patients had nasal polyps and were atopic. Eight of the patients had allergic rhinitis and three had asthma. Mean total IgE at surgery was 1,385 IU/mL. A fungus was isolated from allergic mucin of eight patients. All patients demonstrated fungal-specific IgE. Mean allergic mucin eosinophil cationic protein levels obtained at surgery were significantly higher in patients with allergic fungal sinusitis than in four patients not meeting strict diagnostic criteria, and in 16 control patients having sinus surgery for other indications. There was no significant difference in serum eosinophil cationic protein levels between the three groups. Serial allergic mucin eosinophil cationic protein levels appeared to correspond with disease activity in some allergic fungal sinusitis patients. CONCLUSIONS: Eosinophils in allergic mucin are activated. Measuring eosinophil cationic protein may be useful in diagnosis of allergic fungal sinusitis and in monitoring response to therapy.

Adolescent↗

Allergenic fungi in allergic fungal sinusitis.

BACKGROUND: Patients with allergic fungal sinusitis demonstrate skin test reactivity to many fungal extracts. Various fungi have been isolated from the characteristic allergic mucin. OBJECTIVE: This study was designed to identify allergens in allergic mucin and to compare them to those found in commercial fungal extracts. METHODS: Allergic mucin was collected during functional endoscopic sinus surgery from 11 patients meeting strict diagnostic criteria for allergic fungal sinusitis and from three allergic rhinitis patients with chronic sinusitis. A portion was solubilized in saline and centrifuged. To identify allergens, proteins in allergic mucin and fungal extracts were separated by SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose and immunostained using patient sera and enzyme-labeled anti-human IgE. RESULTS: All patient sera recognized numerous bands ranging from 18 to 90 kD. In mucin, bands were consistently found in the 35 to 50 kD range. Corresponding bands in fungal extracts were found in only 1/11 patients with allergic fungal sinusitis. Sera from 4/11 patients detected an 18-kD protein in allergic mucin, but sera from all patients with allergic fungal sinusitis recognized an 18-kD protein in commercial fungal extracts. Sera from selected patients with allergic fungal sinusitis detected human epithelial proteins in the 35 to 50 kD range. CONCLUSIONS: Fungal allergens were not detected in allergic mucin of all patients with allergic fungal sinusitis. The 18-kD allergen appears to be shared by many fungi, and may be a fungal panallergen. The source of the apparent allergens in the 3550-kD range warrants further study.

Allergens↗

Fungal sinusitis: an update.

OBJECTIVE: To review the classification of fungal sinusitis as well as discuss current approaches to diagnosis and management. DATA SOURCES: A MEDLINE literature search was performed using the index terms sinus infection, fungal, diagnosis, radiology, microbiology, and treatment. The search was restricted to the English language and human subjects. With one exception the references were restricted to the last 10 years. Clinical data from studies performed at our institution were also included. RESULTS: Fungal sinusitis can be divided into four primary categories: (1) acute/fulminant (invasive), (2) chronic/indolent (invasive), (3) fungus ball, and (4) allergic fungal sinusitis. Each subtype has unique immunologic, pathologic, and clinical features. Allergic fungal sinusitis is the most recently described and most common form. The treatment and prognosis of fungal sinusitis varies significantly among the four different categories. CONCLUSION: Recent advances in endoscopy and computed tomography have enhanced the understanding of fungal sinusitis; however, they remain diseases surrounded by controversy. New insights into the etiology and pathogenesis of these diseases along with advances in diagnosis and treatment will lead to improved medical therapy.

Humans↗

Laboratory evaluation of a commercial immunoassay for fire ant allergen-specific IgE antibodies.

BACKGROUND: In vitro testing for fire ant sensitization would be useful for research purposes and in special clinical situations. METHODS: Laboratory performance of a commercial assay (Pharmacia CAP System, [PCS]), for specific IgE to Solenopsis invicta whole body extract was studied in 46 persons. Assay results were compared with those of venom skin testing, RAST, and ELISA. The manufacturer's global cutoffs were compared with cutoffs set by using methods derived from analytical detection limit theory. RESULTS: Thirty-two study subjects had positive skin test results, and 14 had negative results. Raw PCS data demonstrated a high level of correlation with RAST (rho = 0.941) and ELISA (rho = 0.931), and showed good correlation with skin testing (rho = -0.769). Analysis of binormal receiver operating characteristic curves, using skin test results as the reference standard, demonstrated no difference in performance among the three assays. The fixed global quantitative cutoff of 0.35 kUa/L was relatively insensitive. Use of the manufacturer's qualitative alternate scoring method cutoff substantially increased sensitivity without loss of specificity, as did lower limit of detection set by use of diluent. CONCLUSIONS: In situations in which skin testing for fire ant sensitization is not feasible, PCS appears to be an acceptable in vitro alternative method for determination of fire ant allergen-specific IgE.

Allergens↗

Use of standardized and conventional allergen extracts in prick skin testing.

This study examined whether commercially available conventional and standardized allergen extracts differ enough in potency to affect routine prick skin test results. Extracts of white oak, timothy, Bermuda, Russian thistle, short ragweed, sagebrush, Alternaria, and cat dander were examined in allergic patients and in nonatopic subjects with no personal or family history of asthma, rhinitis, or eczema. Conventional nonstandardized extracts (1:10 or 1:20 wt/vol) from two sources were compared with three concentrations (100,000, 10,000, and 1000 AU/ml) of a single standardized extract. Preparations were compared in the allergic patients with computerized planimetry, and in all patients and subjects with a conventional skin test grading system. Skin test area for the conventional extracts generally fell between the 10,000 and 100,000 AU/ml concentrations of the standardized extract. Skin test reactivity to at least one allergen extract occurred in 31% of the nonatopic subjects; there was no difference between the number of 3+ and 4+ reactions for conventional and standardized extracts. Results indicate that standardized and conventional extracts are frequently similar, but are not directly interchangeable.

Adolescent↗

The histamine content of allergen extracts.

The histamine content of 108 inhalant, food, and venom extracts from four commercial sources was measured by chemical (glass fiber-based) and immunologic (competitive RIA) methods. Histamine was present in 64 of 76 inhalant extracts (range, 0.005 to 7.4 micrograms/ml), 20 of 26 food extracts (range, 0.16 to 23 micrograms/ml), and six of six venoms, 100 micrograms/ml (range, 1.0 to 38 micrograms/ml). Histamine was removed by treatment with diamine oxidase or dialysis of an extract. Repeat assay of selected extracts after addition of known amounts of histamine in the glass fiber-based method produced additive results, and glycerin- or phenol-extract preservatives did not affect assay performance. Timed extractions of dried-pollen grains demonstrated maximal histamine content at 30 seconds, suggesting that histamine is an inherent component of at least some pollens. Histamine found in some allergen extracts could, under extreme circumstances, produce false-positive results in skin testing and in basophil histamine release assays, and could affect the result of research that uses intact pollen or allergen extracts.

Allergens↗

Aerobiology of the Colorado Rockies: pollen count comparisons between Vail and Denver, Colorado.

Pollen patterns were compared between Vail, CO (8,200 feet elevation), Aspen, CO (7,900 feet) and Denver, CO (5,280 feet) from 1984 through 1988. Counts were obtained at all sites with a volumetric intermittent cycling rotating impaction sampler. Aspen and Denver were compared in 1984, and Vail and Denver from 1985 through 1988. While counts were generally lower in the mountain sites than Denver, certain pollens, especially trees, were quite high. Ragweed was essentially absent from Aspen and Vail, and chenopod-amaranth counts were very low. Cedar, pine, and aspen frequently pollinated despite active snowfall.

Air Pollution↗

Immunoassay of specific IgE: low level assays require measurement of allergen specific assay background.

The allergen-specific backgrounds of a paper disk radioimmunoassay (RIA) system and a cellulose sponge fluorescent enzyme immunoassay (FEIA) system were evaluated using three inhalants, timothy, short ragweed, and cat, with reagents obtained from the same manufacturer. Radioimmunoassay was performed with Phadebas RAST reagents by the modified RAST method, and FEIA by the Pharmacia CAP System. Horse serum, 5% HSA, assay diluent, and serum pools from nonallergic and allergic patients, were assayed. The lower limit of detection (LLD) was defined using both the Z distribution (as is conventional) and the t distribution. The solid phases, analytes, and assays differed (p less than .001) in background results. For RIA, background was lowest for timothy and highest for cat; for FEIA, background was lowest for cat and highest for short ragweed. For RIA, background assessed with the allergic serum pool was higher than the other analytes; for FEIA, responses of the five analytes did not differ. For timothy and short ragweed, background of RIA was lower than FEIA. For FEIA, the highest LLD calculated using the Z distribution was 11 SD lower than the manufacturer's recommended quantitative cutoff; for RIA, the highest LLD calculated was 2.5 SD higher than the recommended analytic cutoff. The analytic false positive rate for RIA may differ between allergic and nonallergic patient populations. Laboratories reporting results near either assay's background should set LLD based on assay of allergen-specific negative controls in each assay run.

Allergens↗

Immunoassay of specific IgE: use of a single point calibration curve in the modified radioallergosorbent test.

Interest in immunoassay standardization has prompted development of specific IgE assays reporting results related to the international IgE reference. To examine the single point calibration curve employed in the modified RAST assay (MRT) to convert MRT counts to IgE units, independent dilutions of a 25 kU/L total IgE reference and nine allergic sera (three each for short ragweed, cat, and timothy) were made in horse serum and assayed. In a log-log plot, the single point curve was, by definition, linear over its entire range; the dilution curve was curvilinear because of reagent system saturation, which was at 7 kU/L. Curves were not parallel (P less than .001). Allergen-specific dilution curves showed saturation points at values similar to or less than the total IgE system. The linear portions of these curves paralleled the total IgE dilution curve but not the single point curve. This lack of parallelism would have resulted in varying magnitudes of error in estimation of IgE antibody levels in the upper and lower assay ranges, and would imply a lower detection limit for IgE than that which the assay actually has. Modified RAST assay is not appropriate in research or a clinical situation in which accurate quantitative results are needed. Modified RAST assay would furthermore be an inappropriate means of assigning units to proposed reference preparations for standardization.

Allergens↗

Comparison of skin testing and three in vitro assays for specific IgE in the clinical evaluation of immediate hypersensitivity.

A new assay, Pharmacia CAP System (PCS), for allergen-specific IgE (sIgE) was evaluated in 198 new patients presenting with respiratory symptoms to an urban allergy practice. An experienced allergist examined each patient and clinically assessed sensitivity to timothy, short ragweed, Alternaria tenuis, cat, or D. farinae. Puncture and selected intracutaneous skin tests (ST) with these inhalant extracts were then performed. The physician again rated the likelihood of clinical sensitivity to each inhalant, and serum was obtained for sIgE measurements by Phadebas RAST, modified RAST, and PCS. Results of the three in vitro tests (IVT) correlated well with each other and generally agreed with physician assessments and ST results. Individual differences for extracts and assay methods were identified. A few patients with negative ST had positive IVT, but more patients with positive ST were negative by IVT. Modified RAST had greater sensitivity but less specificity than the other two IVT. Analysis of receiver operating characteristic curves showed that sensitivity of the three assays when compared at the 95% level of specificity, did not differ. This result suggests that the cutoff criterion for a positive modified RAST result is too low and should be reevaluated. Skin tests remain the most sensitive and specific test available. The Pharmacia CAP System is a clinically useful assay for sIgE and appears to be a clear advancement for IVT technology.

Allergens↗

Assessment of nasal patency by rhinoscopic measurement of cross sectional nasal airway area: correlation with subjective nasal symptoms.

Assessment of nasal patency by the recording of nasal symptom scores was compared with an objective method of determining nasal airway area using a fiberoptic rhinoscope. Sixty patients with active allergic rhinitis and nasal congestion requiring treatment were studied. Nasal symptoms were recorded and nasal airway area was measured before and at fixed time intervals after administration of either pseudoephedrine or oxymetazoline. Both methods detected a decongestant response to both drugs, and the symptom of congestion correlated with the measured nasal airway area. Rhinoscopic measurement of cross sectional nasal airway area is an objective method that may be used to complement other methods for evaluation of nasal patency.

Adult↗