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Biomedical subjects

J Savolainen

Publications and source records attributed to J Savolainen.

At least 19 recordsLinked to original sources

alpha-Chymotrypsin-catalyzed degradation of desmopressin (dDAVP): influence of pH, concentration and various cyclodextrins.

Desmopressin [1-(mercaptopropanoic acid)-8-D-arginine vasopressin; dDAVP] is a vasopressin analogue with a selective antidiuretic effect. The oral bioavailability of desmopressin is limited due both to its high hydrophilicity leading to a low intestinal permeability and to low enzymatic stability. The degradation of desmopressin was investigated in aqueous buffer solutions (pH 6.00-9.00) containing the enzyme alpha-chymotrypsin at a concentration of 0.50 mg/ml at 37 degrees C. The degradation of desmopressin was also studied in solutions containing alpha-chymotrypsin in the concentration range 0.10-1.00 mg/ml (pH 7.40 and 37 degrees C). The rate of degradation was shown to be highly dependent on both enzyme concentration and pH. Maximal alpha-chymotrypsin activity was observed in the pH range 7.40-8.00. It was observed that phenylalanine was formed during the degradation of desmopressin. Phenylalanine was formed in the amount of 20% in 120 min. In the same time period 95% of desmopressin was degraded. The formation of phenylalanine can be explained from the substrate specificity of alpha-chymotrypsin. Cyclodextrins are known to stabilize drugs including peptides against both chemical and enzymatic degradation. In this study it was shown that hydroxypropyl cyclodextrins (alpha, beta and gamma) stabilized desmopressin against alpha-chymotrypsin-catalyzed degradation. The stabilization was by a factor of 3, 9 and 8 at the concentration 12.5% (w/v) for hydroxypropyl-alpha-cyclodextrin, hydroxylpropyl-beta-cyclodextrin and hydroxypropyl-gamma-cyclodextrin.

Chymotrypsin

Human isometric force production and electromyogram activity of knee extensor muscles in water and on dry land.

This study was designed to determine trial-to-trial and day-to-day reproducibility of isometric force and electromyogram activity (EMG) of the knee extensor muscles in water and on dry land as well as to make comparisons between the two training conditions in muscle activity and force production. A group of 20 healthy subjects (12 women and 8 men) were tested three times over 2 weeks. A measurement session consisted of recordings of maximal and submaximal isometric knee extension force with simultaneous recording of surface EMG from the vastus medialis, vastus lateralis and biceps femoris muscles. To ensure identical measurement conditions the same patient elevator chair was used in both the dry and the wet environment. Intraclass correlation coefficients (ICC) and coefficients of variation (CV) showed high trial-to-trial (ICC = 0.95-0.99, CV = 3.5%-11%) and day-to-day reproducibility (ICC=0.85-0.98, CV=11%-19%) for underwater and dry land measurements of force and EMG in each muscle during maximal contractions. The day-to-day reproducibility for submaximal contractions was similar. The interesting finding was that underwater EMG amplitude decreased significantly in each muscle during maximal (P < 0.01-P < 0.001) and submaximal contractions (P < 0.05-P < 0.001). However, the isometric force measurements showed similar values in both wet and dry conditions. The water had no disturbing effect on the electrodes as shown by slightly lowered interelectrode resistance values, the absence of artefacts and low noise levels of the EMG signals. It was concluded that underwater force and EMG measurements are highly reproducible. The significant decrease of underwater EMG could have electromechanical and/or neurophysiological explanations.

Adult

Local reactions and IgE antibodies to pertussis toxin after acellular diphtheria-tetanus-pertussis immunization.

Local reactions and pertussis toxin specific immunoglobulin E antibodies (PT-IgE) were investigated in healthy children following primary and booster immunization with a combined diphtheria tetanus acellular pertussis vaccine (DTPa) including pertussis toxin, filamentous haemagglutinin and pertactin. A primary series of DTPa was administered to 150 infants, and 104 of them received a booster dose of DTPa combined with inactivated polio vaccine at 2 years of age. PT-IgE was measured in serum samples from 72 children using a modified nitrocellulose RAST. Primary immunization was associated with low incidence of local reactions (1%-5%). After the booster dose 21% of children had a local reaction >/=20 mm. Local reactions after the booster dose tended to be more common in children who had experienced reaction at primary immunization. PT-IgE was detected in 18% and 86% of children following primary and booster vaccinations, respectively. Allergic and non-allergic children did not differ in PT-IgE responses. After primary immunization, elevated PT-IgE levels were found more often in children with a family history of allergy than in those without known allergy in the family. Children with local reactions had significantly higher pre- and post-booster PT-IgE levels and median post-booster pertactin IgG and diphtheria-IgG levels than children without local reactions. Conclusion Acellular pertussis immunization induces IgE antibodies to pertussis toxin, especially after booster vaccination. The higher median pre- and post-booster levels of pertussis toxin specific immunoglobulin E and post-booster levels of IgG to pertactin and diphtheria in children with local side-effects reflect a multifactorial immunological mechanism of such reactions.

Child, Preschool

Drug release from a porous ion-exchange membrane in vitro.

The effect of environmental ionic strength on the rate of drug release from a cation exchange membrane was evaluated. Cationic propranolol-HCl, timolol, sotalol-HCl, atenolol and dexmedetomidine-HCl and neutral diazepam were adsorbed onto a porous poly(vinylidene fluoride) (PVDF) membrane that was grafted with bioadhesive poly(acrylic acid) chains (PAA-PVDF). Despite its porosity, the PAA-PVDF membrane acted as a cation exchange membrane. The release of adsorbed drug from the PAA-PVDF membrane was investigated by using a USP rotating basket apparatus. Adsorption of cationic drugs onto the PAA-PVDF membrane tended to increase with increasing lipophilicity of the drug. A decrease in the ionic strength of the adsorption medium increased the amount of the cationic drugs adsorbed onto the membrane, but had no effect on diazepam adsorption. The release of cationic drugs from the PAA-PVDF membrane was greatly affected by the ionic strength of both the adsorption medium and the dissolution medium, while ionic strengths did not affect diazepam release. Our results suggest that the ionic strength of both the adsorption and dissolution media substantially affects the release rate of a drug that has been adsorbed onto the ion exchange membrane, primarily via electrostatic interactions, while ionic strength has no effect on the release of a drug which has been adsorbed onto the membrane via non-electrostatic forces.

Acrylic Resins

Cross-reacting IgE and IgG antibodies to Pityrosporum ovale mannan and other yeasts in atopic dermatitis.

Atopic dermatitis (AD) patients often demonstrate positive skin prick test results and serum IgE antibodies to a range of different yeasts. This has been thought to be due to cross-reactivity. In this study, the cross-reactivity of IgE and IgG antibodies between mannan and crude antigens of Pityrosporum ovale, Candida albicans, and Saccharomyces cerevisiae and crude antigens of Cryptococcus albidus and Rhodotorula rubra was examined by RAST and ELISA inhibition with two serum pools of AD patients. We found cross-reacting IgE and IgG antibodies. In the IgE response, the main cross-reacting pattern was the mannan region, although inhibition could be achieved also with crude antigens of C. albicans, S. cerevisiae, and, to some extent, C. albidus. P. ovale was the most potent inhibitor of IgE-binding components, and against it the highest IgE antibody levels were detected in AD serum pools. In contrast, C. albicans was found to be the most important inducer of IgG antibodies, since the IgG level against P. ovale mannan in both AD serum pools was very low. Cross-reacting antibodies were also seen in ELISA inhibition with both crude and mannan antigens, but since the IgG antibody level of P. ovale mannan in AD serum pools was low, further studies are needed to confirm the IgG results.

Antibodies, Anti-Idiotypic

Candida albicans mannan- and protein-induced humoral, cellular and cytokine responses in atopic dermatitis patients.

BACKGROUND: The cytokine observed most often in atopic dermatitis (AD) is IL-4, but a role for IL-5 and IFN-gamma in the late and delayed phase reactions has been suggested. In AD with head, neck and shoulder distribution, hypersensitivity to saprophytic yeasts is an important pathogenetic factor. The yeast allergens include both the mannan polysaccharides and the proteins. Mannans are major cross-reacting allergens likely to be involved in the pathogenesis of AD. OBJECTIVE: To characterize the humoral, lymphoproliferative and cytokine (IL-2, 4, 5 and IFN-gamma) responses of peripheral blood mononuclear cells (PBMCs) induced by Candida albicans mannan and protein antigens in AD. METHODS: Fifteen AD patients and seven healthy controls were included. Ficoll-isolated PBMCs were stimulated by PHA and laboratory-generated mannan and protein extracts of C. albicans. Lymphocyte proliferation was measured and cytokine production was studied by ELISA. The antigen-specific IgG and IgE antibodies were analysed by ELISA and nitrocellulose RAST. RESULTS: In AD mannan (P < 0.005) and protein (P < 0.002), specific IgE levels were higher than in healthy controls. Both mannan and protein-specific lymphoproliferations (both: P < 0.02) were higher in AD than in healthy controls. Mannan, but not protein, induced long lasting IL-2 and IL-4 productions from 24 h lasting up to 66-96 h and IL-5 and IFN-gamma productions with elevated levels at 66 and 96 h. The mannan-induced IL-2 (P = 0.015) and IFN-gamma (P < 0.005) were increased in AD as compared with healthy controls. Significant correlations were seen between the protein-induced proliferation responses and both serum total IgE (r = 0.59, P < 0.01) and protein-specific IgE (r = 0.65, P < 0.005). The mannan-induced IL-2 responses correlated with the specific IgE (r = 0.62, P < 0.01) and proliferation (r = 0.51, P < 0.02) and S-IgE level (r = 0.71, P < 0. 002). Mannan-induced IL-4 and IFN-gamma productions also correlated (r = 0.43, P < 0.05). CONCLUSIONS: C. albicans mannan induced elevated IL-2 and IFN-gamma responses in AD patients. The correlations of the cytokine responses with mannan-induced IgE and proliferation responses suggest that C. albicans mannan induced TH1 type cytokine responses are involved in AD.

Adult

Muscle capillary supply in hind limb and diaphragm of the common shrew (Sorex araneus).

Shrew species of the subfamily Soricinae have unusually high metabolic rates when compared to Crocidurinae shrews and other similar-sized mammals. The aim of this study was to clarify whether the high basal metabolic rate of Soricinae shrews is reflected in a high capillary density in their muscles. To this end, the capillary supply of four limb muscles and diaphragm of the common shrew (Sorex araneus) was quantified from cross-sectioned muscles. The capillary densities of the limb muscles were 2575 +/- 329, 3111 +/- 299, 2812 +/- 197 and 2752 +/- 173 capillaries mm-2 fibre area in gastrocnemius lateralis, g. medialis, plantaris and soleus, respectively. Capillary density of the shrew diaphragm (6691 +/- 1057) was double that of the limb muscles. This value is among the highest ever measured in mammals. In general, the capillary supply in the hind limb of the common shrew is about 3-4 times higher than commonly found in the leg muscles of the laboratory rat or other bigger mammals, but similar to those in Crocidurinae shrews and some small rodents. Thus the high resting metabolism of the common shrew is not associated with an extraordinarily high capillary density. The apparent disparity between basal metabolic rate and muscle capillary supply in S. araneus is probably due to the small aerobic scope of shrews in the subfamily Soricinae.

Analysis of Variance

The common cold: effects of intranasal fluticasone propionate treatment.

OBJECTIVE: A double-blind, randomized, placebo-controlled trial was conducted to study the effect of the intranasal corticosteroid, fluticasone propionate (FP), in the naturally occurring common cold. METHODS: One hundred ninety-nine young adults received high-dose FP (200 microg four times daily) or placebo beginning 24 to 48 hours after onset of the common cold for 6 days. All symptoms were recorded on diary cards on days 1 to 20, and clinical examinations were carried out on days 1, 7, and 21. Nasopharyngeal aspirates were collected on days 1 and 7 for detection of rhinoviruses (found in 105 subjects) and Streptococcus pneumoniae, Haemophilus influenzae, or Moraxella catarrhalis (found in 52 subjects) in the nasopharynx. RESULTS: In general, FP treatment had no clinically recognizable effects on the symptoms of the common cold, although it significantly reduced nasal congestion and cough on some study days. After treatment, rhinoviruses were cultured more often in the FP treatment group (37% vs 14%, p < 0.001), but this had no effect on the symptoms of common cold. FP treatment produced no changes in the colonization of pathogenic bacteria in the nasopharynx. Some symptoms of common cold were significantly more severe during days 1 to 10 (p < 0.05) in subjects found to have positive cultures for S. pneumoniae, H. influenzae, or M. catarrhalis in the nasopharynx on day 1 (n = 33). CONCLUSION: FP treatment does not have any marked effects on the symptoms of the common cold. FP treatment induced prolonged shedding of viable rhinoviruses. Some symptoms of the common cold were significantly more severe in subjects with pathogenic bacteria in the nasopharynx.

Administration, Intranasal

Co-administration of a water-soluble polymer increases the usefulness of cyclodextrins in solid oral dosage forms.

PURPOSE: The aim of this study was to investigate the effect of cyclodextrins (beta-CD, HP-beta-CD and (SBE)7m-beta-CD), and co-administration of a water-soluble polymer (HPMC) and cyclodextrins, on the oral bioavailability of glibenclamide in dogs. METHODS: Effects of cyclodextrins on the aqueous solubility of glibenclamide, with and without hydroxypropylmethylcellulose (HPMC), were determined by a phase-solubility method. Solid inclusion complexes were prepared by freeze-drying. Glibenclamide was administered orally and intravenously to beagle dogs. RESULTS: Aqueous solubility of glibenclamide increased as a function of cyclodextrin concentration, showing an AL-type diagram for beta-CD and an Ap-type diagrams for both of the beta-CD derivatives studied. HPMC enhanced the solubilising effect of cyclodextrins, but did not affect the type of phase-solubility diagram. Orally administered glibenclamide and its physical mixture with HP-beta-CD showed poor absolute bioavailability, while orally administered glibenclamide/cyclodextrin-complexes significantly enhanced the absolute bioavailability of glibenclamide. Orally administered glibenclamide/beta-CD/HPMC and glibenclamide/(SBE)7m-beta-CD/HPMC complexes showed similar absolute bioavailability compared to formulations not containing HPMC, even though 80% (in the case of (SBE)7m-beta-CD) or 40% (in the case of beta-CD) less cyclodextrin was used. CONCLUSIONS: The oral bioavailability of glibenclamide was significantly increased by cyclodextrin complexation. HPMC increased the solubilising effect of cyclodextrins and, therefore, the amount of cyclodextrin needed in the solid dosage form was significantly reduced by their co-administration. In conclusion, the pharmaceutical usefulness of cyclodextrins in oral administration may be substantially improved by co-administration of a water-soluble polymer.

Administration, Oral

Stability of Pityrosporum ovale allergens during storage.

BACKGROUND: Pityrosporum ovale is a common saprophyte on the skin capable to induce IgE antibody production in atopic dermatitis. Generally IgE response to P. ovale has been established with skin-prick test, but the stability of P. ovale skin test allergens is not known. OBJECTIVE AND METHODS: In this study we analysed the stability of identified allergens of P. ovale in skin test solutions during storage. The stability was checked with immunoblotting and densitometry after different time periods and at various temperatures. RESULTS: Coomassie Brilliant blue staining clearly showed, that proteins were preserved poorly at + 20 degrees C in 50% glycerol. Even extracts stored at + 4 degrees C during 6 and 12 months in 50% glycerol were degraded. Only extract stored at + 4 degrees C for 1 month in 50% glycerol was quite well preserved. According to densitometry results the 9 kDa band was the most stable main protein allergen. The 20 kDa and especially the 96 kDa bands were far more labile. The stability of low molecular allergens was in general better than the stability of high molecular weight allergens. CONCLUSIONS: The stability of P. ovale extracts stored for more than 1 month was poor even when kept at + 4 degrees C temperature. Reliability of negative skin prick results should be questioned when older extracts are used. In skin test negative cases, the sensitization should be evaluated also with the measurement of specific serum IgE antibodies to P. ovale.

Allergens

Genetic and environmental factors affecting the allergenicity of birch (Betula pubescens ssp. czerepanovii [Orl.] Hämet-ahti) pollen.

BACKGROUND: Environmental variation, such as an increase of mean temperature due to the greenhouse effect, as well as the genetic factors may affect the allergenicity of pollen and thus, the prevalence of allergies. The connection between these factors and the allergen content of pollen is poorly understood. OBJECTIVES: To evaluate the role of environmental and genetic factors on the allergenicity of birch pollen. METHODS: Mountain birch (Betula pubescens ssp. czerepanovii (Orl.) Hämet-Ahti) pollen was studied using SDS-PAGE and IgE-immunoblotting. Pollen samples were collected from the trees of 10 half-sib families. The study trees from each family were reared in two tree line gardens where the daily mean temperatures were different during the growing season. RESULTS: The quantitative analysis of band intensities suggested that the responses of the major birch pollen allergen, Bet v 1, were stronger in the samples collected from the garden with higher daily mean temperature. Half-sib families and individual trees differed in their Bet v 1 content. CONCLUSIONS: Our results show that both genetic and environmental factors have an effect on the amount of Bet v 1. This suggests that breeding for trees low in allergen content may be possible.

Allergens

In-house reference (IHR) preparation of Candida albicans allergen extract. A standardized extraction procedure.

A standardized, controlled procedure for preparation of an in-house reference (IHR) preparation of an allergen extract of Candida albicans is described. The procedure, based on previous studies of allergens of C. albicans, is designed to yield a maximum of allergens in optimum extraction conditions and to provide a reference preparation for further extract production. The SDS-PAGE, IgE-immunoblotting, and crossed radioimmunoelectrophoresis (CRIE) analyses showed that the procedure is reproducible with acceptable batch-to-batch variation. The variation in the content of the most important allergens, namely, proteins with molecular weights of 46, 29, and 27 kDa in the pooled final batches, is acceptable (coeff. of variation < 15%), although in the intermediate batches of different strains, the coefficient of variation may occasionally exceed 20%. A comparison with other C. albicans allergen preparations used in our previous studies is also presented. The resulting extract can be used as a reference in further extract production and also in experimental in vitro and in vivo studies.

Allergens

IgE, IgA, and IgG responses to common yeasts in atopic patients.

This study was undertaken to analyze the differences in exposure and sensitization to five common environmental yeasts. The responses of IgG, IgA, and IgE to Candida albicans, C. utilis, Cryptococcus albidus, Rhodotorula rubra, and Saccharomyces cerevisiae and purified S. cerevisiae enolase were analyzed by immunoblotting (IgE-IB), and the cross-reactivity of their IgE-binding components by IgE-IB inhibition. Twenty atopic subjects, with asthma, allergic rhinitis, or atopic dermatitis were included. In skin prick tests (SPT), 12 of the patients showed simultaneous reactivity to at least two of the five yeasts, four reacted to one of the yeasts, and four had no responses. Antigens run in SDS-PAGE and transferred to nitrocellulose were probed with enzyme-labeled IgA-, IgG-, and IgE-specific antibodies. The IgE immunoblotting revealed most IgE-binding bands in C. albicans (11 bands) followed by C. utilis (eight bands), S. cerevisiae (five bands), R. rubra (five bands), and Cr. albidus (four bands). Six of the IgE-binding bands of C. albicans and C. utilis shared molecular weight, and only two bands shared molecular weight with other yeasts. These were the 46-kDa band, shared by all five yeasts, and a 13-kDa band shared by four yeasts. Prominent IgE binding was seen to a 46-kDa band of C. albicans (seven patients), C. utilis (five patients), and S. cerevisiae (one patient) and to corresponding weak bands of Cr. albidus and R. rubra (one patient). The possible cross-reactivity of the 46-kDa band was analyzed by IgE-IB inhibition and densitometry, revealing clear C. albicans inhibition of C. utilis (80%) and enolase (98%) (autoinhibition 100%). The strongest IgG responses were seen against S. cerevisiae and C. albicans. The responses were mainly against mannans of C. albicans and S. cerevisiae, suggesting that most of the exposure is to these yeasts. Yeasts with different types of exposure, from saprophytic growth on human mucous membranes to exposure by air and food, were shown to cross-react at the allergenic level. Atopic patients primarily sensitized by C. albicans and S. cerevisiae may develop allergic symptoms by exposure to other environmental yeasts due to cross-reacting IgE antibodies.

Adolescent

Effects of air pollution and other environmental factors on birch pollen allergens.

To determine the effects of anthropogenic pollution on water-soluble proteins and specifically allergens in birch (Betula pendula and B. pubescens) pollen, we analyzed extracts of pollen from the pollution gradient around a factory complex (emitting sulfur oxides and heavy metals) by sodium dodecyl sulfate (SDS)--polyacrylamide gel electrophoresis (PAGE) and IgE immunoblotting. In addition, tree density-associated shading of the tree habitat, and quantity and quality of proteins and allergens in pollen of the two birch species were studied. The two studied birch species gave identical allergen profiles even though their protein profiles differed. Distance from the factory did not affect the amount of birch pollen major allergen, Bet v 1 (17 kDa), or of two other strong allergens (23 and 36 kDa). Trees growing in shaded places had significantly stronger responses to Bet v 1 and to the 23-kDa allergen than trees growing in open or half-open environments. Thus, we propose that combined heavy metal and sulfur dioxide pollution does not have an important effect on birch pollen allergens. Instead, other factors, e.g., shading and soil properties of the tree habitat, as well as the genetic background of the tree, may have a stronger influence on the quantity and relative composition of allergens.

Air Pollutants, Occupational

IgE antibody response against Aspergillus umbrosus in farmer's lung disease.

IgG and IgA antibodies against fungus Aspergillus umbrosus have been found in the sera of patients with farmer's lung (FL) disease and healthy exposed farmers in Finland. To determine the IgE response to antigens of A. umbrosus and Candida albicans, sera from 20 patients with FL, 20 healthy farmers and 20 nonfarming controls were tested by nitrocellulose radioallergosorbent test (RAST). The values of RAST indices were low in each group and the only statistically significant difference was found between the groups of FL patients and nonfarming controls against A. umbrosus polysaccharide antigen. Individual IgE responses to polysaccharide antigens of A. umbrosus and C. albicans correlated among FL patients, however, no cross-reacting IgE antibodies could be shown. To conclude, the IgE antibody levels of A. umbrosus polysaccharide and crude antigens were low in FL patients, healthy exposed farmers and nonfarming controls. Neither polysaccharide nor crude antigen-specific IgE antibodies of A. umbrosus have any diagnostic value in FL disease.

Antibodies, Fungal

Respiratory symptoms, pulmonary function and allergy to fur animals among fur farmers and fur garment workers.

OBJECTIVES: This study determined the prevalence of respiratory symptoms and immediate hypersensitivity to fur allergens among fur farmers and fur garment workers and measured the pulmonary function of these groups of workers. METHODS: Fur farmers (N=188) and fur garment workers (N=175) were compared with workers in a factory producing polyvinyl products (N=181) and bank and health center workers (N=118), respectively. The groups were given a self-administered questionnaire, lung function tests (spirometry, diffusing capacity), and skin prick tests to common environmental allergens, and epithelium (hair) and urine of fur animals. RESULTS: Rhinitis symptoms and eye complaints were significantly more common among the fur garment workers than among their referents, but were not associated with atopy. Between the fur farmers and their referents, the symptom prevalence did not differ significantly. Smoking explained the lower forced vital capacity and forced expiratory volume in 1 second of the fur garment workers. The prevalence of positive skin tests did not differ significantly between the exposed group and their respective referents. The skin tests showed cross-reactivity between antibodies to fur and domestic animal allergens. As confirmed by a questionnaire sent to former fur workers, selection took place for both groups of fur workers. CONCLUSIONS: Fur garment workers have an excess of rhinitis and eye symptoms, which primarily appear to be nonimmunologic. Allergy to fur animals forces sensitized workers, especially asthmatics, to leave the trade. A supplementary questionnaire to former workers on pertinent exposures and reasons for leaving a particular job can be recommended for use in prevalence studies.

Adolescent

IgE antibodies to protein and mannan antigens of pityrosporum ovale in atopic dermatitis patients.

BACKGROUND: Pityrosporum ovale is a common saprophyte on the skin capable of inducing IgE antibody production in atopic dermatitis (AD) patients. Allergens of P. ovale have been examined in several studies, but consensus on them is lacking. OBJECTIVE: This study was carried out to obtain more information about the IgE antibody response against P. ovale, including mannan. METHODS: Sera from 64 AD patients and 10 healthy controls were analysed with immunoblotting and the nitrocellulose radio allergosorbent test (RAST) method specifically developed to detect antimannan P. ovale IgE antibodies. RESULTS: In immunoblotting a total of 39 different IgE stained protein bands were seen. A high molecular weight staining was also seen especially in patients who displayed elevated mannan P. ovale RAST values. The most commonly stained protein bands in immunoblotting were 9 and 96 kD bands with antibodies in 73 and 65% of AD patients who had been positive in commercial P. orbiculare RAST with total serum IgE less than 4000 kU/l. Mannan RAST appeared positive in 77% of them. Positive immunoblotting to either of these bands was seen in 90% and, if added with staining with the 20 kD band, in 100% of these AD patients. A combination of 9 kD IgE staining and mannan P. ovale RAST was positive in 92% of the patients and 96 kD and mannan P. ovale RAST in 85% of the patients. CONCLUSION: It is evident that P. ovale has several allergens, the 9, 96 and 20 kD regions being the most important. According to our results mannan is also an important allegen of P. ovale.

Adolescent