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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 24 (1994), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A two-site solid-phase enzymimmunoassay was used for the quantification of group V allergen in grass pollen extracts in mass units. The assay is based on the monoclonal antibodies (MoAbs), 1D11 and 3B2 which recognize different epitopes on the standard Phl p V. The MoAb-ELISA is very sensitive (15 100ng ml Phl p V)and highly specific for group V allergens. Six pollen extracts of different grasses demonstrated parallel binding curves. The group V content ranged between 73 and 673 μg/ml in the extracts. The International Standard of Phleum pratense (IS 82 520) contains 400 μg/nil Phl p V. A good correlation was observed between the group V content and RAST inhibition, with the exception of Poa pratensis.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 23 (1993), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A two-site monoclonal antibody (MoAb) ELISA has been developed for the quantification of the Phleun pratense major allergen, Phl p V. The assay is based on two MoAbs which recognize different non-overlapping epitopes on the Phi p V molecule; one antibody (1D11) was immobilized on the solid phase and the other (3B2) was biotinylated. An affinity-purified Phi p V preparation (purity of 95%) was used as standard. The assay has a sensitivity of 10 ng/ml of allergen and is suitable for the detection of group V allergen in aqueous grass extracts. The specificity of the assay was investigated with 14 grass pollen and five non-grass pollen extracts. Different levels of group V allergen were delected in extracts of grasses, but not in non-grasses. The assay gives a good correspondence with allergenic activity of extracts as determined by ELISA inhibition using serum pool of allergic patients. The results indicate that the two-site MoAb ELISA could be very useful in the standardization of allergenic extracts from grass pollen.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science, Ltd
    Clinical & experimental allergy 31 (2001), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Previous studies have shown that the risk for allergic sensitization is lower in children who grew up on farms and in young adults who were exposed to dogs in early childhood. A higher microbial exposure in general and in particular to endotoxin in early childhood might contribute to this lower risk of atopy.Objective We examined whether the presence of pets or vermin in the home is associated with higher endotoxin concentrations in settled house dust.Methods House dust was sampled in a standardized manner on the living room floors of 454 homes of German children aged 5–10 years (participation rate 61%). Endotoxin was assessed with a quantitative kinetic chromogenic Limulus Amebocyte Lysate (LAL) method. Associations between endotoxin levels, pets and vermin are presented as ratios of the crude and confounder adjusted geometric means (means ratios) in the category of study vs. a reference category using multiple linear regression models.Results Endotoxin concentrations in living room floor dust sampled in homes without pets and vermin were lower (1246 ng per square meter, 1519 ng endotoxin/g dust, n = 157) than those sampled in homes with pets or vermin (2267 ng per square meter, 2200 ng endotoxin/g dust, n = 296). After adjustment for city of residence, season of dust sampling, age of the building and story of the dwelling, means ratios for endotoxin expressed per gram of dust were statistically significantly increased for dog (1.64, 95% CI 1.09–2.46), for cat (1.50, 95% CI 1.03–2.18) and for cockroach (3.01, 95% CI 1.37–6.60), whereas no major statistically significant associations were found for other pets, ants and mice.Conclusion Keeping a dog or a cat in the home is consistent with higher exposure to endotoxin and might therefore contribute to the lower risk of atopy in later life.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 23 (1993), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Monoclonal antibodies (MoAb) to defined allergens of P. pratense were raised. Five of them were selected for detailed studies by means of immunoblotting after SDS–PAGE and IEF of extracts from P. pratense and L. perenne. Three antibodies (1D11, 3B2, 2D1) recognize structures with mol. wt of 29 and 34 kD and pI of 4.4-7.7, corresponding to group V allergens. Two other MoAbs (2D8, 3C4) are directed against strong basic structures with a mol. wt of 50 kD and pI of 7.7-9.9 according to group IV allergens. The specificity of antibodies was supported by direct ELISA with purified group V and IV allergens. The isolated allergens were characterized before by SDS-PAGE and CIE and that allergenicity was detected with sera of patients with allergic rhinitis. Using our selected MoAbs crossreactive epitopes on group V and IV allergens have been excluded. Our antibodies have been used to detect crossreactivity in 14 grass pollen extracts. The evaluation of the pollen extracts has been performed by enzyme immunoassay (EIA) inhibition. One MoAb (3C4) is able to recognize group IV allergens in all grass species analysed whereas the MoAb 2D8 seems to identify group IV structures in selected grasses only. Binding to conserved structures of group V has been proved for MoAb 1D11. Other group V specific MoAbs (2D1, 3B2) identify similar, however incomplete, spectra. These results have been confirmed also by the dot immunobinding assay.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford BSL : Blackwell Science Ltd
    Clinical & experimental allergy 30 (2000), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: It is widely known and accepted that grass pollen is a major outdoor cause of hay fever. However, it is of virtual importance for grass pollen allergic patients with symptoms all the year round to know the concentration of grass pollen allergens in their homes.〈section xml:id="abs1-2"〉〈title type="main"〉ObjectiveThe main objective of this study was to quantify the amount of grass pollen allergen in mass units (μg Phl p 5) in dust settled indoors and to detect the distribution of allergenic activity in different sampling locations of homes. Furthermore, we studied the seasonal fluctuation of allergen content in dust samples.〈section xml:id="abs1-3"〉〈title type="main"〉MethodsWe adapted the two site binding assay for detection of group 5 grass pollen allergens in samples from randomly selected homes in Hamburg (n = 371), Erfurt (n = 396), Hettstedt (n = 353), Zerbst (n = 289) and Bitterfeld (n = 226), Germany. Dust samples were collected from floor of living room (LR), bedroom (BR) or children's room (CR) and mattress (MA) during period of June 1995 to August 1998. The amount of the major grass group 5 allergens was detected in μg/g dust.〈section xml:id="abs1-4"〉〈title type="main"〉ResultsPhl p 5 was detected in 67% of the samples analysed (n = 4760). The range was between undetectable (〈 0.03 μg/g dust) and 81 μg/g dust. Phl p 5 levels were significantly higher in the dust from LR (geometric mean 0.117 μg/g dust) or BR/CR floors (geometric mean 0.098 μg/g dust) than in mattresses (geometric mean 0.043 μg/g dust). We observed seasonal fluctuation of indoor Phl p 5 levels with peak in June but also annual differences. Thus Phl p 5 content indoors reflects also the different quantities of pollen counts of annual courses. During pollination period we found two times higher Phl p 5 levels (0.172 μg/g dust, P 〈 0.001) than outside of grass pollination season (0.095 μg/g dust). The indoor Phl p 5 levels outside of season seem to be independent of pollination before. We suppose that settled pollen grains or allergenic material from outdoor particles carried indoors via footwear and clothes accumulates in house dust.〈section xml:id="abs1-5"〉〈title type="main"〉ConclusionAlthough we not known how the allergens in settled dust are equilibrated with those in the air, the considerable high level of Phl p 5 in indoor dust even during periods when no grass pollen is present in the atmosphere may be an important cause of pollen-allergy symptoms outside of season.
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 33 (2003), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background  Studies investigating the effect of exposure to indoor allergens and endotoxin on human respiratory health usually take dust samples only at one single point of time and consider them to be representative for the indoor biocontaminant burden during a time period of several years. This singly measured biocontaminant concentration is related to adverse respiratory outcomes such as asthma, wheeze or others.Objective  We analysed two repeated measurements of mite and cat allergens in mattress and living room floor dust as well as endotoxin concentrations in living room floor dust. The repeated samples were taken over a time period of about 6 years. We investigated the repeatability over time of their concentrations by determining correlation coefficients and computing within- and between-home variance components.Methods  Our analysis was based on the population of a study on Indoor Factors and Genetics in Asthma (INGA) being carried out in 1995/96 and followed up in 2000/01. Complete data were available from 152 participants.Results  The measured allergen concentrations were low and a considerable percentage of the values was below the limit of detection. The crude Spearman's rank correlation coefficients between the two measurements taken in 1995/96 and 2000/01 ranged from 0.32 to 0.61 for the dust mites allergens, from 0.21 to 0.44 for cat allergen and from 0.35 to 0.51 for endotoxin. Correlations were higher if measurements were performed on the same floor or the same mattress at both sampling time-points. The within-home variance for all measured biocontaminant concentrations was of about the same order as the between-home variance.Conclusion  For studies like ours with low allergen and endotoxin concentrations, the repeatability of these concentrations over a time period as long as 6 years is low and a single measurement does not accurately reflect the true long-term exposure in the homes.
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  • 7
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background: Allergic reaction to guinea pig has been recognized as a problem in domestic settings and work environments for many years. Until recently, limited information was available on the properties of guinea pig allergen(s). In this study the major allergen Cav p 1 was characterized and the N-terminal amino-acid sequence was determined.Methods and results: Sera from 40 patients with IgE-mediated allergy to guinea pigs were investigated by means of immunoblotting using extracts prepared from guinea pig hair and urine. Three major allergens were identified within both sources with molecular weights (MW) of 8 kDa, 17 kDa and 20 kDa, respectively. From aqueous hair extracts the 20 kDa allergen (Cav p 1) was purified to homogeneity by anion exchange chromatography and reverse-phase HPLC and the N-terminal amino-acid sequence was determined. On the basis of the 15 residues, 57% identity was obtained from computer search with a sub-sequence of MUP (major urinary protein), a member of the lipocalin superfamily. Allergenic relationships among guinea pig allergens derived from various sources (hair and urine) or different animal species (mouse, rat, cat) were studied by ELISA inhibition assays. Neither urine of mouse, rat and cat, nor hair extracts of rat and cat produced appreciable inhibitions in guinea pig ELISA.Conclusion Although the physicochemical characteristics of isolated Cav p 1 are very similar to those for other rodent allergens and furthermore partial sequence identity with Mus m 1 was found, it is clearly shown here to be an immunologically independent major allergen.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Munksgaard International Publishers
    Allergy 60 (2005), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background:  Moisture is vitally important for house dust mites and they cannot survive in cold or hot-dry climates.Aims of the study:  To investigate the influence of two extraordinarily cold and dry winters in 1995/1996 and 1996/1997 on house dust mite levels in German homes.Methods:  Dust samples were collected between June 1995 and December 2001 on the mattresses of 655 adults and 454 schoolchildren living in five different areas of Germany. We compared house dust mite allergen Dermatophagoides pteronyssinus (Der p 1) levels before and during the winters of 1995/1996 and 1996/1997 with levels after these winters.Results:  D. pteronyssinus (Der p 1) levels in samples taken after the cold winters of 1995/1996 and 1996/1997 were approximately two times lower than Der p 1 levels in dust samples collected before or during these respective winters (Geometric means: Erfurt 89 vs 33 ng/g; Hamburg 333 vs 219 ng/g; Bitterfeld, Hettstedt, and Zerbst 296 vs 180 ng/g). Except for Hamburg, the decrease in Der p 1 levels was statistically significant. D. pteronyssinus levels measured in dust samples collected in 2001 (i.e. 3 years after the two cold winters) show a statistically non-significant increase (Geometric means: Erfurt 33 vs 39 ng/g; Hamburg 219 vs 317 ng/g), suggesting that it may take a long time for mite allergen levels to increase again after a sudden decrease.Conclusion:  We conclude that Der p 1 levels in German mattress dust samples have been approximately reduced by a factor of three to four by the two consecutive cold winters of 1995/1996 and 1996/1997.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background: Guinea pigs are important sources of inhalant allergens in home and working environments. However, little is known about the molecular characteristics and the relevant epitopes of guinea pig allergens. Recently, several allergens have been identified in hair extract and urine, and the major allergen Cav p 1 (20 kDa) has been characterized.Objective: The aim of the present study was to isolate and to characterize a further major allergen from guinea pig hair with 17 kDa.Methods: Guinea pig hair extract was fractionated using anion exchange chromatography and reverse-phase high performance liquid chromatography. Analyses were carried out by sodium dodecyl sulphate–polyacrylamide gel electrophoresis, 2D-PAGE, immunoblotting, immunoblot inhibition, glycoprotein detection, and N-terminal amino acid sequencing.Results: The nonglycosylated 17 kDa allergen, which was named Cav p 2, was purified to homogeneity. On the basis its 15 N-terminal residues, there was 69% identity with a sequence of Bos d 2, an allergenic protein from cow dander belonging to the lipocalin family. The 2D-immunoblotting analyses of guinea pig hair extract demonstrated that Cav p 2 and Cav p 1, contained several isoforms with pI values ranging from 3.6 to 5.3. The 2D-immunoblot inhibition disclosed cross-reactive IgE epitopes on the allergens Cav p 2 and Cav p 1. Furthermore, Cav p 1 can form both monomers (20 kDa) and dimers (40–42 kDa).Conclusion: These studies provide important information on the isoallergen character of two relevant guinea pig allergens Cav p 1 and Cav p 2 as well as on their cross-reactive properties.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Allergy 54 (1999), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The background of this study is the finding of several studies that the frequency of respiratory allergies was significantly higher in the former West Germany than the former East Germany. The present study investigated the levels of allergens of house-dust mite (Der p 1 and Der f 1), cat (Fel d 1), and cockroach (Bla g 2) in the household dust of 201 homes in Hamburg (West Germany) and 204 homes in Erfurt (East Germany), and examined the factors that affect these levels. The characteristics of homes were assessed by a questionnaire. The allergen levels were studied in dust from living rooms (LR), bedrooms (BR), and mattresses (MA). We detected in samples from Hamburg significantly higher allergen concentrations than in Erfurt: three times higher Der p 1, five times higher Der f 1, and three times higher Fel d 1. For Bla g 2, no comparison was possible because the concentrations were below the detection limit in 93% of the samples. Most of the differences could be explained by differences in housing and living characteristics between both cities. The mean ratio of Der p 1 levels in mattress dust between Hamburg and Erfurt decreased from 4.1 to 1.54 (NS) after adjustment for season, building material, age of the house, story of the dwelling, type of heating, age of carpet/mattress, presence of dogs, and indoor climate (temperature, humidity). The mean ratio of Der f 1 levels decreased from 6.9 to 2.78 (P〈0.05) after adjustment for these factors. The mean ratio for Fel d 1 in mattress dust decreased fom 4.03 to 1.65 (P〈0.05) after adjustment for season, building material, story of dwelling, size of dwelling, ventilation, cleaning routines, and pets. A similar reduction was seen for floor dust (LR plus BR). Our results indicate that the differences between the concentrations of mite and cat allergens found in Hamburg and Erfurt are explicable mainly, but not completely, by different building characteristics (age of houses, building material, story, and size of the dwelling) which affected the indoor climate, as well as by differences in other individual living habits (keeping of pets, age of carpets or mattresses, and cleaning routines).
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