bluuebunny/crossref_metadata_2025 huggingface dataset:
Survey of gastrointestinal reactions to foods in adults in relation to atopy, presence of mucus in the stools, swelling of joints and arthralgia in patients with gastrointestinal reactions to foods
Authors: BENGTSSON U. , HANSON L. Å. , AHLSTEDT S.
Published: April 2006

Background
Food intolerance in adults is mostly associated with vague symptoms and not clearly related to atopy and food allergy. A combination of different pathogenetic mechanisms may be responsible for the symptoms.

Objective
The aim of this study was to describe patients with a history of food‐related gastrointestinal symptoms in relation to the presence of mucus in the stools, joint swelling and arthralgia and to determine whether or not there is an association between the presence of these parameters, atopic disease and the presence of immune complexes in serum.

Methods
Fifty‐eight patients consecutively referred to our clinic with food‐related gastrointestinal symptoms were investigated.

Results
Thirty‐five patients (60%) had mucus in their stools, 24 patients (41%) complained about joint swelling and 41 patients (71%) had arthralgia. There were no correlations between these parameters and atopy according to Phadiatope test or skin prick test (SPT). No correlations were found between the occurrence of mucus in the stools, arthralgia and joint swelling. There were significantly higher levels of circulating immune complexes in patients with a history of arthralgia compared with patients with no such history (P < 0.03) and the number of individuals with the presence of such immune complexes was higher among the patients with arthralgia than among the patients without. In general the patients did not relate the exposure to certain foods to symptoms like joint swelling, arthralgia and presence of mucus in the stools. However, there were significant positive correlations between food‐related gastrointestinal symptoms in the following instances: chocolate‐induced gastrointestinal symptoms and mucus in the stools (P= 0.006), vegetable‐induced gastrointestinal symptoms and mucus in the stools (P= 0.002) and meat‐induced gastrointestinal symptoms and mucus in the stools (P= 0.003). In a group of individuals without foodrelated symptoms investigated separately, a very low frequency of mucus in the stools, joint swelling and arthralgia was seen (none, two and three individuals of the 20 subjects, respectively). Of 41 patients with immediate onset of gastrointestinal symptoms, 20 were atopic according to Phadiatope and SPT, Of 11 patients with late onset of symptoms 10 were negative in Phadiatope and SPT (P < 0.05). The most frequently involved foods were fruits, vegetables, milk, fish and meat.

Conclusion
The results suggest the involvement of different inflammatogenic mechan isms in food‐related gastrointestinal symptoms.
A role for the sodium, potassium adenosine triphosphatase (Na+, K+ ATPase) enzyme in degranulation of rat basophilic leukaemia cells
Authors: GENTILE D. A. , SKONER D. P.
Published: April 2006

Background
A circulating inhibitor of the sodium, potassium adenosine triphosphatase (Na+, K+ ATPase) enzyme has been described in allergic subjects. Recent studies have suggested that the Na+, K+ ATPase enzyme may be involved in the signal transduction pathways of various cell types and that inhibition of its activity can modulate histamine release from basophils and mast cells.

Objective
The purpose of this study was to determine if modulation of Na+, K+ ATPase activity alters degranulation in the 2H3 subline of rat basophitic leukaemia cells (RBL‐2H3), a mucosal mast cell model bearing high‐affinity Fc receptors for IgE.

Methods
Degranulation was measured by the release of both exogenous serotonin and endogenous histamine. Na+, K+ ATPase activity was assessed by ouabain‐sensitive [86rubidium] uptake ([86Rb] uptake) and ex situ enzyme activity.

Results
Ouabain‐sensitive [86Rb] uptake and degranulation increased in parallel and in a dose–response fashion with increasing Fc receptor cross‐linking. Additionally, incubation with ouabain, a known inhibitor of Na+, K+ ATPase activity, decreased both anti‐IgE and calcium ionophore‐induced degranulation, but increased spontaneous degranulation, each in a dose‐response manner. Moreover, the effect of ouabain on degranulation was reversed by rinsing and mimicked by other known inhibitors of Na+, K+ ATPase activity. Finally, in the absence of anti‐IgE or calcium ionophore, stimulation of ouabain‐sensitive [86Rb] uptake by the sodium (Na+) ionophore monensin was associated with a corresponding dose–response increase in ouabain‐sensitive degranulation. These experiments demonstrate that ouabain‐sensitive [86Rb] uptake increases following IgE receptor cross‐linking in RBL‐2H3, and that factors which modulate Na+, K+ ATPase activity in these cells may also regulate degranulation.

Conclusion
The results of this study suggest an important role for Na+, K+ ATPase activation in the signal transduction pathway of stimulated RBL‐2H3.
Allergenic and antigenic determinants of latex allergen Hev b 1: peptide mapping of epitopes recognized by human, murine and rabbit antibodies
Authors: CHEN Z. , VAN KAMPEN V. , RAULF‐HEIMSOTH M. , BAUR X.
Published: April 2006

Background
The rubber elongation factor in Hevea rubber (Hev b 1) is one of the most important latex allergen and is leading cause oflatex type 1 hypersensitivity in children with spina bifida.

Objective
The aim of this study was to define the allergenic and antigenic epitopes of Hev b 1.

Methods
The immunoglobulin‐ (Ig)E and IgG antibody binding sites on Hev b 1 allergen were delineated by enzyme linked immunosorbent assay (ELISA) using synthetic overlapping peptides covering the whole Hev b 1 sequence. In order to improve the binding capacity and specificity all peptides were biotinylated at the N‐terminal end via a 6‐aminohexanoic acid as spacer and then adsorbed to streptavidin pre‐coated microtitre plates. Fine mapping to define the essential amino acid residues for the antibody binding was achieved by using overlapping peptides with one amino acid offset.

Results
It was demonstrated that the IgE epitopes were located in different regions of Hev b 1 including the C‐terminal segment (121–137) and the segments with amino acid residues of 30–49 and 46–64. Two monoclonal antibodies (MoAbs) II2F3 and II4G9 raised against purified Hev b 1 recognized the C‐terminal segment only. The results of epitope mapping with three rabbit antisera revealed that five positive peptides, including the epitope peptides 31–49, 46–64 and 121–137, were involved in the antibody‐binding sites. Eine mapping on the segments 46–64 and 121–137 showed that the two MoAbs reacted with the peptide 125–134 in the C‐terminal region, whereas the peptide with amino acids 124–134 was essential for recognition by human IgE antibodies. Epitopes to rabbit polyclonal IgG and human IgE were also found to be involved in the amino acid residues of 47–59.

Conclusion
Our results indicate that the most allergenic/antigenic portions of Hev b 1 allergen are the C‐terminal region and the region with amino acid residues of 31–64. In both regions, the minimal IgE‐binding epitope is almost identical with the IgG‐binding epitope.
Stability of cereal allergens
Authors: VARJONEN E. , BJÖRKSTÉN F. , SAVOLAINEN J.
Published: April 2006

Background
Ingested cereals and inhaled cereal dusts can cause IgE‐mediated allergy. Testing for these allergies yields variable, often undependable results, which could be due to the use of unsuitable or degraded testing materials.

Objective
We studied the effect of storage media, time and temperature on the stability of protein allergens extracted from wheat, rye, barley and oats flour. We also examined the effect of heat processing similar to that used when preparing foods on the stability of allergenic proteins present in wheat flour.

Methods
After storage experiments proteins in allergen extracts were separated using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and then visualized using immunoblotting or staining with Coomassie brilliant blue. After heat processing wheat flour proteins were extracted and their IgE‐binding capacity was estimated.

Results
Extracted proteins from wheat, rye and barley flour appeared stable for at least 21 months when stored at 4°C in an aqueous solution containing 50% glycerol and 0.45% sodium chloride. In a medium with 0.4% phenol and 0.9% sodium chloride degradation of many proteins became apparent already after 1 month of storage. Oats extracts lost their IgE‐binding capacity more rapidly and the maximal storage time appeared to be 10 months at 4°C in a solution containing 50% glycerol and 0.45% sodium chloride. The IgE‐binding capacity of the allergenic proteins decreased as heat‐processing temperature and heating time increased. No processing conditions entirely abolished IgE‐binding.

Conclusion
Cereal allergen preparations can be stored either in lyophilized form or at 4°C in 50% glycerol. Unheated materials should be used also when testing for food allergy, as heating reduces allergenicity and thus sensitivity at testing.
Allergenic components in three different species of Penicillium: crossreactivity among major allergens
Authors: SHEN H.‐D. , LIN W.‐L. , TSAI J.‐J. , LIAW S.‐F. , HAN S.‐H.
Published: April 2006

Background
Penicillium species have been considered as important causative agents of extrinsic bronchial asthma. However, little is known about the allergens of these ubiquitous airborne fungal species.

Objective
This study cotnpares the allergenic profiles and allergenic crossreactivity among allergens of three prevaletit airborne Penicillium species.

Methods
IgE‐binding Penicillium components were identified by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE)‐immunoblotting using sera from 67 asthmatic patients. The presence of allergenic crossreactivity was analysed by immunoblot inhibition.

Results
Among the 67 serum samples tested, 15, 14 and 11 samples showed IgE reactivity to components of P. citrinun, P. notatum and P. brevicompactum, respectively. All 15 P. citrinum‐positive serum samples showed IgE‐binding to a 33 kDa extract component of this species. Thirteen (93%) of the 14 P. notatum‐positive serum samples and 10 (91%) of the 11 P. brevicompactum‐positive sera also showed IgE reactivity to components with a molecular weight of about 33 kDa in individual Penicillium species. All of the 10 P. brevicompactum 33 kDa component‐positive serum samples showed IgE reactivity to the 33 kDa components of the other two Penicillium species tested. Dose‐dependent inhibition of IgE‐binding to these major allergens was observed when the positive serum sample was absorbed with different amounts of individual allergenic extract as well as with different amounts of extracts prepared from the other two Penicillium species.

Conclusion
Although different allergenie profiles were observed in the three different Penicillium species tested, results showed that there was an IgE crossreactivity among the 33 kDa group major allergens of P. citrinum, P. notatum and P. brevicompactum.
Analysis of the allergenic composition of rat dust
Authors: GORDON S. , TEE R. D. , NEWMAN TAYLOR A. J.
Published: April 2006

Background
Allergy to rats is an important occupational health problem. The allergens of rat urine have been well defined but those in rat room dust, a potentially important source of inhalant exposure, have not.

Objective
To describe the allergens present in rat room dust and to identify a suitable marker protein which may be used to quantify airborne rat allergen. Methods Dust collected from the air‐conditioning system (bulk dust, ‘bd’) and with an air sampler (airborne dust, ‘ad’) were analysed by radioallergosorbent test (RAST) inhibition, immunoblotting and immunoblot inhibition techniques and comparisons made with hair and urine extracts prepared from adult male Wistar rats. Results Extensive crossreactivity was found between the extracts by RAST inhibition under different experimental conditions. Dust was more potent as an inhibitor than other extracts. The immunoblotting patterns of both dusts were similar although ‘ad’ contained an allergen at 29kDa not found in ‘bd’. Forty‐two sera from rat allergic subjects were used to identify 18 allergens in ‘bd’. Three ‘major’ allergens were found; 100% of subjects had immunoglobulin (Ig)E to a 44 kDa allergen and 74% and 88% of subjects had IgE with bound to the 20.5 and 17 kDa allergens respectively. Immunoblot inhibition experiments identified the 17 kDa dust allergen as α2u‐globulin (Rat nI). Conclusions Rat dust is a complex allergenic source. The 17 kDa dust allergen has immunological identity with Rat nl and is a suitable marker protein for the quantitation of airborne rat allergen.
Induction of hyperresponsiveness in human airway tiss by neutrophils — mechanism of action
Authors: ANTICEVICH S. Z. , HUGHES J. M. , BLACK J. L. , ARMOUR C. L.
Published: April 2006

Background
The two main features of asthtna are bronchial hyperresponsiveness and inflammation. The inflammatory response in asthma consists of infiltration and activation of a variety of inflammatory cells including neutrophils. Our previous studies have shown that stimuhited neutrophil supernatants cause hyperresponsiveness of human bronchial tissue in vitro.

Objective
To investigate the effect of the sensitization status of the tissue and the albumin concentration used to prepare supernatants on the response of human bronchial tissue to stimulated neutrophil supernatants.

Methods
Neutrophil supernatants were prepared from human isolated blood in the presence of varying concentrations of albumin (0%, 0.1% and 4%), Neutrophil supernatants were added to sensitized and non‐sensitized human isolated bronchial tissue which was stimulated with electrical field stimulation (EFS) (20 s every 4min). Receptor antagonists specific for the prostaglandin and thromboxane (10−7M GR3219I), platelet activating factor (10−6M WEB 2086), leukotriene D4 (10−6M MK‐679) and neurokinin A (10−7M SR48968) receptors were used to identify neutrophil products responsible for the effects observed in the bronchial tissue. Results In non‐sensitized human bronchial tissue, stimulated neutrophil supernatants induced a direct contraction in the presence of 0% and 0.1 % but not 4% albumin. This contraction was due to leukotriene D4 as MK‐679 completely inhibited the contraction. In contrast, stimulated neutrophil supernatants increased responsiveness of sensitized human bronchial tissue to EFS. The increased responsiveness was observed only in the presence of 0.1% albumin, with the site of modulation likely to be prejunctional on the parasympathetic nerve. The increased responsiveness was not inhibited by any of the antagonists tested.

Conclusion
Sensitization status of the tissue and albumin concentration effect the responsiveness of human bronchial tissue to stimulated neutrophil supernatant. Our results suggest a possible role for neutrophils in hyperresponsiveness.
In vitro primary sensitization of hapten‐specific T cells by cultured human epidermal Langerhans cells—a screening predictive assay for contact sensitizers
Authors: KRASTEVA M. , PEGUET‐NAVARRO J. , MOULON C. , COURTELLEMONT P. , REDZINIAK G. , SCHMITT D.
Published: April 2006

Background
The need to develop predictive tests which could identify potential allergens has been recognized for many years. There is as yet no accepted in vitro method for the assessment of contact sensitizers.

Objective
We have tested the ability of a range of contact allergens to induce in vitro primary sensitization of autologous T cells.

Method
T‐cell proliferation induced by haptens using 2‐day cultured human Langerhans cells as antigen‐presenting cell was assessed by 3H thymidine incorporation. Antigen specific stimulation was calculated as stimulation indexes. Results Strong allergens induced in vitro a primary T‐cell response in all (trinitrophenyl, TNP: 13/13) or in the majority (fluorescein isothiocyanate, FITC: 7/10) of experiments. An irritant, sodium dodecyi sulfate (SDS). failed to generate a significant T‐cell proliferation in any of the experiments (0/10). We obtained a significant lymphoproliferative response to weak sensitizers only in a limited number of experiments: (coumarin: 1/12, citronellal: 0/10, hydroxycitronellal: 2/8). p‐Phenylenediamine (PPDA), a prohapten and highly sensitizing chemical in vivo, generated primary sensitization in vitro in only one of six experiments, while Bandrowski's base (BB), a metabolization product of PPDA induced a significant T‐cell response in all six experiments.

Conclusion
The present in vitro model allows discrimination between two groups of substances: strong contact sensitizers (TNP, FITC. BB) on the one hand and weak sensitizers (coumarin, citronellal and hydroxycitronellal) and irritants (SDS) on the other hand. It could be used as a screening in vitro assay to eliminate strong contact allergens before further predictive animal tests have to be performed.
Allergy and the risk of sudden infant death syndrome
Authors: FORD R. P. K. , SCHLUTER P. J. , TAYLOR B. J. , MITCHELL E. A. , SCRAGG R. ,   THE MEMBERS OF THE NEW ZEALAND COT DEATH STUDY GROUP
Published: April 2006

Background and Objective
There are several sources that suggest that there is a link between allergy and sudden infant death syndrome. We endevavoured to look for evidence of an association between allergic disease and the risk of sudden infant death syndrome (SIDS).

Methods
A nationwide case‐control study covering a region with 78% of all births in New Zealand during 1987–90. Interviews were completed with the parents of 393 (81.0% of total) infants who died from the sudden infant death syndrome (SIDS), and 1592 (88.4% of total) control families who were a representative sample of all hospital births in the study region.

Results
Eczema was reported in 13.9% control infants compared with only 8.0% of the SIDS infants, univariate odds ratio for this in terms of risk for SIDS was 0.56 (95% confidence interval 0.37, 0.84) for infants with eczema compared with those without. This lesser risk for SIDS was unchanged when adjusted for potential confounding factors. The risk of SIDS was not associated with reported cow's milk reactions or a family history of allergic symptoms once adjustments were made for possible confounding factors. Conclusion Infants with skin disorders identified by their parents as eczema had a low risk for SIDS. Families can be reassured that atopy is not a risk factor for SIDS.
Evidence of antibodies to erythromycin in serum of a patient following an episode of acute drug‐induced hepatitis
Authors: GÓMEZ‐LECHÓN M. J. , CARRASQUER J. , BERENGUER J. , CASTELL J. V.
Published: April 2006

Background
A patient who had taken erythromycin orally developed mild signs of hepatocellular damage and appeared to became sensitized. After a fortuitous oral intake of the antibiotic, 18 months later, the patient developed an extensive intrahepatic cholestasis.

Objective
To confirm the allergic nature of this form of hepatitis by demonstrating the presence of serum antibodies directed at the drug.

Methods
Primary cultured human hepatocytes were used to investigate whether erythromycin can covalently bind to hepatocytes; and to demonstrate the presence of drug‐directed immunoglobulins in the serum of the patient.

Results
As a consequence of its metabolism by hepatocytes, a small percentage of erythomycin remained bound to hepatocyte proteins. The extent of covalent binding varied among hepatocytes from different donors, and was not observed in metabolically inactive hepatocytes. Antibodies able to bind erythromycin were detected in the serum of the patient. When this serum was added to a human hepatocyte culture that had been previously incubated with the drug, binding of immunogiobulins was observed around the cell membrane and close to pseudo‐bile canaliculi, thus indicating the presence of antibodies able to recognize erythromycin‐labelled hepatocytes. Conclusions The existence of serum antibodies directed at erythromycin haptens in this patient strongly suggests an allergic mechanism for this drug‐induced hepatitis.
Role of mast cells, eosinophils and IL‐5 in the development of airway hyperresponsiveness in sensitized mice
Authors: NAGAI H. , YAMAGUCHI S. , MAEDA Y. , TANAKA H.
Published: April 2006

Background and objective
In order to study the role of mast cells and IL‐5 in allergen‐induced airway hyperreactivity in mice, airway responsiveness in WBB6F1‐W/Wv mice (mast cell deficient) and the effects of anti‐IL‐5 monoclonal antibody (NC‐17) and three anti‐allergic drugs (N‐556, ketotifen and amlexanox) on airway hyperreactivity in Balbc mice were studied.

Methods
Mice were immunized with an antigen (ovalbumin; OA) at intervals of 12 days. OA was inhaled 10 days after the secondary immunization. Twenty‐four hours after the last inhalation, airway reactivity to acetyleholine was measured and broncho‐alveolar lavage fluid (BALF) was obtained.

Resutls
Three inhalations of OA caused an increase in leucocytes (including eosinophils). accompanied by increases in IL‐5 in BALF, and airway hyperreaetivity to acetylcholine in Balb/c and WBB6F1‐ +/+ mice. In WBB6F1‐W/Wv mice, antigen inhalation resulted in increases in leucocytes and IL‐5 in BALF but did not result in airway hyperreactivity. NC‐17 at doses between 10 and 20μg (intratracheal injection) inhibited the antigen‐induced eosinophilia but did not affect airway hyperreaetivity in Balb/c mice. Three ‘anti‐allergie’ drugs clearly inhibited antigen‐induced increases in IL‐5 levels and the number of eosinophils in BALF, but did not alTect airway hyperreactivity in Balb/c mice.

Conclusions
These data suggest that mast cells play an important role in the onset of airway hyperreactivity but do not play a role in the production of IL‐5 and eosinophilia. Furthermore., indicate that the inhibition of IL‐5 is not always associated with a reduetion in antigen‐induced airway hyperreactivity in mice.
Low‐level exposure to house dust mites stimulates T‐cell responses during early childhood independent of atopy
Authors: BJÖRKSTÉN B. , HOLT B. J. , BARON‐HAY M. J. , MUNIR A. K. M. , HOLT P. G.
Published: April 2006

Background
The imtnune responses which underlie the expression of allergic symptotns in childhood are believed lo be initiated in infancy and early childhood. The kinetics of this response have hardly been researched.

Objective
To analyse, in an environment with low house dust mite (HDM) exposure levels, the relationship between house dust mite (HDM)‐specific T‐cell reactivity as expressed by in vitro proliferation of blood mononuclear cells.

Methods
The study comprised a prospective analysis of patterns of allergen‐specific T‐cell reactivity in a cohort of 19 children, from whom blood samples were obtained in the spring during their second and third years of life. Blood mononuclear cell cultures were established in 200 μL AIM‐V serum free medium. Crude house dust mite (HDM) and purified Der p 1 and Der p 2 extracts were used at optimal concentrations, i.e. 100μg/mL for HDM and 30μg/mL for the purified allergens. Tetanus toxoid (0.5 μglrnL) and ovalbumin (10 μg/mL) served as positive controls. A clinical diagnosis of allergy was verified with skin‐prick tests. Dust samples were collected from a mattress and/or carpet or sofa in homes, day care centres and day care homes. Major mite allergen levels (Der p 1/Der f 1) in dust were analysed by an enzyme linked immunosorbent assay (ELISA).

Results
Specific T‐cell responses were seen in the majority of the children against house dust mite (crude HDM extract. Der p 1 and Der p 2). The levels of the house dust mite allergens Der p 1 and Der f I were low, i.e. < 0.68 μg/g fine dust in the homes of the children and the day care centres that they were attending. This indicates that doses of mite antigen well below the suggested sensitization threshold level of 2 μg/g dust can induce mite‐specific T‐cell responses in young children. None of them showed clinical reactivity to house dust mites as indicated by negative skin‐prick tests.

Conclusions
The findings suggest that active immunological recognition of environmental allergens and the ensuing initiation of allergen‐specific T‐cell responses, is a normal part of the ‘education’ of the immune system in early childhood and can occur even at very low exposure levels. Priming per se does not imply clinically significant sensitivity, however.
Investigating paediatric airways by non‐bronchoscopic lavage: normal cellular data
Authors: HEANEY L.G. , STEVENSON E. C. , TURNER G. , CADDEN I.S. , TAYLOR R. , SHIELDS M. D. , ENNIS M.
Published: April 2006

Background
Bronchoscopic bronchoalveolar lavage in children to investigate bronchia disorders such as asthtna has both ethical and procedural difficulties.

Objective
The aim of this study was to establish a standardized non‐bronchoscopic method to perform bronchoalveolar lavage in children attending for elective surgery to obtain normal cellular data.

Methods
Bronchoalveolar lavage was performed on normal children (n= 55) by infusing saline (20 mL) through an 8 FG suction catheter passed after endotracheal intubation. Oxygen saturation, heart and respiratory rate were monitored during the bronchoalveolar lavage procedure. Cellular analysis and total protein estimation of the lavage fluid were performed. Epithelial lining fluid volume was calculated (n = 15) using the urea dilution method.

Results
The procedure was well tolerated by all children. Total cell count and differential cell count for children (macrophages 70.8 ± 2.3%, lymphocytes 3.8 ± 0.6%, neutrophils 5,7 ± 1.0%, eosinophils 0.14 ± 0.03%. epithelial cells 19.6 ± 2.1%, mast cells 0.21 ± 0.02%) were similar to those reported for adults. Age and sex comparisons revealed no differences between groups. The mean total protein recovered in the cell free supernatant was 49.72 ± 4.29 mg/L and epithelial lining fluid volume was 0.82 ± 0.11% of return lavageate.

Conclusion
This method allows bronchoalveolar lavage to be performed safely and quickly on children attending for routine elective surgery. Using this method and taking the ‘window of opportunity’ of elective surgery, the presence or absence of airway inflammation could be studied in children with various patterns of asthma during relatively asymptomatic periods.
Design of an optimally‐diagnostic skin test solution for diagnosis of sensitivity to timothy grass (Phleum pratense) pollen
Authors: WHEELER A. W. , JESSBERGER B. , DRACHENBERG K. J. , RAKOSKI J.
Published: April 2006

Background
Although most of the common allergen extracts that are used for diagnosis of type 1 hypersensitivity are now well standardized, this gives no assurance that they are within the concentration range that gives the best chance of a true diagnosis.

Objective
The objective of this study was to identify the most appropriate concentration range of timothy grass pollen Phleum pratense extract to diagnose sensitivity to this pollen correctly through skin‐testing.

Methods
Dilutions of a well‐standardized extract were made and used to skin test ‘true’ positive and ‘true’ negative populations of subjects as identified by case history, challenge tests and radioallergosorbent test (RAST). Weal diameters were measured and the data were submitted to receiver operating characteristics (ROC) analysis. For any particular weal size cut–off, the optimal diagnostic concentration (ODC) range was thus calculated.

Results
A 3 mm weal diameter cut‐off was chosen as an appropriate size for routine diagnosis. Therefore the ODC range at this diameter was used to establish a product target concentration and specification for formulation of the diagnostic reagent. This method of allergen extract standardization can lead to a true‐biological unitage that can be used for labelling purposes.

Conclusion
The optimun concentration range at which to formulate an allergen extract, in terms of an in vitro immunologically based assay, can be determined by carrying out ROC analysis of the results of clinical studies as described in this communication. Diagnostic units (DU), are now used by us for labelling of such final formulations which conveys the information that the product is at the most appropriate concentration for diagnosis.
Comparison of basophil histamine release, eosinophil cationic protein and non‐specific airway responsiveness between mite‐sensitive asthmatic and non‐asthmatic children and non‐allergic controls
Authors: TAKEDA K. , SHIBASAKI M. , IMOTO N. , SHIMAKURA Y. , TAKITA H.
Published: April 2006

To understand the relevance of allergy to the development of asthma in children, we examined basophil hislamine release (HR) with Df antigen, blood cosinophii counts, serum eosinophil cationic protein (ECP) levels, and bronchial responsiveness to methacholine (PC20) in three groups of children, including 36 asthmatics with high RAST titre for Df (group 1), 36 non‐asthmatics with similarly high RAST titre for Df (group 2) and 21 non‐asthmatics with negative RAST titre for Df (group 3). The amount of Df antigen inducing 50% HR from basophils did not vary significantly between group 1 and 2 (P>0.05), while none of the cells responded to higher concentrations of Df in group 3. The mean number of blood eosinophils and level of serum ECP were highest in group 1, and lowest in group 3, with group 2 being intermediate, and the differences were significant between all three groups (P<0.01). The mean PC20 value was the lowest in group 1, intermediate in group 2, and the highest in group 3, and the differences were significant between all three groups (P<0.01). While correlation studies showed that PC20 values of group 2 subjects signficantly correlated with their eosinophil numbers (r=–0.48, P<0.01) and ECP levels (r =–0.49, P < 0.01), such correlations were not found in group 1 subjects. These results suggest that the degree of the eosinophilic inflammation caused by the allergic reaction to mites is an important factor in determining the clinical expression of asthma in atopic subjects.
Inhalative occupational and ingestive immediate‐type allergy caused by chicory (Cichorium intybus)
Authors: CADOT P. , KOCHUYT A.‐M. , DEMAN R. , STEVENS E. A. M.
Published: April 2006

We report a first case of occupational allergy to chicory (Cichorium intybus) in a vegetable wholesaler. Symptoms occurred after oral, cutaneous or inhalatory exposure. The patient also reported reactions after ingestion of botanically related endive (Cichorium endivia) and lettuce (Lactuca sativa.) We identified the responsible allergen by SDS‐PAGE and immunoblot to be a 48‐kDa protein, confined to the non‐illuminated parts of the plants. No cross‐reactivity was found with mugwort (Artemisia vulgaris) ryegrass (Lolium perenne) and birch (Betula verrucosa) pollen, which suggests that the vegetable is the primary allergenic material.
Determination of the stability of diluted allergen extracts using a concentration step prior to EAST inhibition
Authors: NIEMEIJER N. R. , MEIJER G. , KAUFFMAN H. F. , DE MONCHY J. G. R.
Published: April 2006

Background
Generally the stability of diluted allergen extracts, as used for skin testing, provocation testing und immunotherapy can not be tneasured using a normal enzyme allergosorbent test (EAST) inhibition method.

Objective
The aim of this study was to determine the stability of diluted allergen extracts using an ultrafiltration step prior to the standard EAST inhibition procedure, in which the allergen extract was concentrated 100‐fold.

Methods
This eoncentration procedure was validated for Dermatophagoides pteronyssinus, timothy pollen, birch pollen and cat dander extraets and used in a stability study in which three batches were stored for 1 year al 6° C and 25° C.

Results
There was no difference in relative potency before and after concentration of birch and timothy pollen extracts. D. pteronyssinus and cat dander extracts showed a significant decrease of 25% and 35% of the relative potency after concentration. The mean coeffieient of variation of 12 determinations of the stability study was 11.8%.

Conclusion
For all allergens the 30BU/mL or approximately 0.00025 mg/mL solution was stable for 12 months at both temperatures, except for D. pteronyssinus which declined rapidly at 25°C.
Airway hyperresponsiveness to acetylcholine induced by aerosolized arachidonic acid metabolites in guinea‐pigs
Authors: SATO T. , IWAMA T. , SHIKADA K. , TANAKA S.
Published: April 2006

Background and objective
To investigate the role of arachidonic acid (AA) metabolites in airway hypersensitivity.

Method
We studied the change in airway responsiveness to acetylcholine (ACh) after inhalation of some AA metabolites in guinea‐pigs.

Results
Exposure to prostaglandin (PG) D2, thromboxane (TX) A2 mimetic U‐46619, leukotriene (LT) D4 or LTE4 at concentrations which did not influence bronchial tone and blood pressure dose‐dependently caused airway hyperresponsiveness. However, the change was not observed after challenge with a high concentration of PGF2α Furthermore, PGD2 and U‐46619 induced an acute and short‐lived increase in responsiveness, while LTD4 and LTE4 induced a slow‐onset and longer‐lived increase. In the tachyphyiaxis study, although the tachyphylaxis for airway hyperresponsiveness provoked by PGD2 and U‐46619 was not observed, airway hyperresponsiveness induced by the second LTD4 tended to decrease, and the second LTE4‐induced airway hyperresponsiveness obviously diminished. In the study using antagonists and inhibitors, TX‐receptor antagonist BM‐13177 inhibited PGD2‐ and U‐46619‐, but not LTD4‐ and LTE4‐induced airway hyperresponsiveness. TX synthase inhibitor OKY‐046 had no effect on PGD2‐, U‐46619‐ and LTD4‐induced airway hyperresponsiveness, while LTE4‐induced airway hyperresponsiveness tended to be inhibited by these inhibitors. However, the LT‐receptor antagonist ONO‐1078 inhibited both LTD4‐ and LTE4‐, but not PGD2‐ or U‐46619‐induced airway hyperresponsiveness. 5‐lipoxygenase inhibitor AA‐861 tended to prevent LTE4‐induced airway hyperresponsiveness, but had no effect on PGD2‐, U‐46619‐ and LTD4‐induced enhanced responses.

Conclusion
These findings indicate that the local existence of PGD2, TXA2, LTD4 and LTE4 in the guinea‐pig airway may act as an airway hyperresponsiveness‐ inducing factor rather than as a bronchoconstrictor. In addition, PGD2/U‐46619 may stimulate the TX receptor to induce an acute and short‐lived airway hyperresponsiveness, and LTD4 and LTE4, which may involve secondary mediator release, may act at the LT receptor to induce a slow‐onset and longer‐lived airway hyperresponsiveness, which may be associated with the induction, the development and the long duration of airway hyperreactivity.
Oxygen radical production by blood eosinophils is reduced during birch pollen season in allergic patients
Authors: WOSCHNAGG C. , RAK S. , VENGE P.
Published: April 2006

Background
The eosinophil granulocyte takes part in allergic inflammatory diseases, like asthma and rhinitis. The aim of this study was to investigate the oxidative metabolism of the blood eosinophils and neutrophils from birch pollen allergic patients, during and after exposure to their allergen.

Methods
Thirteen birch pollen allergic patients, with seasonal symptoms of rhinitis, with or without asthma, were followed. The cells were purified using a percoll gradient and the MACS system. The eosinophil purity in all samples was >95%. The oxidative metabolism of both the PMN and the eosinophils was measured by means of a chemiluminescenee (CL) assay, with luminol or lucigenin as the amplifier, and using PMA (16nM) as the activator.

Results
In relative terms, the lucigenin CL by the eosinophils was 5‐10‐foId higher than that of the PMN, P= 0.002. The eosinophils of the birch pollen allergic patients produced less oxygen radicals during the season, compared to the reference group, measured both with luminol CL and lucigenin CL, P=0.01 and P= 0.02 respectively. Out of season, there was no difference. There was also no difference, during either period, between patients and references in PMN CL. Separating the eosinophils into hypodense and normodense fractions, showed a decreased oxidative metabolism by the hydrodense eosinophils.

Conclusion
We conclude that the eosinophils of birch pollen allergic patients have reduced production of oxygen radicals when the patients are exposed to their allergen, which could depend on higher amounts of hypodense eosinophils in the blood during season.
Specific T cell lines for ovalbumin, ovomucoid, lysozyme and two OA synthetic epitopes, generated from egg allergic patients' PBMC
Authors: HOLEN E. , ELSAYED S.
Published: April 2006

Background
Proteitis of hen egg white are common ingredients of food and difficult to eliminate. Allergens of egg while induce allergic symptoms among relatively high numbers of palients suffering from food allergy. B cell epitopes to hen egg white tnajor allergens have been reported. Considering that IgE antibody formation is mostly T cell dependent, the study of T cell epitopes is essential for both T cell dependent and independent IgE response.

Objectives
Little information on T cell epitopes recognizing food allergens has been reported. T cell responses to hen egg white allergens and two synthetic OA peptides located at amino acid residues No. 105–122 and 323–339 were investigated.

Methods
Peripheral blood mononuclear cells from hen egg allergic patients were investigated. Various allergens of hen egg white were used for stimulation. Primary proliferation responses were detected followed by the generation of long‐term cultures which were examined for their specificity, phenotype, cytokine profile and IgE production. The allergen specific T cell lines were mapped using a panel of 13 synthetic peptides of ovalbumin.

Results
Human T cells recognizing ovomucoid, lysozyme and ovalbumin epitope 105–122 are reported for the first time. The cell lines were enriched CD4+/CD8+ T cells (CD2+ 95%). Ovomucoid and ovalbumin induced IgE synthesis by a small fraction of B cells (1%) present in the ovalbumin and ovomucoid specific T cell lines.

Conclusions
Human T cells recognized several egg white allergens and epitopes within the ovalbumin molecule. Specific IgE was produced in cultures stimulated with ovalbumin and ovomucoid. OA peptides 105–122 and 323–339 have no affinity to the specific IgE of the two patients; an observation which could be of particular interest regarding the mechanisms of peptide‐based immunotherapy.
Ethanol metabolite acetic acid as causative agent for type‐I hypersensitivity‐like reactions to alcohohc beverages
Authors: BOEHNCKE W.‐H. , GALL H.
Published: April 2006

Background
Adverse reactions to alcoholic beverages are common and more frequently mediated by immunological mechanisms than previously thought.

Objective
To elucidate relevant allergens in this context we studied patients with an informative medical history.

Methods
This report describes a comprehensive allergological approach in a patient exhibiting type‐I hypersensitivity‐like reactions towards beverages and medication containing alcohol, and salad dressings with acetic acid.

Results
The ethanol metabolite acetic acid was found to yield positive prick test results in concentrations not eliciting reactions in healthy and atopic controls.

Conclusion
Among other ethanol metabolites, acetic acid is a potential allergen in the context of hypersensitivity towards alcoholic beverages.
Nasal polyps: medical or surgical management?
Authors: HOLMBERG K. , KARLSSON G.
Published: April 2006

Nasal polyposis is considered to be a non‐IgE‐mediated inflammatory condition of the nose and sinuses, often associated with chronic non‐allergic rhinitis, aspirin intolerance and non‐allergic asthma. The aetiology of nasal polyposis is unknown. The main symptoms are nasal obstruction and disturbance of the sense of smell. Occlusion of the nasal passage by a few large polyps could be treated by simple polypectomy to help lhe patient breathe through the nose. Polypectomyper sedoes not worsen asthma. Other surgical procedures can be performed, depending on the degree of the disease. The aim of surgery is to restore the physiological properties of the nose by making the nose as free from polyps as possible. and to allow drainage of infected sinuses. Complementary medical treatment of polyphonies is always necessary, as surgery cannot treat the inflammatory component of the mucosal disease. In this respect, topical corticosteroids have long been the drugs of choice to reduce the size of polyps, to prevent recurrence after surgery, and are often the main treatment for the disease in many patients. Fluticasone propionate has now been shown to be at least as effective as beclomethasone dipropionate as a medical tool in the management of polyposis. Short‐term treatment with systemic corticosteroids is an alternative method of inducing remission and controlling nasal polyps. However, in mosl patients with nasal polyps, treatment consists of both medical and surgical management.
House dust mite immunotherapy results in a decrease in Der p 2‐specific IFN‐γ and IL‐4 expression by circulating T lymphocytes
Authors: O'BRIEN R. M. , BYRON K. A. , VARIGOS G. A. , THOMAS W. R.
Published: April 2006

Background Allergen‐specific immunolherapy (IT) can be an important adjunctive therapy in the treatment of allergic disorders. Although it has now been used for over 80 yr. the precise mechanism of action remains unclear. Recently a number of studies have shown that cytokine production may be modified by IT, but different protocols have been used and different results obtained.

Objectives The aims of the present study were: (1) lo document the allergen‐specific expression of intcrleukin‐4 (IL‐4) and interferon‐γ (IFN‐γ) by peripheral blood cells in both untreated house dust mite (HDM) allergic patients(non‐lT) and following at least 10 months of HDM‐specific IT (post‐IT); and (2) to determine whether alterations in these critical regulatory cytokines correlated with the clinical outcome of IT.

Methods IT was undertaken with nine fortnightly subcutaneous injections of increasing amounts of a Dertnatophagoides pteronyssinus (Dpt) extract, reaching a final dose of 10000 PNU. This was followed by 6‐ to 8‐monthly maintenance injections of 5000 PNU. For cytokine measurement, mononuclear cells were separated from peripheral blood and stimulated with the major Dpt allergen, Der p 2. for 18 h, after which mRNA was isolated and IL‐4 and IFN‐γ cDNA were amplified by polymerase chain reaction (PCR). The presence of the particular cytokine was determined by visualization following clcctrophoresis on an agarose gel. The study was observational in nature being open and without a placebo group.

Results Fifteen Dpt‐sensitive patients who had not received HDM IT (non‐IT), and 16 who had, were studied. In the non‐IT group. 80% expressed IL‐4 and 75% expressed IFN‐γ. In those post‐IT, only 12.5% expressed IL‐4 and 19% IFN‐γ. The two patients still expressing IL‐4 post‐IT had had very little clinical response. Six patients were studied both pre‐ and post‐IT. Prior to IT, three were positive for both cytokines, two positive for IL‐4 alone and one for IFN‐γ. Post‐IT, all six were negative for IL‐4 and five were negative for IFN‐γ.

Conclusion Allergen‐specific IT results in a reduction in expression of the critical eytokines IL‐4 and IFN‐γ in circulating lymphocytes. It is possible that this is a contributary mechanism in the beneficial effect of IT.
Feather mites are potentially an important source of allergens for pigeon and budgerigar keepers
Authors: COLLOFF M. J. , MERRETT T. G. , MERRETT J. , McSHARRY C. , BOYD G.
Published: April 2006

BackgroundPrevious studies on allergy to feathers have not adddressed whether orgatiisms living on feathers (mites. lice, moulds) are a source of allergens.

ObjectiveTo investigate whether feather mites produced allergens of clinical relevance to bird keepers.

MethodsWe examined serum IgE responses of 96 pigeon breeders to an extract of feather mites from pigeons (predominantlyDiplaegidia columbae). using Western blotting, specific IgE assay using AlaSTAT EIA and RAST inhibition.

ResultsFeather mites are a major source of soluble proteins derived from feathers, accounting for up to 10% of the total weight of the feather. Forty‐three sera had a negative score (0) for anti‐feather mite IgE. 27 were weakly positive (1–2) and 26 had strongly positive scores (3–4). Fewer pigeon breeders with scores ± 3 were asymptomatic than those with negative scores (12 versus 40%). more had late onset symptoms (with or without early onset symptoms; 77% versus 44%) and had IgE antibody against house dust mite (89% versus 23%). Western blotting of eight sera against the extract ofDiplaegidia columbaerevealed 20 IgE‐binding components ranging from 22 to 200 kDa. A high diversity of components was recognized by each serum: arithmetic mean 7 (range 2 14). RAST inhibition indicated feather mites had species‐specific epitopes as well as ones that cross‐reacted withDermatophagoides pteronyssinus.

ConclusionStrongly‐positive AlaSTAT scores to pigeon leather mite were associated with allergic symptoms of late onset in pigeon breeders. We conclude that feather mites are a major source of clinically‐relevant allergens for pigeon breeders.
Detection of snow‐crab antigens by air sampling of a snow‐crab production plant
Authors: MALO J.‐L. , CHRÉTIEN P. , MCCANTS M. , LEHRER S.
Published: April 2006

Background and objective We previously assessed the prevalence of occupational asthtna (OA) to snow‐crab in production plant workers. We also showed that this type of OA is related to immediate immunological reactivity as demonstrated by skin reactivity and increased specific IgE antibodies. However, we did not show that snow‐crab antigens causing immunological reactivity and OA could be found in the air sampling in the plants atmosphere. This was the purpose of the current work. Material and methods Area air samples worn by workers at four different worksites of a snow‐crab producing plant were obtained on PVC filters with an SKC pump run at 1.5 L/min for 2h. Snow‐crab was being boiled and processed during the air sampling periods. Filters were analysed by RAST inhibition in a blind manner (i.e. without knowledge of the worksite where the filter originated).

Results Eluate from one of the four sites (#2) had the highest protein concentration and yielded the highest per cent inhibition of RAST — 13% inhibition with the snow‐crab meat, 23% and 28% inhibition with the snow crab water RAST in two separate assays. An eluate taken from a filter at another site (#1) showed borderline reactivity (1% and 10% inhibition in two assays) whereas the two other ones and a control filter were negative. The two filters that contained snow‐crab proteins were the ones nearest the boiling process, site #2 being the nearest followed by site #1. It was estimated that a 28% inhibition corresponded to ∼8.6 μg of proteins and to ∼1.5 μg of allergens on the filter.

Conclusion This study suggests that airborne snow‐crab‐derived proteins, released during the boiling process, arc the cause of immunological reactivity and of OA to snow‐crab.