bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ribonuclease‐gold ultrastructural localization of heparin in isolated human lung mast cells stimulated to undergo anaphylactic degranulation and recovery in vitro
Authors: Dvorak  , Morgan  
Published: March 2003

BackgroundHuman mast cells are a rich and unique source of heparin, which is stored in cytoplasmic secretory granules and accounts for metachromasia, a staining property used to identify mast cells by light microscopy.ObjectiveThe sub‐cellular locations of heparin in secretory human mast cells and human mast cells recovering from secretion are not known. Acquisition of this knowledge requires ultrastructural imaging of well‐preserved cells with a visible probe which binds to heparin. We sought to develop this knowledge regarding human mast cell secretion by using a labelling method for heparin that depends on the well‐known property of ribonuclease inhibition by heparin.MethodsHuman lung mast cells were isolated, partially purified, either stimulated or not stimulated to secrete with anti‐IgE, and recovered 20 min or 6 h later for routine electron microscopy. Histamine secretion was also determined. A previously developed post‐embedding, enzyme‐affinity‐gold electron microscopic technique to image ribonucleic acid (RNA) with ribonuclease‐gold (R‐G), which also binds to the enzyme inhibitor, heparin, was employed to determine the sub‐cellular locations of heparin in non‐secretory and secretory mast cells as well as in mast cells recovered from short‐term cultures after secretion. Specificity controls for the novel use of this method and quantification of granule labelling in these controls were performed.ResultsHeparin was labelled by R‐G in electron‐dense granules within non‐secretory human lung mast cells (HLMCs), in electron‐dense granules that persisted in secretory HLMCs at the maximum histamine secretion time (20 min), and in electron‐dense granules within recovering HLMCs. Specificity controls showed that gold alone did not label HLMCs and that absorption with heparin significantly reduced or abrogated HLMC granule staining with R‐G, but that RNA absorption did not. Heparin stores were absent in newly formed, electron‐lucent intracytoplasmic degranulation channels in secretory HLMCs. Electron‐dense granule matrices in the process of extrusion to the cell exterior still retained heparin at the instant of cellular secretion. Non‐granule heparin stores bound R‐G in recovering HLMCs. These locations included resolving degranulation channels, as newly emergent granules partitioned and condensed within them, and electron‐dense content‐containing vesicles and progranules within synthetic mast cells. Ultimately, all known ultrastructural patterns of HLMC granules developed in recovering cells, and each of them contained heparin.ConclusionHeparin was secreted from HLMCs which were stimulated by anti‐IgE, and heparin was recovered by a combination of conservative and synthetic mechanisms in HLMCs after a secretory event.
Anti‐inflammatory effect of roxithromycin in patients with aspirin‐intolerant asthma
Authors: SHOJI  , YOSHIDA  , SAKAMOTO  , HASEGAWA  , NAKAGAWA  , AMAYASU  
Published: March 2003

BackgroundFourteen‐membered macrolides, such as roxithromycin, have been reported to exhibit other pharmacological activity including anti‐asthmatic effects, besides antibiotic activity.ObjectiveThis study was designed to investigate the protective effect of roxithromycin on airway responsiveness to the sulpyrine provocation test and to investigate whether this protective activity is associated with a reduction in aspirin‐induced excretion of urinary leucotriene E4 (u‐LTE4), a marker of cysteinyl leucotriene overproduction that participates in the pathogenesis of aspirin‐intolerant asthma. Also, the present study was designed to examine whether or not its anti‐asthmatic activity was associated with a reduction in eosinophilic inflammation.MethodsFor 8 weeks before analysis, subjects received 150 mg of roxithromycin or matching placebo twice daily. We assessed the effects of pretreatment with roxithromycin on bronchoconstriction precipitated by inhalation of sulpyrine in 14 adult patients with mild or moderate aspirin‐intolerant asthma; those who were in stable clinical condition and were hyperresponsive to sulpyrine provocation test were allocated to this study. A double‐blind, randomized, crossover design was used. Urinary LTE4 was measured by a combined reverse‐phase high‐performance liquid chromatography (rp‐HPLC) enzyme immunoassay on sulpyrine provocation testing day. Blood and sputum samples were taken in the morning on the sulpyrine provocation testing day. Eosinophil counting and measurement of eosinophilic cationic protein (ECP) were performed.ResultsAfter the 8 weeks of treatment with roxithromycin, patients' symptoms, blood eosinophils, serum ECP, sputum eosinophils, and sputum ECP were significantly decreased. On the other hand, values of PC20‐sulpyrine did not improve after roxithromycin at all. Furthermore, although challenge with sulpyrine caused a significant increase in u‐LTE4, pretreatment with roxithromycin or placebo did not affect excretion of u‐LTE4.ConclusionAlthough roxithromycin does not have antileucotriene effects, it has an antibronchial inflammatory effect associated with eosinophilic infiltration. This study raises further interesting therapeutic possibilities and warrants further trials of new approaches to the treatment of aspirin‐intolerant asthma.
Cross‐reactivity between milk proteins from different animal species
Authors: RESTANI  , GAIASCHI  , PLEBANI  , BERETTA  , CAVAGNI  , FIOCCHI  , POIESI  , VELONÀ  , UGAZIO  , GALLI  
Published: March 2003

BackgroundCow's milk allergy is quite frequent in the first years of human life. When breast‐feeding is not possible, a cow's milk substitute must be provided for allergic subjects. Different alternatives to cow's milk have been suggested as protein sources (soy, hydrolysed proteins, goat's milk, etc.), but all these dietetic solutions are not without risks for polyallergic or more sensitive subjects.ObjectiveTo obtain new information on the suitability of other mammalian milks for allergic children, we evaluated the cross‐reactivity between milk proteins from different animal species.MethodsMilk samples were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE). To detect antibody–antigen complexes, immunoblotting was performed by using sera from children allergic to cow's and ewe's milk (RAST class ≥ 4) and monoclonal antibodies (MoAb) specific for bovine proteins (caseins and β‐lactoglobulin).ResultsIgEs from children allergic to cow's milk are capable of recognizing most part of milk proteins from mammals bred in European countries (ewe, goat, buffalo), while no serum used in this study contains IgEs reacting with camel's milk proteins. Camel's milk was also not recognized from circulating IgEs from a child specifically allergic to ewe's milk. Specific antibovine monoclonal antibodies cross‐reacted with proteins from other mammalian species, apart from those of camel.ConclusionsHomologies in amino acidic composition could justify the cross‐reactivity observed between proteins from different animal species. On the other hand, the phylogenetic difference could be responsible for the failed recognition of camel's proteins by circulating IgEs and monoclonal antibodies.
Fel d 1 and Can f 1 in settled dust and airborne Fel d 1 in allergen avoidance day‐care centres for atopic children in relation to number of pet‐owners, ventilation and general cleaning
Authors: WICKMAN  , EGMAR  , EMENIUS  , ALMQVIST  , BERGLIND  , LARSSON  , VAN HAGE‐HAMSTEN  
Published: March 2003

BackgroundSpecial day‐care centres for atopic children have been established in Sweden. Objective To study concentrations of cat (Fel d 1) and dog (Can f 1) allergens in settled dust and airborne cat allergen in day‐care centres in relation to pet ownership among children and staff, ventilation and general cleaning. Methods Twelve allergen avoidanceday‐care centres and 22 conventional day‐care centres were included in the study. Settled dust was collected and analysed with ELISA. Airborne cat allergen levels were measured in eight allergen avoidance and seven conventional centres with a personal air sampler and analysed with an amplified ELISA. Air change rate per hour (ACH) was measured. A questionnaire which focused on keeping of cat and dog among staff and children and frequency of general cleaning was used. Results In the allergen avoidance day‐care centres neither children nor staff reported ownership of cats or dogs, compared with 21/22 of the conventional centres in which children and staff kept furred animals. Fel d 1 and Can f 1 were found in settled dust in all day‐care centres. In the allergen avoidance compared with the conventional centres the concentrations of Fel d 1 and Can f 1 were lower, Fel d 1: median 0.64 μg/g vs 5.45 μg/g and Can f 1: 0.39 μg/g vs 2.51, both P < 0.001, and airborne Fel d 1 was also lower in the allergen avoidance centres compared with the control centres, 1.51 ng/m3 vs 15.8 ng/m3, P = 0.002. A correlation was found between airborne and settled Fel d 1, rs = 0.75, P < 0.001. Furthermore, a correlation was found between increased ACH and decreased levels of Fel d 1 in the air in the day‐care centres with no cat‐owners, rs = – 0.86, P = 0.007. No relation was found between levels of cat or dog allergen and amount of general cleaning. Conclusion Not keeping pets seems to reduce children’s exposure to pet‐allergen in their ‘working environment’. Additionally, appropriate ventilation seems to reduce Fel d 1 in the air in day‐care centres.
Mast cell tryptase as a mediator of hyperresponsiveness in human isolated bronchi
Authors: BERGER  , COMPTON  , MOLIMARD  , WALLS  , N'GUYEN  , MARTHAN  , TUNON‐DE‐LARA  
Published: March 2003

BackgroundAlthough the role of mediators and cytokines produced by mast cells is well established in asthmatic bronchial inflammation, the contribution of mast cell‐derived proteases to the development of hyperresponsiveness remains unclear. There have been reports indicating that tryptase alters the mechanical activity of animal airway smooth muscle or spontaneously sensitized human isolated airways.ObjectiveThe aim of this study was to analyse the effect of purified mast cell tryptase on non‐sensitized human isolated bronchi.MethodsBoth central and peripheral bronchi, dissected from lung specimens obtained at thoracotomy, were studied in terms of both mechanical activity i.e. isometric contraction in response to a variety of agonists and distribution of inflammatory cells i.e. immunohistochemistry.ResultsIn both proximal and distal bronchi, the reactivity to histamine was significantly increased by a previous incubation in the presence of 1 μg/mL of tryptase (increase in maximal force, ΔFmax was 12.1 ± 3.8%, and 8.8 ± 3.1%, respectively). This effect of tryptase on histamine‐induced contraction was completely abrogated in the presence of the protease inhibitor benzamidine (100 μmol/L). Histological examination of specimens exposed to tryptase demonstrated an increase in mast cell number within the subepithelial tissue whereas mast cell numbers in the epithelial layer concomittently decreased.ConclusionThese results indicate that human mast cell tryptase alters the contractile response of non‐sensitized human isolated bronchi and that this alteration is accompanied by a change in the mast cell distribution within the airway wall.
Eicosapentaenoic acid inhibits prostaglandin D2 generation by inhibiting cyclo‐oxygenase‐2 in cultured human mast cells
Authors: Obata  , Nagakura  , Masaki  , Maekawa  , Yamashita  
Published: March 2003

BackgroundEicosapentaenoic acid (EPA) is catalysed by cyclo‐oxygenase (COX), as is arachidonic acid, and is a competitive inhibitor of arachidonate metabolism.ObjectivesWe examined the effect of EPA on prostaglandin (PG) D2 generation in the cultured human mast cells with IgE‐anti‐IgE challenge incubation.MethodsCultured human mast cells were incubated with EPA (1 μmol/L) for 20 h, then challenged with anti‐IgE incubation after treatment with IgE. At the same time, COX inhibitors were tested to identify COX‐1 and COX‐2 activity. PGD2 synthetic activity was also assayed in a cell‐free homogenate of cultured mast cells with COX inhibitors and EPA. Histamine in the culture medium and in cells was assayed with the HPLC‐fluorescent method. PGD2 and PGD3 were assayed with gas chromatography‐mass spectrometry and the stable isotope dilution method.ResultsAlthough EPA incubation did not affect histamine release by cultured human mast cells in response to IgE‐anti‐IgE challenge incubation, it did decrease PGD2 generation by inhibiting the COX‐2 pathway. In contrast, in the cell‐free homogenate of cultured human mast cells, EPA inhibited both COX‐1 and COX‐2 activities.ConclusionPre‐incubation with EPA primarily affects the COX‐2 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE‐anti‐IgE challenge incubation. These findings suggest that COX‐1 and COX‐2 have different substrate flow systems in mast cells. They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti‐inflammatory substrate in human mast cells.
Genetic linkage of HLA–class II locus to mite‐specific IgE immune responsiveness
Authors: Stephan  , Kuehr  , Seibt  , Saueressig  , Zingsem  , Dinh  , Moseler  , Wahn  , Deichmann  
Published: March 2003

BackgroundIgE response to common inhalant allergens seems to be the major determinant of the development of atopic rhinitis and asthma but it has been difficult to demonstrate genetic control of the IgE response.ObjectiveTo investigate genetic linkage between specific IgE reactions to purified aero‐allergens (grass, birch, cat, mite) and the HLA–class II locus.MethodsDNA‐based HLA–class II typing was performed for determination of DRB1, DQB1 and DPB1 alleles. Linkage was studied by the affected sibpair method and the extended transmission disequilibrium test in 100 children from 40 nuclear families selected from a homogeneous population in south‐western Germany.ResultsSignificant linkage of mite‐specific IgE response to HLA–DPB (P = 0.00001), HLA–DRB (0.02) and HLA–DQB (P = 0.001) was revealed by sibpair analysis of MHC class II alleles and confirmed by the extended transmission disequilibrium test for HLA–DRB (P = 0.01) and HLA–DPB (P = 0.04). No consistent significant linkage between the HLA–class II locus and IgE response to grass pollen, birch pollen, and cat dander could be demonstrated.ConclusionThe findings are consistent with the existence of one or more genes in the HLA–class II region modifying the IgE immune response to common environmental allergens.
Peels of Rosaceae fruits have a higher allergenicity than pulps
Authors: Fernández‐Rivas  , Cuevas  
Published: March 2003

BackgroundIt is not uncommon that patients allergic to fruits such as apple, pear, and peach, refer adverse reactions after the ingestion of the whole fruit, but subsequently tolerate the pulp.ObjectiveThis study aimed to compare the allergenicity of peels and pulps of apple, peach, and pear in 33 patients allergic to these fruits.MethodsClinical reactivity to the ingestion of whole fruit (peel + pulp) and pulp was established by medical history. Peels and pulps were tested separately in skin prick tests (SPTs), histamine release tests (HRTs) and RASTs. Cross‐allergenicity between peel and pulp of apple and peach was studied by RAST inhibition.ResultsAdverse reactions appeared more frequently and were more severe when the whole fruit was eaten. More than 40% of patients allergic to apple and pear tolerated the ingestion of the pulp of these fruits, and reactions were only elicited by the intake of the whole fruit. Peels induced higher SPTs, HRTs and RASTs than pulps. An important cross‐allergenicity was found between the peel and pulp of apple and peach, although the amount of the shared allergenic epitopes seemed to be higher in peels.ConclusionOur results suggest that peels of Rosaceae fruits such as apple, peach, and pear, have a higher allergenicity than pulps, which is clinically relevant. This aspect should be considered in the evaluation of patients allergic to Rosaceae fruits, and in the production of diagnostic materials.
Submucous turbinectomy decreases not only nasal stiffness but also sneezing and rhinorrhea in patients with perennial allergic rhinitis
Authors: Mori  , Fujieda  , Igarashi  , Fan  , Saito  
Published: March 2003

BackgroundTurbinate surgery, in which mucous epithelium is resected, has often been used for patients with perennial allergic rhinitis, since the mucous epithelium is the principal site for immunoglobulin (Ig)E‐mediated allergic reactions and chronic inflammation.ObjectiveTo assess the effect of submucous turbinectomy on allergic rhinitis.MethodsSixty patients with severe perennial allergic rhinitis underwent submucous turbinectomy and were followed‐up for 1 year. Nasal symptoms were evaluated with a standard symptom score. Rhinometry was used to evaluate nasal congestion, and nasal provocation testsin vivowere performed to evaluate allergic reactions. In 16 cases, biopsies from the nose were also available for immunohistochemical analysis. These examinations were performed before and after submucous turbinectomy.ResultsThe mean total nasal symptom score (7.2 ± 1.7, mean ± sdbefore surgery) was significantly reduced after surgery (1.2 ± 1.4,P < 0.0001), and the effect of the surgery on nasal symptoms continued for at least 12 months (1.9 ± 1.8,P < 0.0001). Submucous turbinectomy reduced both nasal discharge and sneezing, as well as nasal stiffness. Histopathological examination following surgery revealed that the lamina propria was occupied by fibrous tissues, and that the number of vessels, nasal glands, eosinophils and infiltrating IgE+cells decreased in the turbinate. There were no significant differences in the levels of either house dust mite‐specific or non‐specific IgE in the serum between before and after surgery.ConclusionSubmucous turbinectomy preserving the ciliary epithelium is a powerful strategy for improving nasal symptoms induced by allergic reaction via the reduction in the number of allergy‐related cells in the nose.
Exhaled monoxides in asymptomatic atopic subjects
Authors: Horváth  , Barnes  
Published: March 2003

BackgroundAtopy is a genetically determined condition and some atopic people develop airway hyperresponsiveness and sometimes asthma later in life. Since airway inflammation may be present before the onset of clinical symptoms of asthma, early and noninvasive detection of inflammation would be useful in atopic subjects. Mediators produced by activated inflammatory cells may lead to induction of inducible nitric oxide synthase producing nitric oxide (NO) and inducible heme oxygenase releasing carbon monoxide (CO) in the airways. Both monoxides are present in exhaled air and their levels are elevated in asthma reflecting airway inflammation.ObjectiveWe have measured exhaled NO and CO levels in atopic and nonatopic healthy non‐smoking subjects to determine whether inflammation is present in the airways.MethodsExhaled NO was measured by a chemiluminescence analyser and exhaled CO electrochemically and NO in asymptomatic atopic and age‐matched nonatopic normal subjects.ResultsExhaled NO and CO levels were both significantly elevated in 15 atopic subjects compared with 40 nonatopic individuals (means ± sem: 18.3 ± 3.0 p.p.b. vs 6.3 ± 0.3 p.p.b., P < 0.0001 and 4.7 ± 0.3 p.p.m. vs 2.8 ± 0.2 p.p.m., P = 0.0005, respectively).ConclusionIncrease in exhaled monoxide levels may be an early and noninvasive marker of airway inflammation in asymptomatic atopic subjects.
Association of FcεR1‐β polymorphisms with asthma and associated traits in Australian asthmatic families
Authors: Palmer  , Rye  , Gibson  , Moffatt  , Goldblatt  , Burton  , Cookson  , Lesouëf  
Published: March 2003

BackgroundAsthma is a genetically complex disease, and is characterized by elevated serum immunoglobulin E (IgE) levels, elevated blood eosinophil counts and increased airway responsiveness. Polymorphisms in the β subunit of the high affinity receptor for IgE (FcεR1‐β) have been previously associated with these phenotypes and with an increased risk of asthma.ObjectiveTo investigate the association of all known bi‐allelic polymorphisms in FcεR1‐β to asthma and quantitative traits associated with asthma in a selected sample of Australian asthmatic children and their nuclear families.MethodsAustralian Caucasian nuclear families (n = 134 subjects) were recruited on the basis of a child proband with current, severe, symptomatic asthma. The quantitative traits assessed included serum levels of total IgE and specific IgE to house dust mite and mixed grass, blood eosinophil counts and the dose–response slope of the forced expiratory volume in 1 s to histamine provocation.ResultsNeither the Leu181 nor the E237G mutations were detected in this population. Allele B of RsaI intron 2 (RsaI_in2*B) was significantly associated with physician‐diagnosed asthma (ever) (P = 0.002). Alleles of both the RsaI_in2 and RsaI exon 7 (RsaI_ex7) polymorphisms were significantly associated with loge total serum IgE levels and the combined RAST index. RsaI_ex7 was also associated with loge blood eosinophil counts. These associations were independent of age, sex and familial correlations.ConclusionThis study supports a role for the FcεR1‐β gene or a nearby gene in the pathogenesis of asthma.
Are anaphylactic reactions to snake bites immunoglobulin E‐mediated?
Authors: Reimers  , Weber  , Müller  
Published: March 2003

BackgroundBites by poisonous European snakes of the genus Vipera lead to local tissue damage and systemic symptoms such as generalized oedema, hypotension, gastrointestinal symptoms, haemolysis and renal dysfunction. Not rarely anaphylactic symptoms like urticaria, localized angioedema and asthma are observed.ObjectiveTo look for snake venom‐specific immunoglobulin (Ig) E antibodies in patients with a history of bites by European vipers and for cross‐reactions with Hymenoptera venoms, that have a similar composition.MethodTen patients with a history of bites by Vipera aspis or Vipera berus were investigated. Three patients had been bitten only once, and two of these had developed only local reactions. Four reported previous allergic reactions to Hymenoptera stings. All patients, 10 Hymenoptera venom‐allergic and five nonallergic individuals who served as controls underwent i.c. skin test endpoint titration with snake (V. aspis, V. berus) and Hymenoptera venoms (honey bee, Vespula spp.) and were investigated for specific serum IgE antibodies to the same venoms.ResultsSeven of the eight patients with systemic snake bite reactions had both positive skin tests and serum IgE antibodies to snake venoms, while these tests were negative in the two patients with only local reactions to snake bites and all controls. Seven of the eight patients with systemic snake bite reaction also had positive skin tests and specific IgE with one or both Hymenoptera venoms. By RAST‐inhibition with sera of four patients with high IgE to both Vipera and Hymenoptera venoms, partial cross‐reactivity could be demonstrated in one.ConclusionsAnaphylactic reactions following snake bites may be IgE‐mediated, especially in patients with repeated bites.