bluuebunny/crossref_metadata_2025 huggingface dataset:
Local side‐effects of inhaled corticosteroids in asthmatic children: influence of drug, dose, age, and device
Authors: Dubus J. C. , Marguet C. , Deschildre A. , Mely L. , Le Roux P. , Brouard J. , Huiart L. ,   Réseau de Recherche Clinique en Pneumologie Pédiatrique
Published: March 2003

Background: The objective was to investigate the local side‐effects of inhaled corticosteroids (ICS) in daily life in asthmatic children, particularly the younger ones, by an observational prospective cross‐sectional cohort study.

Methods: Asthmatic children (n=639, 75.9±48.9 months, 61.3% boys), treated with beclomethasone dipropionate (BDP) (721.0±287.3 µg per day) or budesonide (BUD) (835.5±684.9 µg per day) for at least 1 month, were recruited at the time of a scheduled visit. Local side‐effects were researched by questionnaire (cough during inhalation, hoarseness, dysphonia, and thirsty feeling) and clinical examination (perioral dermatitis, oral candidiasis, and tongue hypertrophy).

Results: Exactly 63.3% of the children aged under 6 years and 59.5% of the older ones reported one local side‐effect. Cough (39.7%) was dependent on young age, use of BDP, and mainly use of spacer device, with an OR of 4.7 (95% CI: 2.7–8.2). Thirsty feeling (21.9%) and hoarseness (14.1%) occurred in children using ICS and long‐acting β2‐agonists. Dysphonia (11.1%) was favored by high doses of BDP and BUD, and by inhalation from spacer devices or nebulizers. No factor favored oral candidiasis (10.7%). Perioral dermatitis (2.9%) and tongue hypertrophy (0.1%) were associated with nebulization.

Conclusions: Local side‐effects of ICS are common in asthmatic children of all_ ages, and the device used constitutes the most influential factor.
Occupational asthma and rhinitis due to glutaraldehyde: changes in nasal lavage fluid after specific inhalatory challenge test
Authors: Pałczyński C. , Walusiak J. , Ruta U. , Górski P.
Published: March 2003

Background:Glutaraldehyde (GA) is a known respiratory sensitizers, and some studies have reported occupational asthma in exposed workers. Specific changes in nasal lavage fluid (NLF) induced by high‐molecular‐weight allergen provocation in sensitized subjects were described previously. The purpose of this study was to evaluate the changes in cytogram, protein content, eosinophil cationic protein (ECP), and mast‐cell tryptase concentrations in NLF after GA inhalation challenge in patients with a positive history of GA‐induced asthma and late or dual asthmatic response due to exposure to low‐level GA.

Methods:A single‐blind, placebo‐controlled study was performed on 11 health workers with occupational asthma and rhinitis due to GA. The control groups comprised 10 atopic subjects with perennial asthma and rhinitis and 10 healthy ones. A “nasal pool” technique was used to evaluate the examined parameters in nasal washings before and 30 min, 4 h, and 24 h after the inhalatory provocation with GA and placebo.

Results:There was a significant increase in eosinophil number and percentage, and albumin, ECP, and tryptase concentrations in NLF from patients with occupational asthma and rhinitis when compared to controls.

Conclusions:The results indicate the immunologic mechanism of GA‐induced asthma and the applicability of the “nasal pool” technique as the diagnostic procedure in GA‐induced airway allergy.
Increased lymphoid MxA expression in acute asthma exacerbation in children
Authors: Itazawa T , Adachi Y , Imamura H , Okabe Y , Yamamoto J , Onoue Y , Adachi Y. S. , Miyawaki T , Murakami G
Published: March 2003

Background: Although the association between acute asthma exacerbation and viral infection has been well documented, virus identification rates vary. It has recently been reported that the expression of MxA protein in lymphocytes, inducible by type I interferons, can serve as a sensitive marker for viral infection in the host. The objective was to determine the contribution of viral infection to precipitation of asthma attacks in children.

Methods: We studied 186 asthmatic children, aged 0–12 years, over a 1‐year period to evaluate MxA protein levels in peripheral blood lymphocytes by using a flow cytometric analysis in whole blood.

Results: Of all the subjects, 80 (47%) exhibited significantly elevated levels of MxA expression in lymphocytes, presumably indicating the states of viral infection. The association of viral infections with acute asthma exacerbation seemed to be marked in younger children: enhanced MxA expression was seen in 73.3% of infants (aged 0–1 year), 49.5% of toddlers (aged 2–5 years), and 26% of schoolchildren (aged 6–12 years). Seasonal changes in the frequency of viral infection associated with deterioration were also observed.

Conclusions: Flow cytometric assay of MxA protein expression in whole blood appears to be an easy and useful method to evaluate viral infections in acute asthma exacerbation.
B‐cell epitopes of the allergen Chi t 1.01: peptide mapping of epitopes recognized by rabbit, murine, and human antibodies
Authors: Van Kampen V. , Liebers V. , Sander I. , Chen Z. , Baur X. , Raulf‐Heimsoth M. , Falkenberg F. W.
Published: March 2003

Background: Chi t 1.01, a hemoglobin of the midge Chironomus thummi thummi, is a widespread environmental and occupational allergen. The aim of the present investigation was to identify and compare peptides involved in B‐cell epitopes of Chi t 1.01 recognized by 15 human IgE sera, six murine monoclonal antibodies (mAbs), and a polyclonal rabbit antiserum.

Methods: Synthetic peptides 19–21 amino acids long covering the whole Chi t 1.01‐sequence were covalently coupled to activated paper disks as well as adsorbed to wells of immunoplates and used for enzyme‐linked immunosorbent assay. For fine epitope mapping, we used overlapping synthetic octapeptides with one amino‐acid offset.

Results: Peptides containing the amino acids 13–17, 23–29, and 40–50 were recognized by three of the mAbs, while three other mAbs reacting with none of the peptides obviously recognized conformational epitopes. Binding sites for rabbit antibodies and for human IgE antibodies were scattered over the whole molecule. The peptide 80–100 seemed to comprise at least one important IgE epitope. Depending on the method of antigen binding to the solid phase, differing results were obtained.

Conclusions: Several linear epitopes in Chi t 1.01 are recognized by human IgE antibodies, by mAbs, and by polyclonal rabbit antibodies. In addition, the results indicate the presence of conformational epitopes.
Recruitment of CD1a+ Langerhans cells to the nasal mucosa in seasonal allergic rhinitis and effects of topical corticosteroid therapy
Authors: Till S. J. , Jacobson M. R. , O'Brien F. , Durham S. R. , KleinJan A. , Fokkens W. J. , Juliusson S. , Lowhagen O.
Published: March 2003

Background: Local antigen presentation may be necessary for both primary and recall T‐cell responses to grass pollen in hay fever patients. We examined the effect of seasonal allergen exposure on nasal mucosal antigen‐presenting cell (APC) populations and the effects of topical corticosteroid therapy.

Methods: Nasal biopsies were collected from 46 grass pollen‐sensitive seasonal rhinitis patients before the grass‐pollen season. A second biopsy was collected during the pollen season, when patients had received 6 weeks' treatment with either fluticasone propionate (200 µg, twice daily) or placebo. Cell populations in biopsy sections were quantified by immunocytochemistry.

Results: Significant increases in submucosal and epithelial CD1a+ Langerhans cells, but not CD68+ macrophages or CD20+ B cells, were observed during the pollen season. Seasonal increases in CD1a+ Langerhans cells were inhibited by corticosteroid therapy.

Conclusions: Recruitment of CD1a+ Langerhans cells to the nasal mucosa during natural seasonal allergen exposure may contribute to local T cell responses. Topical corticosteroids may act, at least in part, by inhibiting effective allergen presentation to T cells through inhibition of recruitment of Langerhans cells to the nasal mucosa.
Validation of a new questionnaire on asthma, allergic rhinitis, and conjunctivitis in young adults
Authors: Kilpeläinen M. , Terho E.O. , Helenius H. , Koskenvuo M.
Published: March 2003

Background: Epidemiologic data on asthma and allergies among adults are mainly based on questionnaires: this study validates the questions on asthma, allergic rhinitis, and conjunctivitis of a new Finnish questionnaire.

Methods: To validate questions used in a country‐wide study among university students aged 18–25 years, we examined 150 subjects who had ever reported asthma or wheezing, and 140 without asthma symptoms. Questions were validated in relation to current diseases including

1) symptoms detected during the preceding year at the physician's interview

2) objective measurements, such as methacholine challenge, skin prick tests, and specific IgE.

Data were adjusted for original proportions of “asthmatics” and “nonasthmatics” in the questionnaire study.

Results: Questions on “reported asthma” and “doctor‐diagnosed asthma” had good positive predictive value (PPV) and specificity in diagnosing current asthma. The question on “attacks of shortness of breath with wheezing”, and especially the question on “cough with wheezing” were most sensitive. Questions on “allergic nasal symptoms” and “allergic eye symptoms” that were “related to pollen or animals” were sensitive, but a further question on doctor's diagnosis yielded higher specificity and PPV.

Conclusions: Diagnosis‐based questions were found suitable for risk‐factor studies, because of their good specificity and PPV, and symptom‐based questions for screening, because they were highest in sensitivity.
Changes over 13 years in skin reactivity to histamine in cohorts of children aged 9–13 years
Authors: Ronchetti R. , Pia Villa M. , Ciofetta G. , Barreto M. , Falasca C. , Martella S.
Published: March 2003

Background: Several studies report substantial differences in the prevalence of skin test reactivity to allergens in children from adjacent geographic areas; others report an increased prevalence over time. To find out whether these differences depend on variations in skin reactivity to histamine, we determined the time trend of histamine wheal sizes in successive cohorts of unselected children living in the same area (Viterbo, Italy).

Methods: We conducted three epidemiologic surveys, each including children aged 9 and 13 years. The 1983–7 study investigated 170 children (150 were tested twice); the 1992 study, 158 children; and the 1996 study, 208 children.

Results: In both age groups, the mean diameter of the wheal induced by histamine skin prick tests (10 mg/ml) increased significantly over time (9‐year‐olds: 3.25 mm in 1983, 4.68 in 1992, and 5.89 in 1996; 13‐year‐olds: 3.89 mm in 1987, 5.18 in 1992, and 6.50 in 1996) (P<0.001 between subsequent studies). The distribution of the wheal diameters for both ages showed a trend to a right shift in the three successive studies (P<0.001). The dose‐response curves for three histamine concentrations (0.2, 1, and 10 mg/ml) had significantly steeper slopes in 1996 than in 1983–7 (P<0.001).

Conclusions: The marked time‐related increase in the size of the histamine wheals could help to explain the trend toward an increased prevalence of positive allergen skin test reactions reported during the past years. The causes of increased skin reactivity to histamine remain conjectural.
Basophil activation during pollen season in patients monosensitized to grass pollens
Authors: Saporta M. , Kamei S. , Persi L. , Bousquet J. , Arnoux B.
Published: March 2003

Background: Basotest® is a new basophil‐activation test based upon the expression of CD63 (gp53) in the presence of allergens. It is an effective diagnostic test for pollen‐allergic patients. However, it is not known whether Basotest results differ during the pollen season.

Methods: We examined the activation of basophils by Basotest in 13 patients sensitized only to grass pollen, before and during the pollen season, in order to assess whether Basotest could be used as a diagnostic test during the pollen season. Dose‐response curves with 10‐fold increasing concentrations of timothy grass pollen (10−4 to 100 AU) were carried out.

Results: Basophils were not activated spontaneously during the pollen season since the CD63 expression was below detectable levels before in vitro cell activation. A decreased percentage of activated basophils at the peak of activation was found in comparing the pre‐ and in‐season tests, but all patients had a positive test. When basophil activation was at its peak, the allergen concentration was similar during the two periods. Moreover, the median area under the curve was significantly (P<0.02) reduced during the season as compared to before the season.

Conclusions: Basotest can therefore be used as a diagnostic test during the pollen season, but the allergen exposure needs to be characterized if quantitative studies are performed.
A well‐tolerated grass pollen‐specific allergy vaccine containing a novel adjuvant, monophosphoryl lipid A, reduces allergic symptoms after only four preseasonal injections
Authors: Drachenberg K. J. , Wheeler A. W. , Stuebner P. , Horak F.
Published: March 2003

Background: We present data showing that a Th1‐inducing adjuvant can reduce the number of injections required for allergy vaccination. Allergy vaccination is the only treatment for type 1 hypersensitivity that can alter the underlying disease process. A switch of specific T‐cell activity from Th2>Th1 to Th1>Th2 is believed to be an important change seen after long‐term vaccination therapy. An immunologic adjuvant that enhances such a switch could be used to reduce the number of injections required. This would improve compliance with the treatment and provide pharmacoeconomic advantages. Such an adjuvant is 3‐deacylated monophosphoryl lipid A (MPL® adjuvant, Corixa).

Methods: A multicentre, placebo‐controlled, randomized, double‐blind clinical study was performed with a new standardized allergy vaccine comprising a tyrosine‐adsorbed glutaraldehyde‐modified grass pollen extract containing MPL® adjuvant. Four subcutaneous injections of the active product were given preseasonally to 81 grass pollen‐sensitive subjects, and 60 received placebo injections (tyrosine alone). Diary cards were used to record symptoms and medication taken during approximately 30 days of the grass pollen season.

Results: There was a statistical advantage in favour of the active treatment for nasal (P=0.016) and ocular (P=0.003) symptoms and combined symptom and medication scores (P=0.013). Titrated skin prick testing revealed a significant reduction of skin sensitivity in the active group compared to placebo (P=0.04). Grass‐pollen‐specific IgG antibody was raised by active treatment (P<0.01). A rise in IgE antibody was seen in the placebo group during the season (P<0.01). The first year's treatment rise of IgE was not seen in the active group, and no rise occurred during the pollen season. More local adverse events were seen in the active group. There was no difference in generalized adverse events.

Conclusions: A new, well‐tolerated allergy vaccine, incorporating a Th1‐inducing adjuvant, MPL®, was efficacious and after only four preseasonal injections produced antibody changes normally associated with long injection schedules. This may encourage wider application of allergy vaccination. The vaccine is now available in a number of countries as Pollinex Quattro®.
IgE‐mediated anaphylaxis to Thiomucase®, a mucopolyssacharidase: allergens and cross‐reactivity
Authors: Caballero T. , Alonso A. , De Miguel S. , Martín‐Esteban M. , Varga B. , Pascual C. Y. , López‐Serrano M. C.
Published: March 2003

Background: Thiomucase® is a mucopolysaccharidase obtained from ovine tissues mainly used to facilitate the diffusion of local anaesthetics and in the treatment of cellulitis. A patient with an anaphylaxis in relation to the intramuscular administration of Thiomucase® is reported.

Objective: To investigate Thiomucase® allergens and their possible relationship with dander allergens and animal albumins.

Material and methods: Skin prick tests (SPT) and serum‐specific IgE were performed with Thiomucase® and danders. Thiomucase® SDS‐PAGE immunoblotting was performed in order to study allergens. RAST/CAP inhibition and SDS‐PAGE immunoblotting inhibition were carried out to study the cross‐reactivity.

Results: Skin prick tests (SPT) were positive to Thiomucase®, animal dander (cat, dog, sheep, other), bovine serum albumin (BSA), and echinococcus. Specific IgE was also positive to Thiomucase®, animal dander (cat, dog, sheep, other), BSA and echinococcus. In the RAST‐CAP inhibition assays BSA was nearly completely inhibited by Thiomucase®, Thiomucase® was partially inhibited by BSA and cat and Echinococcus granulosus was partially inhibited by sheep and Thiomucase®. In the Thiomucase® SDS‐PAGE immunoblotting several proteins fixed IgE, ranging from 20 kDa to > 94 kDa, the strongest with 43 kDa. The IgE fixation to BSA, cat and sheep in the SDS‐PAGE immunoblotting was completely inhibited by the preincubation of the serum with Thiomucase®.

Conclusions: An IgE‐mediated anaphylaxis to Thiomucase® is documented. Multiple allergens are recognized in Thiomucase® by the patient serum, the main with 43 kDa. Partial cross‐reactivity with BSA, cat dander and sheep dander is documented.
Comparison of cetirizine, ebastine and loratadine in the treatment of immediate mosquito‐bite allergy
Authors: Karppinen A. , Kautiainen H. , Petman L. , Burri P. , Reunala T.
Published: March 2003

Background:  People frequently experience whealing and delayed papules from mosquito bites. Whealing is mediated by antisaliva immunoglobulin (Ig)E antibodies and histamine. Cetirizine, ebastine and loratadine have earlier shown effects on mosquito‐bite reactions but no comparative studies exist.

Methods:  A double‐blind, placebo‐controlled, cross‐over study was performed with cetirizine 10 mg, ebastine 10 mg and loratadine 10 mg in 29 mosquito‐bite‐sensitive adults exposed to Aedes aegypti mosquito‐bites. The size of the bite lesion and the intensity of pruritus (visual analog scale) were measured at 15 min and 2, 6 and 24 h.

Results:  Cetirizine and ebastine, but not loratadine, decreased significantly the size of whealing (P < 0.01) and accompanying pruritus (P < 0.001) compared to placebo. Cetirizine was most effective on pruritus but caused more often sedation than ebastine or loratadine. The delayed bite symptoms remained too faint for any statistical comparison.

Conclusion:  This comparative study in mosquito‐bite‐sensitive adults shows that cetirizine and ebastine decrease significantly whealing and accompanying pruritus, and that cetirizine seems to be the most effective against pruritus.
Development of new IgE specificities to allergenic components in birch pollen extract during specific immunotherapy studied with immunoblotting and Pharmacia CAP System™
Authors: Movérare R. , Elfman L. , Vesterinen E. , Metso T. , Haahtela T.
Published: March 2003

Background: New IgE sensitizations to proteins in allergen extracts have been shown to occur during allergen‐specific immunotherapy (IT). However little is known about the kinetics of the changes in antibody reactivities.

Methods: Twenty‐four allergic children and adults were treated with birch pollen rush IT (RIT). Fifteen matched patients served as allergic controls. Sera were obtained at regular intervals for up to three years and analyzed with immunoblotting and Pharmacia CAP System™ with recombinant (r) birch pollen allergens (rBet v 1, rBet v 2, and rBet v 4).

Results: All birch‐allergic patients had specific IgE to the major birch pollen allergen Bet v 1, but only three had IgE to rBet v 2 and/or rBet v 4 at the beginning of the study. New IgE sensitizations developed in 65% of the birch RIT‐treated patients when studied by immunoblotting. Twenty‐nine percent of the patients developed new sensitizations to rBet v 2 and/or rBet v 4 during RIT as measured by Pharmacia CAP System. Generally, new specific IgE reactivities occurred after at least one year of RIT, and only at low levels (< 1 kUA/l).

Conclusions: Sensitization to additional allergenic pollen components frequently occurs during prolonged birch RIT. However, the IgE levels are low and the clinical relevance is not known.
Passive sensitization of human airways induces mast cell degranulation and release of tryptase
Authors: Berger P. , N′guyen C. , Buckley M. , Scotto‐Gomez E. , Marthan R. , Tunon‐de‐Lara J. M.
Published: March 2003

Background: This study was designed to examine the effect of passive sensitization (PS) on human bronchial mast cells. PS with asthmatic serum induces a hyper‐responsiveness to nonspecific agonists, and immunoglobulin (Ig)E binding mainly on mast cells.

Methods:  Bronchi dissected out from 19 lung specimens were incubated in normal or asthmatic serum. Immunohistochemistry was performed using monoclonal antibodies (MoAbs) directed against tryptase, chymase, or c‐kit. Mast cells were classified as fully granulated (type I), partly (type II) or largely degranulated (type III). Tryptase was measured in supernatant using ELISA. Contractile response was recorded in a separated set of experiments using an organ bath system.

Results:  PS decreased both tryptase positive cells (47.9 ± 10.0 vs. 26.7 ± 4.8 cell/mm2, P = 0.003) and chymase positive cells (26.1 ± 3.3 vs. 14.9 ± 1.8 cell/mm2, P = 0.01), but did not alter the number of c‐kit positive cell. PS decreased the proportion of type I (55.4 vs. 28.9%, P < 0.0001) and, concomitantly increased that of types II (23.2 vs. 41.0%, P < 0.0001) and III (21.4 vs. 30.1%, P = 0.04). Following PS, tryptase concentration significantly increased and the magnitude of histamine response, was correlated with the amount of type II mast cells.

Conclusion:  PS of human isolated bronchi induces a mast cell degranulation related to in vitro hyper‐responsiveness, along with a tryptase release.
Cross‐reactivities between date palm (Phoenix dactylifera L.) polypeptides and foods implicated in the oral allergy syndrome
Authors: Kwaasi A. A. A. , Harfi H. A. , Parhar R. S. , Saleh S. , Collison K. S. , Panzani R. C. , Al‐Sedairy S. T. , Al‐Mohanna F. A.
Published: March 2003

Background: Date fruit and pollen antigens share a number of cross‐reactive epitopes. Date pollen has been shown to cross‐react with antigens from Artemisia, cultivated rye (Secale cereale), Timothy grass (Phleum pratense), Sydney golden wattle (Acacia longifolia) and Bermuda grass (Cynodon dactylon) pollen. The present study was carried out to examine any cross‐reactivities between date palm polypeptides and antigens of some common foods and vegetables that have been implicated in the oral allergy syndrome (OAS). Because most of such cross‐reactivities in other allergens are attributable to the presence of carbohydrate chains and profilin, their role was also investigated.

Methods: Fresh extracts of 20 common fruits and vegetables were prepared. Putative date profilins were isolated by affinity chromatography using a poly L‐proline column. Date fruit extracts were digested by various endoglycosidases and the immunoglobulin (Ig)E binding of the postdigest products was assessed in immunoblots. Rabbit antisera to whole date fruit extracts, Timothy grass profilin and putative date profilins, as well as human sera from date sensitive individuals were used in immunoblotting, ELISA and in inhibition experiments.

Results: IgG, ELISA and immunoblot results with the different rabbit antisera and date‐sensitive atopic sera showed several antigenic cross‐reactivities and similar cross‐reactivities were seen with birch, date and timothy grass profilins. IgE, ELISA and immunoblot experiments with pooled date sensitive human sera showed a range of cross‐reactivities with some food extracts. A number of the IgE cross‐reactivities could be inhibited after preabsorption of pooled sera with date extracts. Sixty‐six percent of individual date hypersensitive human sera bound IgE in putative date fruit profilin and their pooled sera bound IgE in birch pollen profilin. IgE‐binding of the endoglycosidase digested date fruit extracts to atopic serum pool was restricted to only a very low molecular weight band of 6.5–8 kDa.

Conclusion: These results indicate that date palm polypeptides share cross‐reactive IgG and IgE epitopes with a number of foods implicated in the oral allergy syndrome, bind to birch and Timothy grass profilins and bind IgE through glycosyl residues. The clinical relevance of these cross‐reactivities needs to be further elucidated.
Characterization of the human T cell response to rye grass pollen allergens Lol p 1 and Lol p 5
Authors: Burton M. D. , Papalia L. , Eusebius N. P. , O'Hehir R. E. , Rolland J. M.
Published: March 2003

Background:  Knowledge of dominant T cell epitopes of major allergens recognized by allergic individuals is required to improve efficacy and safety of allergen immunotherapy. Rye grass pollen (RGP) is the most important source of seasonal aeroallergens in temperate climates and Lol p 1 and Lol p 5 are the two major IgE‐reactive allergens. This study aimed to characterize the T cell response to these allergens using a large panel of RGP‐sensitive individuals.

Methods:  Short‐term RGP‐specific T cell lines (TCL) were generated from 38 RGP‐sensitive subjects and stimulated with Lol p 1 and/or Lol p 5 allergens and synthetic 20‐mer peptides. Proliferative responses were determined by 3H‐thymidine uptake and IL‐5 and IFN‐γ in culture supernatants analysed by ELISA.

Results:  Of 17 subjects tested for reactivity to both allergens 16 (94%) responded to Lol p 1 and/or Lol p 5, establishing these as major T cell‐reactive allergens. Sites of T cell reactivity were spread throughout the allergen molecules but regions of high reactivity were found. For Lol p 1 these spanned residues 19–38, 109–128, 154–173, 190–209, and for Lol p 5 37–56, 100–119, 145–164, 154–173, 190–209, 217–236 and 226–245. IL‐5 and IFN‐γ were produced by T cells cultured with proliferation‐inducing peptides.

Conclusions:  T cell responses to RGP major allergens have been extensively characterized, providing fundamental information for developing T cell‐targeted immunotherapy for RGP allergy.
Skin test reactivity to natural and recombinant Blomia and Dermatophagoides spp. allergens among mite allergic patients in the UK
Authors: Simpson A. , Green R. , Custovic A. , Woodcock A. , Arruda L. K. , Chapman M. D.
Published: March 2003

Background: Many asthmatics in tropical and subtropical areas have positive skin prick tests to both Dermatophagoides spp. and to the mite Blomia tropicalis. This may be due to recognition by IgE of cross‐reactive allergens between the different mite species or because of sensitization to species‐specific allergens. A 14‐kDa Blomia tropicalis allergen, Blo t 5, has been cloned and shows 40% sequence homology with Der p 5. The aim of this study was to investigate reactivity to B. tropicalis in patients known to be sensitized to D. pteronyssinus and to assess allergenic activity and cross‐reactivity of recombinant (r) Group 5 allergens amongst these patients, who live in the UK and who are not exposed to B. tropicalis in their homes.

Methods: Patients (n = 19) with asthma and/or rhinitis were selected based on clinical history and a positive skin prick test to D. pteronyssinus extract and were compared with non‐allergic skin test negative controls (n = 10). IgE antibody responses to Blomia tropicalis, Dermatophagoides pteronyssinus, rDer p 5 and rBlo t 5 were compared by quantitative intradermal skin testing using serial 10‐fold dilutions of each allergen. End point titre was the highest dilution giving an 8 × 8 mm wheal at 15 min. IgE antibodies to Blomia tropicalis, Dermatophagoides pteronyssinus, rDer p 5 and rBlo t 5 were measured using RAST, CAP and RIA, respectively.

Results: All 19 patients had positive skin tests to D. pteronyssinus at concentrations of 0.001 to 1 AU/ml and 10 were skin test positive to rDer p 5 at concentrations of 10−4 to 5 µg/ml. Positive intradermal tests to Blomia tropicalis were seen in 12/19 patients at concentrations of 0.002 to 2 µg/ml. However none of the patients had positive skin tests to rBlo t 5. Non‐allergic controls were all skin test negative at the highest concentration of each allergen tested. All subjects had quantifiable specific IgE to D. pteronyssinus, but only two had IgE to B. tropicalis. IgE to Der p 5 was found in six patients, but no patients had IgE to Blo t 5.

Conclusions: This study of patients naturally exposed to D. pteronyssinus but not to Blomia tropicalis, provides evidence for IgE mediated cross‐reactivity between allergens produced by both mite species. The results suggest that the Group 5 allergens of D. pteronyssinus and B. tropicalis are species‐specific.
Role of complement in xenotransplantation
Authors: Mollnes T. E. , Fiane A. E.
Published: March 2003

The xenotransplantation research is driven by the increasing gap between the number of patients with end‐stage organ failure on waiting lists for transplantation and the supply of allografts. The lack of success in developing suitable artificial organs for permanent treatment of organ failure has further strengthened the need for xenotransplantation research. Pigs are now generally accepted to be the source animal of choice. Transplantation of pig organs to humans faces several barriers which have to be overcome before it comes to clinical application: (1) anatomical and physiological conditions; (2) immunological rejection mechanisms; (3) molecular compatibility between signal molecules of the two species; (4) risk of transmission of microorganisms, particularly pig endogenous retroviruses; and (5) legal and ethical aspects both with respect to the animal and the recipient. Here we will focus on the role of the complement system in the rejection of immediately vascularized pig‐to‐primate xenografts. The hyperacute rejection occurring within minutes after transplantation is mediated by binding of natural antibodies to the Galα(l‐3)Gal epitope on the endothelial cells with subsequent complement activation. Whereas inhibition of complement activation protects against hyperacute rejection, the role of complement in the later rejection phases is less clarified.
The 18 kDa peanut oleosin is a candidate allergen for IgE‐mediated reactions to peanuts
Authors: Pons L. , Chery C. , Romano A. , Namour F. , Artesani M. C. , Guéant J.‐L.
Published: March 2003

Background : Peanut allergy is one of the five most frequent food allergies in children and in adults. Recently, we purified and evaluated the allergenicity of peanut oleosins, a family of small‐sized proteins involved in the formation of peanut oil bodies.

Methods:  Allergenicity of the purified native protein and of the recombinant protein was tested by Western blot and by IgE‐RIA.

Results:  We found IgE‐binding with oleosin in 3 of 14 sera of patients who had suffered an allergic reaction to peanuts. Two sera reacted weakly against 16–18 kDa proteins corresponding to oleosin monomers, in Western blot. The main reacting bands had a molecular size estimated at ≈34 kDa, ≈50 kDa and ≈ 68 kDa and could therefore correspond to oleosin oligomers. IgE reactivity was higher in extracts from roasted peanuts. The same phenomenon occurred with crude soybean oil fraction, with two bands of 16.5 and 24 kDa corresponding to monomers, and two bands of 50 kDa and 76 kDa corresponding to dimers and trimers, respectively. The 18 kDa band was observed in the 3 Western blots of a membrane‐enriched fraction of recombinant oleosin produced in the Sf9‐baculovirus expression system (performed with the 3 patient sera).

Conclusions:  We have characterized a new peanut allergen which belongs to the oleosins, a family of proteins involved in the formation of oil bodies. The protein may be involved in some of the allergic cross‐reactions to peanuts and soybeans.
Mexican geneticists' opinions on disclosure issues
Authors: Lisker Rubén , Carnevale Alessandra , Villa Jose Antonio , Armendares Salvador , Wertz Dorothy C
Published: March 2003

Here we discuss the opinion on disclosure issues (respect for confidentiality) of Mexican professionals working in clinical genetics. The information was obtained from a comprehensive survey on genetics, ethics and society organized by Wertz and Fletcher in 1993, in which geneticists from 37 nations participated. The responses to 18 questions from 64 of 89 (72%) geneticists invited to participate were analyzed for this paper. The questions were directed to investigate: a) disclosure of clinically relevant information in several situations such as testicular feminization syndrome, mosaic de novo balanced translocation in a fetus, or accidental discovery of non‐paternity; b) the protection of patient's privacy from the spouse or blood relatives; and c) confidentiality in cases with high genetic risk to relatives. The results reveal significant disparity between general responses related to disclosure of all clinically relevant information and specific responses in particular cases. In addition, professionals appear to give higher priority to protecting the spouse, blood relatives or public safety, rather than the patient's confidentiality.
Evaluation of the facioscapulohumeral muscular dystrophy (FSHD1) phenotype in correlation to the concurrence of 4q35 and 10q26 fragments
Authors: Köhler Jutta , Röhrig Dorothee , Bathke Klaus D , Koch Manuela C
Published: March 2003

Probe p13E‐11 (locus D4F104S1) detects two highly homologous polymorphic loci on chromosomes 4q35 and 10q26. Previous reports in the literature have described a correlation of shortened 4q35‐specific fragments and facioscapulohumeral muscular dystrophy (FSHD1). We have identified 30 FSHD1 families (46 patients) carrying one short 4q35 and one short 10q26 fragment. The clinical data of these patients were compared with those of 47 families (131 patients) showing a single short 4q35 fragment, in order to evaluate a potentially modifying influence of shortened 10q26 fragments on the phenotype. According to our results, the polymorphic locus on 10q26 does not modify the FSHD1 phenotype. The normal population (14%) and our FSHD1 population (13%) did not significantly differ in the overall frequency of short polymorphic 10q26 fragments. The specificity of the p13E‐11/EcoRI–BlnI test for FSHD1 was 100%.
A proven case of materno‐foetal transfusion determined by cytogenetic and DNA analysis
Authors: Paoloni‐Giacobino A , Dutoit Mh , Morris Ma , Dahoun Sp
Published: March 2003

We report a case of materno‐foetal transfusion in a phenotypically normal male foetus after death in utero at the 35th week of gestation. We have used cytogenetic and polymerase chain reaction (PCR) microsatellite analysis to determine the presence of maternal cells in foetal blood collected by intracardiac puncture. In the intracardiac blood sample, maternal cells were estimated to comprise between 5 and 10% of nucleated foetal blood cells. When there is a suspicion of foetal genetic pathology, it is necessary to be aware that the foetal blood karyotype may be misrepresentative, as the analysed blood cells can indeed be of maternal origin.
Possible interaction of genotypes at cystathionine β‐synthase and methylenetetrahydrofolate reductase (MTHFR) in neural tube defects
Authors: Speer MC , Nye J , McLone D , Worley G , Melvin EC , Viles KD , Franklin A , Drake C , Mackey J , George TM ,   NTD Collaborative Group
Published: March 2003

Neural tube defects are a common, complex disorder with genetic and environmental components to risk. We investigated the previously reported interaction between homozygosity for the thermolabile variant at the methylenetetrahydrofolate reductase and heterozygosity for the 844ins68 allele at the cystathionine β‐synthase loci in cases with lumbosacral myelomeningocele and their parents. Using control allele frequencies from our sample pooled with those published in the literature, we confirm a marginally significant interaction at these two loci. This finding suggests that additional, larger studies are warranted to investigate this possible interaction in more detail.
Neural stem cells – a versatile tool for cell replacement and gene therapy in the central nervous system
Authors: Ourednik V , Ourednik J , Park Ki , Snyder Ey
Published: March 2003

In recent years, it has become evident that the developing and even the adult mammalian central nervous system contains a population of undifferentiated, multipotent cell precursors, neural stem cells, the plastic properties of which might be of advantage for the design of more effective therapies for many neurological diseases. This article reviews the recent progress in establishing rodent and human clonal neural stem cell lines, their biological properties, and how these cells can be utilized to a correct variety of defects, with prospects for the near future to harness their behaviour for neural stem cell‐based treatment of diseases in humans.
Partial CFTR genotyping and characterisation of cystic fibrosis patients with myocardial fibrosis and necrosis
Authors: Żebrak Jerzy , Skuza Barbara , Pogorzelski Andrzej , Ligarska Renata , Kopytko Ewa , Pawlik Jacek , Rutkiewicz Ewa , Witt Michal
Published: March 2003

Myocardial necrosis and fibrosis is a rare complication of cystic fibrosis (CF) causing sudden and unexpected death in infancy due to cardiac arrest. Characteristic morphological lesions are recognisable post‐mortem. The 18 CF patients with this complication had varied clinical features including mild pulmonary involvement, early onset severe pancreatic insufficiency, and profound electrocardiogram (ECG) changes. In this group of patients, 5 were ΔF508 homozygotes, 1 was ΔF508/N1303K and 1 was a ΔF508/M compound heterozygote. A pair of affected siblings (ΔF508 homozygotes) were fully concordant for myocardial involvement and for the general course of the disease. The co‐existence of a genetic predisposition to myocardial lesions resulting most probably from severe cystic fibrosis transmembrane (CFTR) genotypes (such as ΔF508/ΔF508, ΔF508/N1303K) and deficiency of certain trophic factors necessary for metabolism of the myocardium, are postulated to cause myocardial complications in CF leading to circulatory failure and early death.
Genetic evaluation of pervasive developmental disorders: the terminal 22q13 deletion syndrome may represent a recognizable phenotype
Authors: Prasad Chitra , Prasad Asuri N , Chodirker Bernard N , Lee Christine , Dawson Angelika K , Jocelyn Leslie J , Chudley Albert E
Published: March 2003

The evaluation of mental retardation is always a challenge to clinicians. The recognition of specific physical or behavioral characteristics can vastly improve diagnostic yield. Several genetic disorders have been identified to have certain behavioral characteristics, such as Williams syndrome, Smith–Magenis syndrome, and the velocardiofacial syndrome (VCFS). The deletion affecting the chromosome 22q in the most distal band (22q13) appears to define yet another neurobehavioral phenotype. In addition to our report, there are about 17 other cases published of this particular deletion syndrome. We describe three children who share features of developmental delay and pervasive behaviors in addition to normal to advanced growth patterns. Results of cytogenetic analysis suggest that the 3 patients share a deletion affecting the terminal 22q13 region. Two were found to have a cryptic deletion, in the third it was detected by conventional cytogenetics. The cryptic deletions were demonstrated using fluorescent in situ hybridization (FISH), where the control probe for the DiGeorge/VCFS region was deleted.  While there remain gaps in our understanding of this particular deletion syndrome, we propose that patients with normal or advanced growth, significantly delayed speech, deviant development and pervasive behaviors, with minor facial dysmorphism, be screened for this deletion.
Genotype–phenotype analysis in Apert syndrome suggests opposite effects of the two recurrent mutations on syndactyly and outcome of craniofacial surgery
Authors: Von Gernet S , Golla A , Ehrenfels Y , Schuffenhauer S , Fairley Jd
Published: March 2003

Apert syndrome is an autosomal dominant condition characterized by craniosynostosis and severe syndactyly, caused by two recurrent mutations in the fibroblast growth factor receptor 2 gene (FGFR2). The genotype–phenotype correlations of 21 patients with Apert syndrome were analysed as to the craniofacial appearance following surgery and the degree of syndactlyly. The craniofacial appearance following craniofacial surgery was better in patients with the P253R mutation, whereas these patients showed a more pronounced severity of the syndactyly.
Germline mutation screening of the STK11/LKB1 gene in familial breast cancer with LOH on 19p
Authors: Chen Jindong , Lindblom Annika
Published: March 2003

The recently cloned STK11/LKB1 on chromosome 19p has been shown to be a new tumor suppressor gene. Mutations in the LKB1/STK11 gene on chromosome 19p account for most cases of Peutz–Jeghers syndrome (PJS), in which intestinal hamartomas are associated with elevated risks of several cancer types, including breast cancer. A previous study revealed that familial breast cancer is associated with loss of heterozygosity (LOH) on 19p. To establish whether germline mutations of STK11/LKB1 account for familial breast cancer, 22 patients from 14 breast cancer families with LOH on 19p and one PJS family were selected for screening for germline mutations of LKB1/STK11. A combination of polymerase chain reaction (PCR)‐heteroduplex, single‐strand conformational polymorphism (SSCP) analyses, Southern blot analysis and direct sequencing were used for mutation detection. No mutations were identified. Germline mutations of LKB1/STK11 did not contribute to breast cancer in these families.
Imprinting centre deletions in two PWS families: implications for diagnostic testing and genetic counselling
Authors: Buiting K , Färber C , Kroisel P , Wagner K , Brueton L , Robertson ME , Lich C , Horsthemke B
Published: March 2003

Prader–Willi syndrome (PWS) is a complex genetic syndrome involving imprinted genes on chromosome 15. It is usually sporadic, and very few affected siblings have been described. Here, we report the clinical and molecular findings in two families with a microdeletion affecting the chromosome 15 imprinting centre (IC). Carrier males have a 50% risk of having children with an imprinting defect leading to PWS, and in one of the two families, a father has two affected daughters. In the other family, diagnostic testing was confounded by the presence of a neutral microdeletion close to the IC. The silent transmission of PWS IC deletions through the female germline and the occurrence of neutral microdeletions close to the IC can impose considerable problems on diagnostic testing and genetic counselling in affected families.
Velo‐cardio‐facial phenotype and deletion of 22q11.2 in Hungarian children
Authors: Morava É , Czakó M , Melegh B , Kosztolányi G
Published: March 2003

Velo‐cardio‐facial syndrome is a developmental disorder characterized by heart defects, specific facial features, cleft palate and learning disability. Most patients have a 3‐Mb deletion in chromosomal region 22q11.2. This microdeletion has also been found in patients with isolated conotruncal malformations. Although no significant ethnic variability has been reported in the frequency 22q11.2 deletions, some recent studies question the high frequency of this as the underlying cause of velo‐cardio‐facial syndrome in Anglo‐American populations. A screening program was initiated, including a detailed clinical assessment, followed by fluorescence in situ hybridization studies for microdeletion 22q11.2 in 24 children with congenital cardiac malformations referred consecutively to our genetics clinic. We found a high ratio of associated findings including cleft palate and developmental delay in our patient group. The clinical diagnosis of velo‐cardio‐facial syndrome was established in 8 patients. However, the common deletion was detected in only two children. We conclude that, although the ‘velo‐cardio‐facial phenotype’ appears to be common in Hungarian children with congenital cardiac malformations, many patients may have different etiologies other than del(22)(q11.2).
Polymorphisms in the CAG repeat – a source of error in Huntington disease DNA testing
Authors: Yu S , Fimmel A , Fung D , Trent RJ
Published: March 2003

Five of 400 patients (1.3%), referred for Huntington disease DNA testing, demonstrated a single allele on CAG alone, but two alleles when the CAG+CCG repeats were measured. The PCR assay failed to detect one allele in the CAG alone assay because of single‐base silent polymorphisms in the penultimate or the last CAG repeat. The region around and within the CAG repeat sequence in the Huntington disease gene is a hot‐spot for DNA polymorphisms, which can occur in up to 1% of subjects tested for Huntington disease. These polymorphisms may interfere with amplification by PCR, and so have the potential to produce a diagnostic error.
Identification of a common variant in the lipoprotein lipase gene in a large Utah kindred ascertained for coronary heart disease: the −93G/D9N variant predisposes to low HDL‐C/high triglycerides
Authors: Samuels ME , Forbey KC , Reid JE , Abkevich V , Bulka K , Wardell BR , Bowen BR , Hopkins PN , Hunt SC , Ballinger DG , Skolnick MH , Wagner S
Published: March 2003

Defects in the lipoprotein lipase (LPL) gene are associated with dyslipidemia in the general population. Several rare mutations in the gene, as well as two common coding region polymorphisms, D9N and N291S, exhibit deleterious effects on circulating lipid levels. Using a linkage‐based approach, we have identified a large Utah kindred segregating the D9N variant in the LPL gene. The kindred was ascertained for premature coronary heart disease and was expanded based on familial dyslipidemia. A genomic scan identified a region of linkage including LPL, and mutation screening identified the segregating variant. In the kindred, the variant shows high penetrance for a hypoalphalipoproteinemia phenotype, but is also associated with hypertriglyceridemia and elevated insulin levels. The strength of linkage was dependent on the combination of phenotype definition and model parameters, favoring the use of a MOD score approach. Most other studies of LPL have proceeded by mutation screening of randomly chosen individuals or selected affected probands; this is the first example identifying a segregating LPL mutation using direct linkage.
Epidemiology of myotonic dystrophy in Italy: re‐apprisal after genetic diagnosis
Authors: Siciliano G , Manca ML , Gennarelli M , Angelini C , Rocchi A , Iudice A , Miorin M , Mostacciuolo ML
Published: March 2003

Before the discovery of the myotonic dystrophy (DM) gene, the DM epidemiological rates could not be accurately estimated. The aim of this study was to calculate the DM prevalence rates in Padova (North‐East Italy) and in four provinces of North‐West Tuscany (Central Italy) and, as of 30 June 1999, to do so using molecular genetic testing.  A minimum prevalence rate of 9.31×10−5 inhabitants was found, consistent with epidemiological rates worldwide, and more than two times as high as those of two previous studies conducted in the same areas during the era prior to molecular genetic testing.  This study, the first in Italy since the discovery of the DM gene, underlines the importance of direct genetic diagnosis of DM, especially in detecting mildly affected patients, a fundamental step in correctly estimating the risk of disease transmission in affected families.