bluuebunny/crossref_metadata_2025 huggingface dataset:
Dextran sedimentation induces a difference in the percentage of hypodense eosinophils in peripheral blood between children with allergic asthma and healthy controls
Authors: HOEKSTRA M. O. , BERENDS C. , DIJKHUIZEN B. , GERRITSEN J. , KAUFMAN H. F.
Published: April 2006

Considerable differences in the percentage of hypodense eosinophils in the peripheral blood of asthmatics have been reported by different investigators. In these previous studies dextran sedimentation was used for removal of erythrocytes prior to density centrifugation. We hypothesized that the sedimentation procedure might induce the presence of hypodense eosinophils in the peripheral blood of asthmatic patients. In order to test this hypothesis, we compared eosinophil density profiles from peripheral blood of children with asthma and of age‐matched healthy controls, using different procedures. In the first method (direct method) blood samples were directly layered on a discontinuous Percoll gradient. Erythrocytes were removed by isotonic lysis. In the second method (dextran sedimentation) erythrocytes were removed by sedimentation with dextran prior to gradient centrifugation. Results of the direct method show no significant difference in percentage of hypodense eosinophils between children with asthma and healthy controls (9.19% and 6.84% respectively). However, after dextran sedimentation, children with asthma had a significantly higher percentage of hypo‐dense eosinophils than healthy controls (15.40% and 8.84% respectively; P < 0.05). The percentage of hypodense eosinophils was correlated with the number of eosinophils and with the lung function, measured as the Tiffeneau index (FEV1/VC), in the whole group of subjects when the direct method was used. We conclude that an increased percentage of hypodense eosinophils is not present in the circulation of children with asthma, but can be induced in vitro by dextran sedimentation. Therefore, in vitro generation of hypodense eosinophils in the blood of patients with asthma seems to be related with the primed state of eosinophils.
Estimation of Der p and Der f I quantities in the reference preparations of Dermatophagoides mite extracts
Authors: YASUEDA H. , SAITO A. , AKIYAMA K. , MAEDA Y. , SHIDA T. , SAKAGUCHI M. , INOUYE S.
Published: April 2006

A monoclonal antibody‐based enzyme‐linked immunosorbent assay (MoAb‐ELISA) was developed to measure the major Dermatophagoides mite allergens, Der p I and Der f I, The assay was highly species‐specific and sensitive. Using this assay system, the absolute mass unit of Der p I and Der fI in the reference preparations of the extracts was estimated. The primary standards used were the purified Der p I and Der f I preparations. The reference preparations of the D. pteronyssinus and D. farinae extracts (92‐Dp) and 92‐Df), which had been prepared from the same amount of mite bodies of both species, were found to contain the same levels of the Der I allergens, 10.1 μg/ml of Der p I and 10.0 μg/ml of Der f I, respectively. A histamine release assay with leucocytes from mite‐allergic donors showed that the total allergenic potency of 92‐Dp and 92‐Df was comparable. This result indicates that the estimated Der I levels in these extracts seem to be valid, at least, in the balanece between the two species, although further comparisons of the absolute quantities by several different laboratories are needed. The Der I levels in the WHO/IUIS international reference preparation of D. pteronyssinus and the CBER standard mite extracts, E4‐Dp and E5‐Df, were also estimated using this assay system. They were found to contain 4.4 μg/viaI and 13.3 μg/ml of Der p I and 9.5 μg/ml of Der f I, respectively.
Tryptase and histamine release due to a sting challenge in bee venom allergic patients treated successfully or unsuccessfully with hyposensitization *
Authors: EBERLEIN‐KÖNIG B. , ULLMANN S. , THOMAS P. , PRZYBILLA B.
Published: April 2006

Background: Hyposensitization with bee venom leads to full protection in most, but not all patients with IgE‐mediated systemic reactions to bee stings.

Objective: To determine the relationship of clinical reactivity to the release of mediators and to changes of antibody concentrations in the peripheral circulation at a bee sting challenge test.

Methods: Blood was sampled before (0 min) and at 15, 60 and 180 min after a sting challenge from 19 patients on hyposensitization. Of these, six still reacted and 13 were protected. Histamine, mast cell tryptase, bee venom‐specific IgE and IgG in the serum, and histamine release from peripheral blood leucocytes (PBL) upon exposure to bee venom were determined.

Results: Tryptase above the detection level was found only at 15 (60)min in 4/6 (1/6) patients who reacted. After the sting challenge there was a significant increase of the histamine levels in patients who reacted at 15 min (P < 0.05) and in patients who did react at 60 and 180 min (P < 0.01). The total histamine content of PBL was significantly decreased after 15 and 60 min in patients who reacted (P < 0.01) and in those that did not (P < 0.05). Bee venom‐induced histamine release was significantly reduced in patients reacting and those that did not at 15 min (P / 0.05), and was significantly decreased in reactors also at 60 and 180 min (P < 0.05/0.01). Specific IgG antibodies showed a minor decrease (P < 0.05) after the sting challenge in both groups, whereas specific IgE did not change significantly.

Conclusion: These results indicate that bee venom anaphylaxis is associated with the release of mediators from both mast cells as well as basophils. Successful hyposensitization does not induce a state of immunological non‐reactivity, but rather alters the magnitude and the pattern of mediator release.
Soluble ICAM‐1 as a regulator of nasal allergic reaction under natural allergen provocation
Authors: KATO M. , HATTORI T. , KITAMURA M. , BEPPU R. , YANAGITA N. , NAKASHIMA I.
Published: April 2006

Background: Intercellular adhesion molecule‐1 (ICAM‐1) plays a key role in the early stage of the signal cascade leading to cellular extravasation and the development of an inflammatory response. Recently, it has been reported that the soluble form of this adhesion molecule is present in human sera, possibly mediating biological actions.

Objective: The purpose of this study was to investigate levels of soluble ICAM‐1 (sICAM‐1) and its receptors in patients with allergic rhinitis, and to discuss sICAM‐1's biological function.

Methods: The levels of sICAM‐1 in sera and nasal epithelial lining fluids (ELF), the percentage of CD11a‐positive lymphocytes in the peripheral blood, and scores of subjective symptoms from 14 patients with pollinosis (allergic group) were measured from pre‐ to post‐season, results were compared with those from 10 non‐allergic subjects (control group).

Results: The levels of sICAM‐1 in sera and ELF were upregulated, and CD11a‐positive lymphocytes were downregulatcd during the in‐season in the allergic group. In addition, levels of sICAM‐1 in sera from the allergic group remained high during the post‐season, when levels of other parameters (symptoms, blood eosinophil counts, sICAM‐1 in ELF and CD11a‐positive lymphocytes) had roughly returned to the initial pre‐season levels.

Conclusions: We demonstrate systemic and local upregulation of sICAM‐1 and systemic downregulation of LFA‐1 positive lymphocytes in patients with seasonal allergic rhinitis under natural allergen provocation, suggesting that sICAM‐1 plays a role in regulating seasonal allergic inflammation.
The decay of house dust mite allergens, Der p I and Der p II, under natural conditions
Authors: BOER R. DE , VAN DER HOEVEN W. A. D. , STAPEL S. O.
Published: April 2006

Background: Fluctuations in the level of mite allergens in domestic house dust are the result of changes in the balance between synthesis, removal and decay. Purely physical forces as well as enzymatic degradation, mediated by house dust inhabiting microbes, may contribute to the decay of allergens in domestic dust. Knowledge about the speed of decay is essential for an understanding of the dynamics of allergen levels.

Objective: The present study is a quantitative assessment of the speed of decay at nine combinations of temperature (15°C, 20°C and 25°C) and relative humidity (33%, 55% and 75%).

Methods: Samples of mite infested material of an old rug were stored at these temperature/relative humidity‐combinations for 6, 12 or 18 months, after the mites were killed by cither a freezing treatment or an acaricide (lindane). The microbes living in the rug presumably survive these treatments. Concentrations of Der p I and Der p II + Der f II. in extracts of the rug material, were measured by a radio immunoassay.

Results: No significant changes in the levels of Der p I and Der p II +Der f II, could be detected even after 11/2 year at a high temperature and humidity.

Conclusion: These findings incidate that mite allergens can be extremely stable under normal domestic circumstances.
Peripheral eosinophil counts as a marker of disease activity in intrinsic and extrinsic asthma
Authors: ULRIK C. S.
Published: April 2006

Background: Recently it has been suggested that the bronchospasm and hyperresponsiveness phenomena observed in asthma are secondary to the actions of the eosinophils; the purpose of this study was to evaluate the relationship between the peripheral number of eosinophils and various markers of disease activity in a group of asthmatics examined in childhood (mean age 10 years) and early adulthood (mean age 21 years).

Methods: The relationship between eosinophil count and pulmonary function (FEV1), respiratory symptoms, bronchial responsiveness to histamine and diurnal variation in peak expiratory flow rate (PEF) was studied in 70 subjects with bronchial asthma, of whom 24 had intrinsic and 46 extrinsic asthma. Self‐reported symptoms of asthma were graded on a scale from 0 to 5, where 0 = no symptoms within the preceding 12 months and 5 = daily including nocturnal symptoms, and histamine responsiveness was analysed by means of the dose‐response slope (DRS).

Results: In both childhood and adulthood, a direct correlation was found between blood eosinophil count and symptom score (r= 0.69, P< 0.001 and r= 0.58, P < 0.001, respectively), whereas inverse correlations were observed between number of eosinophils and FEV, % predicted (r= .0.75, P < 0.001 and r= 0.80, P < 0.001. respectively). Furthermore, in adulthood, eosinophil count was found to be significantly correlated to hisiamine responsiveness (log DRS) (r= 0.65, P < 0.001) and diurnal PEF variation (r= 0.81, P < 0.001); these correlations were also noted after dividing the subjects into intrinsic and extrinic asthmatics. In both groups of subjects a significant inverse correlation was also found between histamine responsiveness and pre challenge FEV1% predicted. The eosinophil count in childhood was weakly correlated to the symptom score in adulthood (r= 0.29, P < 0.02).

Conlusions: This study showed a relationship between eosinophil count and seventy of asthmatic symptoms, level of pulmonary function, histamine responsiveness and diurnal variation in PEF in both intrinsic and extrinsic asthma; suggesting that the peripheral eosinophil count reflects asthmatic activity, and possibly the degree of inflammation in the airways, in both children and adults. Furthermore, a low number of eosinophils in childhood might be related to a relatively favourable prognosis with regard to symptoms of asthma in early adulthood.
Effects of IL‐4 and IL‐13 on total and allergen specific IgE production by cultured PBMC from allergic patients determined with recombinant pollen allergens
Authors: DOLECEK C. , STEINBERGER P. , SUSANI M. , KRAFT D. , VALENTA R. , BOLTZ‐NITULESCU G.
Published: April 2006

Background: Interleukin (IL)‐4 and IL‐13 have been shown to be potent switch factors for IgE synthesis in human B cells.

Objective: In this study we investigated the effects of recombinant human IL‐4 and IL‐13 on total and allergen specific IgE synthesis by peripheral blood inononuclear colls (PBMC) from pollen allergic patients and healthy control individuals.

Methods: Peripheral blood mononuclear cells (PBMC) from allergic patients were investigated for their capacity to produce allergen specific IgE in vitro. Total protein extracts from birch pollen and timothy grass pollen as well as purified recombinant birch pollen allergens, Bet v I, birch profiling (Bet v II) and recombinant timothy grass pollen allergens. Phi p I, Phi p II, and Phi p V were used to measure specific IgE‐antibody synthesis in cell culture supernatants by IgE‐immunoblot and ELISA.

Reults PBMC obtained from allergic patients spontaneously secreted allergen specific IgE in the culture supernatants. Addition of Interleukin 4, Interleukin 13 and anti‐CD40 antibody to the cultures alone or in combinations significantly induced total IgE production whereas allergen specific IgE production was not affected.

Conclusion: Our results indicate that the peripheral blood of allergic individuals contains long lived allergen specific B cells which have already switched to IgE production and which are not sensitive to IL‐4 and IL‐13 treatment. These results may have implications on attempts to use cytokines or cytokine antagonists in therapy of Type I allergy.
Exudative hyperresponsiveness of the airway microcirculation in seasonal allergic rhinitis
Authors: SVENSSON C. , ANDERSSON M. , ALKNER L. GREIFF. U. , PERSSON C. G. A.
Published: April 2006

Background
Mucosal exudation of plasma is a non‐injurious, physiological response of the airway microcirculation to different inflammatory processes. The exudative response is similar in the nose and bronchi and exudation occurs in both allergic asthma and rhinitis. The educative response is a specific end‐organ function of the mucosal microcirculation that may be altered in airway diseases.

Objective
This study examines the hypothesis of altered responsiveness of the superficial airway microcirculation to vascular permeability‐increasing challenges in sustained allergic inflammation.

Methods
Fourteen patients with birch‐pollen induced allergic rhinitis were studied for 7 weeks during a Swedish birch‐pollen season. Nasal symptoms (itching, sneezing. blockage, and discharge) were recorded and the occurrence of pollen was determined. The plasma exudation response was examined by topical histamine challenges at the end (May) and well out of (December) the season. Challenge and lavage were carried out concomitantly using a‘nasal pool’ ‐device. The unilateral nasal cavity was filled for consecutive 10 minute periods with saline and two concentrations of histamine (80μg/ mL and 400μg/mL). The lavage fluid levels of different‐sized plasma proteins (albumin‐66 000 D. fibrinogen‐340000 D, and α2‐macroglobulin‐725000D) were determined.

Results
The pollen season was mild resulting in only minor nasal symptoms. Histamine produced exudation of all plasma proteins across the microvascular epithelial barriers with particularly strong correlation between the levels of albumin and α2‐macroglobulin (r =0.98; P< 0.001). The exudative response to histamine was concentration‐dependent (P<0.05) and, furthermore, it was significantly greater late into the season compared with outside the pollen season (albumin: P < 0.05. tibrinogen: P<0.05. α2‐macroglobulin: P<0.01).

Conclusion
We conclude that histamine produced concentration‐dependent nasal airway exudation of bulk plasma in subjects with seasonal rhinitis and that this response is abnormally great during the pollen season. Whether angiogenesis or increased responsiveness of the mierovascular endothelium may explain this phenomenon now remains unknown. We suggest that a mierovascular exudative hyperresponsiveness may characterize allergic airway disease.
Effect of intranasal fluticasone proprionate on the immediate and late allergic reaction and nasal hyperreactivity in patients with a house dust mite allergy
Authors: DE GRAAF‐IN'T VELD C. , GARRELDS I. M. , JANSEN A. P. H. , VAN TOORENENBERGEN A. W. , MULDER P. G. H. , MEEUWIS J. , VAN WIJK R. GERTH
Published: April 2006

Background
Patients with perennial allergic rhinitis develop nasal symptoms not only after allergen exposure, but generally also after non‐specific stimuli.

Objective
To evaluate the effect of 2 week's treatment with fluticasone propionate aqueous nasal spray (FPANS) on the nasal clinical response, inflammatory mediators and nasal hyperreactivity.

Methods
Twenty‐four rhinitis patients allergic to house dust mite (HDM). participated in a douhle‐blind. placebo‐controlled crossover study. After 2 week's treatment with placebo or 200 μg FPANS twice daily, patients were challenged with HDM extract. Symptoms were recorded and nasal lavages were collected for up to 9.5 h after challenge. Nasal hyperreaclivity was determined by histamine challenge 24 h later. Results Because of a carry‐over effect for the immediate symptom score, for this variable only the data from the first treatment period were used. FPANS treatment resulted in a significant decrease of nasal symptoms with 70%. 69% and 63% after 100. 1000 and 10000 Biological Units (BU)/mL of HDM extract respectively. Active treatment resulted in a 76% decrease of the late‐phase symptoms. FPANS treatment significantly reduced albumin influx after HDM 1000 BU/mL with 62% and tended to reduce tryptase release after HDM 1000 BU ml. (P 0.0629). During the late phase FPANS treatment reduced albumin influx with 67% and eosinophil cationic protein (ECP) release with 83%. No effect of FPANS was seen on histamine levels. FPANS significantly decreased histamine‐induced symptom score with 34%, secretion with 32%, and sneezes with 41%.

Conclusion
FPANS significantly inhibits the immediate and late allergic response, and nasal hyperreactivity, probably by suppressing mast cells and eosinophils in the nasal mucosa.
Anaphylactic reaction to lychee fruit: evidence for sensitization to profilin
Authors: FÄR J. , WÜTHRICH B. , VIETHS S.
Published: April 2006

Background and Objective
Due to the increasing popularity of exotic fruits in the Western diet, allergologists are confronted with allergic reactions to substances in these plants. The present report describes an anaphylactic reaction after the consumption of lychee fruit (Litchi sinensis). The atopic patient also suffers from rhinoconjunctivitis due to a sensitization against pollen of the Compositae family as well as from dyspnoea after eating sunflower seeds. Our goals were to determine crossreactivity between antibodies against lychee fruit and other plants and to characterize the allergen.

Methods and results
Specific IgE against lychee fruits were detected by an EAST assay. The allergen was characterized by immunoblot, immunoblot inhibition and KAST inhibition assays. Broad crossreactivity between lychee fruit and other plants was found and profilin identified as the protein responsible for the patient's complex allergy syndrome.

Conclusion
Lychee fruit contains a significant amount of profilin. Consumption of this exotic fruit can cause severe anaphylactic reactions in patients being sensitized against the plant pan‐allergen profilin.
Introduction to positional cloning
Authors: PAPADOPOULOS N.
Published: April 2006

The most common procedure for isolating genes that predispose to a certain disease is positional cloning. This process is initiated by mapping the responsible gene to its location on a chromosome linked to the disease. Successive cloning of the region results in the isolation of candidate genes. Finally, identification of mutations by mutational analysis points to the correct gene.
Oral antihistamine/decongestant treatment compared with intranasal corticosteroids in seasonal allergic rhinitis
Authors: NEGRINI A. C. , TROISE C. , VOLTOLINI S. , HORAK F. , BACHERT C. , JANSSENS M.
Published: April 2006

This international, multicentre, randomized, double‐blind, double‐dummy, parallel‐group trial was undertaken to compare the efficacy and tolerability of once‐daily astemizole‐D (10mg astemizole plus 240 mg pseudoephedrine) with beclomethasone nasal spray (0.05mg/ml) two puffs/nostril administered twice daily in a total of 204 patients with seasonal allergic rhinitis. Treatment duration was 4 weeks, Although investigator assessments of symptom severity were generally comparable in the two treatment groups throughout the trial, statistically significant differences in favour of astemizole‐D for sneezing and ocular symptoms were apparent at the end of the 4‐week treatment period (P < 0.05). Patient diary data support these findings, with significant differences in favour of the antihistamine/decongestant combination reported for ocular symptoms after 2 weeks of treatment (P < 0.05) and non‐significant trends for sneezing after 2 weeks and ocular symptoms over the entire treatment period (P= 0.07). Use of rescue medication for ocular symptoms was also significantly lower in the astemizole‐D treatment group (P < 0.05). A wide range of adverse experiences were reported, however, there were no statistically significant differences in the type or incidence of those between the two treatment groups. In conclusion, astemizole‐D appears to be at least as effective and well tolerated as intranasal beclomethasone in the treatment of seasonal allergic rhinitis, providing at least comparable relief from all nasal symptoms including congestion and significantly greater relief from ocular symptoms than the topical steroid.
Chronic asthma and chiropractic spinal manipulation: a randomized clinical trial
Authors: NIELSEN N. H. , BRONFORT G. , BENDIX T. , MADSEN F. , WEEKE B.
Published: April 2006

The purpose of this randomized patient‐ and observer‐blinded cross‐over trial was to evaluate the efficacy of chiropractic treatment in the management of chronic asthma when combined with pharmaceutical maintenance therapy. The trial was conducted at the National University Hospital's Out‐patient Clinic in Copenhagen, Denmark. Thirty‐one patients aged 18–44 years participated, all suffering from chronic asthma controlled by bronchodilators and/or inhaled steroids. Patients, or who had received chiropractic treatment for asthma within the last 5 years, who received oral steroids and immunotherapy, were not eligible. Patients were randomized to receive either active chiropractic spinal manipulative treatment or sham chiropractic spinal manipulative treatment twice weekly for 4 weeks, and then crossed over to the alternative treatment for another 4 weeks. Both phases were preceded and followed by a 2‐week period without chiropractic treatment. The main outcome measurements were forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), daily use of inhaled bronchodilators, patient‐rated asthma severity and non‐specific bronchial reactivity (n‐BR). Using the cross‐over analysis, no clinically important or statistically significant differences were found between the active and sham chiropractic interventions on any of the main or secondary outcome measures. Objective lung function did not change during the study, but over the course of the study, non‐specific bronchial hyperreactivity (n‐BR) improved by 36% (P= 0.01) and patient‐rated asthma severity decreased by 34% (P= 0.0002) compared with the baseline values. The results do not support the hypothesis that chiropractic spinal manipulative therapy is superior to sham spinal manipulation in the management of pharmaceutically controlled chronic asthma in adults when administered twice weekly for 4 weeks.
Pollen allergy in the Bilbao area (European Atlantic seaboard climate): pollination forecasting methods
Authors: ANTÉPARA I. , FERNÁNDEZ J. C. , GAMBOA P. , JAUREGUI I. , MIGUEL F.
Published: April 2006

Forecasting pollination can help the allergist to establish the appropriate treatment and advice for patients. Based on previous studies, we have related the climate variables with the grass pollen counts in the search for pollination predictors. By relating the meteorological data of the temperature recorded every 6h and of the rainfall in hourly periods, together with the daily pollen counts obtained by the Hirst volumetric system, over a period of 3 years, we have tried to predict the start, duration and severity of the grass pollination, as well as the days of peak pollination. We have established a relationship by means of a polynomic regression originating from the mean cumulated temperature higher than 9°C [R2= 0.927 (P= 0.0001)], with the pollination season starting from 300°C and the maximum peak at 356°C, in the 3 years of the study. During the days of pollination, peaks higher than 50 grains/m3 coincide with average daily temperatures of 18.7±3 and lower than 50 grains/m3 with temperatures of 16.8 ± 3 (significant to 95%). The duration of the pollination is influenced by the cumulated average temperatures (from 800 to 900°C) and especially by precipitation at the start of and during pollination. In order to forecast grass pollination, the cumulated average temperatures are useful, starting from a basal (9°C), pollination begins when this sum is greater than 300°C, whereas when 800°C is reached and depending on the rainfall during the season, pollination will end. The total severity of pollination seems to depend on the rainfall prior (in the same year) to the start. During pollination, the days with an average temperature of 18.7 ± 3°C and an absence of rainfall between 4 and 12 h, will be the days with a presence of more than 50 grains/m3. Thus, it is possible to forecast and estimate risk factors in the long‐term (by means of the rainfall prior to pollination) and in the short‐term (for the following day, if a weather forecast is available) so as to offer it to those members of the population affected by pollinosis.
Immunotherapy with Alpare® in patients with respiratory allergy to Parietaria pollen: a two year double‐blind placebo‐controlled study
Authors: D'AMATO G. , KORDASH T. R. , LICCARDI G. , LOBEFALO G. , CAZZOLA M. , FRESHWATER L. L.
Published: April 2006

Allergy to Parietaria judaica pollen causes significant morbidity in many areas of the world. In addition to rhinitis, patients who are allergic to this pollen have a high incidence of asthma. The pollinating season is long, making this particular allergy challenging for clinicians to treat. This study was designed to determine if immunotherapy with an alum adsorbed partially purified Parietaria extract (Alpare Parietaria) containing a targeted maintenance dose of 12 500 BUs was effective in decreasing rhinitis symptoms in patients allergic to Parieiaria. Using a double‐blind placebo‐controlled technique 36 patients received placebo or active extract for 2 years. Twenty (11 placebo and nine active) completed the 2 year study. Efficacy of treatment was evaluated by determining changes in skin reactivity, visual analog scores, diary symptom scores and end of study assessments. Reactions were monitored as well. Skin‐test suppression was marginally significant in the actively treated group after 1 year and showed even more significant suppression after the second year. Nasal block, rhinorrhoea and sneezing all were significantly decreased in the active group. The nasal provocation test did not show a significant change after 1 year, in either group, but after 2 years of treatment the active group did show significant improvement. Although almost all patients in the actively treated group experienced local reactions, the incidence of systemic reactions was not different between the two groups. In conclusion, immunotherapy with this extract at this dose was effective in ameliorated rhinitis symptoms in patients allergic to Parietaria judaica.
Substance P induces histamine release from human pulmonary mast cells
Authors: HEANEY L. G. , CROSS L. J. M. , STANFORD C. F. , ENNIS M.
Published: April 2006

Substance P elicits histamine release from human skin and rodent mast cells. Since neuropeptide‐mediated reflexes may be important in asthma, we examined the ability of substance P to stimulate human mast cells obtained at bronchoalveolar lavage (BAL). BAL samples were obtained at routine bronchoscopy from 35 non‐preselected patients. Histamine release experiments were performed in a standard manner using substance P and the calcium ionophore A23187. Both substance P (50 μM) and A23187 caused histamine release (median 26.7%, range 6.2–62.8% and 32.1%, 7.7–56.8% respectively) which was significantly greater (P < 0.0001) than the spontaneous release (median 15.6%, range 4.1–33.4%), i.e. that in the absence of any stimulus. Substance P induced histamine release was via an energy dependent process and was blocked by preincubation with antimycin A. A significant correlation was observed between substance P induced release and spontaneous release but was not observed with A23187 induced release. Mast cell counts correlated significantly with substance P induced release but not with spontaneous or A23187 induced release. The substance P induced histamine secretion was elicited at similar concentrations to those used with rodent and human skin mast cells. Asthma is associated with increased numbers of mast cells which have both increased spontaneous and stimulated secretory responses. Thus, in vivo, the bronchoconstrictor action of substance P may in part result from activation of mast cells in the bronchial lumen.
Mast cells and T lymphocytes in chronic urticaria
Authors: BARLOW R. J. , ROSS E. L. , MACDONALD D. M. , BLACK A. KOBZA , GREAVES M. W.
Published: April 2006

Mast cells are considered to be the primary effector cells in urticaria but it is possible that lymphocytes contribute to the formation of weals by secreting histamine releasing factors. The aim of this study was to examine the population of mast cells and to quantify the T cell subsets and their activation status in delayed pressure urticaria (DPU), chronic idiopathic urticaria and normal controls. Three biopsies were obtained from each of four patients with chronic idiopathic urticaria but not DPU. Three biopsies were taken from each of 13 patients with DPU, from a combination of unchallenged skin and at 0, 2, 6, 24, 48 and 120 h after weighted steel rods (diameter 1.5 cm) had been applied to the thighs. Three biopsies were similarly obtained from each of four normal controls before an identical pressure challenge and at 6, 24 and 48 h afterwards. The chloracetate esterase stain was used to demonstrate mast cells and an alkaline phosphatase anti‐alkaline phosphatase immunohistochemical technique to assess the phenotypic and activation characteristics of the T cell infiltrate. The mast cell count did not differ significantly between unchallenged skin from DPU patients and normal controls. Following a pressure challenge to the DPU patients, the number of stainable mast cells decreased significantly to a level comparable with that in spontaneous weals of chronic idiopathic urticaria. Investigation of T cell subsets showed a preponderance of CD4+ cells over CD8+ cells. There was no evidence of T cell activation in chronic idiopathic urticaria or DPU when compared with normal controls. These data support the view that mast cell degranulation occurs in chronic idiopathic urticaria and suggest that it may also play a role in the pathogenesis of DPU. There was no evidence that lymphocyte activation occurs in either condition.
Molecular cloning of cDNA coding for the 68 kDa allergen of Penicillium notatum using MoAbs
Authors: SHEN H‐D. , LIAW S‐F. , LIN W‐L. , RO L‐H. , YANG H‐L. , HAN S‐H.
Published: April 2006

To characterize the 68 kDa allergen of Penicillium notatum (also known as P. chrysogenum), a molecular antibody (MoAb) (P40) was previously generated. For cDNA cloning, three more MoAbs (3F, 5A3, 5G2) were generated in the present study. A mixture of all the four MoAbs was used in cloning of the gene coding for the 68 kDa allergen from a λgt11 cDNA library of P. chrysogenum. A cDNA clone (A6) with DNA insert of about 0.5 kb which encodes for the 3′‐terminal nucleotide sequence of the 68 kDa allergen was obtained. The cloned sequence contained two putative N‐glycosylation sites. The reduction in molecular weight from 68 to 62 kDa in immunoblotting after treatment of the crude extract of P. notatum with N‐glycosidase F indicates that the 68 kDa allergen is a glycoprotein. Nucleotide sequence determination showed that 188 (54%) of the 348 nucleotides of the cDNA sequence obtained were identical to the same region of the nucleotide sequence of the beta‐N‐acetylglucosaminidase gene of Candida albicans. Although the cDNA clone obtained did not encode the full‐length gene of the 68 kDa allergen, polypeptide expressed from the A6 cDNA showed positive immunological reactivities to all four MoAbs used in the cloning experiment and to IgE antibodies in sera of asthmatic patients. There was a loss of immunoblotting activity to the 68 kDa component after absorption of MoAb P40‐containing culture supernatant with filters blotted on plaque lawns of cDNA clone A6. Moreover, the immunoblotting activity remained when the MoAbs affinity‐purified with filters containing polypeptides encoded by the cDNA insert of clone A6 were used. These two observations indicate that clone A6, which encodes 117 amino acid residues of a putative 560‐residue polypeptide, is a cDNA clone of the 68 kDa component of P. notatum. In conclusion, results obtained from cloning and characterization of a partial cDNA clone described in this study suggest that the 68 kDa allergen of P. notatum is a beta‐N‐acetylglucosaminidase.
Benzalkonium chloride in a decongestant nasal spray aggravates rhinitis medicamentosa in healthy volunteers
Authors: GRAF P. , HALLÉN H. , JUTO J.‐E.
Published: April 2006

A randomized double‐blind parallel study with 20 healthy volunteers was performed to research the effect of a preservative in a decongestant nasal spray on the development of rhinitis medicamentosa. Ten subjects received oxymetazoline nasal spray with benzalkonium chloride and the others used oxymetazoline nasal spray without the preservative three times daily for 30 days. Before starting the course of treatment and after its conclusion, recordings of the mucosal surface positions were made with rhinostereometry followed by histamine challenge tests. Symptoms of nasal stuffiness were estimated on visual analogue scales (0–100) in the morning and the evening just before using the nasal spray. After 30 days, rebound swelling and nasal stuffiness were found in both groups. In the group receiving oxymetazoline nasal spray with benzalkonium chloride the mean rebound swelling was 1.1 mm and the estimated mean evening symptom score for nasal stuffiness was 43. In the group without benzalkonium chloride the corresponding variables were significantly less marked, with a mean rebound swelling of 0.5 mm (P<0.05) and a mean evening symptom score of 25 (P<0.05). The increase in histamine sensitivity in both groups was interpreted as a sign of nasal hyperreactivity. A new type of nasal spray bottle was used that has been shown to prevent bacterial contamination. In conclusion, the long‐term use of benzalkonium chloride in oxymetazoline nasal spray accentuates the severity of rhinitis medicamentosa in healthy volunteers.
Sensitization to Ganoderma lucidum in patients with respiratory allergy in India
Authors: SINGH A. B. , GUPTA S. K. , PEREIRA B. M. J. , PRAKASH D.
Published: April 2006

Although human sensitization to basidiomycete Ganoderma has been reported in New Zealand. North America and Europe, hypersensitivity due to this fungi is not known in India, in spite of its prevalence in the atmosphere. We have studied the atmospheric concentration of Ganoderma in different localities within Delhi and the sensitization level to this fungi amongst the Indian population. Aerobiological sampling, using a Burkard personal slide sampler, was carried out in Delhi for 2 consecutive years (October 1989 September 1991). The sampler was operated at 10 day regular intervals for 10min to trap the spores. The peak season for Ganoderma is recorded from July to September with highest average monthly catch of 336 spores/m3 in September 1991 from south Delhi. Antigenic extracts were prepared from both, the spores and whole body of Ganoderma lucidum. The results of intradermal skin tests conducted on 172 patients revealed that 28.48% and 17.44% of patients showed marked skin reactivity to spore and whole body extracts, respectively. A significant correlation (r= 0.963, P < 0.01) was found between intradermal and skin‐prick tests. More than 80% of the intradermal test positive patients had significantly (P < 0.01) elevated IgE antibodies to the fungi in question. Thus, sensitization to Ganoderma lucidum has been reported for the first time in the atopic population of India.
Influence of the menstrual cycle on skin‐prick test reactions to histamine, morphine and allergen
Authors: KALOGEROMITROS D. , KATSAROU A. , ARMENAKA M. , RIGOPOULOS D. , ZAPANTI M. , STRATIGOS I.
Published: April 2006

The purpose of this study was to examine the possible influence of the phases of the menstrual cycle on dermal reactivity to skin‐prick testing. We studied 15 atopic, menstruating women with seasonal rhinoconjunctivitis and/or asthma, with known sensitivity to olive and parietaria (mean age 25.2 years) and 15 non‐atomic, healthy, female controls (mean age 24.7 years). Skin‐prick tests with histamine, morphine, and in the atopic group with parietaria/and/or olive, were repeated three times during the same menstrual cycle, corresponding to bleeding (day 1–4), midcycle (day 12–16) and the late progesterone phase (day 24–28). None of the patients had received oral antihistamines or exogenous hormones for at least 1 month prior to testing. Results indicate a significant increase in weal‐and‐flare size to histamine, morphine, and parietaria on days 12–16 of the cycle, corresponding to ovulation and peak oestrogen levels. This was observed in both atopic and non‐atopic women. Differences in skin reactivity to histamine and morphine between the groups were not significant. Therefore, in women, the phase of the menstrual cycle is another factor that may influence skin‐test results.
Interleukin‐4 and interferon‐gamma production in atopic and non‐atopic children with ashma
Authors: TANG M. L. K. , COLEMAN J. , KEMP A. S.
Published: April 2006

Previous studies have demonstrated increased production of interleukin‐4 (IL‐4) and reduced production of interferon (IFN)‐γ in stimulated peripheral blood mononuclear cell cultures from children and adults with atopic dermatitis, however, it is unclear whether such an imbalance of cytokine production relates to other childhood atopic diseases such as asthma, and in particular to the presence of the atopic state per se. The production of IL‐4 and IFNγ in phytohaemagglutin‐ (PHA)‐stimulated peripheral blood mononuclear cell (PBMC) cultures from atopic and non‐atopic children with moderately severe chronic persistent asthma, and a group of age‐matched non‐atopic controls who did not have asthma was examined. Atopic children with asthma produced significantly more IL‐4 and less IFNγ than non‐atopic children with asthma and non‐atopic controls who did not have asthma. There was no significant difference in IL‐4 or IFNγ production between non‐atopic children with asthma and controls. These findir. 3 demonstrate that an imbalance of IL‐4 and IFNγ production is present in atopic asthma as previously documented in atopic dermatitis, therefore suggesting that it is a feature of the atopic state per se.
House dust mite fauna in Thailand
Authors: MALAINUAL N. , VICHYANOND P. , PHAN‐URAI P.
Published: April 2006

House dust mites have been shown to be the most important source of allergen in Thailand. The results of our initial mite survey were in conflict with previous data reported 20 years ago. To evaluate the current condition of mite fauna in Thailand, we conducted a larger scale survey of house dust mites in dusts from three different parts of the country. A total of 630 mattress dust samples was collected from various types of housings from central, northern and northeastern parts of Thailand. House dust mites were identified and enumerated by standard techniques and 388 dust samples were analysed further for group I antigen by a two‐site monoclonal antibody enzyme‐linked immunosorbent assay. House dust mites were identified in 555/630 (88.1%) of dust samples collected; 27.6% of these 555 samples contained over 100 mites per gram of fine dust. Dermatophagoides pteronyssinus was the most abundant species found with a mean count of 87.2 mites per gram of dust (range 1–1460). Group I antigen was detected in 380/388 (97.9%) samples analysed and 330 of which (86.8%) contained over 2 μg/gm of group I antigen. The highest density of mites was found in dusts from northern Thailand whereas dusts with highest amount of antigen were those collected from the northeastern part. Some degree of correlation between mite count and mite allergen was observed (r2= 0.33, P< 0.001). No significant difference in the mite count and mite allergen was found among various types of housings. House dust mites are the source of allergens ubiquitously found all over Thailand. Dust from several houses surveyed in this study had significantly high level of group I antigens.
Prevention of asthma with ketotifen in preasthmatic children: a three‐year follow‐up study
Authors: BUSTOS G. J. , BUSTOS D. , BUSTOS G. J. , ROMERO O.
Published: April 2006

Growing morbidity and mortality rates call for research towards more effective methods of preventing asthma. During the last decade several groups have reported the results of natural history and asthma prevention studies. However, the attempt to prevent development of asthma in genetically predisposed children, has not resulted in a generally accepted management scheme. The aim of this study was to evaluate the effectiveness of ketotifen in preventing the onset of asthma in infants considered to be at high risk of developing the disease, but who had no history of respiratory obstruction. These children have been described as preasthmatic. In this double‐blind, placebo‐controlled, parallel study, 100 infants with a family history of major allergy and elevated serum IgE levels, but with no history of bronchial obstruction, were treated with either ketotifen (n= 50) or placebo (n= 50) over a 3‐year period. There were no statistically significant differences between the two groups with regard to age, sex, degree of hereditary allergy, levels of serum IgE upon joining the study, and family smoking habits. At the end of 3 years, only four of the 45 infants who had received ketotifen had developed asthma (9%). Of the 40 children given placebo, 14 had developed asthma (35%) (P= 0.003). These results suggest that ketotifen is effective in preventing the onset of asthma in preasthmatic children.
Cyclic nucleotide phosphodiesterase isoenzyme activities in human alveolar macrophages
Authors: TENOR H. , HATZELMANN A. , KUPFERSCHMIDT R. , STANCIU L. , DJUKANOVIC R. , SCHUDT C. , WENDEL A. , CHURCH M. K. , SHUTE J. K.
Published: April 2006

Background
Alveolar macrophages and their precursors, the monocytes are involved in airway inflammation in asthma. An increase in intraceliular cAMP by PDE inhibitors is known to suppress macrophage and monocyte functions. A comparison of the PDE‐isoenzyine profiles of human alveolar macrophages from normal and atopic donors and of human peripheral blood monocytes might form a basis to differentially affect functions of these cells by PDE inhibitors.

Objective
The study compares the PDE isoenzyme activity profiles of human alveolar macrophages from normal and atopic asthmatic donors and human peripheral blood monocytes. In addition, the effect of in vitro maturation of monocytes on their PDE isoenzyme profile is studied.

Methods
Macrophages were purified (95‐97%) by adherence to plastic, and blood monocytes were purified (88%) by counter‐current elutriation. PDE isoenzyme activity profiles were investigated using isoenzyme selective inhibitors and activators.

Results
In macrophages substantial PDE I activity, which was significantly higher than PDE IIF‐V activity was detected and PDE II was absent. PDE III was membrane‐bound whereas PDE I, IV and V were soluble. No difference was found between alveolar macrophages of normal donors and atopic asthmatics. Monocytes exclusively contained PDE IV but their in vitro maturation led to a PDE isoenzyme profile similar to that of alveolar macrophages.

Conclusion
These results indicate that human monocytes and alveolar macrophages are distinct targets for the effects of selective PDE inhibitors while alveolar macrophages from normal and atopic individuals appear to be equally sensitive.
Variability and repeatability of house dust mite allergen measurement: implications for study design and interpretation
Authors: MARKS G. B. , TOVEY E. R. , PEAT J. K. , SALOME C. M. , WOOLCOCK A. J.
Published: April 2006

Background To interpret individual measurements of house dust mite (MDM) allergen and to design and analyse HDM studies it is necessary to quantify the variability which is inherent in the measurement of this exposure.

Objective To estimate the repeatability of one method of HDM allergen measurement.

Methods We analysed data from one or more HDM allergen measurements in 215 houses included in four previous studies conducted in Sydney (a high allergen environment) and Busselton. Western Australia (a moderate allergen environment). Samples were collected from the bed by vacuuming above and below the sheets and inside the pillow case and from the bedroom and living room floors by vacuuming a I m2 area for 1 min. Extracts from aliquots of fine dust from each sample were assayed for HDM allergen Der p I using a monoclonal antibody enzyme linked immunosorbent assay (IBLISA). The values for HDM allergen were positively skewed and the suitability of a log transformation was established by the resulting normal distribution and stable within‐site variance.

Results The range of single determination (within which the true value lies with 95% certainty) was 3‐1‐fold for samples from the bed and 3.5‐fold for samples from the floor. The coefficient of repeatability (the ratio beyond which a change between two estimates is established with 95% certainty) was 4.9 for the bed and 5.8 for the floor.

Conculsion We estimate that, to detect a twofold difference or change in allergen levels. 35 houses per group will be required in cross‐sectional studies and 30 houses per group in parallel‐group, randomized controlled trials. We recommend that beds be sampled by collecting dust from the layer of bedding below the bottom sheet. A single site within the bedroom floor may be taken as representative of this site but this is not true for the living‐room floor.