bluuebunny/crossref_metadata_2025 huggingface dataset:
The final step of the ethylene biosynthesis pathway in turnip tops (Brassica rapa): molecular characterization of the 1‐aminocyclopropane‐1‐carboxylate oxidase BrACO1 throughout zygotic embryogenesis and germination of heterogeneous seeds†
Authors: Del Carmen Rodríguez‐Gacio María , Nicolás Carlos , Matilla Angel Jesús
Published: April 2004

In a previous report from the present authors, it was shown that the 1‐aminocyclopropane‐1‐carboxylate (ACC) oxidation may play a crucial role during zygotic embryogenesis of turnip tops seeds. The present study was performed to elucidate the contribution of the silique‐wall and seeds in ethylene production during this developmental process. ACC content in the silique wall is only higher than in seeds during the middle phases of zygotic embryogenesis. The ACC‐oxidase (ACO) activity peaks in the silique‐wall and seeds during the onset of embryogenesis, declining gradually afterwards, being undetectable during desiccation period. Using reverse transcriptase‐polymerase chain reaction, one cDNA clone coding for an ACO and called BrACO1, was isolated. The deduced protein for BrACO1 has a molecular weight of 36.8 kDa and a high homology with other crucifer ACOs. The heterologous expression of this cDNA confirmed that BrACO1 is an ACO. The expression of this gene was high during the first phases of silique‐wall development, low during the middle phases and undetectable during desiccation. By contrast, BrACO1 transcript was accumulated only in the earliest phases of seed embryogenesis and may participate in the highest ACO activity and ethylene production by seeds at the beginning of embryogenesis. Finally, in this work a correlation between the heterogeneity of Brassica rapa L. cv. Rapa seeds and the ability to oxidize the ACC to ethylene has been demonstrated.
Identification, cloning and expression analysis of strawberry (Fragaria × ananassa) mitochondrial citrate synthase and mitochondrial malate dehydrogenase
Authors: Iannetta Pietro P. M. , Escobar Nieves Medina , Ross Heather A. , Souleyre Edwige J. F. , Hancock Robert D. , Witte Claus‐Peter , Davies Howard V.
Published: April 2004

Salt‐extractable proteins from the cell walls of immature and ripe strawberry (Fragaria × ananassa Duch. cv. Elsanta) fruit were separated using two‐dimensional polyacrylamide gel electrophoresis. Seven polypeptides (enzymes) were characterized from their N‐terminal sequences: (1) glyceraldhyde‐3‐phosphate dehydrogenase (EC 1.2.1.12); (2) triose phosphate isomerase (TPI; EC 5.3.1.1); (3) mitochondrial malate dehydrogenase (mMDH; EC 1.1.1.37); (4) NADH glutamate dehydrogenase (EC 1.4.1.3); (5) chalcone synthase (ChS; EC 2.3.1.74); (6) mitochondrial citrate synthase (mCS; EC 4.1.3.7); and (7) UDP glucose:flavonoid 3‐O‐glucosyltransferase (UDPG:FGT; EC 2.4.1.91). The sequenced polypeptides identified only cytosolic proteins, two of which (ChS and UDPG:FGT) had already been identified as being up‐regulated in ripening (strawberry) fruit and important contributors to ripe fruit character. Our focus was therefore diverted to the enzymes mMDH and mCS for further molecular characterization as potentially important determinants of fruit flavour via regulation of the sugar : acid balance. Citrate synthase (CS) and malate dehydrogenase (MDH) enzyme activities increased substantially during ripening, as did citrate and malate contents. The increase in CS activity is supported by western blot analysis. One strawberry mCS (Fa‐mCS‐I) and two mMDH (Fa‐mMDH‐I and ‐II) cDNAs were cloned that were 77, 82 and 53% identical (respectively) to sequences from other plant sources. Northern analysis showed that CS and MDH expression did not correlate with enzyme activities and these findings are discussed.
Functional characterization ofSEPALLATA3andAGAMOUSorthologues in silver birch
Authors: Lemmetyinen Juha , Hassinen Minna , Elo Annakaisa , Porali Ilkka , Keinonen Kaija , Mäkelä Hannu , Sopanen Tuomas
Published: April 2004

The development of flowers is regulated by a complex network of transcriptional activators and repressors, many of which belong to the MADS box gene family. In this study, we describe two MADS box genes of silver birch (Betula pendulaRoth),BpMADS1andBpMADS6,which are similar toSEPALLATA3andAGAMOUSinArabidopsis thaliana,respectively. In situ hybridization showed thatBpMADS1was expressed in the inflorescence meristem at a very early stage, but not later. Both genes were expressed in developing carpels, ovules and stamens but not in tepals or scales. Ectopic expression ofBpMADS1inArabidopsisresulted in a reduced number of floral organs or whole whorls and in petaloid or carpelloid sepals, a phenotype reminiscent of that offilmutants. 35S::BpMADS6 caused very early flowering inArabidopsis. In tobacco, both 35S::BpMADS1 and 35S::BpMADS6 accelerated flowering and, in addition, 35S::BpMADS6 caused changes in sepals and petals. In some transgenic birch plants, 35S::BpMADS1 antisense resulted in the development of both male and female organs in the axil of a single bract and in a change of some inflorescences into vegetative shoots. In two plants, either 35S::BpMADS6 sense or antisense constructs resulted in an increase in the number of tepals and in complete lack of stamens in some male inflorescences. These results suggest thatBpMADS1participates both in inflorescence and in flower formation andBpMADS6participates in flower formation and that they are functional homologues toSEPALLATA3andAGAMOUS, respectively.
Effect of reduced arginine decarboxylase activity on salt tolerance and on polyamine formation during salt stress in Arabidopsis thaliana
Authors: Kasinathan Vasuki , Wingler Astrid
Published: April 2004

Polyamines have been suggested to play an important role in stress protection. However, attempts to determine the function of polyamines have been complicated by the fact that, dependent on the conditions, polyamine contents increase or decrease during stress. To determine the importance of polyamine formation during salt stress, we analysed polyamine contents and salt tolerance in two Arabidopsis thaliana mutants, spe1‐1 and spe2‐1 (Watson et al. Plant J 13: 231–239, 1998), with reduced activity of arginine decarboxylase (EC 4.1.1.19), an important enzyme in polyamine synthesis. Polyamines accumulated in wild‐type plants (Col‐0 and Ler‐0) that were pre‐treated with 100 mM NaCl before transfer to 125 mM NaCl, but not in plants that were directly transferred to 125 mM NaCl without prior treatment with 100 mM NaCl. This shows that polyamine accumulation depends on acclimation to salinity. The salt treatment that induced polyamine accumulation in wild‐type plants did not lead to polyamine accumulation in the spe1‐1 and spe2‐1 mutants. Decreased fresh weight, chlorophyll content and photosynthetic efficiency indicated that the spe1‐1 mutant was more severely affected by salt stress than its wild type, Col‐0. In the spe2‐1 mutant decreased salt tolerance compared to its wild type, Ler‐0, became apparent as bleaching under severe salt stress. The present results demonstrate that decreased polyamine formation due to lower arginine decarboxylase activity leads to reduced salt tolerance.
An in vitro comparison of adhesive systems to seal pulp chamber walls
Authors: Ozturk B. , Özer F. , Belli S.
Published: April 2004

Aim  To compare in vitro the sealing properties of five different dentine adhesive materials (Prime&Bond NT (PBNT); Prompt L‐Pop (PLP); Clearfil SE Bond (CSEB); Scotchbond Multi Purpose Plus (SMPP); EBS‐Multi (EBSM)) inside the pulp chamber.

Methodology  Seventy‐five recently extracted human molar teeth were used. The roof of the pulp chambers and roots were removed under water cooling. Pulp tissue was removed, and the canal orifices were sealed. The pulp chambers were then treated with 5% sodium hypochlorite (NaOCl) for 1 min. The teeth were randomly divided into five groups of 15 teeth each. Adhesive systems were applied to the pulp chamber walls according to the manufacturers' instructions. The samples were connected to Plexiglass plates, and a fluid filtration method was used for quantitative evaluation of leakage. Measurements of fluid movement were made at 2‐min intervals for 8 min. The quality of seal of each specimen was measured immediately, after 24 h, 1 week and 1 month. The data were statistically analysed by repeated‐measurements multivariate anova, Friedman test, Wilcoxon signed rank test, Kruskal–Wallis of one‐way anova and Mann–Whitney U‐tests. The pulp chamber wall with and without NaOCl and resin–dentine interfaces of specimens were observed under a scanning electron microscope (SEM).

Results  The leakage values of the materials were significantly different at different measurement periods. In all groups, leakage values decreased with time (P < 0.05). PBNT and PLP had the least leakage during immediate measurements (P < 0.05). After 1 month, leakage of all adhesive systems was not significantly different (P < 0.05). SEM observation of pulp chamber walls demonstrated that the irregular dentine surface without smear layer was present in the nontreated group. However, NaOCl application removed the collagen fibrils leaving the dentine surface smooth. At resin–dentine interfaces of specimens, no hybridization zone was observed.

Conclusions  None of the materials had created a perfect seal to the pulp chamber walls. PBNT and PLP had better sealing over the short term, but over the long term, there were no differences between the materials.
Periapical radiolucency mimicking an odontogenic cyst
Authors: Nary Filho H. , Matsumoto M. A. , Fraga S. C. , Gonçales E. S. , Sérvulo F.
Published: April 2004

Aim  To present a clinical case of a giant cell lesion located in an unusual site, initially misdiagnosed and treated as an odontogenic cyst.

Summary  Periapical radiolucencies often suggest the presence of odontogenic pathosis, usually inflammatory granulomas or cysts. The high frequency of such lesions tends to lead clinicians to arrive at a diagnosis without completing a comprehensive assessment of the patient or carrying out the full range of available diagnostic tests. A case report of a giant cell lesion, which was misdiagnosed and treated initially as an odontogenic lesion because of its unusual location, is presented.

Key learning points

Clinical signs and radiographic appearance are usually sufficient to reach a diagnosis of periapical pathosis.

When traditional treatment does not lead to success, a biopsy should be considered to ascertain the diagnosis and allow the correct treatment to be provided.

Histological examination of soft tissue removed during endodontic surgery is essential.
Estimation of Tuberculosis Risk on a Commercial Airliner
Authors: Ko Gwangpyo , Thompson Kimberly M. , Nardell Edward A.
Published: April 2004

This article estimates the risk of tuberculosis (TB) transmission on a typical commercial airliner using a simple one box model (OBM) and a sequential box model (SBM). We used input data derived from an actual TB exposure on an airliner, and we assumed a hypothetical scenario that a highly infectious TB source case (i.e., 108 infectious quanta per hour) travels as a passenger on an 8.7‐hour flight. We estimate an average risk of TB transmission on the order of 1 chance in 1,000 for all passengers using the OBM. Applying the more realistic SBM, we show that the risk and incidence decrease sharply in a stepwise fashion in cabins downstream from the cabin containing the source case assuming some potential for airflow from more contaminated to less contaminated cabins. We further characterized spatial variability in the risk within the cabin by modeling a previously reported TB outbreak in an airplane to demonstrate that the TB cases occur most likely within close proximity of the source TB patient.
Risk‐Adjusted Mission Value: Trading Off Mission Risk for Mission Value
Authors: Miles Ralph F.
Published: April 2004

This article develops a decision‐theoretic methodology for the risk‐adjusted mission value (RAMV) for selecting between alternative missions in the presence of uncertainty in the outcomes of the missions. This methodology permits trading off mission risk for mission value, something that probabilistic risk analysis cannot do unless it explicitly incorporates both mission value and risk aversion of the project management. The methodology, in its complete implementation, is consistent with the decision theory known as expected utility theory, although it differs from conventional decision theory in that the probabilities and all but one of the utilities are not those of the decision maker. The article also introduces a new interpretation of risk aversion. The methodology is consistent with the elementary management concept concerning division of labor. An example is presented for selecting between discrete alternatives—four landing sites on Mars. A second example is presented for selecting among a set of continuous alternatives—a comet flyby distance. The methodology is developed within the context of scientific missions, but the methodology is equally applicable to any situation requiring outcome value judgments, probability judgments, and risk aversion judgments by different constituencies.
Science‐Based Trade Disputes: A New Challenge in Harmonizing the Evidentiary Systems of Law and Science
Authors: Harlow Siobán D.
Published: April 2004

In November 2001, the University of Michigan hosted one of the first dialogues among international trade law scholars and scientists in the field of risk assessment with the goal of identifying critical areas of misunderstanding between the two fields. This article discusses key issues that need to be addressed in order to better harmonize the scientific and legal systems of evidence within the context of trade disputes and trade law and presents the recommendations that emerged from the Michigan meeting.
The WHO/WTO Study on Trade and Public Health: A Critical Assessment
Authors: Howse Robert
Published: April 2004

This article provides a brief critical examination of a 2002 report jointly issued by the World Health Organization and the World Trade Organization on WTO law and public health. The author argues that the report is in many respects misleading as to the content of WTO law, as interpreted by the Appellate Body of the WTO, its final court. In particular, the WHO/WTO report systematically understates or underestimates the margins of regulatory autonomy that WTO member states have to make regulations that protect public health.
The Asbestos Case: A Comment on the Appointment and Use of Nonpartisan Experts in World Trade Organization Dispute Resolution Involving Health Risk
Authors: Carruth Russellyn S. , Goldstein Bernard D.
Published: April 2004

The World Trade Organization is currently evolving its approach to incorporating scientific and technological evidence into its dispute‐resolution process. In European Communities—Measures Affecting Asbestos and Asbestos‐Containing Products, the Panel was faced with a large amount of complex and conflicting scientific evidence presented by the partisan experts. The Asbestos Panel's solution was to appoint independent, nonpartisan experts to help it understand and evaluate the scientific evidence. While this was far better than trying to unravel the conflicting scientific evidence on its own, two aspects of the Panel's adopted procedure merit scrutiny. First, the expert‐selection process used by the Panel may not assure that the collective expertise of the appointed experts is broad enough when the dispute involves multidisciplinary scientific issues. Second, the process adopted by the Panel for consulting the appointed experts—which involved individual consultation rather than a consensus process—may leave a panel with a distorted or confused picture of the science. A consensus approach is the best means of obtaining scientific advice from appointed experts; it is most calculated to provide a clear and accurate report of the scientific information needed by a panel to make a fair and informed decision on the dispute before it. The underlying principle of world trade agreements is that it is beneficial to all of us to have free trade. Among other things, this requires an effective means of resolving disputes, and increasingly that includes disputes involving complex scientific and technological issues. This can be achieved only if the parties have confidence that their disputes will be decided in a fair and informed manner, based on the best science available. To achieve this goal, we suggest that future WTO panels depart in certain respects from the procedures utilized by the Asbestos Panel.
Does the position of the premature termination codon in COL7A1 correlate with the clinical severity in recessive dystrophic epidermolysis bullosa?
Authors: Ishiko Akira , Masunaga Takuji , Ota Takayuki , Nishikawa Takeji
Published: April 2004

Abstract:  Recessive dystrophic epidermolysis bullosa (RDEB) is an inherited skin disease caused by mutations in the gene encoding type VII collagen (COL7A1). The mutations are highly variable and this greatly complicates the study of the genotype–phenotype relationships. To date, three recurrent mutations, specific to Japanese RDEB patients have been reported. By comparing the phenotypes of RDEB patients with different recurrent mutations, the upstream positions of the premature termination codons (PTCs) showed strong correlation with the RDEB clinical disease severity. However, such correlations have not been supported by patients with mutations that were different from these recurrent Japanese patients mutations. In this study, we report a case of RDEB with a very mild clinical phenotype, who was a compound heterozygote harbouring both a recurrent Japanese mutation and a novel deletion mutation resulting in a more upstream PTC. The patient and his mother were shown to have a recurrent donor splice site mutation within intron 81 (6573 + 1G > C), a recurrent Japanese mutation that activates a cryptic donor splicing site and results in a downstream PTC. The patient and his father shared a single‐nucleotide deletion within exon 64 (5504delA), which causes a downstream frame shift in five amino acids before creating a PTC. Occurrence of the PTCs in mRNA was confirmed by reverse transcription‐polymerase chain reaction (RT)‐PCR. The patient's skin showed reduced immunofluorescence staining for COL7A1 and reduced number of abnormal or short anchoring fibrils by electron microscopy. Although the position of the mutation 5504delA PTC was located upstream of the previous mutations reported in combination with the 6573 + 1G > C mutation, the two mutations together give an apparently milder clinical phenotype. Therefore, genotype–phenotype relationships in RDEB cannot be explained purely by the position of PTC.
A placebo‐controlled evaluation of butterbur and fexofenadine on objective and subjective outcomes in perennial allergic rhinitis
Authors: Lee D. K. C. , Gray R. D. , Robb F. M. , Fujihara S. , Lipworth B. J.
Published: April 2004

Background
There are presently no placebo‐controlled data regarding the effects of butterbur (BB) on subjective and objective outcomes in patients with perennial allergic rhinitis.

Objective
We performed a placebo‐controlled evaluation of the effects of BB and fexofenadine (FEX) on subjective and objective outcomes in patients with perennial allergic rhinitis.

Methods
Sixteen patients with perennial allergic rhinitis and house dust mite sensitization were randomized in double‐blind cross‐over fashion to receive for 1 week either BB 50 mg twice daily, FEX 180 mg once daily and placebo (PL) once daily, or PL twice daily. The peak nasal inspiratory flow (PNIF) response to adenosine monophosphate (AMP) challenge administered as a single 400 mg/mL dose was measured over a 60‐min period after challenge, and domiciliary total nasal symptom score was recorded.

Results
Pre‐challenge values for mean±SEM PNIF (L/min) were not significantly different comparing all groups; BB (138±8), FEX (140±9), and PL (138±8). The maximum % PNIF fall from baseline after nasal AMP challenge was significantly attenuated (P<0.05) compared to PL (46±3), with BB (34±3) and FEX (39±3). The area under the 60‐min time–response curve (%.min) was also significantly attenuated (P<0.05) compared to PL (1734±156), with BB (1052±258) and FEX (1194±161). There was also a significant reduction (P<0.05) in total nasal symptom score with BB (1.8±0.4) and FEX (1.8±0.4), compared to PL (2.8±0.5). There were no significant differences between BB and FEX for any outcomes.

Conclusion
BB and FEX, in comparison to PL, were equally effective in attenuating the nasal response to AMP and in improving nasal symptoms, highlighting a potential role for BB in the treatment of allergic rhinitis.
Comparative effects of desloratadine, fexofenadine, and levocetirizine on nasal adenosine monophosphate challenge in patients with perennial allergic rhinitis
Authors: Lee D. K. C. , Gardiner M. , Haggart K. , Fujihara S. , Lipworth B. J.
Published: April 2004

Background
There are no data directly comparing the relative efficacy of modern H1‐antihistamines in allergic rhinitis using nasal provocation challenge.

Objective
We elected to study the comparative effectiveness of usual clinically recommended doses of desloratadine (DES), fexofenadine (FEX), and levocetirizine (LEV), on nasal adenosine monophosphate (AMP) challenge in patients with perennial allergic rhinitis (PAR).

Methods
16 patients with PAR were randomized in double‐blind cross‐over fashion to receive single doses of DES 5 mg, FEX 180 mg, LEV 5 mg, or placebo (PL), with nasal AMP challenge performed 12 h after dosing. Measurements of peak nasal inspiratory flow (PNIF) were made over 60 min after nasal AMP challenge.

Results
Pre‐challenge values (mean±SEM) for PNIF (L/min) were not significantly different comparing all groups; DES (129±9), FEX (128±11), LEV (128±13), and PL (128±12). The maximum % PNIF fall from baseline over 60 min after nasal AMP challenge was significantly attenuated (P<0.05) compared to PL (50±4), with DES (32±5), FEX (36±4), and LEV (36±4). The area under the 60‐min time–response curve (%.min) was also significantly attenuated (P<0.05) compared to PL (2110±268), with DES (1126±285), FEX (1225±255), and LEV (1261±194). There were no significant differences between the three H1‐antihistamines for any outcomes.

Conclusion
DES, FEX, and LEV were equally effective in attenuating the response to nasal AMP challenge. However, further long‐term studies will be required to study their comparative effects on nasal symptoms, quality of life, as well as on nasal inflammatory cells.
Comparing tacrolimus ointment and oral cyclosporine in adult patients affected by atopic dermatitis: a randomized study
Authors: Pacor M. L. , Di Lorenzo G. , Martinelli N. , Mansueto P. , Rini G.B. , Corrocher R.
Published: April 2004

Background
Atopic dermatitis (AD) is a chronic allergic inflammatory disease, which manifests itself with eczematous skin lesions.

Objective
We compared the clinical efficacy of tacrolimus ointment (0.1%) given twice a day and oral cyclosporine (3 mg/kg) given once daily. Rescue medication for itching included cetirizine 10–20 mg (equal to one or two tables).

Methods
Thirty patients, aged 13–45 years (mean±SD 27.1±10.9), with a history of moderate‐to‐severe AD were randomized to treatments, 15 patients for each treatments. Assessment of efficacy was based on SCORAD, on scores of daily itching, erythema, interference with sleep, due to the skin condition and days without use of cetirizine tablets. SCORAD, measured on a scale (0–103), was evaluated before treatment (0) and at 7, 14, 21, 28, 35 and 42 days after treatment. Similarly, the means of daily symptoms, on a scale (0–3), were evaluated before the treatment (0) and at 7, 14, 21, 28, 35 and 42 days after treatment; finally, on day without use of cetirizine tablets. The safety of the study treatments was assessed through haematologic, biochemical and urinary testing and on systolic and diastolic blood pressures and heart rate measurements.

Results
SCORAD decreased in the two treatment groups 14 days after the beginning of the period study. However, the patients in tacrolimus ointment group reported significantly lower SCORAD than those treated with oral cyclosporine. Overall SCORAD, as assessed by the area under the curve (AUC) day0–42 (score/day), was significantly lower in the tacrolimus ointment group when compared with oral cyclosporine (P<0.001). Similarly, AUC day0–42 (score/day) for itching, erythema and number of nights without interference with the sleep due to skin condition were significantly lower in the group of patients treated with tacrolimus compared with those treated with cyclosporine (P=0.003, 0.005 and 0.01, respectively). As regards the use of rescue medication, expressed by median of number of days without use of anti‐H1, it was significantly lower in the group treated with tacrolimus (82.5) than in the cyclosporine group (76.5) (P=0.03). There were no appreciable changes in haematological and biochemical indices, in both treatments groups.

Conclusion
The results of this comparative study demonstrate that tacrolimus ointment twice daily and cyclosporine administered orally once daily are effective on SCORAD, daily symptoms and anti‐H1 rescue. When we compared tacrolimus and cyclosporine there was a faster onset of action in the group treated with tacrolimus. The two drugs presented the same safety. However, these data support the preferential use of topical tacrolimus 0.1% in AD, because cyclosporine has potential side‐effects.
Differential activation of signal transducer and activator of transcription 6 in B cells from allergic children and their non‐allergic siblings
Authors: Deveikaite V. , Fila C. , Laurencikiene J. , Klein E. , Kihlström A. , Lilja G. , Sverremark ekström E. , Severinson E.
Published: April 2004

Background
The germline (GL) ɛ promoter is regulated by IL‐4 and is essential for class switching to IgE. IL‐4‐induced gene expression is largely mediated through activation of latent transcription factor STAT6 (signal transducer and activator of transcription 6).

Objective
We investigated whether increased levels of IgE in allergic individuals may be associated with alteration in the level or activation of STAT6 and subsequent increase in GL ɛ promoter activity.

Methods
Electrophoretic mobility shift assay and Western blotting assays were used to investigate the level of expression and activation of STAT6 in Epstein–Barr virus (EBV)‐transformed B cell lines from children with birch pollen allergy and their non‐allergic siblings. The activity of the GL ɛ promoter was tested in a transient transfection assay.

Results
STAT6 was expressed at the same level in all B cell lines tested. In two out of five sibling pairs STAT6 was activated by IL‐4 more efficiently in the allergic individuals but in the three other pairs the opposite was found. In transient transfections, no difference in IL‐4‐induced GL ɛ promoter function was detected, although basal promoter activity varied between allergic and healthy siblings in two out of five pairs.

Conclusions
We demonstrate for the first time that upon IL‐4 signalling STAT6 transcription factor activation differs in B cells from different individuals. Although we did not find any association between STAT6 activation and allergy, we do not exclude a possibility that stronger activation of this transcription factor is associated with an expression of allergic phenotype.
Efficacy and safety of a recombinant anti‐immunoglobulin E antibody (omalizumab) in severe allergic asthma
Authors: Holgate S. T. , Chuchalin A. G. , Hébert J. , Lötvall J. , Persson G. B. , Chung K. F. , Bousquet J. , Kerstjens H. A. , Fox H. , Thirlwell J. , Cioppa G. Della ,   on behalf of the Omalizumab 011 International Study Group
Published: April 2004

Background
Patients with severe asthma are often inadequately controlled on existing anti‐asthma therapy, constituting an unmet clinical need.

Objective
This randomized, double‐blind, placebo‐controlled trial evaluated the ability of omalizumab, a humanized monoclonal anti‐IgE antibody, to improve disease control sufficiently to enable inhaled corticosteroid reduction in patients with severe allergic asthma.

Methods
After a run‐in period when an optimized fluticasone dose (1000 μg/day) was received for 4 weeks, patients were randomized to receive subcutaneous omalizumab [minimum 0.016 mg/kg/IgE (IU/mL) per 4 weeks; n=126] or matching placebo (n=120) at intervals of 2 or 4 weeks. The study comprised a 16‐week add‐on phase of treatment followed by a 16‐week fluticasone‐reduction phase. Short‐/long‐acting β2‐agonists were allowed as needed.

Results
Median reductions in fluticasone dose were significantly greater with omalizumab than placebo: 60% vs. 50% (P=0.003). Some 73.8% and 50.8% of patients, respectively, achieved a 50% dose reduction (P=0.001). Fluticasone dose reduction to 500 μg/day occurred in 60.3% of omalizumab recipients vs. 45.8% of placebo‐treated patients (P=0.026). Through both phases, omalizumab reduced rescue medication requirements, improved asthma symptoms and asthma‐related quality of life compared to placebo.

Conclusion
Omalizumab treatment improves asthma control in severely allergic asthmatics, reducing inhaled corticosteroid requirements without worsening of symptom control or increase in rescue medication use.
Eosinophil activation and cysteinyl leukotriene production in infants with respiratory syncytial virus bronchiolitis
Authors: Dimova‐Yaneva D. , Russell D. , Main M. , Brooker R. J. , Helms P. J.
Published: April 2004

Background
It has been suggested that acute infantile bronchiolitis associated with respiratory syncytial virus (RSV) may share some pathogenic features with atopic asthma in that virus‐specific IgE is produced and cysteinyl leukotrienes (cLTs) and eosinophil cationic protein (ECP) have been detected in airway secretions. ECP is a specific marker of eosinophil activation although leukotrienes can be released from a variety of cells including mast cells, eosinophils and monocytes.

Objective
To test the association between eosinophil activation and cysteinyl leukotriene production in the upper airway secretions of infants with RSV positive (RSV+ve) bronchiolitis.

Methods
Nasal lavage samples were performed in 78 infants (0.0–11.5 months) admitted to hospital with RSV+ve bronchiolitis soon after admission (0–48 h). Leukotriene C4 (LTC4) was assayed by enzyme immunoassay (EIA) and eosinophil cationic protein (ECP) by fluoroimmunoassay (FIA).

Results
LTC4 was detectable in 51 and ECP in 57 of 78 samples with a significant positive relationship between LTC4 and ECP (r=0.557, P<0.001).

Conclusion
In the majority of our subjects with RSV+ve bronchiolitis ECP and LTC4 were detectable in upper airway secretions and were significantly associated with each other. In this clinical setting much of the detected LTC4 within upper airway secretions is likely to originate from the eosinophil, an observation that may have implications for clinical management and for delineation of the underlying mechanisms associated with this illness.
The hevein domain of the major latex‐glove allergen Hev b 6.01 contains dominant T cell reactive sites
Authors: De Silva H. D. , Gardner L. M. , Drew A. C. , Beezhold D. H. , Rolland J. M. , O'Hehir R. E.
Published: April 2004

Background
Sensitization to natural rubber latex (Hevea brasiliensis) is a major cause of occupational asthma and rhinitis affecting frequent latex‐glove users. Hev b 6.01, a known major latex allergen, is cleaved naturally into hevein (4.7 kDa) and a C‐terminal fragment (14 kDa). Hevein is an abundant protein in latex‐glove extracts. As the immune response to allergens is initiated by activation of allergen‐specific CD4+ T cells, identification of dominant T cell epitopes is crucial for the development of specific immunotherapy.

Objective
To identify dominant T cell epitopes of Hev b 6.01 in latex‐allergic glove users.

Methods
Ten latex‐allergic frequent glove users and six non‐latex‐allergic atopic control subjects were selected, based on clinical symptoms and positive latex‐specific serum IgE. Serum IgE reactivity to glove extract and recombinant Hev b 6.01 (rHev b 6.01) were analysed by ELISA. Latex‐specific short‐term oligoclonal T cell lines were generated from peripheral blood of latex‐allergic subjects. These lines were tested for proliferative responses to overlapping 20‐mer peptides of the Hev b 6.01 molecule. CD4+ T cell intracellular cytokines, IL‐4 and IFN‐γ were assessed following stimulation with immobilized anti‐CD3 in the presence of IL‐2.

Results
All ten of the latex‐allergic patients showed serum IgE binding to glove extract while eight of these also showed IgE binding to rHev b 6.01 by ELISA. Western blotting confirmed reactivity with rHev b 6.01 at around 20 kDa. T cell proliferation assays showed that latex‐specific T cell lines from all subjects responded to one or more peptides, with greatest frequency of reactivity to peptides Hev b 6.01 p(10–29) and Hev b 6.01 p(19–38) in the hevein domain. An allergic‐type cytokine profile with considerable IL‐4 in addition to IFN‐γ was evident from intracellular cytokine staining.

Conclusion
Hevein is an important T cell as well as B cell immunogen and contains dominant T cell reactive sites.
Random outcomes of allergen‐specific responses in atopic families
Authors: Jackola D. R. , Liebeler C. L. , Blumenthal M. N. , Rosenberg A.
Published: April 2004

Background
Allergens are common non‐infectious antigens to which people will mount T cell dependent humoral responses. Among genetically susceptible individuals, an antigen‐specific response results involving the production of allergen‐specific IgE (atopy).

Objective
Determine if this susceptibility is manifested as an inherited, allergen‐specific trait or a random response to allergens among susceptible people.

Methods
We evaluated allergen‐specific outcomes in 1099 members of families with positive atopic history (26 multi‐generation and 112 nuclear families). Each was tested for sensitivity to 14 common allergens by standardized skin prick test (SPT), a marker of specific IgE production. Over 15 000 individual SPT's were evaluated. Among five randomly selected multi‐generation families (N=163), semi‐quantitative determinations of Amb a 1‐specific IgA1,2 and IgG1–4 were determined in three groups: (A) Amb a SPT+/Amb a 1‐IgE+, (B) Amb a SPT−/Amb a 1‐IgE+, (C) Amb a SPT−/Amb a 1‐IgE−.

Results
By rank correlation statistics, there were no discernible ‘patterns’ of specific SPT outcomes among any of the multi‐generation families, suggesting that environmental exposure rather than allergen‐specific inheritance determined the responses. This was confirmed among the nuclear families since the conditional SPT outcomes among children were independent of the SPT responses of their parents. Among five randomly selected multi‐generation families, the relative proportionate concentrations of the Amb a 1‐specific IgA and IgG subclasses were comparable, regardless of atopic sensitization to the ragweed allergen Amb a.

Conclusion
While the general propensity for atopy may be inherited, an individual's specific atopic outcome is a random variable independent of familial sensitization patterns.
Reduced levels of soluble CD14 in atopic children
Authors: Zdolsek H. A. , Jenmalm M. C.
Published: April 2004

Background
A reduced microbial stimulation has been reported as a reason for the increasing prevalence of atopic diseases in industrialized countries. Antigen‐presenting cells (APC), responding to microbial signals by pattern recognition receptors such as CD14, have an important role in the development of the Th1/Th2 balance.

Objective
We hypothesized that atopic children have a lower expression of CD14 on monocytes and lower soluble CD14 levels (sCD14).

Methods
Seventy‐six children were followed prospectively from birth and signs of atopic disease were evaluated. The expression of CD14 on monocytes was analysed with flow cytometry at 0, 3, 6, 12 and 18 months. Circulating levels of sCD14 were analysed by ELISA and total IgE was analysed by fluoroenzymo immunoassay at these ages, and in a subgroup, followed up at 7 years.

Results
Levels of sCD14 were reduced at 7 years both in children with a current or a cumulative history of atopy compared to non‐atopic children with P=0.002 and 0.001, respectively. Sensitized children with atopic symptoms had lower sCD14 at 3 and 18 months and at 7 years of age than non‐atopic non‐sensitized children with P=0.023, 0.039 and 0.008, respectively.

Conclusion
The lower levels of sCD14 observed in atopic children may be a consequence of an atopic family heredity and/or atopic disease, but it may also reflect a reduced capacity to respond to microbial signals.
Relevance of Ara h1, Ara h2 and Ara h3 in peanut‐allergic patients, as determined by immunoglobulin E Western blotting, basophil–histamine release and intracutaneous testing: Ara h2 is the most important peanut allergen
Authors: Koppelman S.J. , Wensing M. , Ertmann M. , Knulst A.C. , Knol E.F.
Published: April 2004

Background
A number of allergenic proteins in peanut has been described and the relative importance of these allergens is yet to be determined.

Objectives
We have investigated the relevance of previously identified peanut allergens in well‐characterized peanut‐allergic patients by in vitro, ex vivo and in vivo assays.

Methods
Thirty‐two adult peanut‐allergic patients were included based on careful and standardized patient history and the presence of peanut‐specific IgE. The diagnosis peanut allergy was confirmed using double‐blind placebo‐controlled food challenges in 23 patients. Major peanut allergens Ara h1, Ara h2 and Ara h3 were purified from peanuts using ion‐exchange chromatography. IgE immunoblotting was performed and IgE‐cross‐linking capacity was examined by measuring histamine release (HR) after incubating patient basophils as well as passively sensitized basophils with several dilutions of the allergens. Intracutaneous tests (ICTs) using 10‐fold dilution steps of the purified allergens and crude peanut extract were performed.

Results
Ara h2 was recognized most frequently (26 out of 32) in all tests and induced both positive skin tests and basophil degranulation at low concentrations, whereas Ara h1 and Ara h3 were recognized less frequently and reacted only at 100‐fold higher concentrations as analysed with HR and intracutaneous testing (ICT). Next to the three tested allergens, proteins with molecular weights of somewhat smaller than 15 kDa were identified as a IgE‐binding proteins on immunoblot in the majority of the patients (20 out of 32).

Conclusion
We conclude that Ara h2 is, for our patient group, the most important peanut allergen, and that previously unidentified peanut proteins with molecular weights of somewhat smaller than 15 kDa may be important allergens as well. ICT in combination with basophil–HR and IgE immunoblotting provides insight in the patient specificity towards the individual peanut allergens.
Effect of ingestion of cow's milk hydrolysed formulas on whey protein‐specific Th2 immune responses in naïve and sensitized mice
Authors: Peng H.‐J. , Su S.‐N. , Tsai J.‐J. , Tsai L.‐C. , Kuo H.‐L. , Kuo S.‐W.
Published: April 2004

Background
For genetically predisposed atopic infants, cow's milk protein hydrolysed formulas have been widely used.

Objective
Whether hydrolysed formulas can induce oral tolerance to whey proteins will be extensively studied in naïve and sensitized mice.

Methods
Antigenicity of hydrolysed formulas was first studied using immunoblotting. Naïve mice fed hydrolysed formulas for 1–4 weeks were sensitized with whey allergens. In contrast, mice sensitized with whey allergens were fed hydrolysed formulas continually for 12 weeks.

Results
Whey allergens were found in Nan and Neoangelac FL. Large whey peptides with antigenicity were found in Nan‐HA. Profound suppression of IgE, IgG1 and IgG responses to whey allergens were induced in those fed Nan for 1 week, or Nan‐HA for 4 weeks. IgE responses to whey allergens were suppressed in those fed Neoangelac FL for 4 weeks, or Nan‐HA for 1–2 weeks. In contrast, those fed extensively hydrolysed formulas for 1–4 weeks failed to show decreased responses. On the other hand, IgE responses to β‐lactoglobulin, but not to bovine serum albumin or α‐lactalbumin, were decreased in sensitized mice fed Nan for 12 weeks. There was no suppression in sensitized mice fed hydrolysed formulas.

Conclusion
Suppression of IgE responses to whey proteins was readily induced in naïve mice fed Nan or Nan‐HA for 1 week. In contrast, it was hardly induced in sensitized mice even after prolonged feeding of Nan for 12 weeks, let alone hydrolysed formulas.
Are maternal psychosocial factors associated with cord immunoglobulin E in addition to family atopic history and mother immunoglobulin E?
Authors: Lin Y.‐C. , Wen H.‐J. , Lee Y.‐L. , Guo Y. L.
Published: April 2004

Background
Atopy in maternal and family histories is known to be a risk factor for elevated umbilical cord immunoglobulin E (cIgE). However, the association between cIgE and psychosocial factors remains under investigation.

Objective
To explore whether psychosocial factors in addition to atopy contribute to elevated cIgE.

Methods
Four private maternity hospitals fitting the quantile levels of SO2 in 2000 cooperated with us by recruiting participants for this study: pairs of mothers and neonates living within 3‐km catchment areas of air‐monitoring stations. We used a questionnaire to collect exposure data, and the Pharmacia UniCap IgE assay test system to determine the levels of IgE in gravidas and cord blood.

Results
Between July 2001 and March 2003, 334 mother and neonate pairs participated in this study. The frequencies of sensitization, serum IgE (sIgE)>100 IU/mL, or cIgE0.35 IU/mL were not different between the four different hospitals. By multi‐variate logistic regression analysis adjusted for environmental factors, genetic factors, and psychosocial factors, the risk factors for elevated cIgE were being a male neonate (odds ratio (OR)=3.5, 95% confidence interval (CI)=[1.5, 8.5]), carpets at home (OR=3.0, 95% CI=[1.02, 8.4]), maternal allergy to dog dander (OR=9.7, 95% CI=[1.2, 98.8], maternal total serum IgE>100 IU/mL (OR=5.1, 95% CI=[2.2, 12.8]), maternal regularly/mostly/often self‐reported nervousness (OR=4.0, 95% CI=[1.3, 12.8]), family income $11 574–17 361/year (OR=3.7, 95% CI=[1.3, 11.5]), incense burning (OR=4.0, 95% CI=[1.4, 13.3]), and atopy in maternal grandparents (OR=4.8, 95% CI=[1.7, 14.0]). By principle component analysis and logistic regression, psychosocial stress (β±standard error=0.26±0.13, P=0.04) was associated with increased cIgE.

Conclusion
Psychosocial factors are potentially important risk factors for elevated cIgE.
Component‐resolved diagnosis of house‐dust mite allergy with purified natural and recombinant mite allergens
Authors: Pittner G. , Vrtala S. , Thomas W. R. , Weghofer M. , Kundi M. , Horak F. , Kraft D. , Valenta R.
Published: April 2004

Background
Mites belong to the most frequent and potent allergen sources. Immunotherapy with mite allergen extracts is frequently performed if allergen avoidance is not possible or successful. However, highly controversial results have been reported for mite‐specific immunotherapy.

Objective
The aim of this study was to develop diagnostic concepts that may contribute to an improved selection of patients for immunotherapy with Der p allergen extracts and that may be used for immunological monitoring of patients undergoing this treatment.

Methods
The IgE reactivity profiles to Der p extract were determined in a Middle European mite‐allergic population by IgE immunoblotting and by using a panel of seven purified natural or recombinant Der p allergens (nDer p 1, nDer p 4, rDer p 2, rDer p 5, rDer p 7, rDer p 8, rDer p 10). Furthermore, we investigated the sensitization and cross‐reactivity to house‐dust‐ and storage‐mite allergen extracts by CAP FEIA measurements and by IgE competition studies.

Results
More than 95% of the patients could be diagnosed with a combination of nDer p 1 and rDer p 2. With the methods used, we could discriminate mite‐allergic patients who were mainly sensitized to the major Der p allergens (Der p 1, Der p 2) from patients with a broad sensitization profile, including highly cross‐reactive allergens (e.g. Der p 10: tropomyosin) as well as reactivity to storage mites.

Conclusions
Diagnostic tests containing the major mite allergens (i.e. Der p 1, Der p 2) and highly cross‐reactive mite allergens (e.g. Der p 10) may improve the diagnostic selection of patients for immunotherapy with Der p extracts. These tests may also be used for the immunological monitoring of patients undergoing immunotherapy.
Different cytokine production and activation marker profiles in circulating cutaneous‐lymphocyte‐associated antigen+ T cells from patients with acute or chronic atopic dermatitis
Authors: Antúnez C. , Torres M. J. , Mayorga C. , Cornejo‐García J. A. , Santamaría‐Babi L. F. , Blanca M.
Published: April 2004

Background  Atopic dermatitis (AD) is an inflammatory skin disease whose lesions can have two stages: acute and chronic. In skin biopsies a biphasic pattern of cytokine expression has been shown, Th2 in acute lesions and Th1 in chronic AD lesions.

Objective  We investigated the expression of an activation marker and a homing receptor, as well as cytokine production, in different peripheral blood T cell subpopulations from AD patients with chronic (Group A) and acute lesions (Group B) and controls.

Methods  We evaluated 26 adult AD patients (12 Group A, 14 Group B) and 14 non‐atopic controls. IgE was measured by immunoassay. CD4, CD8, cutaneous‐lymphocyte‐associated antigen (CLA) and human leucocyte antigen (HLA)‐DR expression, and cytokine production (IL‐2, IL‐13, IFN‐γ, TNF‐α, IL‐10, IL‐4) were analysed in mononuclear cells by flow cytometry.

Results  In Group B there was a significant increase in eosinophil levels and a non‐significant increase in IgE. In Group A we found an increase in CLA+CD4+ cells (8.19±1.84) compared with controls (4.83±0.53) (P<0.05) and CD4+HLA‐DR+ cells in the CLA+ subpopulation (45.54±15.40) compared with controls (30.49±6.07) (P<0.05). In the CLA+CD4+ subpopulation, there was a significant increase in IL‐4, IL‐13 and TNF‐α production in Group B (12.46±7.7, 11.26±5.97, 43.92±15.55) compared with controls (5.34±3.50, 4.54±1.78, 19.29±9.97) with no differences in Group A.

Conclusion  Greater immunological differences were detected in peripheral blood from patients with acute compared with chronic lesions, especially in the circulating T cell‐subset with skin tropism that preferentially responded to cutaneous allergens. This is the first demonstration of phenotypic changes in circulating CLA+ T cells between AD patients with acute and chronic lesions.