Genetic variants of the IL13 and IL4 genes and atopic diseases in at-risk children.

We studied a cohort containing 368 children at high risk of developing atopy and atopic disorders and 540 parents of those children to investigate whether the IL13 Arg130Gln and C-1112 T polymorphisms were associated with these outcomes. We also investigated whether haplotypes consisting of any two polymorphisms of IL13 Arg130Gln, IL13 C-1112 T and IL4 C-589 T were associated with these phenotypes. In 288 white children, the IL13 130Gln allele was associated with atopy (RR=1.9, P=0.047), and with atopic dermatitis (RR=2.5, P=0.014). The associations were confirmed using a family-based test of association (P=0.027 and 0.030, respectively) in all subjects. In white subjects there were associations of haplotypes consisting of IL13 Arg130Gln and IL4 C-589 T with atopic dermatitis (P=0.006) and with atopy (P=0.009). Our data suggest that the IL13 Arg130Gln polymorphism and haplotypes consisting of IL13 Arg130Gln and IL4 C-589 T were associated with the development of atopy and atopic dermatitis at 24 months of age.

Crystallization and rhenium MAD phasing of the acyl-homoserinelactone synthase EsaI.

Acyl-homoserine-L-lactones (AHLs) are diffusible chemical signals that are required for virulence of many Gram-negative bacteria. AHLs are produced by AHL synthases from two substrates, S-adenosyl-L-methionine and acyl-acyl carrier protein. The AHL synthase EsaI, which is homologous to the AHL synthases from other pathogenic bacterial species, has been crystallized in the primitive tetragonal space group P4(3), with unit-cell parameters a = b = 66.40, c = 47.33 A. The structure was solved by multiple-wavelength anomalous diffraction with a novel use of the rhenium anomalous signal. The rhenium-containing structure has been refined to a resolution of 2.5 A and the perrhenate ion binding sites and liganding residues have been identified.

The association between respiratory viruses and symptoms in 2-week-old infants at high risk for asthma and allergy.

OBJECTIVES: The role of viral respiratory tract infections in the onset of childhood asthma and allergy is controversial, partly because of limited understanding about postnatal viral exposures. We investigated the prevalence of 3 common respiratory viruses and associated respiratory symptoms in 2-week-old infants at high risk for having asthma and allergy. STUDY DESIGN: Frozen nasal specimens from 2-week-old children at high risk (n = 495) underwent reverse transcription-polymerase chain reaction (RT-PCR) for picornavirus-, parainfluenza-, and respiratory syncytial virus-specific nucleic acid. RT-PCR findings were related to respiratory symptoms (cold, cough, and wheeze) and to characteristics implicated with increased risk for asthma and allergy. RESULTS: Viral RT-PCR was positive in 199 (40.2%) of 495 specimens examined, with picornavirus and parainfluenza significantly associated with respiratory symptoms. Viral prevalence was significantly higher in children born during the winter and summer months. CONCLUSIONS: A high percentage (40.2%) of infants at high risk for asthma and allergy had been exposed to common respiratory viruses at 2 weeks of age. RT-PCR is a powerful diagnostic method that can be used in epidemiologic studies examining the role of viral respiratory tract infections in the pathogenesis of pediatric asthma and allergy.

Polymorphisms of the IL-4, TNF-alpha, and Fcepsilon RIbeta genes and the risk of allergic disorders in at-risk infants.

Polymorphisms in the TNF-alpha (A-308G), IL-4 (C-589T), and Fcalpha RIbeta (E237G) genes have been associated with asthma and related phenotypes. To determine the predictive value of these polymorphisms we have assessed their relative risk (RR) for the development of atopy, asthma, and rhinitis in a high-risk infant population that is being followed longitudinally from birth. DNA was extracted and genotyped for 373 infants and 572 parents for each polymorphism. Phenotypic data were collected for atopy and allergic diseases in the infants at 12 mo of age. The prevalence of these phenotypes in the 281 white infants was compared in each genotypic group. There were no differences in the prevalence of any phenotype between genotypes of the TNF-alpha and Fcalpha RIbeta polymorphisms. However, we found that the IL4-589*T allele was associated with "probable" asthma (RR = 4.1) and that homozygotes for the IL4-589*T allele had an increased risk for the development of rhinitis (RR = 2.4). Using the transmission disequilibrium test, an association of IL4-589*T with atopy was found. We conclude that IL-4-589*T, but not TNF-alpha-308*2 or Fcalpha RIbeta*G, is a risk factor for the development of atopy, asthma, and rhinitis by 12 mo of age.

Visual and Infrared Assessment of Root Colonization of Apple Trees by Phymatotrichopsis omnivora.

Root systems of 5-year-old, trellised apple trees (Malus domestica Borkh) on cv. M.7a root-stocks were assessed for the presence of fungal strands of Phymatotrichopsis omnivora (Duggar) Hennebert in two orchards in central Texas. Fungal advance within each orchard was assessed in five directions. Pathogen growth (P < 0.01) occurred beyond symptomatic trees along and perpendicularly across rows. In one orchard, 80% of the first asymptomatic trees were infected along rows, followed by 60% infection perpendicularly across rows. In the other orchard, there was 100% infection of the first asymptomatic trees along rows and 60% infection perpendicularly across rows. No growth was observed diagonally across rows in either orchard. Infrared readings of canopy temperature and differences between canopy temperature and air temperature were significant (P < 0.01) for predicting infection of asymptomatic, infected trees in one orchard. Trees were shown to have extensive taproot decay and infection of lateral roots before canopy symptoms began to develop. Root diameter appeared to have no effect on the growth of the fungus.

Phosphatidylinositol 3-phosphate recognition by the FYVE domain.

Recognition of phosphatidylinositol 3-phosphate (Ptdlns(3)P) is crucial for a broad range of cellular signaling and membrane trafficking events regulated by phosphoinositide (PI) 3-kinases. PtdIns(3)P binding by the FYVE domain of human early endosome autoantigen 1 (EEA1), a protein implicated in endosome fusion, involves two beta hairpins and an alpha helix. Specific amino acids, including those of the FYVE domain's conserved RRHHCRQCGNIF motif, contact soluble and micelle-embedded lipid and provide specificity for Ptdlns(3)P over Ptdlns(5)P and Ptdlns, as shown by heteronuclear magnetic resonance spectroscopy. Although the FYVE domain relies on a zinc-binding motif reminiscent of RING fingers, it is distinguished by ovel structural features and its ptdlns(3)P-binding site.

Breast-feeding and environmental tobacco smoke exposure.

BACKGROUND: Exposure to environmental tobacco smoke is associated with adverse effects in infants and children. OBJECTIVE: To explore whether an increase in urinary cotinine fumarate level is caused by ingested nicotine and cotinine in breast-feeding infants. METHODS: We studied newborns at risk for developing asthma and allergies based on a strong family history. We measured urinary cotinine levels in the infants as a measure of environmental tobacco smoke exposure and cotinine levels in the breast milk of breast-feeding mothers. RESULTS: Of 507 infants, urinary cotinine levels during the first 2 weeks of life were significantly increased in infants whose mothers smoked. Breast-fed infants had higher cotinine levels than non-breast-fed infants, but this was statistically significant (P<.05) only if mothers smoked. Urinary cotinine levels were 5 times higher in breast-fed infants whose mothers smoked than in those whose mothers smoked but did not breast-feed. CONCLUSIONS: Mothers should be encouraged to not smoke, and parents must be advised of the potential respiratory and systemic risks of environmental tobacco smoke exposure to their child, including the potential for future addiction to smoking.

Elderly patients' perceptions of care in the emergency department.

OBJECTIVE: Patients' perceptions and satisfaction are areas of growing concern in health care research, but little has been reported from the perspective of elderly persons. The purpose of this study was to describe elderly patients' perceptions of care in the emergency department. METHODS: A qualitative, descriptive study design was used. Twelve elderly people were interviewed following a treatment episode in 1 of 3 emergency departments in the western United States and data were submitted to content analysis according to qualitative, interpretive methodology. FINDINGS: The following 5 themes emerged from the analysis: "needs for information," "observations of waiting time," "perceptions of professional competency and caring service," "concerns about process and facility design," and "personal tolerance." DISCUSSION: Findings support some aspects of existing literature and offer additional information regarding care of elderly persons in the emergency department. Results also support the need for more research in the area of the actual experience of elderly patients in the emergency department.

Fel d 1 peptides: effect on skin tests and cytokine synthesis in cat-allergic human subjects.

We tested peptide immunotherapy in cat-allergic humans, using a formation of two synthetic peptides, IPC-1 and IPC-2, each of which is 27 amino acids long and contains T cell-reactive regions of Fel d 1, the major cat allergen. In this exploratory, randomized, double-blind, parallel-group study, 42 subjects received s.c. injections of treatment peptides 250 micrograms or placebo weekly for four consecutive weeks. Changes in immediate- and late-phase skin test reactivity, and in antigen-driven cytokine synthesis were assessed. Epicutaneous (end-point titration) and intradermal tests were performed with cat extract (ALK SQ Cat Hair) containing Fel d 1, before the first injection, then 2, 6 and 24 weeks after the fourth and last injection of peptides or placebo. IL-4, IL-10 and IFN-gamma expression by circulating peripheral blood mononuclear cells (PBMC) in response to cat extract was measured using short-term bulk culture of PBMC and short-term limiting dilution analysis. Subjects who received peptide immunotherapy did not tolerate significantly more cat extract containing Fel d 1 in the skin tests 2, 6 or 24 weeks after the last injection than they did at baseline, and their late-phase responses did not decrease significantly compared to baseline. Substantial IL-4, IL-10 and IFN-gamma responses were observed following primary culture of cat antigen-stimulated PBMC; however, the intensity of cytokine synthesis and the IFN-gamma: IL-4 ratio were unchanged in peptide- and placebo-treated groups 6 and 24 weeks after the last injection. A few hours after the injections, subjects receiving peptides reported more allergic rhinitis and asthma symptoms and more pruritus than those receiving placebo. We conclude that under the conditions tested, peptide immunotherapy did not reduce immediate- or late-phase skin reactivity to cat extract containing Fel d 1 or modify cat antigen-specific cytokine production significantly.

Pharmacokinetics of the orally administered decongestants pseudoephedrine and phenylpropanolamine in children.

BACKGROUND: There is little published, objective information about pseudoephedrine and phenylpropanolamine in the treatment of children. Our goal was to determine the pharmacokinetics of these medications in young subjects. METHODS: In two sequential double-blind, parallel-group, single-dose studies, 21 children received either pseudoephedrine, 30 or 60 mg, or placebo, and 20 children received either phenylpropanolamine, 20 or 37.5 mg, or placebo. Before dosing and at intervals up to 7 hours after dosing, serum pseudoephedrine or phenylpropanolamine concentrations were measured, and pulse and blood pressure were recorded. In two children receiving each drug, these tests were also performed at 12 and 24 hours, and urine was collected from 0 to 12 and from 12 to 24 hours. RESULTS: In children, the mean (+/-SEM) terminal elimination half-life values for pseudoephedrine, 3.1 +/- 0.5 hours, and for phenylpropanolamine, 2.6 +/- 0.6 hours, were significantly shorter than those found by other investigators in adults. Pharmacokinetics were not dose dependent in the dose ranges studied. CONCLUSION: Further studies of pseudoephedrine and phenylpropanolamine should be performed in children with the use of objective measurements. The widespread use of these medications in young subjects should be reevaluated.

Nasal patency in children with allergic rhinitis: correlation of objective and subjective assessments.

BACKGROUND: Children with allergic rhinitis may have difficulty with self-assessment of nasal symptoms. OBJECTIVE: To correlate objective and subjective assessments of nasal stuffiness in children with seasonal allergic rhinitis. METHODS: Children, aged 6 to 12 years, with seasonal allergic rhinitis recorded their degree of nasal obstruction on two separate occasions using a nasal stuffiness score and a visual analogue scale. Physicians also assessed the degree of nasal obstruction using a visual analogue scale. Anterior rhinometry was performed and saccharin transient time was also measured. Correlations between subjective scores and objective measurements were calculated using Spearman correlation coefficients. RESULTS: Patient's nasal stuffiness scores correlated with their visual analogue assessment (r = .45, P = .0001). Patients visual analogue assessments did not correlate with anterior nasal airflow (r = -.12, P > .05). Physicians' visual analogue assessment correlated better with nasal airflow than childrens' assessment (r = -.41, P = .0001). Saccharin transit time was not helpful in assessment of degree of nasal obstruction. CONCLUSION: Children appear to have difficulty in self-assessment of nasal symptoms, and to be poor judges of the presence or severity of nasal obstruction. In studies of allergic rhinitis in children, objective measurements should be performed, if possible, to facilitate more accurate interpretation of data.

Food allergy in children.

The diagnosis of hypersensitivity reactions to foods in infancy and childhood requires the use of clinical skills and laboratory diagnostic methods to identify suspect foods. Patients and parents occasionally may need to keep food/symptom diaries to explore the association of foods and adverse reaction. Skin testing or RAST may shorten the list of potential allergens because of their excellent negative predictive value. Except for obvious serious allergic reactions after ingestion of a single food, confirmation of the reaction may be ideally confirmed by a DBPCFC, especially if the reported symptoms are subjective in nature. Equivocal responses should be repeated. Although many in vitro and in vivo diagnostic methods have been developed to potentially improve the diagnosis of food allergy in children, no test has been able to predict the results of the DBPCFC with any greater accuracy than skin tests or RAST. The "perfect" test with excellent positive and negative predictive values has yet to be developed.

Effect of nebulized ipratropium bromide on intraocular pressures in children.

STUDY OBJECTIVE: To evaluate the effects of nebulized ipratropium bromide on intraocular pressures and pupillary responses in children with asthma. DESIGN: A double-blind, randomized, crossover study. SETTING: Children's Hospital of Winnipeg, University of Manitoba. PATIENTS OR PARTICIPANTS: Age 6 to 17 years with asthma. INTERVENTION: Nebulized ipratropium bromide added to albuterol sulfate, albuterol alone, or saline solution was given by face mask and nebulizer. Before and 0.5 h after nebulization, intraocular pressures (mm Hg), pupillary size (mm), and pupillary responses were measured. In a subsequent open study, patients who had been admitted to hospital with acute asthma who were treated with nebulized ipratropium bromide were recruited for measurement of intraocular pressures, pupillary size, and pupillary responses. MEASUREMENTS AND RESULTS: Twenty patients completed the double-blind study, and 26 patients completed the open study. There were no changes in intraocular pressures, pupillary size, or pupillary response after any treatment on any study day in either the double-blind or the open studies. CONCLUSION: In children with asthma, who have no pre-existing ocular abnormalities, the risk of an adverse reaction to nebulized ipratropium bromide delivered by face mask inadvertently absorbed in the eye is extremely small.

Cetirizine pharmacokinetics and pharmacodynamics in primary biliary cirrhosis.

The new H1-receptor antagonist, cetirizine, is eliminated primarily unchanged by renal excretion and is thus potentially useful for relief of pruritus in patients with hepatic dysfunction, in whom many H1-receptor antagonists are contraindicated. The authors studied the elimination of cetirizine in six patients with primary biliary cirrhosis. In contrast to data obtained in healthy adults with normal hepatic function reported in the medical literature, they found that the mean serum elimination half-life value of cetirizine, 13.8 +/- 1.8 hours, was longer, and the mean clearance rate, 0.44 +/- 0.10 mL/min/kg, was lower (P < .05). The mean peak serum cetirizine concentration, 498 +/- 118 ng/mL, was higher, the mean area under the curve, 6438 +/- 1621 ng/mL/hr, was larger, and the mean fraction of the dose excreted as unchanged cetirizine in the urine, .32 +/- .14, was lower (P < .05). The duration of action of cetirizine was prolonged, as evidenced by significant suppression of the histamine-induced wheal and flare for 48 and 72 hours, respectively, after a single dose. Cetirizine elimination was impaired in patients with hepatic dysfunction.

Pharmacokinetics and pharmacodynamics of ebastine in children.

Ebastine is a new piperidine-containing, relatively nonsedating second-generation H1-receptor antagonist. In a double-blind, parallel-group study of a single 5 mg or 10 mg dose of ebastine syrup used to treat allergic rhinitis in 20 children aged 6 to 12 years, we tested the hypothesis that the medication would have a duration of action of at least 24 hours. We measured plasma concentrations of carebastine, the pharmacologically active metabolite of ebastine, and the wheals and flares produced by epicutaneous tests with histamine phosphate, 1.0 mg/ml. Ebastine was absorbed well; peak carebastine concentrations occurred approximately 3 hours after dosing. Mean plasma elimination half-life values of carebastine ranged from 10 to 14 hours. The pharmacokinetics of carebastine were linear and dose independent in the dosage range studied. After the 5 or 10 mg dose, there were no significant differences between mean plasma elimination half-life values, mean oral clearance values, or mean apparent volumes of distribution. Mean peak plasma carebastine concentrations and mean areas under the plasma carebastine concentration-time curve after the 10 mg dose were 1.93 and 1.76 times, respectively, the values obtained after the 5 mg dose. Both doses significantly reduced the histamine-induced wheal-and-flare areas for up to 28 hours compared with predose values. The differences in effect between the doses generally were not statistically or clinically significant. No adverse effects were noted. We conclude that ebastine, an effective H1-receptor antagonist with a prompt onset of action and a long duration of action, is suitable for once-daily administration to children.

Treatment of allergic rhinitis with intranasal corticosteroids in patients with mild asthma: effect on lower airway responsiveness.

The effect of treatment of allergic rhinitis with intranasal corticosteroids on lower airway responsiveness was assessed in a randomized, double-blind, placebo-controlled, crossover study. Twenty-one young patients with perennial allergic rhinitis and asthma, with documented lower airway hyperresponsiveness (PC20 methacholine < 8 mg/ml), were treated with intranasal aqueous beclomethasone dipropionate and placebo, each given for 4 weeks. Patients recorded rhinitis and asthma symptom scores and monitored peak expiratory flow rates every morning and evening. Patients recorded global assessment of rhinitis and global asthma symptom scores at the beginning and end of each treatment. PC20 methacholine was performed at baseline and at the end of each treatment period. Intranasal beclomethasone dipropionate significantly reduced global rhinitis symptom scores (p = 0.05) after 4 weeks of treatment. Global asthma scores did not change significantly (p = 0.2). Geometric mean PC20 methacholine improved significantly after 4 weeks of intranasal beclomethasone, but not after placebo (p = 0.04). Daily morning and evening rhinitis symptom scores were lower in patients treated with intranasal corticosteroids over the first 4 weeks of treatment, but carryover effect of steroids precluded comparative analysis of the second 4-week block (morning p = 0.06, evening p = 0.03). Morning asthma scores tended to decrease (p = 0.07). Evening asthma scores were significantly decreased at weeks 2 and 3 (p = 0.001, p = 0.02, respectively). No change in peak expiratory flow rate was seen. This study confirms that treatment of inflammation in the upper airways indirectly improves asthma symptoms and decreases bronchial hyperreactivity. Ignoring inflammation in the upper airway may lead to suboptimal results in asthma treatment.

Bronchodilator and bronchoprotective effects of salmeterol in young patients with asthma.

BACKGROUND: In adults with asthma, the selective beta 2-adrenergic agonist salmeterol has a prolonged bronchodilator and bronchoprotective effect. To date, there are few published studies of salmeterol in children. METHODS: We compared the bronchodilator and bronchoprotective effects of salmeterol, 25 and 50 micrograms, with salbutamol, 200 micrograms, and with placebo, administered via metered-dose inhaler, in a randomized, double-blind, within-patient, four-way crossover, single-dose study in 20 children. RESULTS: Mean baseline forced expiratory volume in 1 second (FEV1) and PC20 methacholine were not significantly different (p > 0.05) on the 4 study days, and did not change significantly after placebo. FEV1 increased significantly from 5 to 30 minutes after salbutamol, and from 5 minutes to 12 hours after 25 micrograms or 50 micrograms salmeterol, compared with placebo. After 25 micrograms or 50 micrograms salmeterol, FEV1 was significantly lower than after salbutamol at 5 and 10 minutes, did not differ from salbutamol at 30 minutes, and was significantly greater than after salbutamol from 3 to 12 hours. No significant difference occurred between the effect of 25 micrograms salmeterol and the effect of 50 micrograms salmeterol on FEV1. After salbutamol, there was a significant increase in PC20 only at 30 minutes. After 25 micrograms or 50 micrograms salmeterol, PC20 increased significantly from 30 minutes to 12 hours. Salmeterol, 25 micrograms and 50 micrograms provided significantly greater bronchoprotection than salbutamol from 3 to 12 hours and from 30 minutes to 12 hours, respectively. Salmeterol, 50 micrograms, provided significantly better bronchoprotection than 25 micrograms salmeterol from 30 minutes to 12 hours. The amount of change in PC20 accounted for by change in FEV1 varied from 14% to 28%, indicating that protection against bronchoconstriction was not entirely dependent on bronchodilation. CONCLUSIONS: Salmeterol is a potent, long-acting bronchodilator, with a slower onset of bronchodilation than salbutamol. It provides significantly greater and longer-lasting protection against bronchoconstriction than salbutamol.

Continuous complete collection of uncontaminated urine in conscious rodents.

We describe a novel technique for complete and continuous bladder urine collection in conscious unrestrained rats. After urethral ligation, a silastic catheter is placed through a flexible steel tether, tunneled subcutaneously from the posterior neck area to a suprapubic incision, and inserted and fixed into the exposed urinary bladder. The catheter drains by gravity into a dependent collecting vessel protected from contamination by feces or food.