Salt chamber treatment is ineffective in treating eosinophilic inflammation in asthma.

BACKGROUND: We have shown that salt chamber treatment reduces airway hyper-responsiveness as an add-on therapy in adult asthmatics on inhaled corticosteroids. METHODS: We assessed whether this effect is due to the suppression of eosinophilic airway inflammation. Thirty-nine adult asthmatics on inhaled corticosteroids were randomized to receive active salt chamber treatment with low-salt treatment 6.6 mg/m(3) (n = 14), high-salt treatment 10.8 mg/m(3) (n = 15) or placebo 0.3 mg/m(3) (n = 10) 10 times in a 2 weeks' period in a double-blind manner. RESULTS: The level of induced sputum eosinophilic cationic protein µg/l, was 3070 before and 4651 after the low-salt treatment period, on average. In the high-salt treatment group, it was 12 192 µg/l vs 11 803 and in the placebo group 3942 vs 4144, respectively. Salt chamber treatment had no effect on sputum eosinophil or neutrophil cell numbers. CONCLUSIONS: The reduction in hyper-responsiveness observed in the previous study is probably not due to the effect on eosinophilic inflammation.

Development and inter-laboratory validation of the VISAGE enhanced tool for age estimation from semen using quantitative DNA methylation analysis.

The analysis of DNA methylation has become an established method for chronological age estimation. This has triggered interest in the forensic community to develop new methods for age estimation from biological crime scene material. Various assays are available for age estimation from somatic tissues, the majority from blood. Age prediction from semen requires different DNA methylation markers and the only assays currently developed for forensic analysis are based on SNaPshot or pyrosequencing. Here, we describe a new assay using massively parallel sequencing to analyse 13 candidate CpG sites targeted in two multiplex PCRs. The assay has been validated by five consortium laboratories of the VISible Attributes through GEnomics (VISAGE) project within a collaborative exercise and was tested for reproducible quantification of DNA methylation levels and sensitivity with DNA methylation controls. Furthermore, DNA extracts and stains on Whatman FTA cards from two semen samples were used to evaluate concordance and mimic casework samples. Overall, the assay yielded high read depths (> 1000 reads) at all 13 marker positions. The methylation values obtained indicated robust quantification with an average standard deviation of 2.8% at the expected methylation level of 50% across the 13 markers and a good performance with 50 ng DNA input into bisulfite conversion. The absolute difference of quantifications from one participating laboratory to the mean quantifications of concordance and semen stains of remaining laboratories was approximately 1%. These results demonstrated the assay to be robust and suitable for age estimation from semen in forensic investigations. In addition to the 13-marker assay, a more streamlined protocol combining only five age markers in one multiplex PCR was developed. Preliminary results showed no substantial differences in DNA methylation quantification between the two assays, indicating its applicability with the VISAGE age model for semen developed with data from the complete 13-marker tool.

Prevalence of asthma, aspirin intolerance, nasal polyposis and chronic obstructive pulmonary disease in a population-based study.

BACKGROUND: Remarkable overlap exists in symptoms between asthma and chronic obstructive pulmonary disease (COPD), and the symptoms of the patients with mild asthma are often falsely thought to be caused by smoking. The objective of the study was to determine the prevalence of doctor-diagnosed asthma, asthmatic symptoms and doctor-diagnosed COPD in an adult population. The prevalence and relation to asthma of aspirin intolerance, nasal polyposis, allergic rhinitis and smoking habits were also examined. METHODS: Postal questionnaire survey of a population-based random sample (4300) of adult women and men aged 18-65 years served by the Päijät-Häme Central Hospital in southern Finland (a region with 208 000 inhabitants) was performed. RESULTS: The non-response-adjusted prevalence (Drane's linear method) of doctor-diagnosed asthma was 4.4% (95% CI: 3.3-5.5%) and of COPD 3.7% (95% CI: 2.7-4.8%). The prevalence of allergic rhinitis was 37.3% (95% CI: 33.3-41.2%), and of overall aspirin intolerance 5.7% (95% CI: 4.4-7.1%). The observed prevalence of aspirin intolerance causing shortness of breath or attacks of asthma was 1.2% and it was higher in patients with doctor-diagnosed asthma than without (8.8% versus 0.8%, relative risk [RR] = 11.4, P < 0.0001), and higher in those with allergic-like rhinitis than without (2.6% versus 0.3%, RR = 7.7, P < 0.0001). The prevalence of nasal polyposis was 4.3% (95% CI : 2.8-5.8%). CONCLUSIONS: The current prevalence of doctor-diagnosed asthma among adults is 4.4%, and allergic rhinitis, nasal polyposis and aspirin intolerance are associated with an increased risk of asthma. There is also association between aspirin-induced asthma and allergic-like rhinitis.

A rapid dosimetric methacholine challenge in asthma diagnostics: a clinical study of 230 patients with dyspnoea, wheezing or a cough of unknown cause.

The rapid methacholine challenge test using a pocket turbine spirometer (Micro Spirometer) and the Spira Elektro 2 dosimeter was performed with 230 consecutive patients who had dyspnoea, wheezing or a prolonged cough of unknown cause. Patients with previous asthma diagnoses as well as those who had used inhaled steroids during the preceding 4 weeks were excluded. Seventy-eight patients (34%) were methacholine positive (PD20FEV1 < or = 6900 micrograms) 47 (60%) of whom had a final diagnosis of American Thoracic Society (ATS) criteria fulfilling bronchial asthma. One hundred and fifty-two patients (66%) were methacholine negative (PD20FEV1 > 6900 micrograms) 14 (9%) of whom had bronchial asthma according to clinical evaluation. Increased bronchial responsiveness was strongly associated with ATS criteria fulfilling asthma (P < 0.0001). When PD20FEV1 was used, 47 (77%) of the asthmatic patients were hyper-responsive (range 40-6900 micrograms) compared to 31 (18%) of the non-asthmatic patients (range 160-6900 micrograms). When using PD15FEV1, 51 (84%) of the asthmatic patients (range 28-6900 micrograms) and 52 (31%) of the non-asthmatic patients (range 100-6900 micrograms) were hyper-responsive. The level of bronchial responsiveness measured by both PD20FEV1 and PD15FEV1 differed significantly between asthmatic and non-asthmatic patients (P < 0.0001). Hyper-responsiveness was associated with an increased daily variation in peak expiratory flow (PEF) (P < 0.0001) and an increased number of blood eosinophils (P < 0.0001). Hyper-responsiveness was also associated with decreased levels of FEV1 and percentages of predicted FEV1 (P = 0.04 and P < 0.0001, respectively). Stepwise logistic regression analysis showed that the number of positive prick results (OR = 1.15, 95% CI 1.01-1.31), blood eosinophils (1.004, 1.00-1.01), level of FEV1 (0.56, 0.36-0.87) and current smoking (2.36, 1.00-5.59) were factors significantly associated with the probability of hyper-responsiveness. Age, gender, atopy, pets and a history of ex-smoking were not significantly associated with hyper-responsiveness, neither in univariate nor in multivariate analyses. The Bayesian analysis was used to investigate the diagnostic value of the rapid methacholine challenge test. A receiver operator characteristic curve demonstrated that PD20FEV1 separated asthmatic and non-asthmatic patients better than PD15FEV1. The best cutoff value of PD20FEV1 was 6000 micrograms, but the difference from 6900 micrograms was minimal. The best results of the test using a PD20FEV1 cutoff point of 6900 micrograms (PPV: 0.80, NPV: 0.79) were obtained when the pre-test probability was 0.48. The interval security of the test was established by a pre-test probability between 0.19 and 0.78. Maximal positive (0.34) and negative (0.31) final gains were achieved when pre-test probabilities were 0.33 and 0.65, respectively. The cutoff level of 150 micrograms gave 100% of specificity and predictive value of a positive test for clinical asthma diagnosis. The Bayesian analysis approach demonstrated that the test is useful in asthma diagnostics if not performed on patients with lowest or highest probabilities of asthma.

Serum ECP and MPO, but not urinary LTE4, are associated with bronchial hyper-responsiveness.

A random population-based sample of 131 subjects was used to assess the value of serum eosinophil cationic protein (ECP), serum myeloperioxidase (MPO), and urinary leukotriene E4 (LTE4) in predicting bronchial hyper-responsiveness measured by methacholine challenge. Special interest was focused on the history of aspirin intolerance and on smoking as contributing factors. The mean serum ECP and MPO were higher in hyper-reactive [provocational dose causing a 20% fall in forced expiratory volume in 1 sec. (PD20) < or = 6900 micrograms] than in non-hyper-reactive subjects (22.3 vs. 13.2 micrograms l-1, P < 0.001 and 377 vs. 278 micrograms l-1, P = 0.001, respectively). This was also seen in current smokers vs. never smokers (17.2 vs. 12.9 micrograms l-1, P = 0.03 and 372 vs. 286 micrograms l-1, P = 0.04, respectively). There were no differences in baseline urinary excretion of LTE4 between hyper-reactive and non-hyper-reactive subjects. During the 2 h after methacholine challenge, urinary LTE4 excretion increased from 53.8 and 69.0 ng mmol-1 creatinine in non-hyper-reactive subjects, but there was no change in hyper-reactive subjects (non-hyper-reactive vs. hyper-reactive, P = 0.06). The increase was greatest in subjects with aspirin intolerance causing urticaria or angioedema but not aggravation of asthma (from 58.5 to 87.2 ng mmol-1 creatinine), probably due to extrapulmonary leukotriene production. Our results indicate that serum ECP and MPO, but not urinary LTE4 (even in subjects with a history of aspirin intolerance), predict bronchial hyper-responsiveness to methacholine. The subject's smoking history must be taken into account when these parameters are considered.

The bronchoprotective efficacy of salbutamol inhaled from a new metered-dose powder inhaler compared with a conventional pressurized metered-dose inhaler connected to a spacer.

The aim of this study was to compare the efficacy of 100 micrograms of salbutamol inhaled from a new metered-dose powder inhaler (MDPI, Leiras Taifun, Finland) with that of a same dose of salbutamol inhaled from a conventional pressurized metered-dose inhaler with a large volume spacer (pMDI + S) in protecting against methacholine (Mch) induced bronchoconstriction. This was a 3 day, randomized, cross-over, partly blinded, placebo-controlled multicentre study where the pMDI + S was used as an open control. Twenty-six asthmatic outpatients with a baseline FEV1 > or = 60% of predicted and with bronchial hyperreactivity (PD20 FEV1 < or = 890 micrograms of Mch) were studied. On each study day the patients underwent an Mch provocation 30 min after inhaling placebo from the MDPI or a dose of 100 micrograms of salbutamol from the MDPI and from the pMDI + S. PD20 FEV1 and dose-response slope [DRS; maximal change in FEV1 (%)/dose of Mch (mumol)] were used to evaluate efficacy. The median values of PD20 FEV1 were 250, 622 and 1737 micrograms after placebo MDPI, salbutamol pMDI + S and salbutamol MDPI, respectively. The corresponding DRS values were -11.0%, -4.5% and -2.0% mumol-1. With both parameters, all differences were statistically significant (P < 0.05). In conclusion, 100 micrograms of salbutamol inhaled from Leiras Taifun MDPI offers better protection against Mch-induced bronchoconstriction than 100 micrograms of salbutamol from a pMDI connected to a large volume spacer device.

The effect of salt chamber treatment on bronchial hyperresponsiveness in asthmatics.

BACKGROUND: Randomized controlled trials are needed to evaluate the effects of complementary treatments in asthma. This study assessed the effect of salt chamber treatment as an add-on therapy to low to moderate inhaled steroid therapy in asthma patients with bronchial hyperresponsiveness (BHR). METHODS: After a 2-week baseline period, 32 asthma patients who exhibited BHR in the histamine inhalation challenge were randomized: 17 to 2-week active treatment, during which salt was fed to the room by a salt generator, and 15 to placebo. The salt chamber treatment lasted 40 min and was administered five times a week. RESULTS: Median provocative dose causing a decrease of 15% in Fev(1) (PD(15)FEV(1)) [corrected] increased significantly in the active group (P = 0.047) but not in the placebo group. The difference in changes between the active and placebo groups was significant (P = 0.02). Nine patients (56%) in the active group and two patients (17%) in the placebo group exhibited at least one doubling dose decrease in BHR (P = 0.040). Six patients (38%) in the active group and none in the placebo group became non-hyperresponsive (P = 0.017). Neither the peak expiratory flow (PEF) values measured just before and after the treatment, nor FEV(1) values measured before the histamine challenges, changed. The reduction in BHR was not caused by changes in the baseline lung function. CONCLUSIONS: Salt chamber treatment reduced bronchial hyperresponsiveness as an add-on therapy in asthmatics with a low to moderate dose of inhaled steroids. The possibility that salt chamber treatment could serve as a complementary therapy to conventional medication cannot be excluded.

Fluoride concentration in saliva after consumption of a dinner meal prepared with fluoridated salt.

The aim was to determine the fluoride concentration in saliva after intake of a dinner meal prepared with fluoridated salt. The investigation had a randomized cross-over design, and 10 healthy adolescents with natural fluoride content (1.06 ppm) in their drinking water participated after informed consent. After a run-in week, the subjects were served a standardized dinner of spaghetti with minced meat sauce prepared with either fluoridated salt (test arm) or non-fluoridated salt (control arm). The fluoride concentration of the test salt was 250 ppm. Samples of stimulated whole saliva was collected at baseline, directly after eating (0 min) and then after 10, 30 and 180 min. After a 1-week wash-out period, the experimental procedure was repeated with the opposite salt. Fluoride concentration in saliva was measured with a fluoride-specific electrode and the post-ingestion levels were compared with baseline using repeated-measures ANOVA. The mean baseline concentrations were 10.9 and 8.0 microg/l in the test and control arms, respectively. Immediately after the intake, the mean fluoride values increased significantly to 81.6 microg/l in the test arm and to 31.5 microg/l in the control arm (p<0.05). The fluoride levels remained elevated (p<0.05) for 30 min after ingestion of the test meal but not following the control meal. In conclusion, consumption of a dinner meal prepared with fluoridated salt increased the salivary fluoride levels for about 30 min.

Accuracy and repeatability of a pocket turbine spirometer: comparison with a rolling seal flow-volume spirometer.

The accuracy and repeatability of a recently introduced pocket spirometer (Micro Spirometer; Micro Medical Instruments Ltd, Rochester, UK) was evaluated. FEV1 and FVC values obtained with this instrument were compared with those measured with a rolling-seal flow-volume spirometer (CPI 220 with microcomputer) in 31 patients and 11 healthy volunteers. In the whole material, expressed as mean +/- SD, the pocket spirometer recorded 0.44 +/- 0.23 l (13 +/- 7%) smaller values for FEV1 (P < 0.001) and 0.64 +/- 0.48 l (15 +/- 11%) smaller values for FVC (P < 0.001) than the rolling-seal spirometer. The short-term repeatability of the measurements expressed as the coefficient of variation of repeated measurements using the pocket spirometer was 2.2% for FEV1 and 2.3% for FVC in a series of 10 healthy subjects and 10 patients with COPD. It is concluded that the underestimation of FEV1 and FVC of the pocket spirometer was too large and inconsistent for the device to be used interchangeably with conventional spirometers. However, the repeatability of the measurements with the pocket spirometer is close to that reported previously for flow-volume spirometry. Thus the pocket spirometer may be suitable in assessing acute changes of spirometric indices e.g. during provocation tests or during patient follow-up in asthma.

Controlled trial of methotrexate in patients with severe chronic asthma.

The aim of this study was to investigate the efficacy and adverse effects of methotrexate (MTX) in the treatment of severe chronic asthma in 12 patients with severe asthma requiring continuous treatment with oral steroids at the Outpatient Department of Helsinki University Central Hospital. The study was a randomised, double-blind placebo-controlled trial of methotrexate treatment 15 mg weekly on a crossover basis over 24 weeks. During the 2 weeks baseline phase the mean dose of oral steroids administered was 10.9 (3.2-28) mg.day-1, and the mean dose of inhaled steroids administered was 2.3 (1.6-3.2) mg budesonide or beclomethasone. The average dose of oral steroids administered was 12.8 mg.day-1 during the last 2 placebo weeks but only 7.9 mg.day-1 during the last 2 weeks with MTX treatment. The reduction in daily dose of oral steroids was 38%, while daily bronchodilator use was reduced by 22%. During MTX treatment the patients experienced significantly less wheezing, dyspnoea and coughing. Nine out of 12 patients reported better asthma control during MTX treatment. The peak expiratory flow rate (PEF) 1-s forced expiratory volume (FEV1) values did not differ between MTX and placebo treatments. There was no statistical correlation between serum MTX concentration and clinical improvement. No serious adverse effects of MTX were found during the study. It was concluded that low-dose MTX may be beneficial for severe chronic asthma and that this therapy is well tolerated by patients.

Repeatability of a rapid dosimetric method for methacholine challenge using a pocket turbine spirometer for FEV1 measurements.

The repeatability of a rapid dosimetric method for methacholine challenge was evaluated with 11 asthmatic patients. A Spira Elektro 2 dosimeter was used for methacholine delivery and a pocket turbine spirometer (Micro Spirometer) for FEV1 measurements. Methacholine was delivered in four successive, increasing doses ranging from 80 micrograms up to a cumulative dose of 6900 micrograms. The single determination standard deviation was low (12.5%), corresponding to a 95% confidence interval of +/- 0.925 doubling doses. The mean difference (+/- SE) between measurements of log PD20 FEV1 was -0.015 (0.056), and the absolute value of the difference in log PD20 FEV1 was not significantly related to the average log PD20 FEV1 (r = -0.155, P = 0.65). The rapid dosimetric methacholine challenge test, performed with a pocket turbine spirometer, proved to be as reproducible as previous methods. Furthermore, this methacholine challenge is clearly less time consuming than conventional provocations, including bronchodilator aerosol (given to resolve post-challenge bronchoconstriction); the whole test can be performed in 20 min. This is especially valuable in epidemiologic studies, as well as in clinical practice.

Acute asthma during pregnancy.

BACKGROUND: Acute asthma during pregnancy is potentially dangerous to the fetus. The aim of this study was to investigate the effect of an acute attack of asthma during pregnancy on the course of pregnancy or delivery, or the health of the newborn infant, and to identify undertreatment as a possible cause of the exacerbations. METHODS: Five hundred and four pregnant asthmatic subjects were prospectively followed and treated. The data on 47 patients with an attack of asthma during pregnancy were compared with those of 457 asthmatics with no recorded acute exacerbation and with 237 healthy parturients. RESULTS: Of 504 asthmatics, 177 patients were not initially treated with inhaled corticosteroids. Of these, 17% had an acute attack compared with only 4% of the 257 patients who had been on inhaled anti-inflammatory treatment from the start of pregnancy. There were no differences between the groups as to length of gestation, length of the third stage of labour, or amount of haemorrhage after delivery. No differences were observed between pregnancies with and without an exacerbation with regard to relative birth weight, incidence of malformations, hypoglycaemia, or need for phototherapy for jaundice during the neonatal period. CONCLUSIONS: Patients with inadequate inhaled anti-inflammatory treatment during pregnancy run a higher risk of suffering an acute attack of asthma than those treated with an anti-inflammatory agent. However, if the acute attack of asthma is relatively mild and promptly treated, it does not have a serious effect on the pregnancy, delivery, or the health of the newborn infant.

Equivalence of two steroid-containing inhalers: easyhaler multidose powder inhaler compared with conventional aerosol with large-volume spacer.

BACKGROUND AND OBJECTIVES: An equivalence study was conducted in which the efficacy and safety of a daily dose of 800 microgram of beclomethasone diproprionate administered via a multidose powder inhaler, Easyhaler, and via a metered-dose inhaler (MDI) with a large-volume spacer were compared in adult, newly diagnosed, steroid-naive asthmatic patients. Acceptability of the medications was also compared. METHODS: One hundred and forty-four patients were recruited into the double-blind, double-dummy, randomised, parallel-group multicentre study. The study treatment period was 8 weeks. It was preceded by a 2-week run-in period. Morning and evening peak expiratory flow (PEF), numbers of inhalations of a sympathomimetic and asthma symptoms were recorded daily. Spirometry and histamine challenge were performed, and health-related quality of life and morning serum cortisol levels measured during control visits. RESULTS: Criteria indicating treatment equivalence were met. The mean of the primary outcome variable, morning PEF, increased significantly, from 426 to 461 litres/min in the Easyhaler group and from 436 to 467 litres/min in the MDI+spacer group. Similar improvements between groups were also seen in relation to all secondary variables. Changes in serum cortisol levels were minor. In 6 out of 10 questions about device acceptability, the majority of patients rated Easyhaler as better than the MDI+spacer combination. CONCLUSION: It was concluded that the devices tested were equivalent in terms of efficacy and safety.

Improved asthma control with budesonide/formoterol in a single inhaler, compared with budesonide alone.

Budesonide/formoterol in a single inhaler was compared with budesonide alone, and with concurrent administration of budesonide and formoterol from separate inhalers, in patients with asthma, not controlled with inhaled glucocorticosteroids alone. In this 12-week, double-blind, randomized, double-dummy study, 362 adult asthmatics (forced expiratory volume in one second 73.8% of predicted, inhaled glucocorticosteroid dose 960 microg x day(-1)) received single inhaler budesonide/formoterol (Symbicort Turbuhaler) 160/4.5 microg, two inhalations b.i.d., or corresponding treatment with budesonide, or budesonide plus formoterol via separate inhalers. There was a greater increase in morning peak expiratory flow (PEF) with single-inhaler (35.7 L x min(-1)) and separate-inhaler (32.0 L x min(-1)) budesonide and formoterol, compared with budesonide alone (0.2 L x min(-1); p<0.001, both comparisons); the effect was apparent after 1 day (p<0.001 versus budesonide, both comparisons). Similarly, evening PEF, use of rescue medication, total asthma symptom scores and percentage of symptom-free days improved more with both single inhaler and separate inhaler therapy than with budesonide alone, as did asthma control days (approximately 15% more, p<0.001 versus budesonide, both comparisons, with a marked increase in the first week). All treatments were well tolerated and the adverse event profile was similar in all three treatment groups. It is concluded that single inhaler therapy with budesonide and formoterol is a clinically effective and well-tolerated treatment for patients with asthma that is not fully controlled by inhaled glucocorticosteroids alone.