Early supplementation with Lactobacillus rhamnosus HN001 reduces eczema prevalence to 6 years: does it also reduce atopic sensitization?

BACKGROUND: The role of probiotics in prevention of allergic disease is still not clear; efficacy may depend on the timing, dose, duration, and specific probiotic used. Using a double-blind randomized placebo-controlled trial (Australian New Zealand Clinical Trials Registry: ACTRN12607000518460), we have shown that in a high-risk birth cohort, maternal supplementation from 35 weeks gestation until 6 months if breastfeeding and infant supplementation from birth until 2 years with Lactobacillus rhamnosus HN001 (HN001) (6 × 10(9) cfu/day) halved the cumulative prevalence of eczema at 2 and 4 years. Bifidobacterium animalis subsp lactis HN019 (HN019) (9 × 10(9) cfu/day) had no significant effect. OBJECTIVE: To determine whether differences in effects of HN001 and HN019 on eczema persist to age 6 years, and to investigate effects on sensitization. METHODS: Standard procedures were used to assess eczema (The UK Working Party's Criteria), eczema severity (SCORAD), atopic sensitization [skin prick tests (SPT), total and specific IgE] and standard questions used for asthma, wheeze, and rhinoconjunctivitis. RESULTS: HN001 was associated with significantly lower cumulative prevalence of eczema (HR = 0.56, 95% CI 0.39-0.80), SCORAD ≥ 10 (HR = 0.69, 0.49-0.98) and SPT sensitization (HR = 0.69, 95% CI 0.48-0.99). The point prevalence of eczema (RR = 0.66, 95% CI 0.44-1.00), SCORAD ≥ 10 (RR = 0.62, 95% CI 0.38-1.01) and SPT sensitization (RR = 0.72, 95% CI 0.53-1.00) were also reduced among children taking HN001. HN019 had no significant effect on any outcome. CONCLUSION AND CLINICAL RELEVANCE: This study provides evidence for the efficacy of the probiotic L. rhamnosus HN001 in preventing the development of eczema and possibly also atopic sensitization in high risk infants to age 6 years. The absence of a similar effect for HN019 indicates that benefits may be species specific.

Dampness and moulds in relation to respiratory and allergic symptoms in children: results from Phase Two of the International Study of Asthma and Allergies in Childhood (ISAAC Phase Two).

BACKGROUND: Many studies report that damp housing conditions are associated with respiratory symptoms. Less is known about mechanisms and possible effect modifiers. Studies of dampness in relation to allergic sensitization and eczema are scarce. OBJECTIVE: We study the influence of damp housing conditions world-wide on symptoms and objective outcomes. METHODS: Cross-sectional studies of 8-12-year-old children in 20 countries used standardized methodology from Phase Two of the International Study of Asthma and Allergies in Childhood (ISAAC). Symptoms of asthma, rhinitis and eczema, plus residential exposure to dampness and moulds, were ascertained by parental questionnaires (n = 46 051). Skin examination, skin prick tests (n = 26 967) and hypertonic saline bronchial challenge (n = 5713) were performed. In subsamples stratified by wheeze (n = 1175), dust was sampled and analysed for house dust mite (HDM) allergens and endotoxin. RESULTS: Current exposure to dampness was more common for wheezy children (pooled odds ratio 1.58, 95% CI 1.40-1.79) and was associated with greater symptom severity among wheezers, irrespective of atopy. A significant (P < 0.01) adverse effect of dampness was also seen for cough and phlegm, rhinitis and reported eczema, but not for examined eczema, nor bronchial hyperresponsiveness. HDM sensitization was more common in damp homes (OR 1.16, 1.03-1.32). HDM-allergen levels were higher in damp homes and were positively associated with HDM-sensitization, but not wheeze. CONCLUSION: A consistent association of dampness with respiratory and other symptoms was found in both affluent and non-affluent countries, among both atopic and non-atopic children. HDM exposure and sensitization may contribute, but the link seems to be related principally to non-atopic mechanisms.

Supplementation with Lactobacillus rhamnosus or Bifidobacterium lactis probiotics in pregnancy increases cord blood interferon-gamma and breast milk transforming growth factor-beta and immunoglobin A detection.

BACKGROUND: This study explored the effects of maternal probiotic supplementation on immune markers in cord blood (CB) and breast milk. METHODS: CB plasma and breast milk samples were collected from a cohort of women who had received daily supplements of either 6 x 10(9) CFU/day Lactobacillus rhamnosus HN001 (n=34), 9 x 10(9) CFU/day Bifidobacterium lactis HN019 (n=35) or a placebo (n=36) beginning 2-5 weeks before delivery and continuing for 6 months in lactating women. CB plasma and breast milk (collected at 3-7 days, 3 months and 6 months postpartum) were assayed for cytokines (IL-13, IFN-gamma, IL-6, TNF-alpha, IL-10, TGF-beta1) and sCD14. Breast milk samples were also assayed for total IgA. RESULTS: Neonates of mothers who received a probiotic had higher CB IFN-gamma levels (P=0.026), and a higher proportion had detectable blood IFN-gamma levels, compared with the placebo group (P=0.034), although levels were undetectable in many infants. While this pattern was evident for both probiotics, when examined separately only the L. rhamnosus HN001 group showed statistically significant higher IFN-gamma levels (P=0.030) compared with the placebo group. TGF-beta1 levels were higher in early breast milk (week 1) from the probiotic groups (P=0.028). This was evident for the B. lactis HN019 group (P=0.041) with a parallel trend in the L. rhamnosus HN001 group (P=0.075). Similar patterns were seen for breast milk IgA, which was more readily detected in breast milk from both the B. lactis HN019 (P=0.008) and the L. rhamnosus HN001 group (P=0.011). Neonatal plasma sCD14 levels were lower in the B. lactis HN019 group compared with the placebo group (P=0.041). CONCLUSION: The findings suggest that supplementation with probiotics in pregnancy has the potential to influence fetal immune parameters as well as immunomodulatory factors in breast milk.

Asthma and allergic symptoms in relation to house dust endotoxin: Phase Two of the International Study on Asthma and Allergies in Childhood (ISAAC II).

BACKGROUND: Several studies have consistently reported inverse associations between exposure to endotoxin in house dust and atopy. With regard to the association between house dust endotoxin and asthma, the results are inconsistent. OBJECTIVES: To study the association between house dust endotoxin levels and respiratory symptoms and atopy in populations from largely different countries. METHODS: Data were collected within the International Study on Asthma and Allergies in Childhood Phase Two, a multi-centre cross-sectional study of 840 children aged 9-12 years from six centres in the five countries of Albania, Italy, New Zealand, Sweden and the United Kingdom. Living room floor dust was collected and analysed for endotoxin. Health end-points and demographics were assessed by standardized questionnaires. Atopy was assessed by measurements of allergen-specific IgE against a panel of inhalant allergens. Associations between house dust endotoxin and health outcomes were analysed by logistic regression. Odds ratios (ORs) were presented for an overall interquartile range increase in exposure. RESULTS: Many associations between house dust endotoxin in living room floor dust and health outcomes varied between countries. Combined across countries, endotoxin levels were inversely associated with asthma ever [adjusted OR (95% confidence interval (CI)) 0.53 (0.29-0.96) for endotoxin levels per m(2) of living room floor] and current wheeze [adjusted OR (95% CI) 0.77 (0.64-0.93) for endotoxin levels per gram of living room floor dust]. There were inverse associations between endotoxin concentrations and atopy, which were statistically significant in unadjusted analyses, but not after adjustment for gender, parental allergies, cat and house dust mite allergens. No associations were found with dust quantity and between endotoxin exposure and hayfever. CONCLUSION: These findings suggest an inverse association between endotoxin levels in living room floor dust and asthma in children.

The pulmonary and extrapulmonary effects of inhaled beta-agonists in patients with asthma.

The cardiovascular, respiratory, and hypokalemic effects of repeated inhalation of fenoterol, albuterol, and isoproterenol were compared in 12 subjects with stable asthma according to a double-blind, crossover design. Ipratropium bromide served as a control providing bronchodilatation without extrapulmonary effects. Subjects inhaled the beta-agonists on an equal-weight basis (400 micrograms) at 0, 30, 40, and 45 minutes. Measurements of heart rate, blood pressure, total electromechanical systole (measure of inotropic activity), preejection period, QTc interval, plasma potassium levels, and forced expiratory volume in 1 second were made 5 minutes after each dose and again at 60 and 75 minutes. There were no differences in the bronchodilating effect between the beta-agonists. However, both fenoterol and isoproterenol resulted in greater positive inotropic stimulation than did albuterol, and fenoterol caused a greater fall in plasma potassium levels than did the other beta-agonists.

Partial vs full beta-receptor agonism. A clinical study of inhaled albuterol and fenoterol.

STUDY OBJECTIVE: To compare the maximal extrapulmonary effects of the beta-agonists albuterol and fenoterol in eight healthy volunteers. SUBJECTS AND METHODS: In this double-blind study, we have examined the maximum cardiac effects (electromechanical systole [QS2I]--a measure of inotropy, heart rate, BP) and metabolic effects (plasma K+ and cyclic adenosine monophosphate [cAMP]) of repeated inhalation of albuternol and fenoterol. In eight healthy volunteers, 400 microg of each drug was administered every 10 min until QS2I and plasma K+ had reached a plateau (+/- 0.1 mmo l/L for K+, and +/- 10 ms for QS2I). The maximum response (Emax) and the dose of albuterol required to produce 50% of the maximum response to fenoterol (ED50F) were calculated. RESULTS: The Emax for fenoterol was significantly greater than albuterol for plasma K+ (-1.4 vs -1.03 mmol/L; p<0.002), QS2I (-71.8 vs 57.5 ms; p=0.047), and cAMP (33.8 vs 18.1 nmol/L; p<0.002). The dose required to produce the ED50f was significantly greater for albuterol than for fenoterol with potency ratios of 1.75, 1.61, and 2.26 for plasma K+, QS2I, and cAMP, respectively. There were no significant differences between fenoterol and albuterol with respect to heart rate (Emax, 44.9 vs 32.5 beats/min; p=0.19; potency ratio, 1.98; p=0.052). CONCLUSIONS: These findings suggest that albuterol behaves as a partial agonist at beta-receptors when compared with fenoterol, and that when inhaled in doses currently recommended for severe asthma, albuterol will result in lesser maximum cardiac and metabolic effects than fenoterol. These findings are consistent with the hypothesis that the property of full receptor agonism may contribute to the increased risk of death associated with fenoterol.

Whole blood storage solution for erythrocyte sodium and sodium-lithium countertransport rate determination.

A solution consisting of heparinized and buffered isosmolar magnesium chloride is described in which whole blood may be stored for preservation of erythrocyte sodium (Na) and membrane sodium-lithium countertransport (Na-Li CT). Correlation (r) of fresh versus 24-h stored erythrocytes for Na was 0.990 (y = 0.30 + 0.956x) and for Na-Li CT was 0.995 (y = -0.014 + 1.022x). Na-Li CT rate was preserved in the storage solution for up to five days and erythrocyte Na concentration for at least 24 h. The solution should find application in epidemiological studies of erythrocyte Na and Na-Li CT rate in human essential hypertension as the blood specimens require no centrifugation or erythrocyte washing procedures prior to laboratory analysis.

Assay of sodium-lithium countertransport rate by direct measurement of erythrocyte lithium concentrations.

A method is described for the in-vitro measurement of erythrocytic sodium-lithium countertransport rate (SLC) by direct assay of intra-erythrocytic lithium (Li) concentrations in Li loaded cells during controlled Li efflux. Bicarbonate stimulated erythrocytic Li loading to an approximate intracellular concentration of 3 mmol/l can be achieved within 4 min. Maximum SLC rates are achieved at intra-erythrocytic concentrations greater than 2.5 mmol/l. Lithium efflux is linear over 0-60 min, and independent of pH change. The within-batch coefficient of variation for positive displacement pipetting of Li loaded erythrocytes is 2.8%. Storage of erythrocytes in isosmotic MgCl2 for 24 h had no effect on the SLC. Intra-individual coefficient of variation for SLC was less than 6%. The reference range for Caucasian subjects (n = 43) was 0.14-0.49 mmol Li +/l erythrocytes/h.

Diurnal variation of erythrocyte sodium-lithium countertransport rate and intracellular cation concentrations.

Differences in erythrocyte sodium-lithium countertransport rate and erythrocyte potassium concentration were found between 0900 h and 2100 h in ten healthy individuals. Sodium-lithium countertransport rates were lowest at 0900 h (x: 0.34 mmol/l per h; SD: 0.15) and highest at 1200 h (x: 0.42 mmol/l per h; SD: 0.16). Erythrocyte potassium concentrations increased progressively during the day, while erythrocyte sodium concentrations did not change significantly during the day. For comparative and epidemiological studies of erythrocyte sodium-lithium countertransport rate blood samples should be taken at the same time of the day.

Unsuitability of the human erythrocyte as a model for in vivo sodium transport activation by beta 2 adrenergic agonists.

Administration of the beta 2 adrenergic agonists fenoterol, salbutamol and terbutaline to volunteers significantly reduced plasma potassium concentration in a double-blind crossover study. There was no consistent effect of the three active compounds on erythrocyte sodium concentration. Storage of whole blood at 4 degrees C increased erythrocyte sodium concentration by 33% after 2 h; this could explain the differences between this and another study of the effects of beta 2 adrenergic agonists on erythrocyte sodium concentration. We conclude that the human erythrocyte is unsuitable as an indirect in vivo model to demonstrate stimulation of cellular sodium potassium transport mediated by beta 2 adrenergic agonists.

Nebulized beta 2-adrenoceptor agonists do not affect plasma selenium or glutathione peroxidase activity in patients with asthma.

We investigated whether the beta 2-agonists fenoterol and salbutamol decreased plasma selenium and glutathione peroxidase activity in patients with asthma as this may partially explain the findings of reduced selenium status in asthmatic patients. Nine patients with asthma were studied on 3 occasions and inhaled either fenoterol (5 mg), salbutamol (5 mg) or ipratropium bromide (0.5 mg) administered by nebulization using a randomized, double blind, crossover design. Plasma selenium, glutathione peroxidase activity and potassium were measured prior to drug administration and at 15, 30 and 60 minutes after drug. None of the drugs had any effect on plasma selenium or glutathione peroxidase activity. Ipratropium bromide did not affect plasma potassium. Both beta 2-agonists significantly decreased plasma potassium. The mean (SD) maximum decrease was -0.79 (0-18) mmol/l for fenoterol and -0.26 (0-03) mmol/l for salbutamol (both p < or = 0.01) confirming systemic absorption of the drugs. beta 2-agonists are unlikely to be responsible for the reduced selenium status found in some patients with asthma.

A comparison of flurbiprofen with naproxen in ankylosing spondylitis.

In a four week double-blind crossover study, flurbiprofen 200mg daily was compared with naproxen 750mg daily in the management of 30 patients with ankylosing spondylitis. Both treatments were found to be very effective in alleviating pain and stiffness. No significant difference in efficacy was discernible between the two drugs. Side-effects were more frequent with flurbiprofen. A small, but significant, increase in renal excretion of beta-n-acetyl glucosaminidase occurred during treatment with both naproxen and flurbiprofen. Although previous surveys have not shown evidence of renal damage, further surveillance of renal function in patients receiving long term treatment with these preparations to exclude possible renal impairment would be prudent.

Attitudes and concerns amongst laboratory staff regarding handling of HIV positive biological specimens.

AIMS: to ascertain attitudes and concerns of laboratory staff towards handling of HIV positive biological samples. METHOD: an anonymous questionnaire regarding various aspects of biological sample handling was distributed to laboratory staff from the Wellington Area Health Board laboratories. RESULTS: of a total of 133 laboratory staff (response rate: 56.4%), five were seriously considering leaving, and another ten would have chosen another career if given prior information in regard to handling HIV positive biological samples. Of the responders, 22.8% and 38.3% considered that their employer does not provide adequate safety measures and education respectively in regard to HIV. CONCLUSIONS: handling of HIV positive biological samples does not greatly contribute to attrition amongst laboratory staff. Given the concern of laboratory staff for safety and education in regard to HIV, a continuous in-service educational programme would be desirable.

The accuracy of references in Australian and New Zealand Medical Journals.

AIMS: Previous studies have found high error rates in references in biomedical journals. The aim of this paper was to assess the accuracy of references in three Australian and New Zealand general medical journals. METHODS: References from the August 1999 issues of the Medical Journal of Australia, the New Zealand Medical Journal and the Australian and New Zealand Journal of Medicine were assessed for accuracy using PubMed of the National Library of Medicine. RESULTS: This study found a high rate of reference errors in Australian and New Zealand medical journals. The reference error rate ranged from 33.5 to 48.8%. The most frequent errors were in the author's names and in the title. CONCLUSIONS: The reference error rate in Australia and New Zealand medical journals is high and is preventable. Authors should be more diligent and preferably verify cited references against the original article.

The effect of frequent vacuum cleaning on the house dust mite allergen, Der p 1 in carpets: a pilot study.

AIM: To determine the effect of frequent vacuum cleaning of carpets on Der p 1, the major group one allergen of the house dust mite Dermatophagoides pteronyssinus. METHODS: Nine rooms and three hallways in the resident medical officers quarters at Wellington Hospital were regularly vacuum cleaned (daily, except weekends) over a five week period. Dust samples were collected before and at weekly intervals for Der p 1 measurement by a double monoclonal antibody ELISA technique. RESULTS: Der p 1 concentrations progressively declined from an initial geometric mean level (95% CI) of 4.47 micrograms/g fine dust (0.11-21.73) to 1.83 micrograms/g (0.29-9.57) after five weeks, a mean reduction of 48.0% (31.7-65.5). Similarly when expressed per unit area Der p 1 levels declined from 6.35 micrograms/m2 (1.15-35.16) to 1.66 micrograms/m2 (0.33-9.04), a mean reduction of 68.5% (58.6-78.3). CONCLUSIONS: Frequent vacuum cleaning over a short time significantly reduces house dust mite allergen levels in carpets. Larger long term trials are warranted to determine if greater reductions are possible that would be beneficial to house dust mite sensitive individuals.

House dust mite allergen accumulation on sheepskins.

AIM: Sheepskins, which are often used as infant bedding in New Zealand, are known to harbour large quantities of house dust mites and their allergens. In this study we determined the rate of accumulation of the house dust mite allergen, Der p 1 on new sheepskins, and the effects of washing and dry cleaning on its removal. METHODS: New sheepskins were placed on living-room floors (n = 6) and mattresses (n = 6) in six domestic dwellings in Wellington. Sheepskin dust samples were collected by vacuuming before placement, and at two, four and six weeks, after which they were warm-washed. They were replaced for a further six weeks, then dry-cleaned. Dust samples were collected before and after washing and dry-cleaning, and from mattresses and living-room floors. Dust samples were analysed for Der p 1 by double monoclonal antibody ELISA. RESULTS: Der p 1 levels rapidly increased in sheepskins placed on living-room floors and mattresses to a geometric mean level (range) of 9.0 micrograms/g (1.1-102.2) and 29.4 micrograms/g (5.3-131.1) at six weeks, respectively. After warm-water washing and replacement for a further six weeks, these levels were higher at 75.5 micrograms/g (50.1-260.4) and 31.9 micrograms/g (11.1-75.2), respectively. Sheepskin Der p 1 accumulation correlated with mattress (r = 0.78), and living-room floor Der p 1 levels (r = 0.94). Warm water washing and dry-cleaning reduced sheepskins Der p 1 levels by a mean of 79.2% and 95.3%, respectively. CONCLUSIONS: Sheepskins rapidly accumulate house dust mite allergens from the domestic environment. Due to very high levels and rapid accumulation of Der p 1, sheepskins as infant bedding should be discouraged for infants at risk of sensitisation to house dust mites. If they are to be used then it is important that they are washed or dry-cleaned regularly.

Racial differences in platelet counts in New Zealand men.

Healthy Maori males were found to have higher platelet counts (mean 295 x 10(9)/L: SD 46) compared to healthy nonMaori males (mean 253 x 10(9)/L; SD 50). Maori males had higher weights (mean 88.4 kg; SD 12.5) than nonMaori males (mean 81.0 kg; SD 9.0); and the platelet count was significantly correlated with weight (r = 0.30; p = 0.003). After weight adjustment significant differences between Maori and nonMaori males for platelet counts remained. These findings have implications for laboratory reference ranges, and may be of epidemiological significance in cardiovascular disease.

A pilot study to investigate the pulmonary effects of digoxin in patients with asthma.

AIM: A number of studies have shown that increased salt intake is associated with worsening asthma. The aim of this study was to investigate the respiratory effects of digoxin (a potent inhibitor of Na+K+ATPase) in patients with asthma. METHODS: Eight asthmatic patients were given digoxin (0.5 mg/daily) or matching placebo for 8 days. Treatments were assigned using a randomised double blind, crossover design. Bronchial hyperresponsiveness to methacholine, forced expiratory volume is one second (FEV1, serum potassium (K), urinary sodium and K, heart rate, blood pressure and the QTc interval of the ECG were measured on each treatment. RESULTS: When compared to placebo, digoxin significantly decreased FEV1 (p<0.03); the QTc interval (p<0.05), and increased serum K (p<0.02). There was a tendency for digoxin to increase bronchial hyperresponsiveness. CONCLUSION: In this small study digoxin resulted in a decline in spirometry. Further studies in a larger group of patients should be performed to assess this potentially adverse effect of digoxin.

Cat allergen levels in public places in New Zealand.

AIMS: Cat allergen (Fel d 1) is a known risk factor for asthma. Studies have demonstrated Fel d 1 in both public buildings and domestic dwellings where cats have never been. The aims of this study were to measure reservoir Fel d 1 levels in public buildings in New Zealand, to examine determinants of these levels and to compare them with previously measured domestic levels. METHODS: Dust was obtained in two centres (Wellington and Christchurch) from hotels, hospitals, rest homes, churches, primary schools, childcare centres, cinemas, bank head offices and aeroplanes; and from North Island ski lodges. Measurements of temperature and relative humidity were taken. Information was collected on building characteristics. Fel d 1 levels (microg/g of fine dust) for floors (n=203), beds (n=64) and seats (n=24) were expressed as geometric means (95% confidence intervals). RESULTS: Detectable Fel d 1 levels were found in 95% of floor samples, 91% of bed samples and 100% of seat samples. Fel d 1 levels [geometric mean (95% confidence intervals)] were significantly higher on cinema and domestic aircraft seats [36.8 (20.8-65.3) microg/g and 33.3 (28.0-39.7) microg/g respectively] than on floors [3.6 (2.5-5.1) microg/g and 2.4 (1.8-3.0) microg/g respectively]. Floor Fel d 1 levels in the public buildings sampled were lower than those of domestic dwellings without cats [0.9 (0.6-1.4) microg/g vs 1.7 (1.2-2.4)] microg/g in Wellington and [2.0 (1.6-2.6) microg/g vs 4.0 (2.7-6.0] microg/g in Christchurch. After controlling for potential confounders, floor Fel d 1 levels were higher with carpeted floors (p<0.001) and lower in banks and hospitals (p<0.001). CONCLUSION: Fel d 1 levels in public buildings are low in New Zealand public places except for cinema and domestic aircraft seats where all but one sample had Fel d 1 levels potentially high enough to precipitate asthma symptoms in sensitised individuals.