Identification of glucocorticoid receptor in Drosophila melanogaster.

BACKGROUND: Vertebrate glucocorticoid receptor (GR) is an evolutionary-conserved cortisol-regulated nuclear receptor that controls key metabolic and developmental pathways. Upon binding to cortisol, GR acts as an immunosuppressive transcription factor. Drosophila melanogaster, a model organism to study innate immunity, can also be immunosuppressed by glucocorticoids. However, while the genome of fruit fly harbors 18 nuclear receptor genes, the functional homolog of vertebrate GR has not been identified. RESULTS: In this study, we demonstrated that while D. melanogaster is susceptible to Saccharomyces cerevisiae oral infection, the oral exposure to cortisol analogs, cortisone acetate or estrogen, increases fly sensitivity to yeast challenge. To understand the mechanism of this steroid-induced immunosuppression, we identified the closest genetic GR homolog as D. melanogaster Estrogen Related Receptor (ERR) gene. We discovered that Drosophila ERR is necessary for cortisone acetate- and estrogen-mediated increase in sensitivity to fungal infection: while ERR mutant flies are as sensitive to the fungal challenge as the wildtype flies, the yeast-sensitivity of ERR mutants is not increased by these steroids. Interestingly, the fungal cortisone analog, ergosterol, did not increase the susceptibility of Drosophila to yeast infection. The immunosuppressive effect of steroids on the sensitivity of flies to fungi is evolutionary conserved in insects, as we show that estrogen significantly increases the yeast-sensitivity of Culex quinquefasciatus mosquitoes, whose genome contains a close ortholog of the fly ERR gene. CONCLUSIONS: This study identifies a D. melanogaster gene that structurally resembles vertebrate GR and is functionally necessary for the steroid-mediated immunosuppression to fungal infections.

Assessment of exposure to fungi in the heavily contaminated work environment (a solid waste sorting plant) based on the ergosterol analysis.

OBJECTIVES: This paper reports on the results of the study aimed at application of ergosterol as an quantitative indicator of fungal bioaerosol present in the indoor air in occupational environment heavily contaminated with organic dust as well as its comparison with the culturable method. MATERIAL AND METHODS: The study was conducted in the indoor solid waste sorting plant. Using Andersen impactor adapted to 1 plate at the flow rate of 30 l/min, indoor air was sampled in the workers' breathing zone. Ergosterol was sampled using gelatinous filter (1000 l of air) and then analyzed by means of the spectrophotometric method. Fungi were sampled on malt extract agar (MEA) medium (3 replications: 2 l, 7.5 l, 15 l of air) and analyzed by means of the culturable method. Based on ergosterol analyzes, concentration of fungi was calculated. Results were given as the range assuming min. as 5.1 pg ergosterol/spore and max as 1.7 pg ergosterol/spore. RESULTS: The average concentrations of ergosterol in a working room (arithmetic mean (AM), standard deviation (SD); minimum-maximum (min.-max)) were, respectively: 2.16, 0.72; 0.85-2.92 µg/m3; fungi calculated based on ergosterol - 424.1×103-1272.4×103, 140.1×103- 420.4×103, 167×103-1716.5×103 CFU/m3, and culturable fungi - 13×103, 9.7×103, 1.9×103-34×103 CFU/m3). It was revealed that concentrations of calculated fungi were even 2 orders of magnitude higher than culturable fungi. CONCLUSIONS: The quantitative assessment of moldiness by means of ergosterol measurement seems to be a reliable indicator for environments heavily contaminated with organic dust, where viable and non-viable fungi are present in high proportions. Based on that result, more restrictive (as compared to a similar assessment carried out by means of the culturable method) hygienic recommendations, especially those related to the use of preventive measures protecting the employees' respiratory tract, should have been undertaken.

A longitudinal study of sick building syndrome among pupils in relation to microbial components in dust in schools in China.

There are few longitudinal studies on sick building syndrome (SBS), which include ocular, nasal, throat, and dermal symptoms, headache, and fatigue. We studied the associations between selected microbial components, fungal DNA, furry pet allergens, and incidence and remission of SBS symptoms in schools in Taiyuan, China. The study was based on a two-year prospective analysis in pupils (N=1143) in a random sample of schools in China. Settled dust in the classrooms was collected by vacuum cleaning and analyzed for lipopolysaccharide (LPS), muramic acid (MuA), and ergosterol (Erg). Airborne dust was collected in Petri dishes and analyzed for cat and dog allergens and fungal DNA. The relationship between the concentration of allergens and microbial compounds and new onset of SBS was analyzed by multi-level logistic regression. The prevalence of mucosal and general symptoms was 33% and 28%, respectively, at baseline, and increased during follow-up. At baseline, 27% reported at least one symptom that improved when away from school (school-related symptoms). New onset of mucosal symptoms was negatively associated with concentration of MuA, total LPS, and shorter lengths of 3-hydroxy fatty acids from LPS, C14, C16, and C18. Onset of general symptoms was negatively associated with C18 LPS. Onset of school-related symptoms was negatively associated with C16 LPS, but positively associated with total fungal DNA. In general, bacterial compounds (LPS and MuA) seem to protect against the development of mucosal and general symptoms, but fungal exposure measured as fungal DNA could increase the incidence of school-related symptoms.

Muramic acid, endotoxin, 3-hydroxy fatty acids, and ergosterol content explain monocyte and epithelial cell inflammatory responses to agricultural dusts.

In agricultural and other environments, inhalation of airborne microorganisms is linked to respiratory disease development. Bacterial endotoxins, peptidoglycans, and fungi are potential causative agents, but relative microbial characterization and inflammatory comparisons amongst agricultural dusts are not well described. The aim of this study was to determine the distribution of microbial endotoxin, 3-hydroxy fatty acids (3-OHFA), muramic acid, and ergosterol and evaluate inflammatory responses in human monocytes and bronchial epithelial cells with various dust samples. Settled surface dust was obtained from five environments: swine facility, dairy barn, grain elevator, domestic home (no pets), and domestic home with dog. Endotoxin concentration was determined by recombinant factor C (rFC). 3-OHFA, muramic acid, and ergosterol were measured using gas chromatography-mass spectrometry. Dust-induced inflammatory cytokine secretion in human monocytes and bronchial epithelial cells was evaluated. Endotoxin-independent dust-induced inflammatory responses were evaluated. Endotoxin and 3-OHFA levels were highest in agricultural dusts. Muramic acid, endotoxin, 3-OHFA, and ergosterol were detected in dusts samples. Muramic acid was highest in animal farming dusts. Ergosterol was most significant in grain elevator dust. Agricultural dusts induced monocyte tumor necrosis factor (TNF) alpha, interleukin (IL)-6, IL-8, and epithelial cell IL-6 and IL-8 secretion. Monocyte and epithelial IL-6 and IL-8 secretion was not dependent on endotoxin. House dust(s) induced monocyte TNFalpha, IL-6, and IL-8 secretion. Swine facility dust generally produced elevated responses compared to other dusts. Agricultural dusts are complex with significant microbial component contribution. Large animal farming dust(s)-induced inflammation is not entirely dependent on endotoxin. Addition of muramic acid to endotoxin in large animal farming environment monitoring is warranted.

A novel steroid from Elephantopus scaber L. an ethnomedicinal plant with antidiabetic activity.

Acetone extract of Elephantopus scaber, an ethnomedicnal plant, reduced the blood glucose levels in streptozotocin-induced diabetic rats significantly. Acute toxicity studies revealed the non-toxic nature of the crude extract. Fractionation of the acetone extract yielded a new steroid, 28Nor-22(R)Witha 2,6,23-trienolide. Biological testing of the compound demonstrated a significant antidiabetic activity by reducing the elevated blood glucose levels and restoring the insulin levels in streptozotocin-induced diabetic rats. This compound can be a useful candidate to treat diabetes.

Asthmatic symptoms among pupils in relation to microbial dust exposure in schools in Taiyuan, China.

Microbial exposure has been indicated as significant in the development of asthma and allergy among children. The aim of the study was to test whether microbial exposure and allergens in the school environment are associated with asthmatic symptoms in pupils. Data on asthmatic symptoms and respiratory infections were collected through a questionnaire survey among 1993 pupils aged 11-15 yr in 10 randomly selected schools in Taiyuan, China. Settled dust in classrooms was analysed using tandem gas chromatography-mass spectrometry for 3-hydroxy fatty acids, marker of lipopolysaccharide (LPS) from endotoxin, muramic acid (MuA), marker of bacteria and ergosterol (Erg) for fungi, quantifying both culturable and non-culturable microbes. A total of 29.8% reported daytime attacks of breathlessness, 8.4% wheeze and 1.2% had doctor's diagnosed asthma. Generally, MuA was negatively associated with wheeze and daytime attacks of breathlessness, the latter of which was negatively associated with Erg to a weaker extent. Total concentration of LPS was positively associated with daytime attacks of breathlessness, but shorter lengths of LPS, C10, C12 and C14 LPS were negatively associated with either wheezing or daytime attacks of breathlessness. For MuA and C10 and C12 of LPS, the associations were independent of airborne allergens and classroom crowdedness, and even independent of the other two microbial markers for MuA. Microbial exposure indicated by certain chemical markers (e.g. MuA) could be protective for asthmatic symptoms, but for LPS (endotoxin), the picture is more complex, varying by different lengths of fatty acids of LPS.

Changes in indoor allergen and fungal levels predict changes in asthma activity among young adults.

BACKGROUND: Exposures to allergens are thought to be important risk factors for asthma. We conducted a longitudinal study of indoor allergen and fungal levels in Melbourne homes between 1996 and 1998 to examine the effect of changes in allergen exposure upon asthma and associated outcomes. METHODS: Participants were visited at home in 1996 (n=485) and 1998 (n=360), when dust and air samples were collected from their bedrooms and assayed for Der p 1, Fel d 1, ergosterol and fungal propagules. Subjects then attended the lung function laboratory to complete a questionnaire, spirometry, methacholine challenge and skin prick testing. The associations between the change in allergen levels and change in clinical outcomes were examined using multiple logistic and linear regression. RESULTS: Participants whose Cladosporium fungal exposure doubled had 52% greater odds of having had an attack of asthma in the last 12 months. A doubling of fungal exposure was also associated with 53% greater odds of developing atopy. A doubling of Fel d 1 floor levels was associated with 73% increased odds of doctor-diagnosed asthma. A doubling of Der p 1 levels in bed dust was associated with a 64% greater odds of persistent bronchial hyper-reactivity. CONCLUSIONS: These findings provide evidence that changes in indoor levels of fungi and house dust mites can affect the risk of development and persistence of asthma and atopy in adults. Further studies are required to establish any benefit of sustained reductions in indoor allergen exposures, and to determine whether these effects are truly 'allergic' or because of immune stimulation in the airway through other less specific mechanisms.