Chromium tolerance and accumulation in Aspergillus flavus isolated from tannery effluent.

Cr(VI) tolerance in Aspergillus flavus, strain SFL, isolated from tannery effluent was measured and compared with a reference strain of A. flavus, A1120. On solid medium, SFL had a high level of Cr(VI) tolerance (1,600 mg/L), which was 16 times that of A1120 and greater than most previously analyzed fungal strains. When in 100 mg/L of Cr(VI), SFL completely depleted Cr(VI) within 72 h while A1120 depleted 85% of Cr(VI). SFL was more effective in reducing extracellular Cr(VI) than A1120. While A1120 showed greater biosorption of Cr(VI) than SFL, intracellular accumulation was approximately 50% greater in SFL and was more energy-dependent than A1120. Cr(VI) modified the external surface of the hyphae. Cr speciation detected the presence of only Cr(III), corresponding to Cr(OH)3 , which precipitated on the hyphal surface. Cr(VI) bound to the functional groups carboxyl, amine, and hydroxyl in both SFL and A1120. Transmission electron microscopy energy-dispersive X-ray detected Cr on the fungal wall and within membrane-bound organelles of the cytoplasm. In conclusion, the greater tolerance of SFL to Cr(VI) relative to A1120 is due to more effective energy-dependant uptake of Cr(VI) into the cell and increased capacity of SFL to store Cr in intracellular vacuoles compared with A1120.

Outdoor fungal spores and acute respiratory effects in vulnerable individuals.

BACKGROUND: Many outdoor fungal spores are ubiquitous, respirable and possibly allergenic. They may contribute to asthma symptoms; however, little is known about their effects on respiratory function. OBJECTIVE: To investigate if outdoor fungal spore levels were associated with lung function or airway inflammation, and whether fungal sensitization or current asthma modified any associations. METHODS: Cross-sectional associations between same day (Lag0) and cumulative 3-day lagged (Lag0-3) counts of 12 outdoor fungal spore taxa and pre-bronchodilator spirometry (FEV1, FVC, FEF25%-75%), bronchodilator response (BDR) and airway inflammation (fractional exhaled nitric oxide (FeNO) and exhaled breath condensate (EBC) nitrogen oxides (NOx) and pH were investigated in 936 Melbourne Atopy Cohort Study participants during September 2009 to December 2011. Generalized linear models were used to quantify associations with lung function, FeNO and EBC pH; generalized estimating equations for BDR; and ordinal logistic regression for EBC NOx. Models were adjusted for age, sex, height, temperature, relative humidity, grass pollen and sample storage time. Potential effect modification by fungal sensitization and current asthma were examined using interaction terms. RESULTS: Mixed associations were found. Higher levels of Ustilago/smuts were associated with lower lung function at Lag0 (FEV1: 21ml [95%CI -36, -7]; FEF25%-75%: 39ml [-65, -13]) and Lag0-3 (FEV1: 9ml [-14, -4]; FEF25%-75% -18ml [-27, -9]). Positive BDR was associated with Ustilago/smuts (Lag0 OR = 1.1 [1.04, 1.2]; Lag0-3 OR = 1.04 [1.02, 1.07]), Alternaria (Lag0 OR = 1.3 [1.0, 1.6]) and Drechslera (Lag0 OR = 1.1 [1.03, 1.2]). Higher EBC NOx was associated with Cladosporium (Lag0-3 OR = 1.1 [1.0, 1.2]), Alternaria (Lag0-3 OR = 1.1 [1.0, 1.3]). No associations were found with higher FeNO. In those with fungal sensitization, Ustilago/smuts and Drechslera were associated with lower FEV1 and FVC; Cladosporium was associated with increased FEV1, FVC and FEF25%-75% but also with higher FeNO and lower EBC pH. In those with current asthma, Alternaria, Ustilago/smuts and Drechslera were associated with lower FEV1, FVC, FEF25-75% and EBC pH. CONCLUSION: Exposure to outdoor fungal spores may be associated with lower lung function and increased airway inflammation, particularly in those with fungal sensitization and/or current asthma.

Outdoor fungi and child asthma health service attendances.

Asthma is a significant global public health issue. Severe asthma exacerbations can be triggered by environmental factors and require medical care from health services. Although it is known that fungal exposure may lead to allergic sensitization, little is understood about its impact on asthma exacerbations. This review aims to examine whether outdoor fungi play a significant role in child asthma exacerbations. Systematic search of seven electronic databases and hand searching for peer-reviewed studies published in English, up to 31 August 2013. Inclusion criteria were study population aged <18 yr, diagnosis of asthma, attended a health service; outdoor fungi exposure was reported. Quality and risk of bias assessments were conducted. Due to significant heterogeneity, meta-analysis was not conducted. Of the 1896 articles found, 15 were eligible. Findings were not consistent, possibly due to methodological variations in exposure classifications, statistical methods and inclusion of confounders. Cross-sectional studies found no or weak associations. All but one time series studies indicated an association that varied between fungal species. Increasing evidence indicates that asthmatic children are susceptible to asthma exacerbations when exposed to outdoor fungal spores. There is limited understanding of the contributions of different fungal species. Research is needed to investigate interactions of outdoor fungi with pollen, air pollutants and respiratory viruses.

Missense mutations in the copper transporter gene ATP7A cause X-linked distal hereditary motor neuropathy.

Distal hereditary motor neuropathies comprise a clinically and genetically heterogeneous group of disorders. We recently mapped an X-linked form of this condition to chromosome Xq13.1-q21 in two large unrelated families. The region of genetic linkage included ATP7A, which encodes a copper-transporting P-type ATPase mutated in patients with Menkes disease, a severe infantile-onset neurodegenerative condition. We identified two unique ATP7A missense mutations (p.P1386S and p.T994I) in males with distal motor neuropathy in two families. These molecular alterations impact highly conserved amino acids in the carboxyl half of ATP7A and do not directly involve the copper transporter's known critical functional domains. Studies of p.P1386S revealed normal ATP7A mRNA and protein levels, a defect in ATP7A trafficking, and partial rescue of a S. cerevisiae copper transport knockout. Although ATP7A mutations are typically associated with severe Menkes disease or its milder allelic variant, occipital horn syndrome, we demonstrate here that certain missense mutations at this locus can cause a syndrome restricted to progressive distal motor neuropathy without overt signs of systemic copper deficiency. This previously unrecognized genotype-phenotype correlation suggests an important role of the ATP7A copper transporter in motor-neuron maintenance and function.

The perfect storm: temporal analysis of air during the world's most deadly epidemic thunderstorm asthma (ETSA) event in Melbourne.

BACKGROUND: There have been 26 epidemic thunderstorm asthma (ETSA) events worldwide, with Melbourne at the epicentre of ETSA with 7 recorded events, and in 2016 experienced the deadliest ETSA event ever recorded. Health services and emergency departments were overwhelmed with thousands requiring medical care for acute asthma and 10 people died. OBJECTIVES: This multidisciplinary study was conducted across various health and science departments with the aim of improving our collective understanding of the mechanism behind ETSA. DESIGN: This study involved time-resolved analysis of atmospheric sampling of the air for pollen and fungal spores, and intact and ruptured pollen compared with different weather parameters, pollution levels and clinical asthma presentations. METHODS: Time-resolved pollen and fungal spore data collected by Deakin AirWATCH Burwood, underwent 3-h analysis, to better reflect the 'before', 'during' and 'after' ETSA time points, on the days leading up to and following the Melbourne 2016 event. Linear correlations were conducted with atmospheric pollution data provided by the Environment Protection Authority (EPA) of Victoria, weather data sourced from Bureau of Meteorology (BOM) and clinical asthma presentation data from the Victorian Agency for Health Information (VAHI) of Department of Health. RESULTS: Counts of ruptured grass pollen grains increased 250% when the thunderstorm outflow reached Burwood. Increased PM10, high relative humidity, decreased temperature and low ozone concentrations observed in the storm outflow were correlated with increased levels of ruptured grass pollen. In particular, high ozone levels observed 6 h prior to this ETSA event may be a critical early indicator of impending ETSA event, since high ozone levels have been linked to increasing pollen allergen content and reducing pollen integrity, which may in turn contribute to enhanced pollen rupture. CONCLUSION: The findings presented in this article highlight the importance of including ruptured pollen and time-resolved analysis to forecast ETSA events and thus save lives.

Role of glutaredoxin1 and glutathione in regulating the activity of the copper-transporting P-type ATPases, ATP7A and ATP7B.

The copper-transporting P-type ATPases (Cu-ATPases), ATP7A and ATP7B, are essential for the regulation of intracellular copper homeostasis. In this report we describe new roles for glutathione (GSH) and glutaredoxin1 (GRX1) in Cu homeostasis through their regulation of Cu-ATPase activity. GRX1 is a thiol oxidoreductase that catalyzes the reversible reduction of GSH-mixed disulfides to their respective sulfhydryls (deglutathionylation). Here, we demonstrated that glutathionylation of the Cu-ATPases and their interaction with GRX1 were affected by alterations in Cu levels. The data support our hypothesis that the Cu-ATPases serve as substrates for Cu-dependent GRX1-mediated deglutathionylation. This in turn liberates the Cu-ATPase cysteinyl thiol groups for Cu binding and transport. GSH depletion experiments led to reversible inhibition of the Cu-ATPases that correlated with effects on intracellular Cu levels and GRX1 activity. Finally, knockdown of GRX1 expression resulted in an increase in intracellular Cu accumulation. Together, these data directly implicate GSH and GRX1 with important new roles in redox regulation of the Cu-ATPases, through modulation of Cu binding by the Cu-ATPase cysteine motifs.

Assessing the impact of PM2.5 on respiratory disease using artificial neural networks.

Understanding the impact on human health during peak episodes in air pollution is invaluable for policymakers. Particles less than PM2.5 can penetrate the respiratory system, causing cardiopulmonary and other systemic diseases. Statistical regression models are usually used to assess air pollution impacts on human health. However, when there are databases missing, linear statistical regression may not process well and alternative data processing should be considered. Nonlinear Artificial Neural Networks (ANN) are not employed to research environmental health pollution even though another advantage in using ANN is that the output data can be expressed as the number of hospital admissions. This research applied ANN to assess the impact of air pollution on human health. Three well-known ANN were tested: Multilayer Perceptron (MLP), Extreme Learning Machines (ELM) and Echo State Networks (ESN), to assess the influence of PM2.5, temperature, and relative humidity on hospital admissions due to respiratory diseases. Daily PM2.5 levels were monitored, and hospital admissions for respiratory illness were obtained, from the Brazilian hospital information system for all ages during two sampling campaigns (2008-2011 and 2014-2015) in Curitiba, Brazil. During these periods, the daily number of hospital admissions ranged from 2 to 55, PM2.5 concentrations varied from 0.98 to 54.2 µg m-3, temperature ranged from 8 to 26 °C, and relative humidity ranged from 45 to 100%. Of the ANN used in this study, MLP gave the best results showing a significant influence of PM2.5, temperature and humidity on hospital attendance after one day of exposure. The Anova Friedman's test showed statistical difference between the appliance of each ANN model (p < .001) for 1 lag day between PM2.5 exposure and hospital admission. ANN could be a more sensitive method than statistical regression models for assessing the effects of air pollution on respiratory health, and especially useful when there is limited data available.

Links between pollen, atopy and the asthma epidemic.

Pollen allergy has been found in 80-90% of childhood asthmatics and 40-50% of adult-onset asthmatics. Despite the high prevalence of atopy in asthmatics, a causal relationship between the allergic response and asthma has not been clearly established. Pollen grains are too large to penetrate the small airways where asthma occurs. Yet pollen cytoplasmic fragments are respirable and are likely correlated with the asthmatic response in allergic asthmatics. In this review, we outline the mechanism of pollen fragmentation and possible pathophysiology of pollen fragment-induced asthma. Pollen grains rupture within the male flowers and emit cytoplasmic debris when winds or other disturbances disperse the pollen. Peak levels of grass and birch pollen allergens in the atmosphere correlated with the occurrence of moist weather conditions during the flowering period. Thunderstorm asthma epidemics may be triggered by grass pollen rupture in the atmosphere and the entrainment of respirable-sized particles in the outflows of air masses at ground level. Pollen contains nicotinamide adenine dinucleotide phosphate (reduced) oxidases and bioactive lipid mediators which likely contribute to the inflammatory response. Several studies have examined synergistic effects and enhanced immune response from interaction in the atmosphere, or from co-deposition in the airways, of pollen allergens, endogenous pro-inflammatory agents, and the particulate and gaseous fraction of combustion products. Pollen and fungal fragments also contain compounds that can suppress reactive oxidants and quench free radicals. It is important to know more about how these substances interact to potentially enhance, or even ameliorate, allergic asthma.

Altered expression of two zinc transporters, SLC30A5 and SLC30A6, underlies a mammary gland disorder of reduced zinc secretion into milk.

Two cases of zinc deficiency in breastfed neonates were investigated where zinc levels in the mothers' milk were reduced by more than 75 % compared to normal. The objective of this study was to find the molecular basis of the maternal zinc deficiency condition. Significant reductions in mRNA expression and protein levels of the zinc transporters SLC30A5 and SLC30A6 were found in maternal tissue, suggesting a causal link to the zinc-deficient milk. Novel splice variants of the SLC30A6 transcript were detected. No modifications were found in coding regions, or in transcription binding sites of promoter regions or in 5' and 3' untranslated regions of both transporters in lymphoblasts and fibroblasts isolated from both mothers. Altered DNA methylation in SLC30A5 at two CpG sites was detected and may account for the reduced levels of SLC30A5 mRNA and protein in lymphoblasts. Reduced SLC30A6 mRNA and protein levels in lymphoblasts may be secondary to reduced SLC30A5 expression, as they function as a heterodimer in zinc transport. In conclusion, two cases of zinc deficiency are linked to low levels of the SLC30A5 and SLC30A6 zinc transporters. These two zinc transporters have not been previously associated with zinc deficiency in milk.

Legionella pneumophila multiplication is enhanced by chronic AMPK signalling in mitochondrially diseased Dictyostelium cells.

Human patients with mitochondrial diseases are more susceptible to bacterial infections, particularly of the respiratory tract. To investigate the susceptibility of mitochondrially diseased cells to an intracellular bacterial respiratory pathogen, we exploited the advantages of Dictyostelium discoideum as an established model for mitochondrial disease and for Legionella pneumophila pathogenesis. Legionella infection of macrophages involves recruitment of mitochondria to the Legionella-containing phagosome. We confirm here that this also occurs in Dictyostelium and investigate the effect of mitochondrial dysfunction on host cell susceptibility to Legionella. In mitochondrially diseased Dictyostelium strains, the pathogen was taken up at normal rates, but it grew faster and reached counts that were twofold higher than in the wild-type host. We reported previously that other mitochondrial disease phenotypes for Dictyostelium are the result of the activity of an energy-sensing cellular alarm protein, AMP-activated protein kinase (AMPK). Here, we show that the increased ability of mitochondrially diseased cells to support Legionella proliferation is suppressed by antisense-inhibiting expression of the catalytic AMPKalpha subunit. Conversely, mitochondrial dysfunction is phenocopied, and intracellular Legionella growth is enhanced, by overexpressing an active form of AMPKalpha in otherwise normal cells. These results indicate that AMPK signalling in response to mitochondrial dysfunction enhances Legionella proliferation in host cells.

Identification and possible disease mechanisms of an under-recognized fungus, Aureobasidium pullulans.

BACKGROUND: Investigations into the occurrence and health effects of yeast-like fungi in the outdoor air in the US have been limited. We sought to identify a respirable-sized fungus common in the Pasadena air, locate a major source for the emissions and investigate its relevance to allergic disease. METHODS: Yeast-like fungi sampled from the environment were isolated, microscopically examined and sequenced. Pasadena allergy patients were skin tested with commercially available fungal extracts. Patient serum was immunoanalyzed for specific IgE reactivity. Nearby vegetation was analyzed in a controlled emission chamber to find a major source for the aerosols. RESULTS: Hyaline unicellular conidia comprised up to 90% (41,250 m(-3) of air) of total fungal counts and generally peaked at night and during periods of rainfall and ensuing winds throughout the fall and winter. Flowers were determined to be a major source of the emissions. The cellular and colonial morphology of isolates were consistent with Aureobasidium species. The sequence of the D1/D2 region of the 26S ribosomal subunit of isolates from flowers showed identity to two strains of Aureobasidium pullulans (black yeast). Seventeen percent (16/94) of atopic individuals had positive skin testing with A. pullulans extract. Patient sera IgE identified several high molecular weight allergens in Aureobasidium extracts. CONCLUSIONS: Respirable-sized conidia of A. pullulans are emitted from flowers and form high concentrations in the air. They are associated with immediate reactivity on skin tests, bind to patient sera IgE, and might be relevant in allergic upper and lower airway diseases.

Thunderstorm asthma.

Thunderstorms have often been linked to epidemics of asthma, especially during the grass flowering season; however, the precise mechanisms explaining this phenomenon are unknown. Evidence of high respirable allergen loadings in the air associated with specific meteorologic events combined with an analysis of pollen physiology suggests that rupture of airborne pollen can occur. Strong downdrafts and dry, cold outflows distinguish thunderstorm rain from frontal rain. The weather system of a mature thunderstorm likely entrains grass pollen into the cloud base, where pollen rupture would be enhanced, then transports the respirable-sized fragments of pollen debris to ground level where outflows distribute them ahead of the rain. The conditions occurring at the onset of a thunderstorm might expose susceptible people to a rapid increase in concentrations of pollen allergens in the air that can readily deposit in the lower airways and initiate asthmatic reactions.

Release of allergens as respirable aerosols: A link between grass pollen and asthma.

BACKGROUND: Asthma incidence has long been linked to pollen, even though pollen grains are too large to penetrate into the airways where asthmatic responses originate. Pollen allergens found in small, respirable particles have been implicated in a number of asthma epidemics, particularly ones following rainfall or thunderstorms. OBJECTIVE: The aim of this study was to determine how pollen allergens form the respirable aerosols necessary for triggering asthma. METHODS: Flowering grasses were humidified and then dried in a controlled-environment chamber connected to a cascade impactor and an aerosol particle counter. Particles shed from the flowers were analyzed with high-resolution microscopy and immunolabeled with rabbit anti-Phl p 1 antibody, which is specific for group 1 pollen allergens. RESULTS: Contrary to what has been reported in other published accounts, most of the pollen in this investigation remained on the open anthers of wind pollinated plants unless disturbed-eg, by wind. Increasing humidity caused anthers to close. After a cycle of wetting and drying followed by wind disturbance, grasses flowering within a chamber produced an aerosol of particles that were collected in a cascade impactor. These particles consisted of fragmented pollen cytoplasm in the size range 0.12 to 4.67 microm; they were loaded with group 1 allergens. CONCLUSION: Here we provide the first direct observations of the release of grass pollen allergens as respirable aerosols. They can emanate directly from the flower after a moisture-drying cycle. This could explain asthmatic responses associated with grass pollination, particularly after moist weather conditions.