Detection and diagnosis of fungal allergic sensitisation

Airborne fungi are ubiquitous in the environment and human exposure is inevitable. Such fungi differ greatly in their taxonomic, physical, ecological and pathogenic characteristics. Currently, 69 000 species have been taxonomically classified and more than 80 of these are recognised to be aeroallergen sources. Many strategies have evolved to sample, identify and interpret fungal exposure to these species, however no strategy serves all purposes as exposure is a complex and dynamic process confounded by spatial, temporal and geographic variations in airborne counts, in addition to the inadequacies of the immunodiagnostic techniques available. To date, the interpretation of personal exposure and sensitisation to fungal allergens has been restricted to a few select species and the contribution of other genera, airborne hyphae and fragmented conidia to allergic disease are all poorly understood. The aim of the thesis was to utilize the Halogen Immunoassay (HIA) to diagnose fungal allergic sensitisation, to investigate the distribution and factors influencing allergens of fungi in the air and to understand what is actually inhaled in exposure settings. The novelty of the HIA derives from its unique ability to provide allergen sources that are actively secreted by the collected fungal spores and hyphae, which are bound to protein binding membranes (PBM) and then immunoprobed. In Chapter 2, the HIA was compared to the commercial in vitro Pharmacia UniCap assay (CAP) and the in vivo skin prick test (SPT), using 30 sera from subjects SPT positive to Aspergillus fumigatus and/or Alternaria alternata and 30 who were SPT negative to these fungi but sensitised to non-fungal allergens. Sera were analysed by CAP and the HIA against A. alternata, A. fumigatus, Cladosporium herbarum and Epicoccum purpurascens and compared statistically. Between 3% and 7% of SPT negative sera were identified to have specific IgE towards A. fumigatus and A. iv alternata, respectively. For the SPT positive sera, significant associations were found between the HIA and CAP scores for all fungal species tested (P<0.0001). Correlations between the HIA and SPT however, were weakly correlated for A. alternata (rs = 0.44, P<0.05) but not for A. fumigatus. In Chapter 3, personal exposure to indoor fungal aerosols was examined using the HIA to identify the fungal components that people were allergic to. Personal air sampling pumps (PASs) collected airborne fungal propagules onto PBMs for 2.5 hours indoors (n=21). Collected fungi were incubated overnight in a humid chamber to promote the germination of conidia. The membranes were then immunostained with pooled human Alternaria species-positive sera. All air samples contained fungal hyphae that expressed soluble allergens and were significantly higher in concentration than counts of conidia of individual well-characterised allergenic genera. Approximately 25% of all hyphae expressed detectable allergen compared to non-stained hyphae (P<0.05) and the resultant localisation of immunostaining was heterogeneous among hyphae. Fungal conidia of ten genera that were previously uncharacterised as allergen sources accounted for 8% of the total conidia that demonstrated IgE binding. In Chapter 4, the number and identity of fungi inhaled by 34 adults in an outdoor community setting was measured over 2 hour periods by people wearing Intra-nasal air samplers (INASs) and compared to fungal counts made with a Burkard spore trap and filter air samplers worn on the lapel. Using INAS, the most prevalent fungi inhaled belonged to soil borne spores of Alternaria, Arthrinium, Bipolaris, Cladosporium, Curvularia, Epicoccum, Exserohilum, Fusarium, Pithomyces, Spegazzinia, Tetraploa and Xylariaceae species, in addition to hyphal fragments. These results showed that inhaled exposure in most people varied in a 2-fold range with 10-fold outliers. In addition, the INAS and personal air filters agreed more with each other than with Burkard spore trap counts. The analysis was further confounded by different sampling efficiencies, locations of devices and ability to visualise and count fungal propagules. In Chapter 5, a double immunostaining technique based on the HIA was developed and applied to the conidia, hyphae and fungal fragments of A. alternata, A. fumigatus and Penicillium chrysogenum to discriminate between sources of allergens, v using IgE and to identify the fungi, using a fungal-specific antibody. The localisation of immunostaining was heterogeneous between both conidia and the state of germination with greater concentrations of double immunostaining detected following germination for each fungal species (P<0.0001). Fragmented A. alternata hyphae and morphologically indiscernible fragments could be identified for the first time using this technique. In Chapter 6, the factors affecting the release of allergen from the spores of eleven different species were studied. For nine of eleven species, between 5.7% and 92% of spores released allergen before germination. Ungerminated spores of P. chrysogenum and Trichoderma viride did not release detectable allergen. After germination, all spores that germinated eluted allergen from their hyphae. Upon germination there was a significant increase in the percentage of spores eluting detectable allergen (P<0.0001) and the localisation of allergen along the hyphae varied between species. Increased elution of allergen post germination might be a common feature of many species of allergenic fungi following inhalation. Additionally, Chapter 6 explored the extent to which inhaled spores or hyphae germinate after deposition in the nasal cavity and thus cause exposure to allergens. Twenty subjects had their noses lavaged at three separate intervals, (1) at the beginning of the experiment, (2) after one hour indoors and (3) after one hour outdoors. The recovery of spores and hyphal fragments from the nasal cavity varied between individuals and was significantly greater after outdoor exposures. Germinated fungal spores were recovered often in high concentrations for Aspergillus-Penicillium species, however the proportion between ungerminated and germinated spores were much lower for other genera recovered. Conclusions: Our analysis of cultured and wild-type fungi presents a new paradigm of natural fungal exposure, which in addition to commonly recognized species, implicates airborne hyphae, fragmented conidia and the conidia of a much more diverse range of genera as airborne allergens. Exposure is heterogeneous between individuals in the same geographic locality and the spectrum of fungal genera inhaled differs with the method of analysis. Many of the spores inhaled are likely to be allergenic, however upon germination there is an increased elution of allergen and this might be a common vi feature of many fungal species following inhalation. This project also provides novel techniques to diagnose fungal allergy by immunostaining wild-type fungi to which a patient is exposed with the patient’s own serum. Such an immunoassay combines environmental with serological monitoring on a patient specific basis and potentially avoids many problems associated with extract variability, based on the performance of current diagnostic techniques for fungal allergy

Analysis of Airborne Betula Pollen in Finland; a 31-Year Perspective

In this 31-year retrospective study, we examined the influence of meteorology on airborne Betula spp. (birch) pollen concentrations in Turku, Finland. The seasonal incidence of airborne birch pollen in Turku occurred over a brief period each year during spring (April 30 – May 31). Mean peak concentrations were restricted to May (May 5 to 13). Statistically significant increases in the annual accumulated birch pollen sum and daily maximum values were observed over the study period. Birch pollen counts collected in April were retrospectively shown to increase over the duration of the study. Increases in April temperature values were also significantly associated with the earlier onset of the birch pollen season. Furthermore, the number of days where daily birch pollen concentrations exceeded 10 and 1,000 grains/m3 also increased throughout the study period. These data demonstrate that increases in temperature, especially during months preceding the onset of the birch pollen season, favor preseason phenological development and pollen dispersal. Birch pollen derived from other geographical locations may also contribute to the aerospora of Turku, Finland. To date, the public health burden associated with personal exposure to elevated birch pollen loads remains unclear and is the focus of future epidemiological research

Anaerobic methanogenesis in wetland ecosystems: do cut emergent aerenchyma of Typha x glauca facilitate increased atmospheric methane emission?

General EcologyAnaerobic methanogensis by bacteria communities is a major source of global atmospheric methane and therefore an important area of concern regarding climate change. This is particularly relevant to wetland ecosystems whose warm, water-logged, oxygen-poor soil conditions are the ideal environment for the fermentation of methane. In addition to relatively slow diffusion of methane through soil and water, and the sudden ebullition of trapped gas pockets, the aerenchyma (vessel-like tubes composed of mostly airspace that transport essential gases throughout the plant) of plants provide a direct route for gases to reach the atmosphere through the aerenchyma. We tested our hypothesis at a wetland site on the coast of Lake Huron in Cheboygan County, MI called Cheboygan Marsh. Five bucket plots pairs were installed where each pair had one bucket with the Typha stems cut below the water (submerged) and the other had stems cut above the water (emerged). Methane emissions were measured from each plot and we concluded that there was a significantly higher methane flux from emergent Typha than submerged Typha.http://deepblue.lib.umich.edu/bitstream/2027.42/89433/1/Green_Dorey_Pedley_Lu_2011.pd

Mammut pacificus sp. nov., a newly recognized species of mastodon from the Pleistocene of western North America

A new species of mastodon from the Pleistocene of western North America, Mammut pacificus sp. nov. is herein recognized, with specimens identified throughout California and from two localities in southern Idaho. This new taxon differs from the contemporaneous M. americanum in having narrower teeth, most prominently in M3/m3, as well as six sacral vertebrae, femur with a proportionally greater mid-shaft diameter, and no mandibular tusks at any growth stage. All known Pleistocene Mammut remains from California are consistent with our diagnosis of M. pacificus, which indicates that M. americanum was not present in California

Thirdhand Tobacco Smoke: Emerging Evidence and Arguments for a Multidisciplinary Research Agenda

There is broad consensus regarding the health impact of tobacco use and secondhand smoke exposure, yet considerable ambiguity exists about the nature and consequences of thirdhand smoke (THS). We introduce definitions of THS and THS exposure and review recent findings about constituents, indoor sorption-desorption dynamics, and transformations of THS; distribution and persistence of THS in residential settings; implications for pathways of exposure; potential clinical significance and health effects; and behavioral and policy issues that affect and are affected by THS. Physical and chemical transformations of tobacco smoke pollutants take place over time scales ranging from seconds to months and include the creation of secondary pollutants that in some cases are more toxic (e.g., tobacco-specific nitrosamines). THS persists in real-world residential settings in the air, dust, and surfaces and is associated with elevated levels of nicotine on hands and cotinine in urine of nonsmokers residing in homes previously occupied by smokers. Much still needs to be learned about the chemistry, exposure, toxicology, health risks, and policy implications of THS. The existing evidence on THS provides strong support for pursuing a programmatic research agenda to close gaps in our current understanding of the chemistry, exposure, toxicology, and health effects of THS, as well as its behavioral, economic, and sociocultural considerations and consequences. Such a research agenda is necessary to illuminate the role of THS in existing and future tobacco control efforts to decrease smoking initiation and smoking levels, to increase cessation attempts and sustained cessation, and to reduce the cumulative effects of tobacco use on morbidity and mortality

The Fourteenth Data Release of the Sloan Digital Sky Survey: First Spectroscopic Data from the extended Baryon Oscillation Spectroscopic Survey and from the second phase of the Apache Point Observatory Galactic Evolution Experiment

The fourth generation of the Sloan Digital Sky Survey (SDSS-IV) has been in operation since July 2014. This paper describes the second data release from this phase, and the fourteenth from SDSS overall (making this, Data Release Fourteen or DR14). This release makes public data taken by SDSS-IV in its first two years of operation (July 2014-2016). Like all previous SDSS releases, DR14 is cumulative, including the most recent reductions and calibrations of all data taken by SDSS since the first phase began operations in 2000. New in DR14 is the first public release of data from the extended Baryon Oscillation Spectroscopic Survey (eBOSS); the first data from the second phase of the Apache Point Observatory (APO) Galactic Evolution Experiment (APOGEE-2), including stellar parameter estimates from an innovative data driven machine learning algorithm known as "The Cannon"; and almost twice as many data cubes from the Mapping Nearby Galaxies at APO (MaNGA) survey as were in the previous release (N = 2812 in total). This paper describes the location and format of the publicly available data from SDSS-IV surveys. We provide references to the important technical papers describing how these data have been taken (both targeting and observation details) and processed for scientific use. The SDSS website (www.sdss.org) has been updated for this release, and provides links to data downloads, as well as tutorials and examples of data use. SDSS-IV is planning to continue to collect astronomical data until 2020, and will be followed by SDSS-V.Comment: SDSS-IV collaboration alphabetical author data release paper. DR14 happened on 31st July 2017. 19 pages, 5 figures. Accepted by ApJS on 28th Nov 2017 (this is the "post-print" and "post-proofs" version; minor corrections only from v1, and most of errors found in proofs corrected

Performance of the CMS Cathode Strip Chambers with Cosmic Rays

The Cathode Strip Chambers (CSCs) constitute the primary muon tracking device in the CMS endcaps. Their performance has been evaluated using data taken during a cosmic ray run in fall 2008. Measured noise levels are low, with the number of noisy channels well below 1%. Coordinate resolution was measured for all types of chambers, and fall in the range 47 microns to 243 microns. The efficiencies for local charged track triggers, for hit and for segments reconstruction were measured, and are above 99%. The timing resolution per layer is approximately 5 ns

Modern meat: the next generation of meat from cells

Modern Meat is the first textbook on cultivated meat, with contributions from over 100 experts within the cultivated meat community. The Sections of Modern Meat comprise 5 broad categories of cultivated meat: Context, Impact, Science, Society, and World. The 19 chapters of Modern Meat, spread across these 5 sections, provide detailed entries on cultivated meat. They extensively tour a range of topics including the impact of cultivated meat on humans and animals, the bioprocess of cultivated meat production, how cultivated meat may become a food option in Space and on Mars, and how cultivated meat may impact the economy, culture, and tradition of Asia