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

Cultural Resources Survey of the Proposed Moore Park Athletic Complex, Plano, Collin County, Texas

In March 2020, an intensive cultural resources survey of the proposed Moore Park athletic complex in Plano, Collin County, Texas was completed in order to inventory all cultural resources. The project parcel currently lies within an open field at the southwest corner of the intersection of Chaparral Road and Cottonwood Creek. It is bounded by Chaparral Road on the north, Cottonwood Creek to the east, Bright Star Way on the south, and Cloverhaven Way on the west in a densely residential developed area. A North Texas Municipal Water District building is located in the far northwestern corner near Cottonwood Creek. The archeological area of potential effects (APE) covers an area of approximately 103 acres (42 hectares). Because the project is owned and funded by the City of Plano, a political subdivision of the State of Texas, the project is subject to the Antiquities Code of Texas (9 Texas Natural Resources Code [TNRC] 191), which requires consideration of effects on properties designated as—or eligible to be designated as—State Antiquity Landmarks (SALs), which includes archeological resources. The survey was carried out for the City of Plano, Parks and Recreation under Texas Antiquities Permit 9334 by Brett Lang (Project Archeologist) of Cox|McLain Environmental Consulting, Inc. (CMEC). Melissa M. Green was the Principal Investigator. The parcel is situated on an upper terrace above Cottonwood Creek. From the western boundary, an unnamed tributary of Cottonwood Creek flows from the northwest corner to the southwest corner of the project parcel (as the terrace slopes southeastward) into Cottonwood Creek approximately 860 meters or 2821.5 feet away. Cottonwood Creek parallels the eastern boundary of the APE. Ground surfaces within the project area parcel were mostly covered in short, ankle-high prairie grasses used for active cattle grazing allowing for some limited visibility ranging from 20 to 50 percent. A densely wooded section was observed along the eastern boundary and along part of the tributary, allowing for 30 to 80 percent ground visibility. In all, 19 shovel test units were excavated judgmentally across the project area, of which none contained cultural materials. All materials (notes, photographs, administrative documents, and other project data) generated from this work will be housed at the Center for Archeological Studies at Texas State University at San Marcos, where they will be made permanently available to future researchers per 13 Texas Administrative Code 26.16-17. If any unanticipated cultural materials or deposits are found at any stage of clearing, preparation, or construction, the work should cease and Texas Historical Commission personnel should be notified immediately. The Texas Historical Commission concurred with the findings and recommendations of this report on 8 May 2020

Archeological Survey for the Proposed Atmos Energy D9 Natural Gas Pipeline Replacement, Dallas and Denton Counties, Texas

On 23 October 2019, an archeological survey was completed in order to evaluate potential impacts to archeological resources associated with the proposed construction of Atmos Energy Corporation’s (Atmos) proposed D9 natural gas pipeline replacement in Dallas and Denton Counties, Texas. Approximately 5.5 miles of the replacement line is expected to be installed through open-cut trenching with approximately 1.5 miles installed through directional boring; this method will be utilized at all waterways and roadway crossings. Brett Lang (Project Archeologist) of Cox|McLain Environmental Consulting, Inc. (CMEC) carried out the survey for Atmos under Texas Antiquities Permit 9124; Melissa M. Green served as Principal Investigator. Required archeological investigations were conducted per Texas Antiquities Code standards, since the project area is partially located on public lands and no previous survey had been conducted. The approximately 7.0-mile long, 50-foot wide project area, totally 41.53 acres (16.8 hectares), was subject to an intensive survey augmented with shovel test excavations. Shovel tests were excavated where ground visibility was lower than 30 percent, ground disturbance was minimal, or landforms with higher potential for prehistoric or historic archeology were observed. The project area is located in an urban setting of residential or commercial developments, with scattered cleared and open areas extending from just north of Interstate Highway 635 in Dallas County to approximately 7.0 miles (11.3 kilometers) north and terminating just south of the Old Denton Road and Frankfort Road intersection in Denton County. The replacement gas line crosses Farmers Branch, Rawhide Creek, Cooks Branch, Hutton Branch, and Furneaux Creek, as well as several unnamed tributaries. A total of 17 shovel tests were excavated within the proposed project area, with no cultural material observed on the surface or subsurface. CMEC recommends that no further investigation is required, and that the proposed project should be allowed to continue as planned. No new archeological sites were identified, and no artifacts were collected during this survey; therefore, only project records will need to be curated, per TAC 26.16 and 26.17. Project records will be permanently housed at the Center for Archaeological Studies (CAS) at Texas State University. If any unanticipated cultural materials or deposits are found at any stage of clearing, preparation, or construction, the work should cease in that area and Texas Historical Commission (THC) personnel should be notified immediately. During evaluation of the finds and coordination with the THC, clearing, preparation, and/or construction could continue in any other areas along the corridor where no such deposits or materials are observed. The Texas Historical Commission concurred with the findings and recommendations presented in this report on 5 December 2019

Intensive Archeological Survey for Proposed Improvements to Farm-to-Market Road 1735 South of State Highway 49, Titus County, Texas

On April 30 and May 1, 2019, CMEC staff completed an intensive archeological survey augmented with shovel testing to inventory and evaluate archeological resources within the footprint of proposed improvements to Farm-to-Market (FM) 1735 south of State Highway (SH) 49 in Mount Pleasant, Titus County, Texas. The project is identified under Texas Department of Transportation control-section-job number 1226-02-016. The work associated with this archeological survey was carried out under Texas Antiquities Permit 8882 by Brett Lang (Project Archeologist) and Floyd Kent of Cox|McLain Environmental Consulting, Inc. under guidance from Melissa M. Green (Principal Investigator). The area of potential effects (APE) for the proposed project is approximately 2.1 miles (3.4 kilometers) long and is located on approximately 44.57 acres (18.07 hectares) of existing right-of-way and 21.32 acres (8.58 hectares) of proposed right-of-way for a total of 65.89 acres (26.66 hectares). Fourteen shovel tests were excavated within 13.8 acres (5.6 hectares) of privately-owned land in the proposed right-of-way for which access was granted, where subsurface archeological materials might occur, where no obvious impacts or disturbances were observed, where slope (or lack thereof) made it possible, where ground visibility was limited, and where soil moisture was low. A total of 28.9 acres (11.7 hectares) was subjected to pedestrian survey only and a total of 12.5 acres (5.1 hectares) was excluded due to previous surveys. Soil horizons were found to be shallow (generally extending less than 40 centimeters [15.75 inches] below the surface) with subsoil encountered in all of the shovel tests. The shovel tests were negative for cultural material and little cultural material was found on the surface or subsurface of the surveyed areas. One isolated find consisting of a single Gary point was observed on the ground surface in a disturbed pipeline corridor. Results of the survey show that most of the project corridor has been highly disturbed from existing road construction and maintenance, commercial and residential development, installation of buried utilities, and natural impacts such as erosion. No further work is recommended in the APE prior to the proposed improvements to FM 1735. If any unanticipated cultural materials or deposits are found at any stage of clearing, preparation, or construction, the work should cease and the Atlantic District of Texas Department of Transportation should be immediately notified. All materials (notes, photographs, administrative documents, and other project data) generated from this work will be housed at the Center for Archaeological Studies at Texas State University, where they will be made permanently available to future researchers per 13 Texas Administrative Code 26.16- 17. No artifacts were collected and therefore none will be curated. The Texas Historical Commission concurred with the findings and recommendations of this report on June 20, 2019

Intensive Archeological Survey for Proposed Improvements to Farm-to-Market Road 664 from Interstate Highway 35E to Interstate Highway 45, Ellis County, Texas

On March 21, 22, and 28, 2019, an intensive archeological survey was completed in order to inventory and evaluate archeological resources within the footprint of proposed Farm-to-Market Road (FM) 664 roadway improvements from Interstate Highway 35E to Interstate Highway 45 in Ellis County, Texas. The project is identified under Texas Department of Transportation control-section-job number 1051- 01-051. The work associated with this archeological survey was carried out under Texas Antiquities Permit 8817 by Brett Lang (Project Archeologist) and Melissa M Green (Principal Investigator) of Cox|McLain Environmental Consulting, Inc. No archeological sites were recorded during the survey. Results of the survey show that a majority of the project corridor has been highly disturbed from modern residential and commercial development, bulldozing associated with the original roadway construction, installation and repair of buried utilities, and natural impacts such as erosion. The area of potential effects comprises 187.4 acres of existing right-of-way, 164.5 acres of proposed right-of-way, and 7.9 acres of permanent easements for a total of 359.8 acres. The bulk of the 359.8- acre footprint was surveyed via visual examination and photography due to extensive previous disturbance, including previous bulldozing associated with the original roadway construction along FM 664, urban expansion, and Hybrid Potential Archeological Liability Map data indicating lack of archeological potential. Of the 164.5 acres of proposed right-of-way and permanent easements 83.9 acres were not pedestrian surveyed due to existing disturbances from residential/commercial activities, agricultural practices, cattle grazing and based on HPALM data, but were recorded through photography. Access was only allowed on 59.1 of the 164.5 acres, leaving 21.5 acres with no access at the time of this survey. A total of 338.3 acres were investigated (pedestrian surveyed and/or photographed only) during this survey. On the 59.1 acres where access was allowed, 40 shovel test units were excavated where right-of-way was granted; survey was not conducted in areas that were physically inaccessible, previously developed, or in standing water. Soils were found to be shallow (generally extending to 40 centimeters below the surface); clay was encountered on the surface and subsurface in most of the shovel tests. All of the shovel tests were sterile and lacked any cultural material. Four backhoe trenches were excavated on the west banks of a tributary of Long Branch Creek on the flood plain, first terrace, and second terrace. The backhoe trenches lacked cultural materials and showed no evidence of buried soils within the undertaking’s area of potential effects. Access was not available to the west bank of Bear Creek; this area will require trenching once right-of-entry is obtained. Backhoe trenching will not be advantageous in other areas along Bear Creek, Long Branch Creek, or the tributary to Long Branch Creek due to steep slopes and/or locations outside the current area of potential effects. No further work is recommended in the 338.3 acres of the area of potential effects that were investigated/surveyed at this time. When access is available for the 21.5 remaining acres recommended for archeological survey, additional survey, including backhoe trenching near Bear Creek, is warranted. If any unanticipated cultural materials or deposits are found at any stage of clearing, preparation, or construction, the work should cease, and the Texas Department of Transportation should be immediately notified. All materials (notes, photographs, administrative documents, and other project data) generated from this work will be housed at the Center for Archaeological Studies at Texas State University, where they will be made permanently available to future researchers per 13 Texas Administrative Code 26.16– 17. No artifacts were collected; therefore, none will be curated. The Texas Historical Commission concurred with the findings and recommendations of this report on April 10, 2019

Intensive Archeological Survey for the Proposed Enclave at Pecan Creek Residential Development (Swf-2018-00261), Providence Village, Denton County, Texas

On July 3 and 5, 2019, an intensive archeological survey was completed in order to evaluate potential impacts associated with the proposed construction of a residential subdivision in the Town of Providence Village in east-central Denton County, Texas. A total of 162.64 acres (65.81 hectares) were examined with 65.39 acres (26.46 hectares) intensively shovel tested. The project would be situated between Farm-to-Market Road 2931 and Pecan Creek at the intersection of Farm-to-Market Road 2931 and Brewer Road with private properties as the north and south boundaries. Brett Lang (Principal Investigator) of Cox|McLain Environmental Consulting, Inc. carried out the survey in support of a Clean Water Act Section 404 Permit (SWF-2018-00261) for the United States Corps of Engineers, Fort Worth District under Section 106 of the National Historic Preservation Act, as amended. A Texas Antiquities Permit was not required. Based on background review of available data, the potential for intact archeological deposits was considered low for prehistoric and relatively high for historic sites within the archeological area of potential effects. Ground surface visibility varied across the parcel between 0 and 30 percent, depending on the area. Thirty shovel test units were excavated to examine the potential for subsurface archeological deposits in areas around an existing small lake and small pond. The majority of the parcel has been utilized for agricultural practices and grazing in the past and until it was sold in 2018, but urban development of the general area is rapidly encroaching in the area. One historic-age archeological site, 41DN618, was identified during the survey, but lacks integrity and is thus recommended not eligible for listing on the National Register of Historic Places. No further work is recommended within the area of potential effects prior to the construction. No artifacts were collected during the investigation however, all notes, photographs, administrative documents, and other project data will be made permanently available to future researchers via an appropriate public facility. If any unanticipated cultural materials or deposits are found at any stage of clearing, preparation, or construction, the work should cease and Texas Historical Commission personnel should be notified immediately. The Texas Historical Commission and U.S. Army Corps of Engineers concurred with the findings and recommendations of this investigation on August 12, 2019

Archeological Survey For The Austin Industries Proposed Parking Lot Expansion At East Union Bower Road (SWF-2019-00383), Irving, Dallas County, Texas

On December 12, 2019, an intensive archeological survey augmented with shovel testing was completed to evaluate potential impacts associated with the proposed construction of a parking lot expansion in east central Irving in Dallas County, Texas. The overall grade of the area would be converted to a consistent slope toward the unnamed tributary to the Elm Fork of the Trinity River near the center of the parcel and more steeply sloped along the eastern portion of the parcel. Approximately 6.5 acres (2.6 hectares) were examined and shovel tested. The proposed parking lot parcel is undeveloped, heavily vegetated, and surrounded by industrial buildings and parking lots in an urban industrial setting. From East Union Bower Road, the parcel’s main access point is located along an alley and utility corridor on the west side of the Austin Industries equipment yard. An existing sewer line runs along the alley/utility corridor from East Union Bower Road and north of the unnamed drainage that runs southeast across the north part of the parcel. This drainage eventually drains into the Elm Fork of the Trinity River approximately 2,000 feet (609 meters) southeast of the parcel. Other than the buried sewer line and utilities corridor, the parcel has not been utilized, but commercial development of the general area is rapidly encroaching into the area. Brett Lang (Project Archeologist) of Cox|McLain Environmental Consulting, Inc. carried out the survey in support of a Clean Water Act Section 404 Permit (SWF-2019-00383, Commercial Development Bower Road) for the United States Corps of Engineers, Fort Worth District. The project was subject to Section 106 of the National Historic Preservation Act of 1966, as amended. However, a Texas Antiquities Permit was not required for this project. Melissa Green served as Principal Investigator. Based on background review of available data, the potential for intact archeological deposits was considered low for both prehistoric and historic sites within the archeological area of potential effects (APE) due to previous disturbances. Ground surface visibility varied between 0 and 30 percent across the parcel. Four shovel test units were excavated to examine the potential for subsurface archeological deposits in areas around an unnamed tributary of the Elm Fork Trinity River, none of which contained archeological materials. No evidence of historic or prehistoric deposits, materials, or features were identified, and no further work is recommended within the 6.5-acre (2.6-hectare) APE. In addition, there are no extant historic-age buildings or structures and no National Register of Historic Places- (NRHP) or State Antiquities Landmark-eligible (SAL) archeological resources recorded within the 1-mile (1.6-kilometer) buffer around the APE, and no NRHP- or SAL-eligible archeological resources were found during the current survey. Therefore, the proposed project would not cause visual or indirect effects on any above-ground NRHP- or SAL-eligible cultural resources. No artifacts were collected during the investigation however, all notes, photographs, administrative documents, and other project data will be made permanently available to future researchers at the Irving office of CMEC. If any unanticipated cultural materials or deposits are found at any stage of clearing, preparation, or construction, the work should cease and Texas Historical Commission (THC) personnel should be notified immediately. The THC concurred with the findings and recommendations of this report on February 5, 2020

Information spillovers in asset markets with correlated values

We study information spillovers in a dynamic setting with correlated assets owned by privately informed sellers. In the model, a trade of one asset can provide information about the value of other assets. Importantly, the information content of trading behavior is endogenously determined. We show that this endogeneity leads to multiple equilibria when assets are sufficiently correlated. The equilibria are ranked in terms of both trade volume and efficiency. The model has implications for policies targeting post-trade transparency. We show that introducing post-trade transparency can increase or decrease welfare and trading volume depending on the asset correlation, equilibrium being played, and the composition of market participants.Asriyan acknowledges support from the Spanish Ministry of Economy and Competitiveness Grant (ECO2014-54430-P) and the Barcelona GSE Seed Grant. Fuchs gratefully acknowledges support from the ERC Grant 681575